1meckling K A Ed Nutrient Drug Interactions PDF

DK5836_title 4/10/06 9:28 AM Page 1
Nutrient–Drug
Interactions
Edited by
Kelly Anne Meckling
Boca Raton London New York
CRC is an imprint of the Taylor & Francis Group,

an informa business
NUTRITION AND DISEASE PREVENTION
Editorial Advisory Board

CAROLYN D. BERDANIER, PH.D.
University of Georgia, Athens, Georgia, U.S.A.
FRANK GREENWAY, M.D.

Pennington Biomedical Research Center, Louisiana State University,
Baton Rouge, Louisiana, U.S.A.
KHURSHEED N. JEEJEEBHOY, M.D.

University of Toronto Medical School, Toronto, Canada
MULCHAND S. PATEL, PH.D.

The University at Buffalo, The State University of New York,
Buffalo, New York, U.S.A.
KATHLEEN M. RASMUSSEN, PH.D.

Cornell University, Ithaca, New York, U.S.A.
1. Genomics and Proteomics in Nutrition, edited by

Carolyn D. Berdanier and Naima Moustaid-Moussa
2. Perinatal Nutrition: Optimizing Infant Health and Development,

edited by Jatinder Bhatia
3. Soy in Health and Disease Prevention, edited by Michihiro Sugano
4. Nutrition and Cancer Prevention, edited by Atif B. Awad

and Peter G. Bradford
5. Cancer Prevention and Management through Exercise

and Weight Control, edited by Anne McTiernan
6. Nutritional Strategies for the Diabetic/Prediabetic Patient,

edited by Jeffrey I. Mechanick and Elise M. Brett
7. Nutrient–Drug Interactions, edited by Kelly Anne Meckling

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Introduction to Clinical Nutrition: Second Edition, Revised

and Expanded, edited by V. Sardesai
Pediatric Gastroenterology and Nutrition in Clinical Practice,

edited by Carlos Lifschitz
Nutrients and Cell Signaling, edited by Janos Zempleni

and K. Dakshinamurti
Mitochondria in Health and Disease, edited by Carolyn D. Berdanier
Thiamine, edited by Frank Jordan and Mulchand Patel
Phytochemicals in Health and Disease, edited by Yongping Bao

and Roger Fenwick
Handbook of Obesity: Etiology and Pathophysiology, Second Edition,

edited by George Bray and Claude Bouchard
Handbook of Obesity: Clinical Applications, Second Edition,

edited by George Bray and Claude Bouchard
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Nutrient drug interactions / edited by Kelly Anne Meckling.

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Introduction
The idea of food as medicine has been around for millennia. However, the
acceptance of specific foods, food components, extracts, herbals and supplements
as part of conventional medical practice in industrialized countries is still a highly
controversial subject. Western medicine has largely depended on the availability
of pharmacologic agents or “drugs” for the prevention and treatment of human
disease with lesser emphasis on lifestyle and dietary habits as major contributory
factors, particularly with respect to chronic disease prevention. Furthermore, there
are many additional tools in the repertoire of complementary medical practitioners
that go far beyond what is ingested, inhaled or spread on the skin to ward off
disease or treat illness.
The goal of this volume is not to consider the entirety of complementary
medicine, but to focus on food, herbals and their chemical constituents as con-
tributors to human health through control of metabolism, primarily as they relate
to chronic disease development and treatment. More specifically, we will consider
how what is consumed affects response, whether on a population or individual
level, to pharmacologic agents that are the mainstay of chronic disease treat-
ment/prevention around the world. Many of these drugs have their history as
natural compounds isolated from flora and fauna in our environment; however
the public perception has often been that “drugs” come with significant risk and
are less safe than “foods” or “natural health products.” Promulgating this view
are vocal members of the public who perceive a conspiracy by big business and
the pharmacologic giants to protect their billion-dollar stocks by presenting alter-
native medical practice as “flawed.” Furthermore, many complementary practi-
tioners themselves will say that they usually do not consider that adverse events
may occur in response to their therapies, nor do they ask their patients to report
any side effects,1 which exacerbates the problem.
Regardless of the particular approach that is taken, patients must demand of
their practitioners that applied therapies be safe and effective. Thus, evidence-based
medical practice must continue to be the cornerstone of health care delivery for the
foreseeable future. While testing through phase I, II and III clinical trials following
extensive testing in model systems is the standard for determining the safety and
efficacy of pharmacologic agents and synthetic food additives, this has not been
the case for many “natural products.” Assumptions of safety and efficacy for natural
health products have often come from demonstration of historical use over many
years, or possibly centuries, in specific populations with specific lifestyle and
cultural practices. This limited scope of use can hardly be expected to predict the
complicated behaviors that could occur in individuals or populations following very
different lifestyles or using them in combination with other therapies.
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There have been incredible advances in our understanding of how the chem-
icals in foods and herbs we consume interact with natural and synthetic drugs
used to prevent or treat human disease. It is standard practice to examine the
effects of food consumption on the absorption and pharmacokinetics of new
drugs, but the relevant issues have become much greater than “should this med-
icine be taken with or without food.” Long-term use of many medications has
the potential to modify gastrointestinal function and alter the uptake and handling
of both macronutrients, micronutrients and many phytochemicals that we now
know have impacts on nutritional status and overall health. Manipulating the diet
or the administration of supplements, orally or systemically, can help prevent
nutritional deficiencies that could develop during chronic drug administration. A
much more difficult task is to understand the potential for the thousands of
chemical compounds found in food and natural health products to modify drug
bioavailability, biodistribution, toxicity, efficacy and metabolism. In some cases
a single food or supplement could profoundly increase or decrease the toxicity
and/or efficacy of a single drug.
A major emphasis, from a clinical point of view, has been to identify these
substances and remove them from the diets or supplement list of patients under-
going drug therapy. The focus, then, has been almost exclusively on simplifying
the drug treatment plan. An alternative approach would be to use our knowledge
of the mechanisms by which food chemicals modify drug action and to develop
adjuvant therapy protocols that use food (functional foods and nutraceuticals)
and drugs together to optimize treatment outcome. This could result in a decreased
requirement, by the patient, for expensive medications, possibly increased efficacy
with decreased toxicity toward normal or uninfected tissues, and the maintenance
of a food substance with multiple healthful components.
In this collection of reviews, these major issues will be addressed by experts
in their relevant disciplines. The organization of the book will place the focus on
the ailment being treated or prevented and, thus, on the targets of therapy. Within
each section a comprehensive examination of macronutrient, micronutrient and
phytochemical impacts on drug action will be undertaken. Where a given drug
affects nutritional status, the appropriate diet modification or supplement will be
suggested. The major focus of each section will be on the molecular mechanism(s)
by which the food or chemical is thought to modify disease process and drug
behavior. Where specific active chemicals have been identified, their precise
molecular targets, including regulation of gene expression, will be described. The
book will finish with a description of the roles of genetic variation and polymor-
phism in determining nutrient/drug responses, and how individuals might be
“profiled” in future to identify those likely to demonstrate specific interactions
and who would benefit most from adjuvant or complementary therapies.
Kelly Anne Meckling, Ph.D.

DK5836_C000.fm Page vii Monday, May 22, 2006 2:39 PM
REFERENCE
1. Abbott, A., Survey questions safety of alternative medicine. Nature, 436 (7053):
898, 2005.
DK5836_C000.fm Page viii Monday, May 22, 2006 2:39 PM
DK5836_C000.fm Page ix Monday, May 22, 2006 2:39 PM
The Editor
Kelly Anne Meckling, Ph.D., completed an Honors B.Sc. with Distinction in
Biochemistry with a minor in psychology from the University of Calgary in 1984,
and her M.Sc. with Dr. Tony Pawson from the University of British Columbia.
She then transferred to the University of Toronto to complete her Ph.D. in medical
biophysics in 1989, examining the signaling pathways unique to human leukemia
and normal blood cell development.
Dr. Meckling’s travels took her to the University of Alberta in Edmonton in
1989, where she examined the role of growth factors and cancer development on
transport of chemotherapeutic drugs. In 1991, she became a faculty member in
the Department of Nutritional Sciences at the University of Guelph, with no
formal nutrition training. She rapidly acquired this knowledge through reading,
attending undergraduate and graduate lectures and through her teaching activities.
Since that time, she has continued as an active researcher in the area of nutrient
signaling and the prevention/treatment of chronic diseases, including cancer,
cardiovascular disease and obesity.
Dr. Meckling currently is associated with the Natural Sciences Engineering
Research Council of Canada and the Breast Cancer Society, where she carries
out both in vitro studies in model systems and human clinical intervention trials.
She teaches undergraduate biology to 2000 students per year as well as advanced
courses in nutrition and the metabolic control of disease. Also, Dr. Meckling is
academic advisor to students majoring in nutritional and nutraceutical sciences.
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Contributors
Marica Bakovic James B. Kirkland
Department of Human Health and Human Health and Nutritional
Nutritional Sciences Sciences
University of Guelph University of Guelph
Guelph, Ontario, Canada Guelph, Ontario, Canada
Amir Baluch
Department of Anesthesiology Alexandre Loktionov
Louisiana State University Colonix Ltd.
New Orleans, Louisiana Babraham Research Campus
Cambridge, United Kingdom
Matthew Chronowic
Department of Human Health and
Kelly Anne Meckling
Nutritional Sciences
Human Health and Nutritional
University of Guelph
Sciences
Guelph, Ontario, Canada
University of Guelph
Guelph, Ontario, Canada
Ennio Esposito
Istituto di Ricerche Farmacologiche
Mario Negri André J. Scheen
Santa Maria Imbaro, Italy Division of Diabetes, Nutrition and
Metabolic Disorders
Jason M. Hoover Department of Medicine
Department of Anesthesiology Sart Tilman Hospital
Louisiana State University Liège, Belgium
New Orleans, Louisiana
Alan D. Kaye Kevin Wood

Department of Anesthesiology Department of Human Health and
Louisiana State University Nutritional Sciences
Health Sciences Center University of Guelph
New Orleans, Louisiana Guelph, Ontario, Canada
DK5836_C000.fm Page xii Monday, May 22, 2006 2:39 PM
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Contents
Chapter 1
Diabetes, Obesity, and Metabolic Syndrome .......................................................1
André J. Scheen
Chapter 2
Hypolipidemic Therapy: Drugs, Diet, and Interactive Effects ..........................31
Alexandre Loktionov
Chapter 3
Phytochemicals, Xenobiotic Metabolism, and Carcinogenesis..........................63
James B. Kirkland
Chapter 4
Nutrient and Phytochemical Modulation of Cancer Treatment .........................95
Kelly Anne Meckling
Chapter 5
Role of Nutritional Antioxidants in the Prevention and Treatment of
Neurodegenerative Disorders............................................................................129
Ennio Esposito
Chapter 6
Nutrients and Herbals in the Pharmacotherapy of
Unipolar/Major Depression...............................................................................179
Matthew Chronowic
Chapter 7
Supplements and Anesthesiology .....................................................................209
Alan D. Kaye, Amir Baluch, and Jason M. Hoover
Chapter 8
Nutrient- and Drug-Responsive Polymorphic Genes as Nutrigenomic
Tools for Prevention of Cardiovascular Disease and Cancer...........................237
Kevin Wood and Marica Bakovic
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Chapter 9
Nutrigenomics and Pharmacogenomics of Human Cancer:
On the Road from Nutrigenetics and Pharmacogenetics .................................261
Alexandre Loktionov
Index..................................................................................................................319
DK5836_C001.fm Page 1 Monday, May 22, 2006 11:50 AM
1 Diabetes, Obesity, and

Metabolic Syndrome
André J. Scheen
CONTENTS
1.1 Introduction..................................................................................................1
1.2 Diet Intervention..........................................................................................3
1.2.1 Healthy Diet.....................................................................................3
1.2.2 Special Foods and Nutrients ...........................................................4
1.2.2.1 Fibers .................................................................................4
1.2.2.2 Dietary Supplements and Herbal Products.......................5
1.2.2.3 Grapefruit Juice.................................................................5
1.3 Pharmacological Intervention......................................................................6
1.3.1 Glucose-Lowering Agents ...............................................................6
1.3.1.1 Sulfonylureas.....................................................................6
1.3.1.2 Metformin..........................................................................8
1.3.1.3 Thiazolidinediones (TZDs) ...............................................9
1.3.1.4 α-Glucosidase Inhibitors.................................................10
1.3.1.5 Meglitinide Derivatives ...................................................12
1.3.2 Antiobesity Agents ........................................................................13
1.3.2.1 Orlistat .............................................................................13
1.3.2.2 Sibutramine .....................................................................15
1.3.3 Lipid-Lowering Agents .................................................................16
1.3.3.1 Statins ..............................................................................16
1.3.3.2 Fibrates ............................................................................17
1.3.3.3 Resins ..............................................................................18
1.3.3.4 Ezetimibe.........................................................................18
1.3.3.5 Sterols and Stanols..........................................................19
1.4 Conclusions................................................................................................20
References ...........................................................................................................20
1.1 INTRODUCTION
Metabolic disorders, such as diabetes mellitus, overweight/obesity, and the so-
called metabolic syndrome (including atherogenic dyslipidemia) are rapidly
1
2 Nutrient–Drug Interactions
increasing in the westernized world because of poor lifestyle habits favoring fat-
and sucrose-enriched meals and low physical activity or sedentariness.1 Medical
nutrition therapy is an integral component of diabetes mellitus,2 obesity3 and
metabolic syndrome management.4 Therapy should be individualized, with con-
sideration given to the individual’s usual food and eating habits, metabolic profile,
treatment goals, and desired outcomes. When diet, associated with regular phys-
ical exercise and healthy lifestyle habits, is insufficient to control blood glucose,
reduce body weight and/or improve the metabolic profile, pharmacological inter-
ventions may be considered to reach these objectives.5 While insulin therapy is
essential in all patients with type 1 diabetes, drug therapy is now a key element
in the management of type 2 diabetes mellitus and is becoming more and more
sophisticated with the recent launch of new compounds and the increased use of
combined therapy.6,7 In contrast, pharmacological intervention for obesity still
remains a controversial issue because of only modest long-term efficacy and/or
concern about safety.8,9 Finally, the recent recognition of the important health
problem associated with the metabolic syndrome10 has stimulated researchers to
develop drugs aimed at specifically improving insulin sensitivity and correcting
associated metabolic disorders.4,11
Because nutrition therapy and pharmacological intervention are major com-
ponents of the management of metabolic disorders, it is interesting to consider
potential food–drug or nutrient–drug interactions in this particular medical field
(Figure 1.1). An interaction is said to take place when the effects of one drug are
changed by the presence of another drug, food, drink or by some environmental
chemical agent.12 Interactions between food and drugs may inadvertently reduce
or increase the effect of the drug, resulting in therapeutic failure (i.e., hypergly-
cemia in the case of diabetes mellitus) or increased toxicity (i.e., hypoglycemia
in the case of diabetes). The majority of clinically relevant food–drug interactions
are caused by food-induced changes in the bioavailability of the drug.13,14 Indeed,
since the bioavailability and clinical effect of most drugs are correlated, the
bioavailability is an important pharmacokinetic effect parameter. However, in
order to evaluate the clinical relevance of a food–drug interaction, the impact of
food intake on the clinical effect of the drug has to be quantified as well.15 This
may be particularly relevant as far as food-related metabolic diseases are con-
cerned. Thus, in the field of metabolic disorders, where nutrition plays a major
role in the overall treatment, the potential influence of food and nutrient intake
on drug therapeutic effect may be crucial.
Pharmacokinetic interferences often occur as a result of a change in drug
metabolism. Cytochrome P450 system oxidizes a broad spectrum of drugs by a
number of metabolic processes that can be enhanced or reduced by various drugs
(known as inducers or inhibitors, respectively).16 The clinical importance of any
drug interaction depends on factors that are drug-, patient- and administration-
related. As reviewed extensively,17 dietary changes can alter the expression and
activity of hepatic drug metabolizing enzymes, and dietary components may
influence the gastrointestinal metabolism and transports of drugs.18 Athough this
can lead to alterations in the systemic elimination kinetics of drugs metabolized
Diabetes, Obesity, and Metabolic Syndrome 3
TYPE 2 DIABETES
DIGESTIVE ORAL
TRACT BIOAVAILABILITY
F D
O CYP HEPATIC
INTERFERENCES METABOLISM
R
O U
D G
NUTRIENT PHARMACO-
EFFECTS DYNAMICS
OBESITY
METABOLIC SYNDROME
FIGURE 1.1 Potential food–drug or nutrient–drug interactions of clinical interest in

patients with metabolic diseases.
by these enzymes, the magnitude of the change is generally small,13,14 with the
exception of the potentially major influence of grapefruit juice.18 The coadmin-
istration of certain drugs with grapefruit juice can markedly elevate drug bio-
availability and can alter both pharmacokinetic and pharmacodynamic parameters
of the drug.19–21
1.2 DIET INTERVENTION

Type 2 diabetes mellitus,2,22 obesity3 and metabolic syndrome,4,11 especially asso-
ciated dyslipidemia, are clinical entities where diet plays a key role in the overall
management.
1.2.1 HEALTHY DIET

The main objective of a healthy diet in patients with metabolic diseases is to
prevent weight gain, to limit postprandial glucose excursions and/or to reduce
the risk of cardiovascular disease, essentially by improving both lipid profile and
antioxidant status. The National Cholesterol Education Program — Adult Treat-
ment Panel III (NCEP-ATP III) recommendations for diet composition in patients
with metabolic syndrome are consistent with general dietary recommenda-
tions.11,23 Guidelines for healthy antiatherogenic diet call for (1) low intake of
saturated fats, trans fats and cholesterol; (2) reduced consumption of simple
sugars; and (3) increased intake of fruits, vegetables and whole grains. Such
principles also concern diet recommendations for the treatment and prevention
of diabetes mellitus and related disorders,2,22 and of overweight and obesity.3
A number of factors influence glycemic responses to foods, including the
amount of carbohydrates, type of sugar, nature of the starch, cooking and food
processing and food form, as well as other food components. Although it is clear
that carbohydrates do have differing glycemic responses, the data reveal no clear
trend in outcome benefits. If there are long-term effects on glycemia and lipids,
these effects appear to be modest.22
In persons with type 2 diabetes (on weight-maintenance diets), replacing
carbohydrates with monosaturated fats reduces postprandial glycemia and tri-
glyceridemia. However, there is concern that increased fat intake in ad libitum
diets may promote weight gain. Therefore, the contributions of carbohydrates
and monosaturated fat to energy intake should be individualized based on nutrition
assessment, metabolic profiles and treatment goals.2,22
Total energy intake remains the most important factor in dietary interventions
in obese individuals, but strategies based on emerging factors, such as glycemic
index, fiber intake, dietary variety and energy density, may improve efficacy and
tolerability of energy restriction.3
1.2.2 SPECIAL FOODS AND NUTRIENTS

1.2.2.1 Fibers
People are encouraged to choose a variety of fiber-containing foods, such as

whole grains, fruits and vegetables, because they provide vitamins, minerals and
other substances important for good health. Diets that include large amounts of
natural plant fiber delay gastrointestinal digestion and absorption of sugar and
complex carbohydrates, such as starch and dextrins. Early studies show that
ingestion of high-fiber foods improves glycemic control in diabetic patients.24
However, recent studies have reported mixed effects on glycemia and lipids in
patients with type 1 diabetes.22 In subjects with type 2 diabetes, it appears that
ingestion of very large amounts of fiber is necessary to confer metabolic benefits
on glycemic control, hyperinsulinemia and plasma lipids. It is not clear whether
the palatability and the gastrointestinal side effects of fiber in this amount would
be acceptable to most people. Dietary fiber, especially soluble fiber, theoretically
could influence energy balance by slowing gastric emptying, delaying digestion
and absorption of macronutrients, stimulating satiety hormones in the proximal
and distal small intestine, and increasing energy excretion. In intervention trials
in obese individuals, high fiber intake was associated with decreased body weight,
reduced hunger and increased satiety, and decreased short-term food intake.25
By binding water, cations and bile acids, or by forming gels that sequester
mono- or disaccharides, fiber-containing foods could modify the digestive as well
as the absorption process. Reduced absorption of some vitamins and minerals,
including iron, has been reported in subjects on high-fiber diets. However, despite
the recommendation to increase fiber content in individuals with metabolic
diseases, such as obesity, diabetes mellitus and metabolic syndrome, the effects
of fiber content on availability of drugs commonly used for treating such patients
have not been specifically investigated. One trial showed that guar gum decreases
the absorption of metformin in healthy subjects.26
1.2.2.2 Dietary Supplements and Herbal Products
Pharmaceutical research conducted over the past 3 decades shows that natural
products are a potential source of molecules for drug development, including for
diabetes treatment.27 Moreover, the use of botanical products has increased in
recent years. Herbal products are widely used as dietary supplements and are part
of complementary or alternative medicine. Importantly, herbal products are often
used concurrently with prescribed or over-the-counter medications. The use of
herbal products often escapes standard mechanisms for protecting persons from
harmful effects of drugs or drug interactions.28,29 Concurrent use of botanicals
with approved prescription drug products can result in therapeutic failures or
adverse events.30 Interactions of herbs with cytochrome P450 have been exten-
sively described.31 For instance, recent publications have shown that coadminis-
tration of St. John’s wort (Hypericum perforatum) decreases plasma levels of
several concomitant drugs (cyclosporin, indinavir, etc.), and the underlying mech-
anism appears to be induction of the intestinal CYP3A and P-glycoprotein.29,32
The potential effects of herbal products on drugs classically used in patients
with diabetes, obesity or metabolic syndrome are largely unknown. One Japanese
study reported that St. John’s wort treatment (300 mg tid [3 times per day] for
14 days) significantly reduced simvastatin area under the curve (AUC) by 50%,
but did not interfere with pravastatin pharmacokinetics.33 Thus, the potential for
clinically significant drug interactions between St. John’s wort with many
CYP3A4 substrates is a real possibility, one that may not be evident from routine
“screening” for traditional drug–drug interactions.29 The potent effects of herbal
products coupled with their risk of interactions with other commonly used drugs
and the lack of governmental regulation have created a public health dilemma
and encouraged the publication of a conference report.
1.2.2.3 Grapefruit Juice
The incidental discovery in a pilot study of a single volunteer demonstrating that

grapefruit juice dramatically elevates felodipine bioavailability has led to the
publication of numerous articles regarding the interaction between grapefruit juice
and various drugs, focusing on different aspects: interaction mechanisms, grape-
fruit juice constituents that are responsible for the interaction, drugs exhibiting
the interaction and the clinical relevance.19–21,29 An interaction between drug and
food or drink might reveal more difficulties than a drug–drug interaction and a
major difficulty is the question of who is responsible for warning the public.34
The potential interaction of grapefruit juice has been particularly investigated

with lipid-lowering agents (see section 1.3.3, Lipid-Lowering Agents). An exten-
sive review of the international literature does not reveal any specific report on
potential interactions of glucose-lowering agents with grapefruit juice despite the
fact that some recently launched compounds undergo hepatic metabolism at least
partially through CYP3A4.
1.3 PHARMACOLOGICAL INTERVENTION

1.3.1 GLUCOSE-LOWERING AGENTS
Various pharmacological classes can be used to lower blood glucose.6,7,35 Besides
classical sulfonylureas (chlorpropamide, glibenclamide, glipizide, gliclazide,
glimepiride, etc.) and biguanides (metformin), new agents have been launched
during recent years among which α-glucosidase inhibitors (acarbose, miglitol),
meglitinide derivatives (repaglinide, nateglinide) and thiazolidinediones — also
called glitazones — (pioglitazone, rosiglitazone) are included.7,36 Numerous
drug–drug interactions have been described with these compounds.37–39 For
instance, α-glucosidase inhibitors specifically act in the gastrointestinal tract and
potential interferences with intestinal bioavailability of some other drugs have
been reported, while meglitinide analogues and thiazolidinediones are metabo-
lized via the cytochrome P450 system and, thus, subject to metabolic interac-
tions.39 Surprisingly, less is known about the potential influence of food and
nutrients on the pharmacokinetic and pharmacodynamic effects of these glucose-
lowering agents. In a recent extensive review of food–drug interactions, nothing
described effects on oral glucose-lowering agents.15
1.3.1.1 Sulfonylureas
Sulfonylureas were the first oral drugs for treatment of type 2 diabetes in clinical
practice.40,41 They act at the pancreatic β-cell membrane by causing closure of
adenosine triphosphate (ATP)-sensitive potassium channels, which leads to an
enhanced insulin secretion. They are commonly prescribed alone or in combina-
tion with metformin, a thiazolidinedione, or an α-glucosidase inhibitor to reach
the glucose control targets.42 Various sulfonylureas have been, or are currently,
used in clinical practice. Those with the largest experience are tolbutamide and
chlorpropamide (first generation), and glibenclamide, glipizide, gliclazide and
glimepiride (second generation).43
The kinetic-dynamic relations of sulfonylureas are complex. The timing of
food intake, rather than the composition of diet, has been particularly analyzed
with the different sulfonylurea compounds. The objective is to accelerate sulfo-
nylurea gut absorption (leading to short tmax and high Cmax), to maximize meal-
related, early (first phase) insulin secretion and to reduce postprandial hypergly-
cemia.44 The rate of absorption of chlorpropamide or tolbutamide, but not its
extent, is decreased by food.45 Even if concomitant food intake may not delay
the absorption of tolbutamide, its efficacy may be improved if the drug is given
half an hour before meals rather than with meals.46 Chlorpropamide is more
slowly absorbed than tolbutamide and has the longest elimination half-life of all
sulfonylureas. Hence, the timing of the daily dose is less important, and it is
irrelevant whether chlorpropamide is ingested before or with breakfast.43 Prior
food intake has no influence on rate and extent of absorption of the classical form
of glibenclamide.47 Although concomitant food intake does not seem to delay the
absorption of glibenclamide, its efficacy may be increased if given before meals,
at least in the short term. Indeed, 2.5 mg given half an hour before breakfast was
more effective than 7.5 mg given together with breakfast.48
Food significantly delays absorption of gliclazide and peak concentrations
also may be reduced;49 however, it is not known whether intake before meals
makes gliclazide more effective than does intake together with meals.43 The
absorption of glipizide is also retarded by food,50,51 and is more rapidly absorbed
when taken before breakfast than when ingested together with breakfast. Intake
before breakfast is also associated with a more appropriate timing of insulin
release relative to meal, and with enhanced efficacy of the drug.51 Food delays
the absorption of a tablet of gliquidone only insignificantly.52 Glimepiride has an
absolute bioavailability of 100%,53 and there were no significant differences in
pharmacokinetic and pharmacodynamic responses between glimepiride given 30
min before or immediately after a meal in healthy subjects54 or between glime-
piride given 30 min or directly before breakfast in patients with type 2 diabetes.55
In clinical practice, glimepiride is usually prescribed once daily in the morn-
ing.56,57
Gliclazide is a weak acid with a good lipophilicity and a pH-dependent
solubility.58 Gliclazide is practically insoluble in acidic media and its solubility
increases as the pH becomes more alkaline. The absorption rate depends upon
the gastric emptying time and upon the dissolution rate in the small intestine
where the compound is soluble. The variability in absorption of gliclazide could
also be related to an early dissolution in the stomach leading to more variability
in the absorption in the intestine. From this, it can be anticipated that both delay
in gastric emptying and change in pH accompanying food intake might affect the
dissolution of the compound in the gastrointestinal tract. In two studies, food
altered the pharmacokinetics of gliclazide in type 2 diabetic patients.49,59 Two
mechanisms were thought to be involved: (1) delay in the absorption of gliclazide
when given 30 min after breakfast49 and (2) decrease in the extent of absorption
of gliclazide when given 30 min before a meal.59 Some decrease in gastric
emptying is noted; a possible interaction of gliclazide with dietary components,
as well as a change in pH, could explain these observations.
A new modified release (MR) formulation containing 30 mg of gliclazide
was developed to obtain a better predictable release of the active principle and
to allow once-daily dosing regimen at breakfast.60 In a food-effect study in healthy
volunteers, one tablet was given either fasted or 10 min after the start of a
standardized breakfast.61 No significant difference was observed in tmax, t1/2, Cmax
and AUC of gliclazide after administration of the 30 mg MR tablet under fasted
and fed conditions. Thus, the new MR formulation of gliclazide can be given
without regard to meals, i.e., before, during or after breakfast.
Chlorpropamide is less commonly used since the launch of second-generation
sulfonylureas. One reason for this change results from a specific adverse effect
— the chlorpropamide–alcohol flush.62 Indeed, facial flushing after alcohol occurs
much more frequently in type 2 diabetic patients taking chlorpropamide than in
those receiving other sulfonylureas. Both alcohol and chlorpropamide concentra-
tions are critical in determining the chlorpropamide–alcohol flush.63 The mech-
anism that produces the flush is poorly understood. It has been suggested that
chlorpropamide acts as a noncompetitive inhibitor of aldehyde dehydrogenase
resulting in increased acetaldehyde concentrations. There is, however, uncertainty
whether the flush can be explained solely by a rise in acetaldehyde concentrations.
Prostaglandin inhibitors, such as aspirin, have been shown to attenuate the flush
in some patients. Thus, alcohol intake should be avoided in diabetic patients
treated with chlorpropamide.
1.3.1.2 Metformin
Metformin is a biguanide compound that exerts complex metabolic effects.64

Biguanides inhibit absorption of various actively transported substances, such as
hexoses, amino acids, calcium and bile acids.65 This effect has been attributed to
an interaction with transport systems in the intestinal brush border. Enzyme
activities in intestinal epithelium also might be affected.65 However, the antihy-
perglycemic activity essentially results from a reduction of hepatic glucose pro-
duction, which leads to increased insulin action.66,67 Currently, this molecule is
considered as the first-choice drug in obese patients with type 2 diabetes and as
the first adjunctive drug to be added to sulfonylurea therapy in case of failure
with monotherapy.68 It may also be prescribed together with a glitazone, leading
to a clinically relevant additive, glucose-lowering effect.39
The absorption of metformin occurs mainly from the small intestine, and it
has been shown that high concentrations (10 to 100 times plasma levels) accu-
mulate in the walls of the gastrointestinal tract (review in Reference 69). There-
fore, possible pharmacokinetic interferences with food are not excluded. The oral
bioavailability of usual doses of metformin is 50 to 60%.69 Absorption of met-
formin is completed within 6 h. There might be a slight delay and decrease after
food intake,70 although this effect does not significantly influence overall meta-
bolic control in clinical practice. Gastrointestinal side effects are rather frequent
during metformin therapy, especially diarrhea. Such symptoms may be avoided
by gradual dose increases. The symptoms are generally transient and can be
reduced by taking the drug with food. One experimental study showed that the
absorption of metformin is reduced by concomitant intake of guar gum in healthy
subjects.26
The effect of metformin on vitamin B12 absorption has been demonstrated
using conventional Schilling tests, but was not confirmed by a whole-body tech-
nique.64 Serum folic acid concentration might also be affected by metformin,71
although this effect remains controversial.72 Vitamin B12 malabsorption was found
in 30% of diabetic patients during long-term metformin treatment.72,73 Although
it seems to be reversible, it may be persistent.74 Megalobastic anemia due to
vitamin B12 malabsorption associated with long-term metformin treatment has
been reported, but vitamin B12 treatment was successful in correcting such an
abnormality.75 The mechanism of biguanide-induced B12 malabsorption remains
uncertain, but it may be attributed to direct intestinal effects or bacterial over-
growth with binding of the intrinsic factor B12 complex.74
Alcohol potentiates the glucose-lowering and hyperlactatemic effect of bigu-
anides and should be limited in patients on chronic treatment with metformin.67
Alcohol itself decreases lactate oxidation and gluconeogenesis from lactate. The
inhibition of gluconeogenesis by alcohol also promotes hypoglycemia, especially
in the fasting state.
Diet–metformin interactions deserve further evaluation. In the Diabetes Pre-
vention Program,76 individuals with impaired glucose tolerance were randomized
to one of three intervention groups, which included the intensive nutrition and
exercise counseling (“lifestyle”) group or either of two masked medication treat-
ment groups — the biguanide metformin group or the placebo group. The latter
interventions were combined with standard diet and exercise recommendations.
After an average follow-up of 2.8 years, a 58% relative reduction in the progres-
sion to diabetes was observed in the lifestyle group, and a 31% relative reduction
in the progression to diabetes was observed in the metformin group, compared
with controlled subjects (placebo group). Considering these two remarkable pro-
tective effects provided by intensive lifestyle intervention on the one hand and
by metformin on the other hand, it would be interesting to investigate whether a
combination of the two interventions in the same at-risk population is able to
provide a further reduction in the incidence of type 2 diabetes.
1.3.1.3 Thiazolidinediones (TZDs)
TZDs are now widely used as part of antidiabetic treatments.77–79 These agents
act by targeting insulin resistance instead of stimulating insulin secretion. They
interact with the gamma type of the peroxisome proliferator-activated receptor
(PPAR-gamma).79,80 PPAR-gamma, a member of the nuclear receptor subfamily,
stimulates gene expression of proteins involved in glucose metabolism. This
results in an increase in insulin sensitivity in skeletal muscle, and adipose and
liver tissues. Two TZDs are currently commercialized and widely prescribed —
pioglitazone81,82 and rosiglitazone.83,84 Unlike sulfonylureas and metformin, TZDs
are not used commonly as first-line therapy. However, there is considerable
evidence for the incorporation of TZDs into combination regimens, with either
sulfonylureas or metformin, particularly in patients who do not achieve glycemic
goals with conventional regimens.85 Pharmacokinetic characteristics of both
pioglitazone86 and rosiglitazone87 have been described extensively. TZDS are
metabolized through the cytochrome P450 system and, thus, are subject to
pharmacokinetic interactions.88 While some drug–drug interactions have been
reported,39 to our knowledge, clinically relevant food–drug interactions have not

yet been described.
The effect of food on the pharmacokinetics of rosiglitazone was measured in
a single-dose crossover study. Rosiglitazone (2 mg) was administered to healthy
male volunteers either in the fasting state or following a standard high-fat break-
fast.89 Overall exposure to rosiglitazone, as assessed by AUC, was unaffected by
food. Absorption of rosiglitazone in the fed state was more gradual and sustained
than in the fasted state. Cmax was reduced by approximately 20% and tmax was
modestly delayed in the fed state. The absence of alterations in the extent of
absorption of rosiglitazone when administered with food permits rosiglitazone
administration without regard to meals for treatment of type 2 diabetes mellitus.
The effect of food on the pharmacokinetics of pioglitazone was evaluated in
a randomized, two-period crossover study in healthy volunteers.90 Subjects
received a single dose of 45 mg pioglitazone after a 10-h fast and 10 min after
a standardized meal. For fed and fasted states, the mean AUC for pioglitazone
was comparable — the relative bioavailability was 94 to 97% for pioglitazone
administered with food compared to a fasted state. The small differences in the
absorption rate of pioglitazone with and without food are not considered clinically
relevant; pioglitazone, like rosiglitazone, can be taken without regard to meal
timing.
1.3.1.4 α-Glucosidase Inhibitors
α-glucosidase inhibitors are pharmacological agents that competitively and

reversibly inhibit the α-glucosidase enzymes in the brush border of the small
intestine mucosa.91 There they prevent the hydrolysis of complex carbohydrates,
retard the absorption of glucose and limit postprandial hyperglycemia. Acarbose
is the most extensively investigated and widely prescribed α-glucosidase inhib-
itor,92-94 while miglitol95 and voglibose are two other compounds belonging to
this pharmacological class that are available only in a limited number of countries
(Japan for voglibose). While acarbose is an effective monotherapy for type 2
diabetes after diet failure, its distinct and complementary mechanism of action
induces a further improvement in glycemic control when it is used in combination
with other antidiabetic agents.91,96,97
All normal dietary carbohydrates, except for the relatively small amounts of
lactose and monosaccharides, are digested by the intestinal α-glucosidases in
saliva, pancreatic secretions and the enterocyte brush border. Inhibition of the
action of these enzymes by specific inhibitors results in a reduction or even an
abolition of hydrolytic activity. This is of great therapeutic importance for three
reasons. First, the carbohydrate composition of the diet eaten by patients taking
α-glucosidase inhibitors determines the substrate load for the various enzymes.
This matters because the degree of inhibition of the various α-glucosidases is
different for particular inhibitors and is dose-dependent, and the relationship
between the inhibitory profile and the balance of carbohydrate sources in the diet
is critical. This must be considered in assessing clinical response to treatment.
Second, if given in excessively high dosage, α-glucosidase inhibitors could result

in malabsorption of some carbohydrates through inhibition of digestion. Second-
ary to carbohydrate malabsorption, there may also be malabsorption of other
macro- and micronutrients due to intestinal hurry. Malabsorbed nutrients in any
quantity will provoke an osmotic diarrhea and cause associated symptoms. Third,
in the case of severe malabsorption induced by an α-glucosidase inhibitor, the
only carbohydrate that can be absorbed by the intestine must be in a form in
which no enzymatic digestion is required, i.e., monosaccharides and, in particular,
glucose. This has important implications for resuscitation in hypoglycemia
induced by other glucose-lowering agents given with an α-glucosidase inhibitor.
Thus, theoretically, diet composition may influence both the efficacy and tolera-
bility of α-glucosidase inhibitors. Selection of carbohydrates may influence acar-
bose glucose-lowering activity, while adequate nutrition may lead to a reduction
in adverse effects.98
Any use of an α-glucosidase inhibitor to improve diabetic control may depend
on the composition of the prescribed diet.98 In clinical trials, a few patients did
not benefit from treatment with α-glucosidase inhibitors, while most patients
showed a significant reduction in postprandial blood glucose concentrations.
When the dietary intake of the patients was analyzed, it could be demonstrated
that the nonresponders to α-glucosidase inhibitors had ingested very low amounts
of starch and very high amounts of simple sugars. In contrast, the patients who
had an appreciable benefit from treatment with acarbose or miglitol had ingested
reasonable amounts of complex carbohydrates and had preferred starchy foods
to simple sugars. Thus, a diet high in (complex) carbohydrates and low in simple
sugars, as recommended by diabetes associations,2,22 should be maintained by the
patients in order to achieve the greatest possible benefit from treatment with α-
glucosidase inhibitors. A Japanese study has suggested that to obtain the greatest
benefit from acarbose therapy, a strict adherence to high-carbohydrate (>50% of
energy intake) diet is required.99 However, the findings of a Canadian trial do not
support this theory, since the glycemic control improved during the treatment
despite the rather low carbohydrate intake in the acarbose group.100 Nevertheless,
in using these inhibitors, it is essential that consistent dietary guidelines are used,98
which should be in keeping with the general dietary recommendations.2,22
An appropriate diet can also minimize gastrointestinal symptoms that may
occur when fermentable carbohydrates reach the colon. Analysis of individual
dietary intake revealed that patients with persistent adverse effects during treat-
ment with α-glucosidase inhibitors had consumed either very high amounts of
beer or sweets. During treatment with α-glucosidase inhibitors, a reduction in
rapidly absorbable carbohydrates helps to prevent flatulence, abdominal discom-
fort or diarrhea.98 With an adequate diet and individualized acarbose doses (start-
ing with a low dose and followed by individualized titration), malabsorption
should not usually occur and adverse effects will be markedly reduced.
As diet can influence the gastrointestinal tolerance profile of α-glucosidase
inhibitors, one may suspect that the use of these drugs may influence the
composition of the diet in treated patients. Apparently, this is not the case. In a
double-blind, 24-week Finnish study, no significant differences in nutrient intakes

between acarbose-treated and placebo-treated diabetic patients were observed by
analyzing 4-d food diaries at 0, 4, 12 and 24 weeks.101 Also, the energy intake
and energy proportion of fat and carbohydrates remained unchanged in both
groups. Thus, the improvement in metabolic control in patients treated with
acarbose seems not to be a result of concomitant dietary changes. Similar findings
were seen in a pilot study of shorter duration102 and in a Canadian multicenter
trial.103
Acarbose is administrered to patients to modulate the absorption of sugars
and it has caused some degree of compensated carbohydrate malabsorption,
especially at higher dosages. There has been some interest to see whether acarbose
also affects the absorption and metabolism of other nutrients and minerals. In a
double-blind trial involving 24 patients with type 2 diabetes, acarbose 300 mg/d
for 10 weeks induced no significant alterations in the concentrations of important
electrolytes (including calcium and magnesium), iron, vitamin B12 and folic
acid.104 Thus, at least at this point, there is little evidence that acarbose negatively
affects other macro- or micronutrient availability.
1.3.1.5 Meglitinide Derivatives
Meglitinide analogues are nonsulfonylurea agents that are insulin secretagogues

and have a similar mechanism of action to the sulfonylureas, but with different
pharmacokinetic and pharmacodynamic properties.105 Repaglinide106,107 and
nateglinide108,109 belong to a new class of oral agents with a specific action on
early-phase insulin release, thus targeting postprandial hyperglycemia, and a
shorter elimination half-life and duration of action, therefore reducing the risk of
late hypoglycemia. The meglitinide derivatives, especially nateglinide, are ideally
suited for combination use with metformin or with TZDs. The pharmacokinetics
of repaglinide110,111 and nateglinide112,113 are well known. Both compounds are
metabolized through the P450 cytochrome system, which may lead to potential
metabolic interactions. Drug–drug interactions have already been described,39 but
information on possible food–drug interactions is scarce.
Co-administration with food has only minor effects on repaglinide pharma-
cokinetics. Compared with the fasted state, administration of repaglinide (2 mg)
15 min before a high-fat meal, to 24 healthy men, produced a slight reduction in
Cmax (−20%) and AUC (−12%). Conversely, the mean residence time increased
slightly (+21%) and tmax remained unchanged at 0.5 h.114 Therefore, repaglinide
may be administered preprandially without changing the rate of absorption. One
study investigated the effects of repaglinide, 1 mg before each meal, in maintain-
ing glycemic control in type 2 diabetic patients who either missed a meal or had
an extra meal.115 Meal-associated treatment with repaglinide provided good gly-
cemic control and was well tolerated, without significant hypoglycemic events,
irrespective of the number of meals consumed per day.
Results indicate that a synergistic interaction occurs between nateglinide and
elevated mealtime plasma glucose concentrations to stimulate insulin secretion.116
Significantly greater insulin secretion was observed when nateglinide was taken
before a meal compared to when given in the fasted state or in response to merely
a meal. When given before meals, nateglinide produced rapid and short-lived
insulin secretion, effectively reducing mealtime glucose excursions, yet lowering
the risk of hypoglycemic episodes.
When nateglinide was given to healthy subjects 10 min before ingestion of
a high-fat meal, the rate of absorption was increased relative to when it was given
during a continued fast, as evidenced by a 12% increase in Cmax and a 52%
decrease in tmax.117 However, there was no significant effect on the extent of
absorption as assessed by the AUC. Alternatively, when nateglinide was given
after the meal, a food effect was observed that was characterized by a decrease
in the rate of absorption: a 34% decrease in Cmax and a 22% increase in tmax, but
no significant effect on AUC. Regardless of timing, the combination of a meal
and nateglinide produced a larger increase in plasma insulin levels than did
nateglinide alone.
The influence of timing of administration of nateglinide on the glucose profile
and β-cell secretory response to a standardized test meal was studied, as well as
the effect of meal composition on the pharmacokinetic and pharmacodynamic
profile.118 Nateglinide administration 10 min before a meal resulted in a more
rapid rise and a higher peak nateglinide plasma concentration than when it was
given 10 min after the start of the meal, irrespective of the meal composition
(i.e., high in carbohydrates, fat or protein). Preprandial administration of nateg-
linide was more effective in reducing prandial glucose excursions, compared with
post-meal dosing, a consequence of the earlier insulin response. Meal composition
had no effect on the plasma nateglinide profile, which is of particular importance
because nateglinide is a D-phenylalanine derivative and, thus, there was the
possibility that its absorption could be influenced by the presence of high amino
acid concentrations. The findings of this study clearly demonstrated that, even in
the presence of a high-protein meal (32% of calories derived from protein), no
effect on absorption of nateglinide was observed.
1.3.2 ANTIOBESITY AGENTS

During recent years, interest has intensified in two unique antiobesity preparations
— the pancreatic lipase inhibitor, orlistat, and the noradrenaline and serotonin
re-uptake inhibitor, sibutramine.8,9,119
1.3.2.1 Orlistat
Orlistat, a semisynthetic derivative of lipstatin, is a potent and selective inhibitor

of gastric and pancreatic lipases.120 When administered with fat-containing foods,
it partially inhibits the hydrolysis of triglycerides, thus reducing the subsequent
absorption of monoglycerides and free fatty acids. Orlistat treatment results in a
dose-dependent reduction in body weight in obese subjects, with an optimal
dosage regimen of 120 mg tid, and is generally well tolerated, with some intestinal
side-effects during the first few days or weeks of administration.120 Several 1-

and 2-year trials showed that orlistat, when used with a health-promoting, low-
fat and moderately energy-restricted diet, confers some advantages in the long-
term management of obesity.121 Clinical trials performed in obese subjects with
type 2 diabetes demonstrated that orlistat (120 mg tid) slightly increased weight
loss and resulted in a significant reduction in glycated hemoglobin levels in
patients treated with sulfonylureas, metformin or even insulin, as compared to a
placebo.122 In all of these clinicial studies, orlistat proved to be effective only
when its use was combined with a reduced-calorie diet (600 to 800 kcal/day
deficit) containing 30% of calories from fat. Interestingly, treatment with orlistat
plus lifestyle changes resulted in early and sustained improvements in cardiovas-
cular risk factors, including waist circumference, glucose/insulin parameters,
blood pressure and lipids, all parameters belonging to the metabolic syndrome.
A 37% significant reduction in the incidence of new cases of type 2 diabetes was
also reported after 4 years of treatment with orlistat as compared to a placebo in
obese subjects.123 The effects of orlistat on weight and on serum lipids in obese
patients with hypercholesterolemia were investigated in a randomized, double-
blind, placebo-controlled, multicentre study.124 A 10% additional specific choles-
terol-lowering effect of orlistat was demonstrated, independent of weight reduc-
tion, in obese patients already on a lipid-lowering hypocaloric diet.
In normal, healthy volunteers consuming 60 g/day of fat, orlistat (360
mg/day) recipients excreted 21.5 g/day more fecal fat than placebo recipients.125
Approximately 32% of ingested fat was lost in the feces of orlistat recipients vs.
4.4% in placebo recipients, and this percentage does not change significantly
when increasing the dosage of orlistat to 600 mg/day or the amount of dietary
fat from 60 to 76 g/day. Dietary fiber content (28 vs. 10 g/day) did not influence
fecal fat excretion after orlistat (80 mg q3d) in healthy male volunteers.126 Over
an 8-day period, the effect of orlistat (80 mg q3d) on fecal fat excretion was not
influenced by administration time relative to meals in an experimental study in
which the agent was given to 24 healthy volunteers mid-meal or 1 or 2 h after
the meal.127
Adverse events observed with orlistat therapy result from the inhibition of
fat absorption rather than the drug itself. They include abdominal pain, fatty/oily
evacuation and, occasionally, fecal incontinence.120 Orlistat (75 mg/day) was
“better tolerated” by healthy volunteers when the dietary fat content was reduced
from 130 to 110 g/day.125 However, adverse events were similar in placebo and
orlistat recipients when dietary fat was reduced from 130 to 45 g/day and the
agent was administrered 2 h before food (i.e., with little or no fat content in the
stomach).120
Orlistat inhibits the absorption of ingested fats. It is possible, therefore, that
endogenous levels of fat-soluble vitamins (A, D, E and K vitamins) may become
depleted with prolonged use of the agent.120 Orlistat (360 mg/day) for 9 days,
compared with a placebo, decreased the maximum plasma concentrations and
the AUC-time curve values of oral vitamin E acetate supplementation (400 IU
given on Day 4 of orlistat administration) by 43% and 60%, respectively. This
suggests that short-term administration of orlistat may compromise vitamin E

absorption from the gut. Conversely, the pharmacokinetic profile of oral vitamin
A acetate supplementation (25,000 IU given on Day 4 of orlistat administration)
was not significantly affected.128 Small, clinically insignificant decreases in serum
levels of vitamins A and D were seen in obese patients after 12 weeks of treatment
with a placebo or orlistat 30 to 360 mg/day.129 In contrast, vitamin E levels were
significantly reduced with orlistat 180 and 360 mg/day and increased with a
placebo (all, p <0.01). Approximately 60% of a supplemental dose of β-carotene,
a major dietary source of vitamin A, was absorbed when a single 30 to 120 mg
dose was given (on Day 4) to healthy volunteers receiving 360 mg/day orlistat.130
This may be sufficient to achieve physiological levels of β-carotene in obese
patients who may develop reduced levels of this vitamin during orlistat therapy.
Analysis of biological results in obese patients who received 360 mg/day orlistat
or a placebo for up to 2 years in a large European multicenter clinical trial,
revealed that plasma levels of vitamins D and E and β-carotene were significantly
reduced in orlistat compared with placebo recipients; the levels, however,
remained within the normal clinical range.131 Those results were confirmed in
further large, long-term clinical trials performed in Europe and in the U.S. None
of the trials reported bone density or bone mineral changes.121
1.3.2.2 Sibutramine
Sibutramine, a noradrenaline and 5-hydroxytryptamine re-uptake inhibitor, has

been shown to produce a dose-related weight loss in obese subjects, with optimal
doses of 10 to 15 mg/day.132 The multicenter prospective “Sibutramine Trial of
Obesity Reduction and Maintenance” (STORM) clinical study showed that almost
all obese patients who persisted with the management scheme combining
restricted diet and sibutramine (10 mg/day) can achieve at least a 5% weight loss,
and over half can lose more than 10% of body weight within 6 months.133
Furthermore, sustained weight loss was maintained in most patients continuing
therapy with sibutramine for 2 years, whereas weight regain was noticed in most
patients randomized to a placebo, thus demonstrating that sibutramine (10 to 20
mg/day) favors weight maintenance in the long term. The clinical effectiveness
of sibutramine was demonstrated in several other clinical trials in nondiabetic
and diabetic obese individuals.122,134 In all of these trials, patients were prescribed
a 600 kcal deficit diet based on a macronutrient content of <30% fat and 15%
protein.133
Mean total daily energy intake was significantly reduced with sibutramine 10
or 30 mg/day compared with a placebo in obese women who were not attempting
to lose weight.135 After a 2-week treatment period, energy intake was 18.8 and
26.1% lower with sibutramine 10 and 30 mg/day, respectively, than with a pla-
cebo. Correspondingly, food intake was 13.9 and 27.7% lower. With respect to
individual macronutrients, the total daily intake of both fat and protein was
significantly reduced by the two dosages of sibutramine; there was no significant
effect on daily carbohydrate intake. Overall, there was no significant effect on
the percentage of energy from macronutrients consumed during each treatment

phase, although both fat and carbohydrate intake were up to 12.4% lower after
sibutramine than after a placebo.
Similar effects were seen in healthy, nonobese males who received 15 mg/day
sibutramine or a placebo, in a randomized, double-blind, crossover trial.136
Sibutramine significantly reduced total energy, protein, fat and carbohydrate
intake vs. a placebo. Although the overall proportion of macronutrients did not
differ significantly between treatment groups, lunch-time fat and protein, but not
carbohydrate, intake were reduced, and dinner-time carbohydrate, but not fat or
protein, intake was reduced with sibutramine compared with a placebo. However,
in a large group of obese patients on a calorie-reduced diet, carbohydrate and
protein intake were increased by 4.8 and 36%, respectively, from baseline, and
fat intake was reduced by 7.8% after sibutramine (10 mg/day) for 6 months.137
Available published pharmacokinetic data for sibutramine are limited to infor-
mation from abstracts. The bioavailability of the drug over the dose range of 10
to 30 mg was not altered by the presence of food in healthy volunteers.138 The
drug undergoes extensive first-pass metabolism to form pharmacologically active
primary (M1) and secondary (M2) amine metabolites.132 The influence of food
composition on this process remains unknown.
1.3.3 LIPID-LOWERING AGENTS

Most obese individuals (especially those with abdominal adiposity), subjects with
the metabolic syndrome and patients with type 2 diabetes have plasma lipid
disturbances leading to atherogenic dyslipidemia (moderately elevated total and
LDL cholesterol levels, low HDL cholesterol concentration and both fasting and
postprandial hypertriglyceridemia) and increased cardiovascular morbidity and
mortality. Again, healthy diet is essential to improving lipid profile of these
individuals.23 However, lipid-lowering drugs may also be useful and should be
prescribed to numerous high-risk persons in order to reach target lipid levels.139,140
Statins have proven their efficacy in reducing the incidence of cardiovascular
complications in numerous primary and secondary prevention trials.141 The pri-
mary effect of this pharmacological class is a specific inhibition of cholesterol
synthesis in the hepatocytes leading to an impressive reduction in total and LDL
cholesterol levels, although various pleiotropic effects also appear to play a role
in the overall protective cardiovascular effect.142 A recent clinical guideline from
the American College of Physicians recommends lipid-lowering therapy with a
statin in almost all patients with type 2 diabetes.143 Fibric acid derivatives (fibrates)
are also widely prescribed in diabetic patients because these drugs have a positive
impact on diabetic dyslipidemia characterized by low HDL cholesterol and high
triglyceride concentrations.144
1.3.3.1 Statins
The hydroxymethylglutaryl coenzyme A (HMG CoA) reductase inhibitors

(statins) are well tolerated apart from two uncommon but potentially serious
adverse effects: asymptomatic elevation of liver enzymes and skeletal muscle

abnormalities, which range from benign myalgias to life-threatening rhabdomy-
olysis.139 Adverse effects with statins are frequently associated with drug inter-
actions because of their use in patients who are likely to be exposed to polyp-
harmacy. The cytochome P450 enzyme system plays an important part in the
metabolism of most statins (except pravastatin), leading to clinically relevant
interactions with other pharmacological agents.145–148 Alternatively, food interac-
tion may also be suspected, especially an inhibition of metabolism by grapefruit
juice, a potent CYP inhibitor.19–21
The various HMG CoA reductase inhibitors have very different chemical and
pharmacokinetic properties. The bioavailability of lovastatin increases by 50%
when taken with a regular meal; this is reflected in an increased drug effect.149
However, the ingestion of fibers or fruit as part of a lipid-lowering diet may
strikingly reduce the absorption of lovastatin and may increase the risk of treat-
ment failure.150 In contrast to lovastatin, the bioavailability of pravastatin is
reduced by 31% when taken with food; however, since its lipid-lowering efficacy
is unchanged, the interaction is not clinically important.151 For atorvastatin152,153
and fluvastatin,154 bioavailability and lipid-lowering efficacy are unaffected by
food intake. It seems to be also the case with simvastatin and rosuvastatin,
although no published study is available to support this statement.
Excessive ingestion of grapefruit juice increases the bioavailability of lova-
statin, atorvastatin and simvastatin by 1400, 200 and 1500%, respectively. This
may lead to drug accumulation and the possible development of adverse
effects.155–157 Thus, concomitant use of grapefruit juice, at least in large amounts,
and lovastatin or simvastatin should be avoided, or the dose of the statin should
be greatly reduced. However, daily consumption of a glass of regular-strength
grapefruit juice has only a minimal effect on plasma concentrations of lovastatin
(30 to 40% increase) after a 40 mg evening dose of lovastatin.158 The probable
mechanism of this interaction is inhibition of CYP3A4-mediated first-pass metab-
olism of these statins by grapefruit juice in the small intestine and, thus, inhibition
of their first-pass metabolism. Pravastatin and fluvastatin are not metabolized by
CYP3A4 and, consequently, are not subject to drug–grapefruit juice interac-
tions.156,159 No study investigating the possible interaction of grapefruit juice with
rosuvastatin is available, but such interaction appears to be unlikely as rosuvastatin
is only minimally metabolized by the CYP450 enzyme system with no significant
involvement of the 3A4 enzyme.160 Thus, it would be wise to avoid the combi-
nation of certain statins (especially simvastatin and lovastatin) with grapefruit
juice.21 Indeed, the concomitant intake of such statins with CYP3A4 inhibitors
may exacerbate adverse effects, such as rhabdomyolysis and acute renal failure.146
1.3.3.2 Fibrates
The two most commonly used fibric derivatives are gemfibrozil in the U.S.161,162
and fenofibrate163–165 in Europe. Fibrates are metabolized by the hepatic cyto-
chrome P450 (CYP) 3A4 and, thus, are subject to drug interactions.166
Gemfibrozil is rapidly and completely absorbed when orally administered.161

In vitro results suggest that solubility and dissolution of the drug may be increased
in the fed state compared with the fasting state,167 although clinical studies are
needed to verify this. Whereas several important drug–drug interactions have
been reported with gemfibrozil, especially with cerivastatin and repaglinide,39 no
one report is available concerning the potential interactions between food and
gemfibrozil, especially grapefruit juice that contains inhibitors of the CYP P450
system.19–21,159
The Cmax of fenofibric acid occurs within 6 to 8 h after fenofibrate adminis-
tration, and the absorption of fenofibrate is increased when administered with
food.168 With the microcoated tablets, the extent of absorption is increased by
approximately 35% under fed as compared with fasting conditions.169 The fat
content of a meal eaten at the time of fenofibrate administration does not have a
marked effect on pharmacokinetics. When micronized fenofibrate 200 mg cap-
sules were administered with a high-fat meal, the AUC (+15%) and the Cmax
(–2%) were only slightly altered, while the elimination half-life was shortened
(–41%), compared when fenofibrate was taken with a low-fat meal.166 The clinical
relevance of these pharmacokinetic changes remains unclear.
1.3.3.3 Resins
Resins (cholestyramine and colestipol) act by binding bile salts in the gastrointes-
tinal lumen.139 Both compounds are large polymers that are not absorbed system-
ically. They bind bile acids by exchanging them for chloride ions; binding is
irreversible. The bound bile salts are not reabsorbed, which leads to a depletion
of the bile acid pool. The liver compensates by increasing synthesis of bile acids,
which results in intracellular cholesterol depletion leading to increased expression
of LDL receptors on the hepatocyte cell membrane and, thereby, increased LDL
clearance. These drugs are given in the form of a powder, which is usually taken
with meals as a suspension in juice or water. Compliance is the major problem
on the grounds of palatability and adverse gastrointestinal effects. The absorption
of several drugs may be impaired by resins, including statins, digoxin, amio-
darone, thyroxine, warfarin, thiazide diuretics, and beta blockers. Reduced
absorption of folic acid and fat-soluble vitamins may occur, but clinical deficiency
is rare. Resins are less commonly used since the expansion of statin prescription
and the recent launch of ezetimibe, a selective inhibitor of cholesterol intestinal
absorption.
1.3.3.4 Ezetimibe
Ezetimibe is the first of a new class of antihyperlipidemic agents, the cholesterol-

absorption inhibitors. It is indicated for use as monotherapy or in combination
with a statin as an adjunct to dietary treatment for the reduction of elevated total
cholesterol, LDL cholesterol and apo B in patients with primary hypercholester-
olemia.170,171 Although one study found that the oral bioavailability of ezetimibe
did not appear to be substantially affected by the intake of high-fat or nonfat

meals,172 other data suggest that the oral bioavailability of ezetimibe may be
increased by 25 to 35% with food and that plasma concentrations of ezetimibe
may also be increased by up to 38% when the drug is taken with a high-fat
meal.173 Even if food may have a substantial effect on bioavailability of the
compound, this effect is not expected to alter the efficacy or safety profile of
ezetimibe because the drug is not extensively absorbed.170,171 The LDL-lowering
effect of ezetimibe according to dietary fat and cholesterol intake was investigated
in a large multicenter, double-blind, randomized, placebo-controlled study in
1719 men and women (only reported in abstract form, Reference 174). When
stratified according to dietary fat intake or daily cholesterol intake, all ezetimibe
subgroups had significantly greater reductions in LDL-cholesterol levels com-
pared with the respective placebo subgroups. These results suggest that ezetimibe
can reduce LDL-cholesterol levels regardless of dietary fat or cholesterol intake.
1.3.3.5 Sterols and Stanols
Plant sterols have a moderate hypocholesterolemic effect. This is thought to stem

from inhibition of the absorption of both exogenous dietary and endogenous
biliary cholesterol from the distal small intestine. In recent years, research focus
has shifted from plant sterols to plant stanols for reasons of safety and efficacy.175
Especially, esterification of plant stanols with fatty acids from vegetable oil has
made it possible to produce spreads, other foods and supplements containing
plant stanols. Food preparations that provide 3.4 to 5.1 g/day of plant stanol
esters, of which the plant stanol component is 2 to 3 g/day, significantly reduce
serum total and LDL cholesterol levels without affecting HDL cholesterol or
serum triglycerides, and are well tolerated. As both stanol esters and statins can
be used in combination to accentuate the cholesterol lowering effect, it would be
interesting to investigate the potential interactions between these nutrients and
the pharmacological agent. Plant stanol esters block the absorption of cholesterol,
whereas statins block the synthesis of cholesterol. Both mechanisms work in a
complementary or concerted fashion to amplify LDL receptor activity and the
clearance of LDL cholesterol from the plasma. A randomized, double-blind,
clinical trial compared the effect of plant sterol ester spread (5.1 g/day) with a
placebo spread on cholesterol in patients taking statin therapy, but who still had
elevated LDL cholesterol.176 As compared to the placebo, plant stanol ester spread
induced a greater reduction in total cholesterol (–7%) and LDL cholesterol
(–10%) at 8 weeks. No significant adverse events were noted. Thus, an incre-
mental lowering of LDL cholesterol by about 10% was achieved by adding a
stanol ester spread to the daily diet in persons taking a stable dose of a statin,
and this effect was seen in all four statins (atorvastatin, lovastatin, pravastatin
and simvastatin) included in this study. Preliminary observations confirmed these
results in patients after cardiac transplantation.177 Seventeen stable cardiac trans-
plant recipients, of whom 16 were on statin therapy, used margarine with
stanol/sterol esters. Total cholesterol was lowered by 17% and LDL cholesterol
was reduced by 22%, allowing a reduction in statin dosages in some patients.

The tolerance of the combination was good.
1.4 CONCLUSIONS
Metabolic disorders generally require combined dietary and pharmacological
interventions. While food–drug or nutrient–drug interactions might be of partic-
ular importance in patients with metabolic diseases, only scarce data are available
in the scientific literature, as compared to the overwhelming information regarding
drug–drug interactions. Obviously, food composition may influence the efficacy
of drugs affecting glucose, lipid and energy metabolism. In addition, since meal-
related kinetics is crucial in controlling postprandial blood glucose in diabetic
patents, the influence of food on drug bioavailability may also be important,
especially for drugs that stimulate insulin secretion. Furthermore, meal content
and/or timing may also influence drug gastrointestinal tolerability, as described
with metformin, α-glucosidase inhibitors and orlistat. Finally, as food may inter-
fere with CYP P450, potential food–drug interactions may be suspected in using
pharmacological agents metabolized via the cytochrome system. A clear interac-
tion with grapefruit juice was described with statins metabolized via CYP3A4,
especially lovastatin and simvastatin. From a public health point of view, an
interaction between drug and food or drink might reveal more difficulties than a
drug–drug interaction. It is important that the awareness for this potential
food–drug interaction will increase, and actions must be taken in order to increase
the efficacy of diet–drug combination in metabolic diseases and to prevent
undesired and harmful clinical consequences.
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DK5836_C002.fm Page 31 Monday, May 22, 2006 12:37 PM
2 Hypolipidemic Therapy:
Drugs, Diet, and
Interactive Effects
Alexandre Loktionov
CONTENTS
2.1 Introduction................................................................................................31
2.2 Therapy of CVD with Hypocholesterolemic and
Hypolipidemic Drugs ...............................................................................32
2.2.1 HMG-CoA Reductase Inhibitors (Statins)....................................33
2.2.2 Fibric Acid Derivatives (Fibrates).................................................34
2.2.3 Bile Acid Transport Inhibitors ......................................................35
2.2.4 Niacin (Nicotinic Acid) .................................................................36
2.2.5 Cholesterol Absorption Inhibitors .................................................37
2.2.6 Emerging and Future Directions in Hypolipidemic Therapy .......37
2.3 Dietary Factors in Lipid Homeostasis and Their Interactions with
Hypolipidemic Drugs ................................................................................38
2.3.1 Dietary Ingredients Possessing Natural Hypolipidemic and
Cardioprotective Properties ...........................................................39
2.3.2 Interactions between Dietary Components and
Hypolipidemic Drugs ....................................................................44
2.4 Conclusion .................................................................................................48
References ...........................................................................................................49
2.1 INTRODUCTION
In the past, the relationship between dietary factors and cardiovascular disease
(CVD) was often regarded as primarily and almost exclusively dependent on lipid
(especially saturated fat and cholesterol) consumption and metabolism leading
to increased serum cholesterol; in particular, low density lipoprotein (LDL) levels
resulting in atherosclerotic vascular changes. This idea constituted the core of
the traditional “diet–heart” hypothesis.1 With the development of scientific knowl-
edge on both CVD pathogenesis and bioactive properties of nutrients, the field
is rapidly becoming more and more complicated. The range of known pathways
31
and mechanisms contributing to the development of atherosclerosis and CVD has

considerably expanded.2–5 Indeed, the term CVD is now equally applied to dif-
ferent vascular events occurring in different vascular locations (coronary heart
disease, cerebrovascular disease, peripheral vascular disorders).5 Consequently,
there is a definite trend to regard a complex of multiple factors and conditions
predisposing to atherosclerosis development and its cardiovascular manifestations
(lipid-related and general metabolic disorders, inflammation, high blood pressure,
increased blood coagulation, etc.) as an extended area of active CVD prevention
and treatment. The metabolic syndrome, which is defined as a combination of
such common disorders as obesity, insulin resistance, glucose intolerance, hyper-
tension, and dyslipidemia manifested by hypertriglyceridemia and low high den-
sity lipoprotein (HDL) cholesterol levels,6–8 has already become highly prevalent
in the U.S.9 and is regarded as a warning sign of a forthcoming new CVD
epidemic.7,8
In addition to known links between nutrition and atherosclerosis, it became
clear that all conditions constituting the metabolic syndrome are diet-dependent;
therefore, assessment of CVD risk modulation by food ingredients requires a
thorough consideration of multiple overlapping effects of nutritional influences
at the level of different mechanisms contributing to the development of CVD.
Additional emerging factors in CVD pathogenesis, such as hyperhomocysteine-
mia,5,10,11 inflammation,5,12,13 oxidative stress,5,14–16 and others,5,10 are also affected
by diet, thus the area of dietary modulation of CVD risk looks extremely complex.
The complexity becomes even higher when problems of CVD treatment with
pharmaceutical agents, nutritional preventive/therapeutic approaches, and
drug–diet interactions are considered. For this reason, separate analysis of dif-
ferent aspects of the problem appears to be justified.
This chapter is going to be devoted mostly to the analysis of hyperlipidemia
and dyslipidemia treatment using both medicamentous therapy and dietary mod-
ifications. Emerging problems associated with the introduction of combined prob-
lems are also addressed.
2.2 THERAPY OF CVD WITH HYPOCHOLESTEROLEMIC

AND HYPOLIPIDEMIC DRUGS
Persistent hyperlipidemia and dyslipidemia is regarded as the leading pathoge-
netic factor in the development of atherosclerosis and CVD. Recent progress in
the field of therapeutic management of hyperlipidemia has resulted in the emer-
gence of a number of potent hypolipidemic agents. Currently several main groups
of drugs employed for dyslipidemia treatment can be defined as:
1. 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase

inhibitirs (statins)
2. Fibric acid derivatives (fibrates)
3. Bile acid transport inhibitors
Hypolipidemic Therapy: Drugs, Diet, and Interactive Effects 33
4. Niacin (nicotinic acid)

5. Cholesterol absorption inhibitors17
Their mechanisms of action and clinical characteristics are briefly considered

below.
2.2.1 HMG-COA REDUCTASE INHIBITORS (STATINS)

Suppression of cholesterol biosynthesis by HMG-CoA reductase inhibitors
derived from fungal metabolites was discovered in 1976.18 Since that time, the
family of statins has expanded and now includes natural products of fungal origin
(lovastatin, pravastatin), a semisynthetic derivative (simvastatin), and entirely
synthetic drugs (fluvastatin, atorvastatin, rosuvastatin, pitavastatin).17,19,20 Ceriv-
astatin was withdrawn from clinical practice in 2001 because of adverse effect
risk.21
The main hypolipidemic effect of statins is exerted through competitive
inhibition of the binding of HMG-CoA at the catalytic site of HMG-CoA reduc-
tase, the enzyme catalyzing the conversion of HMG-CoA to mevalonate, a sterol
precursor important for cholesterol synthesis.19 Reduced cholesterol biosynthesis
results in up-regulation of LDL receptors in the liver and stimulation of LDL
clearance from the plasma.22 It has also been reported that statins decrease hepatic
production of very low density lipoproteins (VLDL) and increase catabolism of
VLDL remnants.23 It is, however, apparent now that in addition to direct suppres-
sion of lipid biosynthesis, statins influence multiple pathogenetic mechanisms
involved in the development of atherosclerosis, CVD, and other chronic condi-
tions, especially those related to the metabolic syndrome.23 Figure 2.1 schemat-
ically presents pleiotropic beneficial effects of statins. Detailed consideration of
mechanisms involved in such effects is beyond the scope of this chapter. Interested
readers can be addressed to several recent reviews in the field.15,17,24–26
Statins are presently the most widely used hypolipidemic drugs. Several
clinical trials confirmed their efficiency in both blood lipid lowering and cardio-
vascular risk reduction with regard to coronary heart disease (CHD),27–31 cere-
brovascular disease,30–33 and cardiovascular complications of diabetes melli-
tus.30,31,34,35 Decrease of the progression of atherosclerosis and even regression of
atherosclerotic arterial changes have also been reported.19,26
Drugs of this group are usually well tolerated, although such side effects as
hepatotoxicity, myotoxicity, and nephrotoxicity have been described.19 Among
other statins, cerivastatin was associated with much higher risk of rhabdomyol-
ysis, especially when applied in combination with gemfibrozil.21 Carcinogenicity
of statins in experimental rodents has been described;36 however, no evidence of
this effect in humans is available. Statins are metabolized by enzymes of the
cytochrome P450 (CYP) family. Among them, CYP3A4 is especially important
for several statins. As this enzyme is known to be involved in the metabolism of
many other drugs and some dietary components, a background for interactive
CHOLESTEROL
BYOSYNTHESIS
INHIBITION
(SMALL LDL
ANTIOXIDANT REDUCTION)
PLAQUE
EFFECTS STABILIZATION
(REDUCTION OF
oxLDL)
ANTITHROMBOTIC VASOMOTION
EFFECT
STATINS NORMALIZATION
INCREASE
OF ENDOTHELIAL
NITRIC OXIDE FUNCTION
BIOAVAILABILITY IMPROVEMENT
INHIBITION
OF
INFLAMMATION
FIGURE 2.1 Pleiotropic beneficial effects of statin therapy. (Dotted arrows indicate inter-
active links between different mechanisms.)
effects exists.17 Dietary influences on statin effects are possible and will be
considered later in this chapter.
2.2.2 FIBRIC ACID DERIVATIVES (FIBRATES)

The clinical benefit of the use of fibrates is related primarily to the reduction of
triglyceride levels and, to some extent, the elevation of blood HDL cholesterol
concentration.37–39 The principal mechanism of fibrate action is defined by their
ability to mimic the biological function of free fatty acids (FA) through the specific
binding to peroxisome proliferator-activated receptor α (PPARα). PPARα activa-
tion affects expression of multiple genes, products of which are involved in lipid
metabolism. Hypolipidemic action of fibrate-activated PPARα is exerted through
several regulatory pathways, predominantly acting in the liver, including stimulation
of FA uptake, their cellular retention, and modulation of the expression of coenzyme
A synthetase catalyzing the esterification of FAs. Also, activated PPARα induces
mitochondrial FA uptake and catabolism as well as the activity of lipoprotein lipase
(LPL), while inhibiting the expression of its antagonist apoC-III.39,40
The role of activated PPARα in HDL level modulation is less clear; however,
it is believed that induction of the transcription of ApoA-I and Apo A-II lipopro-
teins in hepatocytes may contribute to the increase of plasma HDL concentrations
and a more efficient reverse cholesterol transport.39,40 Given that PPARs are known
to exert multiple biological effects, it is not surprising that a growing body of
information regarding pleiotropic effects of fibrates now emerges. Antiinflamma-
tory action and favorable effects on both blood coagulation and fibrinolysis
system40 look especially interesting from a clinical point of view. It is also
important to stress that other members of the PPAR family, especially PPARγ,
are deeply involved in lipid metabolism regulation. PPARγ agonists of the thia-
zolidinedione family are already in use for the treatment of diabetes mellitus;
however, their hypolipidemic potential has recently emerged as a promising sign
for the development of new approaches to CVD treatment.41 Perspectives of
developing new drugs combining PPARα and PPARγ agonist activity39 may
provide a new step in this direction.
Four members of the fibrate family are now in wide clinical use: bezafibrate,
ciprofibrate, fenofibrate, and gemfibrosil.37 Drugs of this group are especially
efficient in combination with statins or as monotherapy in patients with normal
LDL cholesterol levels, in particular for the treatment of cardiovascular manifes-
tations of the metabolic syndrome.37,42–44 Although fibrates are demonstrated to
be well tolerated in most cases, a few side effects, including changes of hepatic
function (hepatic transaminase level elevation), gastrointestinal side effects, and
myopathy have been reported.45,46 Some effects have been associated with par-
ticular members of the fibrate family, like rhabdomyolysis with gemfibrozil35,47
or hyperhomocysteinemia with fenofibrate.48 Like statins, fibrates have been
shown to be carcinogenic in rodents;36 however, there is no clinical information
on this potential problem. It has also been repeatedly stressed that combined
hypolipidemic therapy with statins and fibrates may be associated with drug
interactions and higher risk of adverse effects.35,49 Dietary influences should be
taken into account in this context and will be discussed later in this chapter.
2.2.3 BILE ACID TRANSPORT INHIBITORS

This heterogenous group of drugs’ principle of action is based on the fact that
blocking the intestinal reuptake of bile acids substantially increases the loss of
bile acids with feces. The loss is compensated in the liver through conversion of
cholesterol to bile, which results in the depletion of hepatocyte cholesterol and
upregulation of LDL receptors with hypocholesterolemic effect.17
Bile acid-binding resins/sequestrants represent the only clinically important
group of drugs inhibiting bile acid reuptake. Initially introduced agents,
cholestyramine and colestipol, had gastrointestinal side effects, such as bloating,
flatulence, heartburn, and nausea.50–52 For this reason their use is presently
restricted to application of low doses in multidrug treatment schemes. A new
member of this family, colesevelam, is much better tolerated and is now recom-
mended as an effective alternative to the two older drugs.51–53 Colesevelam, which
forms nonabsorbable complexes with bile acids in the gastrointestinal tract, has
been reported four to six times more potent than other bile acid sequestrants, and
especially recommended for combination therapy with statins.53
Another family of new drugs affecting bile acid transport is represented by
agents inhibiting ileal sodium-dependent bile acid transporter (IBAT), which
reduce serum cholesterol level by suppressing the reuptake of bile acids in the
ileum. Results from animal experiments with a few synthetic compounds have
provided encouraging results;54–56 however, drugs of this family have not hitherto
been introduced into clinical practice.
2.2.4 NIACIN (NICOTINIC ACID)

Niacin (nicotinic acid) is one of the oldest hypolipidemic agents in clinical use;
however, its mechanisms of action are still partially unknown. The remarkable
feature of niacin is that it acts beneficially on practically all lipoprotein abnor-
malities.57–59 It is the most effective medication for increasing HDL cholesterol
levels, lowering at the same time atherogenic apolipoprotein B.59 It has been
shown that niacin directly inhibits the activity of hepatic diacylglycerol acyltrans-
ferase-2 (DGAT2), which is involved in the synthesis of triglycerides and apoli-
poprotein B.60 The HDL cholesterol-elevating effect of niacin is probably caused
by selective inhibition of the hepatic removal/uptake of lipoprotein AI particles
leading to their increased retention in the circulation.61 Investigation of the mech-
anisms of niacin action has recently been stimulated by the identification of
nicotinic acid receptors62–65 apparently mediating suppression of adipocyte lypoli-
sis by niacin.62,65
Another recently discovered pathway involves stimulation of prostaglandin
D2 (PDG2) formation by niacin, which interacts with PPAR-controlled processes
through PPAR activation by a major PDG2 metabolite.66 It is also interesting to
note that several single nucleotide polymorphisms (SNPs) have been identified
in the coding regions of niacin receptors defining the five major haplotypes,67
which well may have potential of modifying biological effects of niacin.
Niacin is currently available in three formulations (immediate-release,
extended-release, and long-acting).58 Immediate-release niacin has well known
side effects, including flushing, gastrointestinal symptoms, and hyperglycemia.
Long-acting drugs may cause hepatotoxicity, while extended-release niacin is
associated with fewer flushing and gastrointestinal manifestations without seri-
ously increasing the risk of hepatotoxicity. Lower doses of niacin are often applied
in combinations with other hypolipidemic agents, especially statins.59 Neverthe-
less, recent progress in nicotinic acid receptor identification raises hopes of
developing new specific drugs mimicking the beneficial action of niacin, but free
of its adverse effects.
2.2.5 CHOLESTEROL ABSORPTION INHIBITORS

Wide introduction of statins into clinical practice has resulted in the development
of multiple schemes of combined therapy designed to complement the hypolip-
idemic effects governed by different molecular mechanisms. It became obvious
that one potential way to improving statin therapy efficiency would be to reduce
intestinal cholesterol absorption, which results in upregulation of statin-inhibited
HMGCoA reductase expression.17
Ezetimibe, a synthetic 2-azetidinone, is the first of a new class of drugs that
selectively inhibits the absorption of cholesterol in the intestine without affecting
the absorption of triglycerides. The agent is believed to inhibit a putative choles-
terol transporter of enterocytes located within the brush-border membrane of the
small intestine.68 Ezetimibe reduces overall cholesterol delivery to the liver, thus
stimulating the expression of LDL receptors and increasing the removal of LDL
cholesterol from the serum.
Clinical trials have provided promising results demonstrating efficient cho-
lesterol reduction with the use of the drug as a single agent69 or as a part of
combined therapy with statins17,70,71 or fibrates.72 No serious adverse effects asso-
ciated with the use of ezetimibe have been observed,17,68–72 making the drug a
very attractive component of various schemes of dyslipidemia management.
2.2.6 EMERGING AND FUTURE DIRECTIONS IN HYPOLIPIDEMIC

THERAPY
New approaches to hypolipidemic therapy are being developed along several
directions that often derive from investigation of mechanisms of action of the
clinically approved drugs considered above. Whereas covering this rapidly pro-
gressing area is not among the purposes of this chapter, briefly highlighting some
of the emerging directions appears to be justified.
Active search is going on in the field of lipid metabolism and transport
modulation. It is believed that employment of acyl-coenzyme A cholesterol acyl-
transferase (ACAT) inhibitors affecting two ACAT subtypes may reduce foam
cell formation as well as cholesterol absorption;17,73 however, initial testing of the
first agent of this group (avasimibe) in humans has produced inconsistent
results.17,74 Microsomal triglyceride transport protein (MTP) inhibitors, which
should affect the assembly of VLDL, have been successfully tested in animal
experiments,75,76 but have not reached human trials so far. Human studies are
being performed on cholesteryl ester transfer protein (CETP) inhibitors, which
block the transfer of cholesteryl esters from HDL to apoB-containing lipoproteins,
thus elevating HDL levels. Although promising results have been obtained with
CETP inhibitors JTT-705 and torcetrapib,17,77,78 additional clinical studies are
needed to evaluate both the safety and efficacy of this therapeutic strategy. Another
experimentally tested HDL elevation-directed approach is based upon the use of
adenosine triphosphate (ATP)-binding cassette transporter A1 (ABCA1) agonists
affecting the initial step in reverse cholesterol transport.17,73
Finally, the use of nuclear receptor signaling involved in cholesterol homeo-

stasis control is starting to attract increasing attention with regard to the devel-
opment of hypolipidemic therapy approaches. PPARs have already been discussed
in this chapter; however, several other transcription regulators, such as sterol
regulatory element-binding proteins (SREBPs), liver X receptors (LXRs), farne-
soid X receptor (FXR),73,79 are considered as probable targets for new drugs.
Brief consideration of therapeutic strategies applied to provide efficient man-
agement of CVD-related hyperlypidemia and dyslipidemia has shown consider-
able diversity of drugs applied for this purpose. The second part of the chapter
is devoted to the analysis of the role of dietary factors in lipid homeostasis
regulation and interactions between diet and hypolipidemic drugs.
2.3 DIETARY FACTORS IN LIPID HOMEOSTASIS

AND THEIR INTERACTIONS WITH
HYPOLIPIDEMIC DRUGS
Lipid homeostasis directly depends on dietary lipid supply. Consumption of
food rich in cholesterol and saturated fatty acids of animal origin, a typical
feature of traditional Western diets, is a generally recognized atherogenic factor
constituting the cornerstone of the classic “diet–heart” hypothesis.1 Reduced
intake of saturated fat and cholesterol is regarded as the first element of lifestyle
changes recommended for primary prevention of coronary heart disease (CHD)
and is commonly combined with the use of hypolipidemic drugs for secondary
CHD prevention.80,81 However, these “restrictive” dietary aspects of lipid-low-
ering strategies constitute a separate important area and will not be analyzed
here.
Consideration of the effects of drugs and dietary factors as well as interactions
between food and drugs with regard to hypolipidemic treatment in humans is the
principal subject of this chapter. Beneficial biological effects of numerous food
components can be (and are) used for therapeutic purposes, so sometimes the
boundary at which a food ingredient becomes a drug is not well defined.82
Furthermore, it should be understood that relationships between dietary and
medicamentous treatment of hyperlipidemia comprise at least two dimensions.
Firstly, both pharmaceutical agents and food constituents may have lipid-lowering
properties, thus their combined effects can constitute synergistic interactions of
the “first type.” It is obvious that only those food components that possess natural
hypolipidemic properties will be discussed from this point of view. Secondly,
dietary factors can interfere with the effects of drugs by influencing drug metab-
olism or regulatory pathways important for proper drug action, therefore causing
either synergistic or antagonistic effects — interactions of the “second type.”
These two situations will be considered below.
2.3.1 DIETARY INGREDIENTS POSSESSING NATURAL

HYPOLIPIDEMIC AND CARDIOPROTECTIVE PROPERTIES
Description of dietary ingredients naturally possessing hypolipidemic properties
is not an easy task since the field is extremely dynamic and still full of controversy.
Food constituents with reported hypolipidemic properties are listed in Table 2.1,
but in many instances beneficial action of nutrients, in terms of reducing CVD
risk, has multiple endpoints and involves physiological pathways unrelated to
lipid transport and metabolism. Numerous natural products contain complex
combinations of biologically active components simultaneously acting through
different interacting mechanisms, so comprehensive analysis of such complicated
situations is difficult and sometimes prone to simplistic conclusions. Nevertheless,
several major groups of important dietary factors can be easily identified (see
Table 2.1).
Over the past several decades, the main feature of nutritional recommenda-
tions developed to decrease CVD risk was to limit intake of total fat and saturated
fat.132 It is, however, becoming evident that types of fat consumed rather than
total dietary fat are important in determining CVD risk.80,133,134 Indeed, some
human populations have relatively low CVD risk levels despite traditional con-
sumption of meat-based diets rich in fat.135,136 While there is no doubt in the
association between saturated fat intake and elevated serum LDL levels, increased
consumption of polyunsaturated fatty acids (PUFA) and to some extent mono-
unsaturated fatty acids (MUFA) has been shown to exert beneficial effects (see
Table 2.1). Substitution of unsaturated fatty acids for saturated fat appears to be
a logical way to follow with the purpose of developing cardioprotective diets. At
the same time, there is little clarity regarding the optimal amounts of MUFA and
PUFA in the diet.80,133 One desirable target is achieving a higher ratio between
ω-3 PUFA and ω-6 PUFA by encouraging consumption of foods rich in ω-3
PUFA, such as fish, shellfish, and some plant oils.80,86,87,133
The hypothesis suggesting LDL oxidation as a major mechanism involved in
the development of atherosclerosis137 has led to the idea that antioxidants can
prevent atherosclerosis by limiting this process. Numerous natural antioxidants
have been identified, predominantly in foods of plant origin. Flavonoids and
antioxidant vitamins constitute two main groups of dietary antioxidants. Fla-
vonoids are bioactive plant polyphenols exerting various physiologic effects (see
Table 2.1). Several studies have demonstrated inverse relationship between fla-
vonoid intake and CVD risk;95,96,138 however, other groups failed to detect any
effect.93,139,140 Although hypolipidemic properties of distinct flavonoids are being
actively investigated, it should be noted that disentangling specific action of each
of numerous bioactive components present in most natural foods is hardly pos-
sible at the present level of scientific knowledge.
Attempts to find a simple solution to this problem are well illustrated by the
use of antioxidant vitamin supplements for CVD prevention. Experimental and
epidemiological studies suggested that dietary intake of antioxidant vitamins
(vitamin E, vitamin C, β-carotene) may be associated with the decrease in CVD
TABLE 2.1
Dietary Ingredients Exerting Beneficial Influences on Hyperlipidemic and
Dyslipidemic Conditions
Active Ingredients Dietary Sources Physiological Effects Refs.
Fatty Acids
Monounsaturated fatty Canola, olive, Mild hypocholesterolemic effect, 83–85
acids (MUFA): oleic safflower, possible reduction of oxLDL
acid (OA) sunflower oils, and (additional nonlipid beneficial effects
nuts, olives, possible)
avocado
Omega-6 Corn, safflower, Mild hypocholesterolemic effect 86–88
polyunsaturated fatty sunflower oils (downregulation of LDL production
acids (ω-6 PUFA): and enhancement of its clearance);
linoleic acid (LA) adverse effects on cytokines and
platelet aggregation are not excluded
Omega-3 Canola, flaxseed, Mild hypocholesterolemic effect, serum 81, 86, 88,
polyunsaturated fatty soybean oils, triglyceride lowering effect; additional 89–92
acids (ω-3 PUFA): walnuts (nonlipid) beneficial effects include
linoleic acid (ALA) antiarrhythmic (especially EPA and
DHA) and probably antithrombotic
action.
Eicosapentaenoic acid Fish, shellfish
(EPA) and
docosahexaenoic acid
(DHA)
Flavonoids
Flavonols: quercetin, Apple, onion, kale, Mild hypocholesterolemic effect; 93–98
kaemprefol, broccoli, cabbage, atioxidant action is believed to protect
myrocetin, fisetin cherries, berries, from oxLDL accumulation; additional
tea, red wine (nonlipid) effects include
antiinflammatory action, reduction of
platelet aggregation, improved
vascular reactivity, weak estrogenic
activity (only isoflavones)
Flavanols (catechins): Green and black
catechin, epicatechin, tea, red wine,
epicatechin 3-gallate, apples, black
epigallocatechin, chocolate, cocoa
epigallocatechin 3-
gallate, gallocatechin
Flavones: apigenin, Parsley, celery,
luteolin thyme
TABLE 2.1 (CONTINUED)

Flavanones: Oranges,
naringenin, grapefruit, and
hesperetin, their juices
eriodictyol
Anthocyanins: Berries, cherries,
(anthocyanidins) red wine
Cyanidin,
pelargonidin,
delphinidin, peonidin,
petunidin, malvinidin
Isoflavones: genistein, Soy and soy-
daidzein, dihydro- containing foods,
daidzein, O-desmethy- chick peas
langolensin, equol
Antioxidant Vitamins
α-Tocopherol Vegetable oils Experimental evidence of lipid 99–105
(vitamin E) (sunflower, olive), peroxidation inhibition: controversial
nuts, seeds results regarding protection against
CVD in humans
Ascorbic acid Fruit (especially Experimental evidence of lipid 99, 100,
(vitamin C) citrus), vegetables peroxidation inhibition: controversial 102,
results regarding protection against 104–106
CVD in humans
Carotenoids: Green leafy, root, Antioxidant action was suggested as a 99,
β-carotene and fruiting protective factor against oxLDL 102–105,
vegetables accumulation; antiinflammatory effect 107
possible; controversial results
regarding protection against CVD in
humans
Lycopene Tomatoes, tomato Antioxidant action may provide lipid 108–112
products, peroxidation inhibition; protective
watermelon effect against CVD reported
Lutein Leafy vegetables, Antioxidant effect may influence lipid 113
fruit peroxidation; protective effect against
CVD requires confirmation

Sulfur-Containing
Compounds
Allicin, ajoene, Allium vegetables, Mild hypocholesterolemic effect 114–117
S-allylcysteine, especially garlic repeatedly described in experimental
diallyl disulfide, and its and small human studies, but needs
S-methylcysteine preparations further confirmation; possible
sulfoxide, mechanisms may involve inhibition of
S-allylcysteine hepatic cholesterol synthesis and
sulfoxide antioxidant effect; possible additional
(nonlipid) effects include
enhancement of fibrinolytic activity,
inhibition of platelet aggregation,
blood pressure lowering
Fiber
Soluble fiber Cereals, legumes Well-documented mild (118–123)
(psyllium, pectins, guar hypocholesterolemic effect; LDL
gum, some reduction might be related to intestinal
hemicelluloses) binding of bile acids and/or cholesterol
followed by LDL receptor
upregulation and increased LDL
clearance; additional (nonlipid) effects
include C-reactive protein reduction
and normalization of blood pressure
Protein
Legume protein Soy, legumes Legume proteins, especially those 124–127
derived from soy, appear to have
hypocholesterolemic effects;
mechanisms are not clear, difficult to
separate from isoflavone effects
Plant Sterols and

Stanols
Sitosterol, Tall (pine tree) oil, Well-documented hypocholesterolemic 128–131
campesterol, soybean oil, seeds, effect due to cholesterol absorption
stigmasterol, nuts (especially inhibition
sitostanol, macadamia nut),
campestanol sterol/stanol-
enriched
margarines.
risk.99–101 The idea has been tested in a number of controlled trials investigating
the effects of antioxidant vitamin supplements on CVD risk. It is remarkable that
the results of these studies have been largely disappointing,101,103–105,141 leading to
the conclusion that there is no basis for recommending that patients take vitamin
C or E supplements or other antioxidants for the express purpose of preventing
or treating coronary artery disease.141,142
Likewise, there is some controversy regarding the use of garlic as a potential
lipid-lowering and cardioprotective food component. Commonly used garlic prep-
arations and supplements have varying amounts of allicin, which is believed to
be at the core of garlic bioactivity (see Table 2.1), so making a definite conclusion
on their lipid-reducing efficiency is difficult.114–117 Nevertheless, despite all prob-
lems discussed above, beneficial effects of antioxidant-rich diets based on fruit,
vegetables, and products derived from these sources are evident,97,143–145 and such
diets are widely recommended to reduce CVD risk.141 It appears to be very likely
that consumption of natural products containing different antioxidant components
acting synergistically may be much more efficient in providing cardiovascular
protection than isolated action of food supplements.97
Mild hypocholesterolemic effect of dietary soluble fiber is well documented;
however, precise mechanisms of this phenomenon are not clear and may be
partially related to other components of fiber-rich cereals and legumes.118 Pro-
motion of increased fiber consumption, especially with whole grains, should be
considered as one of the strategic ways of changing overall Western dietary
patterns towards CVD prevention.
The amount and type of dietary protein is closely related to the problem of
fat intake. It was repeatedly shown that substitution of soy for animal protein
results in LDL and triglyceride reduction.124–127 At the same time, the mechanism
of this reduction is unknown and, especially in the case of soy, it is difficult to
exclude an impact of isoflavones.124,126 Overall food composition rather than
protein type appears to be more important for serum lipid balance. It has been
suggested that consumption of animal protein per se may not be harmful, but is
commonly associated with an increased intake of saturated fat.146 To avoid this
unhealthy association, it can be recommended to shift the main sources of dietary
protein by replacing red meat with nuts, soybeans, legumes, poultry, and fish.80
The use of natural cholesterol-lowering properties of plant sterols and stanols
inhibiting intestinal cholesterol absorption has emerged as an attractive strategy
for serum lipid level correction, especially in mildly hypercholesterolemic sub-
jects.130 New polyunsaturated margarines with added sterols and stanols have
recently been made available for wide consumption.129 It should be noticed,
however, that the use of these products can also reduce the absorption of fat
soluble vitamins.129 Possible consequences of long-term sterol/stanol intake are
not entirely known, so caution is needed in the use of this approach.
Food components discussed above are widely used as ingredients of different
diets directed on both CVD prevention and attenuation of hyperlipidemia in CVD
patients. The search for new natural substances with hypolipidemic properties
continues, and such agents as policosanol (a mixture of alcohols isolated from
sugar cane),147 guggulipid (Commiphora mucul extract),148,149 or red yeast rice

(embryonic rice fermented with Monascus ruber yeast)150 are now being tested.
However, further studies are needed to assess their efficiency.
The analysis of food components possessing natural hypolipidemic properties
shows that only mild effects of such dietary treatments can be expected. Nutri-
tional counseling certainly constitutes a major element of healthy lifestyle form-
ing the basis of primary CVD prevention. Specifically composed diets can be
successfully combined with the use of hypolipidemic drugs; however, it should
be noted that interactions between bioactive food components and medicines are
sometimes difficult to predict. These interactions are discussed below.
2.3.2 INTERACTIONS BETWEEN DIETARY COMPONENTS AND

HYPOLIPIDEMIC DRUGS
It is well known that simultaneous treatment of a patient with more than one
pharmaceutical agent has potential risk of altering expected results of the treat-
ment and even producing some adverse consequences. Such situations usually
occur when drugs interact at the level of common regulatory, metabolic, or
transport pathways involved in the realization of their action. Certain dietary
components can behave in a similar way when taken together with drugs. Foods
of plant origin that contain complex mixtures of phytochemicals are especially
likely to produce such inadvertent effects.151–153
The small intestine is the main site for absorption of both nutrients and orally
ingested medicines. Over the past 20 years, it has become obvious that this organ
also has complex metabolic functions hosting Phase I and Phase II xenobiotic-
metabolizing enzymes as well as associated molecular transporters,154,155 which,
acting together, determine oral bioavailability of drugs. It has been established
that enzymes of the CYP3A subfamily are especially important for oxidative
reactions of Phase I xenobiotic metabolism, CYP3A4 being the dominant P450
form expressed in human enterocytes.154,156,157 Numerous drugs are known to be
CYP3A4 substrates and several members of the statin family (see Section 2.2.1
of this chapter) are among them. Expression of some other enzymes of the CYP
superfamily in the small intestine has been reported154; however, information
regarding their significance for drug and dietary component metabolism in the
human gut is limited.
Phase II metabolic reactions in the small intestine provide conjugation of
xenobiotics and their Phase I metabolites resulting in generation of nontoxic and
easily excretable products. These reactions are mostly exerted by enzymes of
glutathione S-transferase (GST), uridine diphosphate glucuronosyltransferase
(UGT), and, to some extent, sulfotransferase families.154 Phase II glucuronidation
reactions have been shown to be important for the metabolism of statins and
fibrate-statin interactions in human hepatocytes;158 however, it is not known
whether such interactions occur in enterocytes as well. In addition to Phase I and
Phase II metabolic pathways, the function of transport proteins greatly contributes
to the protection from exogenous xenobiotics by both preventing harmful
substances from entering the cell and eliminating products of xenobiotic detoxi-
fication reactions.155 Intestinal P-glycoprotein (a member of the multidrug resis-
tance transporter family) expressed in the apical membrane of enterocytes appears
to exert its transporter functions in concert with the metabolic role of CYP3A4,
thus constituting a dynamic subsystem controlling xenobiotic metabolism/elim-
ination rate.159 Even from the brief description given above, it becomes apparent
that disruption of proper functioning of this complex system can have hazardous
consequences.
The problem of diet–drug interactions was suddenly highlighted by an unfore-
seen discovery in 1989 of the effect of grapefruit juice (GJ) on bioavailability of
felodipine, a calcium channel blocker used for hypertension treatment.160 Since
then, multiple studies have shown GJ interactions with various drugs of different
families often lead to enhanced adverse effects (see References 153, 161). GJ
interactions with hypolipidemic drugs (statins) and some other medicines often
prescribed to CVD patients are listed in Table 2.2. Investigation of the mecha-
nisms of GJ–drug interaction has revealed that oral intake of GJ selectively
inhibits CYP3A4 in the small intestine,176–178 resulting in a considerable reduction
of oxidative metabolism of the substrates of CYP3A4, including numerous drugs.
Only intestinal enzyme inhibition was observed, whereas liver CYP3A4 activity
appeared to be unaffected177 as well as pharmacodynamics of drugs metabolized
by this enzyme after their intravenous administration.170,179,180
However, it has recently been reported that consumption of large amounts of
GJ can inhibit hepatic CYP3A4 as well181; thus the effect may be dose-dependent
with a much higher threshold for the enzyme in the liver. The search for the active
GJ ingredients was focused on furanocoumarins (psoralens) and isoflavones as
contributors to CYP3A4 inhibition. GJ furanocoumarins, especially bergamottin
and 6′,7′-dihydroxybergamottin are regarded as major CYP3A4 inhibi-
tors,178,182–184 but binding properties and enzyme inhibition kinetics of the two
furanocoumarins are different,185 suggesting that different inhibition mechanisms
can be involved. In addition, it has been shown that grapefruit flavonoids, in
particular, naringin and its derivative naringenin, can act as competitive inhibitors
of CYP3A4.186 The influence of GJ on the intestinal drug metabolism appears to
be even more complex since it is not limited by the effects on CYP3A4. GJ
components have also been shown to suppress the P-glycoprotein transporter
activity,153,161,187,188 thus further increasing ingested drug bioavailability.
From the point of view of hypolipidemic therapy, interactions between GJ
and HMG-CoA reductase inhibitors are especially important since consumption
of the juice by individuals taking statins predominantly metabolized by CYP3A4
(atorvastatin, lovastatin or simvastatin) can significantly increase the risk of
adverse effects.161,164,166 Interactions are much less likely with fluvastatin, which
is mainly metabolized by CYP2C9,162 or pravastatin, rosuvastatin, and pitavasta-
tin, which are largely metabolized through P450-independent pathways.163–165,189
It is also apparent that considerable interindividual variation in responses to statin
therapy strongly depends on genetic heterogeneity of human populations. Indeed,
numerous polymorphic variants of the CYP3A4 gene have been identified,190–193
TABLE 2.2
Grapefruit Juice Interactions with Drugs Used for the Treatment of
Hyperlipidemia and Other Manifestations of Cardiovascular Disease
Drug Classes, Conditions
Individual Drugs Treated Reported Adverse Effects Refs.
HMG-CoA Reductase
Inhibitors (Statins)
Simvastatin, lovastatin, Hyperlipidemia Myopathy, rhabdomyolysis, 161–166
atorvastatin acute renal failure
Pravastatin, fluvastatin, Little or no influence
rosuvastatin, pitavastatin
Calcium Channel
Blockers
(Dihydropyridines)
Felodipine, nicardipine, Hypertension, Excessive vasodilatation, 153, 161,
nifedipine,nimoldipine, angina pectoris hypotension, tachycardia 167–171
nisoldipine, pranidipine, treatment with
nitrendipine, verapamil verapamil
Amlodipine, barnidipine, Little or no influence
diltiazem
Angiotensin II Receptor 1
Antagonistsa
Losartan Hypertension Reduced therapeutic effect 161, 172
Beta-Blockersa
Carvedilol Hypertension, Increased drug toxicity 161, 172
heart failure
treatment
Celiprolol Decreased drug bioavailability
Antiarrhythmic Agentsa
Amiodarone, quinidine, Arrhythmias Increased drug toxicity 161, 174,
disopyramide, 175
propafenone
a No information on interactions with other drugs of this group.
and some of them have already been shown to affect lipid profiles in patients
receiving statin therapy.194,195 Moreover, potential influence of multiple polymor-
phisms of MDR1 gene encoding P-glycoprotein196,197 or other transporter-encod-
ing genes is impossible to exclude. Interestingly, it has recently been reported
that polymorphisms of organic anion transporting polypeptide C have been
associated with plasma pravastatin concentrations.198 At the same time, organic
anion-transporting polypeptides may be inhibited by GJ,161,199 constituting another
interaction area.
Although GJ effects on bioavailability of statins are relatively well docu-

mented, little is known about potential interactions of this dietary factor with
other hypolipidemic drugs. Among them, fibrates present particular interest since
their metabolism and transport can be related to the systems discussed above.
Fenofibrate has recently been shown to inhibit P-glycoprotein-mediated trans-
port,200 so some pathways can present background for both drug–drug and
diet–drug interactions. Cases of severe adverse effects of statin–fibrate combined
therapy, especially associated with the use of gemfibrozil,21,201 are well known;
however, interaction mechanisms causing these effects are still obscure. Although
there is no information about GJ–fibrate interactions, it is difficult to be sure that
potential risk does not exist, especially in situations of combined therapy.
Presented information highlights the complexity of GJ–drug interactions,
which are not necessarily harmful. On the one hand, citrus flavonoids, especially
naringenin, are believed to be antiatherogenic.202,203 On the other hand, in many
situations dietary modulation of drug bioavailability might be an advantage on
the condition that the consequences of the interaction are perfectly predictable.
Identification of GJ–drug interactions has stimulated interest to possible
effects of other fruit juices. Although the information is less abundant, there are
reports indicating that either CYP3A4 or multidrug transporter proteins can be
affected. Several citrus juices (lime, orange, lemon, pummelo), apple juice, and
juices of several exotic fruits have been shown to influence drug pharmacokinet-
ics.188,199,204–207 As some of the observed effects are attributed to fruit flavonoids,
it is difficult to exclude that other flavonoid-containing products can emerge as
potential causes of food–drug interactions.
It is often difficult to set strict limits between nutrition and consumption of
some natural products for health-related reasons. Health-promoting properties of
numerous herbs have been known for centuries, and herbal remedies rich in
bioactive compounds are traditionally used for unconventional therapy of various
diseases including CVD.208 At the same time, many herbs, widely regarded as
“natural health products,” are easily available over the counter and consumed
without proper medical control. St. John’s wort (Hypericum perforatum) is a
popular herbal product used for the treatment of depression. It has been shown
that St. John’s wort affects both intestinal and hepatic xenobiotic metabolism and
transport, and action of its bioactive substances (hypericin and hyperforin) appears
to be opposite to that of GJ. St. John’s wort induces both CYP3A4 and P-
glycoprotein expression,209–211 thereby reducing bioavailability of drugs depen-
dent on this metabolic/transport system. This action may be potentially important
for the efficiency of hypolipidemic therapy with statins. It has already been
reported that St. John’s wort significantly decreases plasma concentrations of
simvastatin, but not of pravastatin.212
These results are not surprising in view of the metabolic differences between
the two drugs described earlier in this chapter. Although no information regarding
other interactions between herbal medicines and hypolipidemic drugs is available,
such interactions are not excluded since the problem of consequences of simul-
taneous intake of herbal and conventional medicines has attracted close attention
only recently. Potentially harmful effects of natural remedies (echinacea, ephedra,

garlic, ginkgo, ginseng, kava, St. John’s wort, valerian) interfering with the action
of preoperative anesthetics have already been described,213 and several of these
natural agents have been shown to be associated with modulation of drug transport
systems.214 Herb–drug interactions in a wider aspect of cardiovascular pharma-
cotherapy have been discussed in a recent review.215
Examples discussed above show that the problem of possible interactions
between food components and hypolipidemic drugs certainly needs further inves-
tigation. Nevertheless, many situations when adverse drug effects are provoked
by consumption of certain foods, food supplements or natural health products
are easily preventable by appropriate medical advice. Moreover, both efficiency
of hypolipidemic therapy and the potential of adverse effects of different thera-
peutic schemes can potentially be individualized if more information is available
on individual genetic backgrounds. The latter option is not immediately available;
however, rapid growth of understanding of genetic influences on the relationship
between nutrition, CVD pathogenesis, and action of hypolipidemic drugs216–218
allows one to expect significant progress in this direction.
2.4 CONCLUSION
Serum lipid lowering is a pivotal component among measures applied in order
to prevent CVD or delay its progression. Dietary approaches can be successfully
used for primary prevention of cardiovascular conditions, especially coronary
heart disease. General dietary strategy should include recommendations to change
fat intake habits (substitute polyunsaturated and monounsaturated fats for satu-
rated fats, increase intake of ω-3 PUFA) and to consume food rich in fruits,
vegetables, nuts, and whole grains (i.e., products rich in natural antioxidants,
soluble fiber, vegetable protein). It appears that natural products containing com-
plex mixtures of bioactive ingredients acting synergistically provide more bene-
ficial effects compared to most presently available artificial dietary supplements
and additives.
Secondary CVD prevention directed to delaying disease progress in individ-
uals with existing conditions is now based upon application of combined thera-
peutic strategies including hypolipidemic drug therapy and additional dietary
measures. The development of new lipid-lowering agents is progressing very fast
through introduction of new potent drugs often providing beneficial pleiotropic
effects affecting different mechanisms involved in CVD pathogenesis. HMG-CoA
reductase inhibitors (statins) have emerged as the central element of hypolipi-
demic therapy, especially in cases with seriously increased LDL levels. Avail-
ability of other types of hypolipidemic medicines that can be combined with
statins considerably widens therapeutic options, allowing development of indi-
vidualized treatment schemes. Successful combination of statin therapy with the
use of fibrates, bile acid transport inhibitors, niacin, and cholesterol absorption
inhibitors provides an expanding arsenal of therapeutic approaches. Dietary rec-
ommendations, which are generally similar to those used for primary CVD
prevention, should be specifically adapted for each individual case. This require-
ment is especially important in view of accumulating evidence of possible inter-
actions between food components and medicines, especially orally ingested drugs.
It appears that in many cases food–drug interactions, if properly assessed and
understood, can potentially be employed to enhance beneficial therapeutic effects.
This understanding, however, requires much better knowledge of individual char-
acteristics of each patient determined by genetic background. “Genetic profiling”
of individuals subjected to hypolipidemic therapy can greatly reduce the risk of
adverse effects and facilitate determination of optimal therapeutic and dietary
schemes applied for CVD treatment.
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DK5836_C003.fm Page 63 Thursday, May 25, 2006 12:52 PM
3 Phytochemicals,
Xenobiotic Metabolism,
and Carcinogenesis
James B. Kirkland
CONTENTS
3.1 Introduction................................................................................................63
3.2 Dietary Carcinogens and Xenobiotic Metabolism....................................64
3.3 Animal-Based Research on Cooked Meat Carcinogens...........................68
3.4 Metabolism of Dietary Secondary Carcinogens .......................................68
3.4.1 Metabolism of NAs .......................................................................68
3.4.2 Metabolism of PAH and HCA Families .......................................69
3.4.3 Metabolism of PAHs .....................................................................69
3.4.4 Metabolism of HCAs ....................................................................70
3.5 Aflatoxin and Food Spoilage.....................................................................70
3.6 Effects of Carcinogen Exposure on Xenobiotic Metabolism...................70
3.7 Diet as a Source of Chemopreventative Agents .......................................73
3.8 Phytochemical Activation of the AhR.......................................................76
3.9 Phytochemical Antagonism of the AhR....................................................77
3.10 Phytochemical Inhibition of the Catalytic Activity of P450 ....................79
3.11 Phytochemical Induction of Phase II Enzymes ........................................80
3.12 Experimental Models for Future Work and Opportunities for
Improving Public Health ...........................................................................81
3.13 Public Perception of Risk and Resulting Behavior ..................................83
3.14 Conclusions................................................................................................85
References ...........................................................................................................86
3.1 INTRODUCTION
Nitrite preservatives in processed meats lead to the formation of carcinogenic
nitrosamines (NAs), while smoking and cooking of meats leads to the formation
of polycyclic aromatic hydrocarbons (PAHs) and heterocyclic amines (HCAs).
Cigarette smoke is also a potent source of NA, PAH, and HCA carcinogens.
Conversely, consumption of vegetables, spices, and green tea are associated with
63
decreased cancer risk, and many phytochemicals from these sources are proven
chemoprotective agents in animal models. NAs, PAHs, and HCAs are stable
secondary carcinogens, which are “bioactivated” by P450-based Phase I metab-
olism to form reactive, ultimate carcinogens. This is also required for effective
metabolism, as it allows them to be conjugated (Phase II metabolism) and
excreted, preventing the accumulation of parent compounds in areas like the
adipose and brain tissue. Thus, the goal in cancer prevention is to find the optimal
balance between Phase I and II activities, which allow clearance, while minimiz-
ing the accumulation of reactive intermediates. Smoking and regular consumption
of cooked meats are negative influences, as P450 enzymes are induced and
bioactivation becomes too rapid for Phase II reactions to keep pace. Chemopro-
tective phytochemicals improve the metabolic balance by antagonizing P450
induction, by competitively inhibiting P450 enzymes, and by inducing the expres-
sion of Phase II enzymes. Thus, phytochemical intake can dramatically alter the
carcinogenicity of a given exposure to dietary carcinogens.
3.2 DIETARY CARCINOGENS AND XENOBIOTIC

METABOLISM
Many of our common cancers are responsive to diet and lifestyle habits, leading
to popular estimates that around 60% of cancers are “avoidable” if the ideal diet
and lifestyle were to be followed.1 The most common cancers in men (worldwide)
are lung, colorectal, stomach, liver, prostate, esophageal, bladder, and oral cavity.2
This pattern is also present in women, with the addition of breast, cervical, and
ovarian cancers as important sites. Many of the common cancers have well-
established negative diet and lifestyle risk factors. Lung and oral cancers are
strongly associated with smoking, chewing of tobacco, and alcohol consumption.1
Colorectal, esophageal, larynx, gastric, bladder, prostate, and breast cancer risks
have been associated with a high intake of cooked meat.3–5 The relationship
between meat intake and increased risk of colorectal cancer is greatly strength-
ened with the use of doneness level as a factor. Sinha et al. found that the enhanced
risk of colorectal cancer increased from 10 to 29% per 10 g/d red meat intake
when the analysis was restricted to well-done meat.6 Knize et al. have reviewed
epidemiological studies on the consumption of well-done meat and cancer risk,
finding that over 80% of studies find significant associations.3 Hematopoietic,7
oral,8 nasopharageal,8,9 esophageal,8,10 stomach,11 colorectal,8 and prostate12 can-
cers have also been linked to preserved meats. Maternal exposure to preserved
meats has been associated with childhood brain tumors as well.13 The association
of cancer risk with cooking and preserving of meats is quite controversial, with
varied research findings in human and animal studies.
Epidemiological studies present difficulties in estimating not only meat
intake, but also cooking methods and the extent of cooking. Survey tools need
to be improved, including databases for carcinogen content by meat type and
cooking method, and visual aids to maximize the accuracy of collected data.14,15
Diets rich in cooked and processed meats tend to have more saturated and higher
Phytochemicals, Xenobiotic Metabolism, and Carcinogenesis 65
FIGURE 3.1 Examples of the three major classes of dietary secondary carcinogens: (A)
Benzo(a)pyrene (BaP), example of a polycyclic aromatic hydrocarbon (PAH), formed
through condensation of small radicals during the incomplete combustion of any burnable
substrate, requiring temperatures above 250°C. (B) 2-amino-3,4-methylimidazo[4,5-
f]quinoline (MeIQ), example of a heterocyclic amine (HCA), formed through condensation
of creatinine, amino acids, and sugars, across a broad range of meat cooking conditions,
enhanced by drying, as seen in the formation of gravy and jerky products. (C) Dimeth-
ylnitrosamine (DMN), example of a nitrosamine (NA), formed through reaction of nitrite
with secondary amines found in preserved meats and seafoods, also central to cigarette
smoke carcinogenesis in the formation nicotine/nitrite reaction products (e.g., NNK).
levels of fat. Animal models making use of practical foods, rather than single
purified carcinogens, will eventually provide unbiased data on these factors. Liver
cancer is associated with alcohol abuse in developed countries and hepatitis B
and aflatoxin exposure in developing countries.
Many of these negative risk factors can be explained by the exposure to
secondary carcinogens. Our major dietary carcinogens are all secondary carcin-
ogens, which are very stable, lipophilic compounds, generally formed during food
processing and cooking.16 The main categories of carcinogens in cooked and
preserved meats are polycyclic aromatic hydrocarbons (PAHs), heterocyclic
amines (HCAs) and nitrosamines (NAs)16 (Figure 3.1). PAHs, like benzo(a)pyrene
(BaP), are formed when material is partially burned, during smoking, grilling,
and barbequing. PAHs will form from any substrate, but flame grilling of high
fat foods is an excellent example.17 PAHs are also formed in the smoke of burning
wood chips and they dissolve into meats during the smoking process.17,18 BaP is
the main focus of regulatory efforts, limited to 5 µg/kg in smoked meats in the
European Community. This is despite the fact that BaP is a relatively small and
variable proportion of total PAH load in cooked or smoked food.18
HCAs are formed when meats are cooked at below carbonization tempera-
tures, as creatinine reacts with various amino acids to form products like 2-amino-
1-methyl-6-phenylimidazo-[4,5-b]-pyridine (PhIP), 2-amino-3,8-dimethylimi-
dazo[4,5-f]quinoxaline (MeIQx), and 2-amino-3-methylimidazo[4,5-f]quinoline
(IQ).3 Sugars may enhance this process when they are added to marinades or
during the processing of beef jerky.19 During the grilling of meat, higher temper-
ature cooking to the same final internal temperature produces significantly higher
levels of HCAs.3 Pan residues used in the production of gravy and food flavor
products tend to have higher levels of HCAs than the meat itself, especially if
the residue dried under heat. This concentrates the reactants and causes large
increases in cooking temperature.20 The recovery of free HCAs from cooked meat
samples is traditionally used to estimate the degree of formation of these carcin-

ogens. However, it is now known that half or more of the total mutagenic capacity
remains covalently bound within the protein structure, to be released during
digestion.21 This is likely due to the participation of amino acid side chains in
HCA formation reactions without being released from the peptide structure, and
it leads to significant underestimations of dietary HCA exposure if not corrected
for.
The preservative nitrite forms reactive species that combine with secondary
amines to form NA end products like dimethylnitrosamine (DMN). These reac-
tions are encouraged by the acidic conditions of the stomach,13,22 and by bacterial
populations in other areas of the digestive tract or in cured, fermented foods.10
N-nitrosamides are produced through the nitrosation of amides, and are unstable
and short-lived.23 They do, however, have an experimental carcinogenic profile
that is more likely to cause leukemias and pediatric brain tumors, and future
research should consider the role of endogenous nitrosamide formation following
cured meat consumption.23 Meats are cured using salt and nitrite to prevent
bacterial growth and food poisoning, especially botulism. Cured, fermented fish
are an excellent source of DMN, as high trimethylamine and dimethylamine levels
and nitrite react with the aid of bacterial metabolism. These food products are
associated with esophageal cancer in certain areas of the world.10 Cured meats,
like hot dogs and cold cuts, contain amino acids, which provide secondary amine
substrates for nitrosamine formation.23
Figure 3.1 shows examples of the three major classes of secondary carcino-
gens. Both PAHs and HCAs are very lipophilic and planar molecules, and they
are metabolized by similar P450 enzymes and have similar effects on gene expres-
sion through the aryl hydrocarbon receptor (AhR). NAs tend to be somewhat more
water soluble and have different activation routes and induction patterns.
Tobacco smoke is a rich source of PAH, HCA, and NA carcinogens. Chewing
tobacco is a more focused source of NAs, specifically the tobacco-specific nitro-
samines, including N′-nitrosonornicotine (NNN) and 4-(methylnitrosamino)-1-
(3-pyridyl)-1-butanone (NNK). These form in a reaction between nitrite
(enhanced by fertilization practices) and nicotine during the curing of tobacco
leaves.24 Thus, tobacco products and cooked meats provide a similar spectrum of
secondary carcinogens, albeit in different concentrations and via different routes
of exposure. Importantly, the risks of contracting cancer from cooked meats and
from cigarette smoke are both related to the balance of Phase I and II metabolic
capacity of the individual, responding to diet, lifestyle and genetic factors.
Secondary carcinogens must be metabolized to prevent them from accumu-
lating in the fashion of polychlorinated biphenyls (PCBs) or methyl mercury.
The main enzyme system that accomplishes this is cytochrome P450, a family
of broad spectrum enzymes that charge substrates with electrons until they
oxidize (Phase I), creating functional groups that can be conjugated by Phase
II reactions.25 Essentially all dietary carcinogens require “bioactivation” by
Phase I reactions to form the ultimate carcinogens that disrupt DNA structure,
leading to mutations and cancer. Phase II reactions, especially glutathione and
FIGURE 3.2 Secondary carcinogen bioactivation and conjugation. Stable secondary car-
cinogens require metabolism to prevent accumulation, leading to the formation of reactive
intermediates that cause DNA damage and cancer. These can be safely metabolized to
water-soluble conjugates, if the balance of Phase I and II enzymes and conjugating agents
is favorable.
glucuronide conjugation, are usually protective in the detoxification of the

reactive products of Phase I reactions. N-acetylation is a notable exception, in
that it aids in the bioactivation of HCA.25 These pathways are summarized in
Figure 3.2.
While dietary carcinogens need to be metabolized by P450 routes to prevent
them from accumulating, they do not need to be metabolized rapidly. We consume
carcinogens in relatively small amounts, and they can be effectively cleared with
low levels of P450 activity. Very high levels of P450 activity will generally
increase cancer risk, as ultimate carcinogens are generated more rapidly than they
can be cleared by Phase II conjugation reactions. This has been demonstrated in
many animal model systems and is supported by human epidemiological findings.
Higher levels of dietary protein enhance liver P450 activity and, in turn, increase
the hepatocarcinogenicity of aflatoxin in rats.26 Similarly, ethanol intake by rats
induces specific forms of P450, increasing the bioactivation of NA compounds.27
This reflects the synergism in human populations between alcohol abuse, cigarette
smoking, and head and neck cancer incidence.28
As expected, a high capacity for Phase II conjugation reactions will generally

decrease cancer risk. This can also be seen in the protective effect of dietary
sulfur amino acids during the treatment of acetaminophen toxicity, where support
of glutathione levels aids in the conjugation of the reactive acetaminophen metab-
olite created by P450 metabolism.29 Thus, diet and lifestyle have a large impact
on the susceptibility to a given dose of secondary carcinogens, through the
expression of genes for P450 enzymes, conjugation enzymes, and the enzymes
that generate conjugating agents.
3.3 ANIMAL-BASED RESEARCH ON COOKED MEAT

CARCINOGENS
It is important to note that well done meat contains a wide variety of HCA and
PAH carcinogens, with additional NAs associated with meat preservative use.
Animal models have followed reductionist approaches in testing potential car-
cinogens individually and usually using high, single-dose models, many of which
are not even based on oral exposure routes. Large numbers of PAHs, HCAs, and
NAs have been shown to be potent multiple organ and transplacental carcinogens
in mice and rats.30 These data are too extensive to cover here. Very little work
has been conducted on the consumption of cooked and preserved meat products
in rodent models, or even chronic low-level dietary exposure to mixtures repre-
senting the different classes of secondary carcinogens. Feeding hot dogs to rats
and mice was shown to cause a progressive increase in NA levels as the diet
moved along the gastrointestinal tract (GIT).31 Feeding a well-cooked beef diet
enhanced 1,2-dimethylhydrazine-induced cancer in rats.32 Conversely, diets con-
taining fried chicken, beef, lamb, pork, and fish were not found to cause mutations
in the Dlb-1 locus in mice,33 and cooked beef was not found to cause PhIP adducts
or genomic instability in the GI tissues of rats.34 A standardized rodent model
for exposure to cooked and preserved meats would allow important work to be
conducted on cancer prevention strategies.
3.4 METABOLISM OF DIETARY SECONDARY

CARCINOGENS
3.4.1 METABOLISM OF NAS
NAs are converted by P450 enzymes to reactive intermediates that break down
to form diazonium ions, which in turn react with DNA bases, leading to adducts
and mutations. DMN is bioactivated most actively by CYP2E1, the P450 enzyme
that is induced by ethanol exposure. The association of head and neck (oral, nasal,
esophageal, larynx, pharynx) cancer risk with ethanol use is likely due, in part,
to the induction of P450 enzymes in these tissues, leading to more rapid bio-
activation of dietary carcinogens. Ethanol exposure causes a rapid induction of
CYP2E1 in gastrointestinal tissues and the liver, via protein stabilization. Ethanol
also has a solvent effect in the transport of dietary carcinogens into epithelial
tissues, and ethanol also causes cytotoxicity and cell division, which may also
increase cancer risk. There is a dramatic synergism between ethanol consumption
and smoking with respect to head and neck cancer risk.28 The smoke-specific
nitrosamine NNK is bioactivated by various P450 enzymes, including CYP2E1
and CYP2A family enzymes.35
NAs are metabolized by P450 to unstable intermediates that break down
spontaneously to form alkyldiazonium ions, which are the ultimate carcinogen
species.36 These are short-lived and Phase II metabolism of these intermediates
is not a popular research area, but they are electrophilic species and the majority
of conjugation occurs with glutathione. Electrophiles are species with a net
positive charge or a local area of positive charge that can react with other
molecules. Glutathione contains a sulfhydryl group with a high electron density
that allows it to react with electrophilic intermediates, preventing them from
alkylating cellular proteins and DNA. This Phase II conjugation is enhanced by
several forms of glutathione-S-transferase, an enzyme with high activity in tissues
that have to metabolize xenobiotics, especially the liver. Oral support of glu-
tathione via N-acetylcysteine supplementation decreased the induction of esoph-
ageal cancer in rats following diethylnitrosamine treatment.37
3.4.2 METABOLISM OF PAH AND HCA FAMILIES

Conditions will tend to produce a mixture of PAHs and HCAs in most well-
cooked meat products. High temperature cooking with flareups and noticeable
black residue will indicate PAH formation. Any combustible substrate that burns
incompletely will form small radicals, which polymerize to create PAHs.25 These
reactions are favored by a high fat content and high cooking temperatures. PAHs
are also formed by the burning of woods chips in food smokers and in cigarette
smoke. PAHs from smoke will diffuse into food products and lung tissue due to
their lipophilic nature.
In contrast, HCAs are favored by slower cooking at lower temperatures,
leading to drying, which concentrates the breakdown products of amino acids
and creatinine, which polymerize to form HCA end products like PhIP and IQ.3
PAH and HCA families are similar in several regards; the parent compounds are
planar and lipophilic and are activated by CYP1A family enzymes. Activation
may require more than one CYP1A oxidation (e.g., BaP) or the participation of
Phase II conjugation (e.g., N-acetylation in the activation of HCAs). Both PAHs
and HCAs interact with the AhR and induce CYP1A family enzymes.
3.4.3 METABOLISM OF PAHS

Most of the research on PAHs metabolism has been conducted on BaP, which is
an excellent substrate for CYP1A family P450 enzymes (CYP1A1 in lung and the
GIT, CYP1A2 in the liver). BaP is converted to a 7,8-epoxide by CYP1A enzymes,
but this is quickly reduced to a 1,8-diol by epoxide hydratase. A second CYP1A
epoxidation occurs at the 9,10 position, and further metabolism of this
electrophilic residue is hindered by the presence of the bay-region ring structure.

This impairs the activity of glutathione-S-transferase (GST) toward this intermedi-
ate, and UDP-glucuronyltransferase (UDPGT) is required to conjugate the 7,8-diols.
In considering the whole family of PAHs metabolites, there will be a mixture of
glutathione conjugation of electrophiles and glucuronic acid conjugation of
nucleophiles38. Many dietary components that act as chemopreventative agents are
found to induce the expression of GST and UDPGT enzymes (see Section 3.11).
3.4.4 METABOLISM OF HCAS

HCAs are also excellent substrates for CYP1A family enzymes and the resulting
products are N-hydroxylated intermediates. These intermediates tend to be N-
acetylated or sulfated in a Phase II reaction that creates the ultimate carcinogenic
metabolites.39 This is one of a small list of cases in which Phase II conjugations
are procarcinogenic. HCAs metabolites are varied in structure and both glu-
tathione and glucuronic acid conjugation appear to play a role in detoxification.
PhIP is the most abundant HCAs in cooked meats, although others are more
potent carcinogens. Reactive PhIP metabolites may be metabolized by conjuga-
tion with both glutathione and glucuronic acid,40,41 while other HCA metabolites
appear to be mainly conjugated via glucuronidation.42 Individuals with low activ-
ity of the glutathione-S-transferase required for PhIP conjugation appear to be at
higher risk for the development of colorectal cancer.40 Conversely, high activity
polymorphisms for N-acetyltransferase (fast acetylators) increase the risk for
occurrence of colorectal cancer when combined with a high intake of red meat .43
3.5 AFLATOXIN AND FOOD SPOILAGE

Aflatoxin is an important hepatocarcinogen in developing countries. It represents
a diversion from the previous examples in that it forms mainly on plant products,
and it forms through spoilage rather than cooking. It is also a secondary carcin-
ogen and its potency is also affected by changes in Phase I and II metabolism.
Aflatoxin is activated to a hepatocarcinogenic epoxide by CYP3A4 and CYP1A2,
and significant protection is provided by glutathione conjugation of this interme-
diate.44 Human liver expression of the most protective form of glutathione-S-
transferase is normally quite low and chemopreventative agents may function by
inducing these enzymes.44 Poorly stored grain products and peanuts are examples
of foods that may be contaminated with aflatoxin.
3.6 EFFECTS OF CARCINOGEN EXPOSURE ON

XENOBIOTIC METABOLISM
When xenobiotics enter a cell, they may interact with two main types of proteins,
namely enzymes and receptors. The primary enzymes will be a variety of P450
gene products, with 57 individual members, with overlapping substrate recogni-

tion. This will usually lead to a catalytic reaction with the formation of oxidized
products of the xenobiotics, possibly leading to the formation of reactive, ultimate
carcinogens. Conversely, the xenobiotic may bind to a receptor protein, forming
a complex that enters the nucleus and drives the expression of new mRNA from
a specific group of genes. This is illustrated in Figure 3.2 and Figure 3.3.
FIGURE 3.3 Cytochrome P450 activity and induction. Cytochrome P450 activity is lim-
ited to the catalytic cycle in the upper left of the diagram. Induction patterns, of both Phase
I and II enzymes, play a critical role in the potency of dietary carcinogens. *Xenobiotics
bind to specific receptors, which then act as transcription factors to induce the synthesis
of new P450 enzymes, usually with a high capacity to metabolize that specific xenobiotic.
**Phytochemicals tend to compete directly at the P450 active site to slow carcinogen
bioactivation. Competition also takes place at the AhR, where phytochemicals can decrease
the induction of P450 expression caused by secondary carcinogens. Large exposure to
phytochemicals can activate the AhR, however, leading to procarcinogenic effects.
***Phytochemicals induce multiple forms of Phase II conjugation enzymes and agents.
As an example, benzo(a)pyrene (BaP) will enter a cell and be metabolized

to reactive intermediates by CYP1A1/1A2 gene products. At the same time, BaP
will also bind the aryl hydrocarbon receptor (AhR), forming a complex that
induces the synthesis of a spectrum of new mRNA sequences, including that of
CYP1A1/2 genes.45 Therefore, with chronic exposure to BaP, there will be a
progressive increase in CYP1A1/2 protein levels, which will enhance the rate of
bioactivation and ultimate carcinogenicity of BaP. This has been shown in numer-
ous animal models and even in human subjects consuming grilled meats.46
This system is logically designed to enhance the metabolic capacity for
xenobiotics, which are present in tissues at a given time. It doesn’t always favor
long-term health, however. From an evolutionary perspective, it is more important
to prevent the accumulation of lipophilic contaminants, which could disrupt fetal
development, rather than prevent the long-term development of cancer. This is
especially true in cultures where the average lifespan was shorter than that
required to observe significant cancer incidence. This would have been true
throughout the majority of evolutionary history, and the recent increase in human
lifespan presents chronic disease conditions that were not a major issue in the
distant past.
Some xenobiotics induce P450 genes that are not effective in metabolism of
the inducing agent. Examples include dioxins and PCBs, which bind to and
activate the AhR but are not metabolized by CYP1A1/2 with any efficiency. A
broad spectrum of oncogenic genes are induced via dioxin-AhR activation, while
other AhR ligands appear to decrease cancer risk.45 These differences may be
based on dose/potency and will be discussed later. Other receptors are present to
control the expression levels of other P450 families, including the steroid and
xenobiotic receptor (SXR) and the constitutive androstane receptor (CAR).47 The
SXR receptor controls the expression of some CYP3A family enzymes, which
are the work horses of xenobiotic metabolism, comprising the majority of intes-
tinal and liver P450 expression. The SXR is activated by binding with a wide
variety of substrates, including endogenous and synthetic steroids, antibiotics, and
various phytochemicals. One potent activator of the SXR is hyperforin, the active
agent in St. John’s wort, an antidepressive herbal product. There have been many
cases of St. John’s wort leading to failure of pharmaceutical regimens, including
immunosuppressant medication and birth control, due to induction of CYP3A4
and decreased circulating drug concentration.47 The CAR was found to mediate
the classical induction of the CYP2B subfamily by phenobarbital, but the response
is now known to be much more complex. CAR, SXR, and the retinoid X receptor
(RXR) heterodimerize and generate complex patterns of P450 expression in
response to xenobiotic and endobiotic exposure.47 Other nuclear receptors that
play a role in P450 expression include the peroxisome proliferator activated
receptor (PPAR), the liver X receptor (LXR), and the farnesoid X receptor (FXR).48
Phase II conjugation enzymes, and the enzymes required to synthesize con-
jugating agents, are also regulated in response to environmental exposure to
xenobiotics. The pattern of regulation is more complex than seen in Phase I genes.
There are significant roles for the AhR, CAR, SXR/PXR, FXR, LXR, and PPAR
receptors in the expression of Phase II enzymes like glutathione-S-transferases

and UDP-glucuronyltransferases.49 It is important to note that these receptors
form a transcription factor after binding with a ligand, and the resulting complexes
have unique actions. A dioxin-AhR complex is an excellent CYP1A inducer,
while an indole-3-carbinol (a known AhR agonist) will induce both Phase I and
II genes, but will antagonize the stronger dioxin-AhR induction of CYP1A
genes.50 There is an additional mechanism that detects oxidant stress through a
complex of Keap1/Nrf2 proteins in the cytosol. Oxidant stress, induced by reactive
oxygen species or electrophilic xenobiotic metabolites, causes the release of Nrf2,
which travels to the nucleus and binds to antioxidant response elements (ARE)
in numerous gene promoters.51 This pathway is active in the induction of glu-
tathione-S-transferase and UDP-glucuronyltransferase genes, and also induces
gamma-glutamyl cysteine synthetase (rate limiting in GSH synthesis) and UDP-
glucose dehydrogenase (last enzyme in UDP-glucuronic acid formation).52
These complex patterns of gene expression in Phase I and II enzymes play
a central role in determining the potency of a given exposure to dietary secondary
carcinogens and are central to the effects of chemopreventative agents. Human
subjects who consumed a diet high in char-grilled meat for 7 days were found
to have a significant induction of intestinal and liver CYP1A activity.46 In a similar
design with pan-fried meat consumption, HCAs alone were shown to induce
hepatic CYP1A2 activity.53 While these experiments did not determine the extent
of DNA damage in target tissues, animal models suggest that the induced state
following cooked meat consumption represents enhanced cancer risk, even if the
exposure to secondary carcinogens was kept constant.
3.7 DIET AS A SOURCE OF CHEMOPREVENTATIVE

AGENTS
Plant-based foods are a good source of almost all of the nutrients required by
humans, but this is a relatively small number of chemicals (40 to 45, depending
on interpretation). When we consume fruits and vegetables, we are exposed to
thousands of additional compounds that may present health benefits or toxico-
logical risks. The important concept is that these are xenobiotics (strange to life),
not nutrients. Very few of these compounds are present for the purpose of being
beneficial to human health, although long-term cultivation of certain plants has
created some examples of this. Most represent chemical defenses (phytoalexins)
designed to counter rodent, insect, or fungal attacks. When humans or experi-
mental animals consume these compounds, the compounds are metabolized by
typical xenobiotic enzymes and often have dramatic effects on Phase I and II
enzyme activities and expression patterns.
In contrast to the effects of meat consumption, the incidence of various GI
cancers54 and lung cancers55 decreases with increased vegetable consumption.
Public attention, popular press, and the research community have focused
attention on the antioxidant roles of nutrients and phytochemicals in fruits and
vegetables. The antioxidant capacity of food extracts may be tested by methods

that determine the ability of compounds to scavenge radicals in an in vitro system
(e.g., FRAP, TEAC, and ORAC methods).56 In these tests, both the phytochem-
icals and radicals are at high concentrations and placed in close proximity, which
does not generally reflect the in vivo situation of tissue distribution and metab-
olism, and subcellular localization of these compounds. Interestingly, some whole
animal and human intervention experiments show that biomarkers of oxidant
stress are decreased and health end points, such as memory and brain function
in aging rats, are improved.57,58 These data are not as clear as it seems, however.
The main defense against reactive oxygen species in mammalian cells depends
on glutathione (GSH) as a reducing agent, and the selenium-dependent enzyme
glutathione peroxidase to detoxify hydrogen peroxide. During chronic oxidant
stress, the pathway for glutathione synthesis is upregulated, namely the limiting
enzyme, gamma-glutamylcysteine synthase.59 If a diet actually protects against
oxidant stress, this enzyme should be downregulated, but it is, in fact, upregulated
with the consumption of a high berry diet,60 and by flavonoids from onions and
other vegetables.61 The apparent explanation is that phytochemicals (which are
xenobiotics) create a low level of oxidant stress that enhances oxidant defense,
leading to improved health. There are many examples of small, manageable stresses
working to strengthen a system and improve health. This is observed in the case
of regular exercise, which increases the formation of reactive oxygen, but enhances
defense systems,59,62 in the end decreasing tissue biomarkers of oxidant stress63 and
delaying the development of chronic disease. This concept of low-level stress
improving cellular function is an important and underappreciated concept in toxi-
cology. The public, and many policy makers, view exposures to one group of
compounds as good, and exposure to another group of compounds as dangerous,
without proper consideration of dose response (further discussion in Section 3.13).
CARET and ATBC experience was an extension of the public and scientific
obsession with oxidant stress and defense as a central theme in the fruit and
vegetable cancer interaction. CARET and ATBC were two randomized human
interventions in which human smokers were treated with large doses (about fivefold
above normal dietary intake) of beta-carotene,55,64–66 based on reasonably strong
epidemiology associating high carotenoid intake with decreased lung cancer inci-
dence. Assuming that oxidant stress was a central force in lung cancer development
and that beta-carotene was an effective radical scavenger, it was hypothesized that
supplementation would decrease lung cancer development. Both large studies found
a significant increase in lung cancer incidence, cardiovascular events, and overall
mortality with double-blinded beta-carotene supplementation, over the relatively
short period of 5 to 8 years. One of the most interesting findings in both the CARET
and ATBC studies was that people with higher baseline carotenoid status were still
protected from lung cancer,55,66 even in the beta-carotene supplemented group where
the baseline carotenoids should have just been adding to the excessive carotenoid
intake. It was clear from these studies that baseline carotenoids were acting as a
proxy for the intake of other nutrients or phytochemicals, which were providing a
significant protection against cigarette smoke-induced carcinogenesis.
One of the basic assumptions driving these clinical trials was that lung cancer
is caused by oxidant stress. It has subsequently been shown that the mutations
present in smoke-induced cancers are not similar to those caused by oxidant
stress. Instead, they are similar to those caused by the two major classes of
secondary carcinogens present in smoke, namely PAHs and smoke-specific nit-
rosamines, like NNK.67 As described earlier in this chapter, PAHs and NAs are
secondary carcinogens that require bioactivation by endogenous P450 enzymes,
and the resulting reactive intermediates are not detoxified by enzymes involved
in reactive oxygen metabolism. Animal models of lung cancer induced by these
classes of chemicals have shown that various phytochemicals, and other agents
that influence Phase I and II metabolism, have potent effects on lung cancer
incidence.68 Conversely, beta-carotene use in animal models has demonstrated
pro-oxidant effects69 and induction of P450 enzymes, which could enhance car-
cinogen bioactivation.70
Similar to the lung cancer situation, the incidence of GI cancers is associated
with the exposure to secondary carcinogens (consumption of well-done and
preserved meats) along with low vegetable and fruit consumption. Experiments
on phytochemical fractions from sources such as vegetables (glucosinolates68),
spices (curcumin71) and green tea (catechins like ECGC72) have shown that these
are very active in the prevention of chemically-induced cancers.73 Several types
of cancers (e.g., GI, lung) are about 50% lower in the upper quartile of vegetable
consumers.74 This relationship is well explained by metabolic interactions
between vegetable phytochemicals and dietary secondary carcinogens.73 Changes
in Phase I and II metabolism largely determine cancer risk from a given exposure
to secondary carcinogens. Chronic exposure to carcinogens alone causes binding
and activation of the aryl hydrocarbon receptor (AhR),45 and other related recep-
tors, which cause a dramatic upregualtion of specific P450 isozymes, like
CYP1A1/246 (Figure 3.2). This increases the rate of formation of reactive inter-
mediates and dramatically increases cancer risk from a given exposure to sec-
ondary carcinogens.25,75 In contrast, the impact of phytochemical exposure on
Phase I and II metabolism is complex, including the following effects:
1. Phytochemicals (alone) may activate AhR, causing a small induction

of P450 activity.76,78
2. In combination with carcinogen exposure, phytochemicals compete
with carcinogens for binding to the AhR, minimizing the large P450
induction seen with chronic exposure to carcinogens alone.78,79
3. Phytochemicals also compete for binding to the active site of P450
proteins, acting as direct competitive inhibitors and noncompetitive
inactivators of CYP1A1/2 and CYP2E1 enzymes.79–84
4. Phytochemicals induce the major Phase II conjugation enzymes (glu-
tathione-S-transferases and UDP-glucuronyltransferases),48,77 and the
enzymes that form the conjugating agents.60,61,85,86
The following sections will expand on these interactions between phytochem-

icals and xenobiotic metabolism.
3.8 PHYTOCHEMICAL ACTIVATION OF THE AHR

Many phytochemicals have been shown to bind to the AhR and/or induce
CYP1A family P450 genes. Perhaps the most studied is indole-3-carbinol
(I3C), which is an atypical breakdown product of a glucosinolate found in
brassica plants like cabbage, Brussels sprouts, and broccoli. Most glucosino-
lates break down to form a mixture of thiocyanates and isothiocyanates, of
which the thiocyanates impair iodine utilization (goitrogens) and the isothio-
cyanates interact with xenobiotic metabolism and have cancer preventative
properties. I3C differs in that the parent indolylmethyl-glucosinolate breaks
down to an isothiocyanate, but this rearranges and decomposes to release a
simple thiocyanate ion and I3C, which is neither isothiocyanate nor thiocyan-
ate in structure. I3C dimerizes to diindolylmethane (DIM) in the acid envi-
ronment of the stomach. I3C and DIM are both capable of binding to and
activating the AhR,45,50,87 although they are considered weak agonists.88 I3C
and DIM will induce CYP1A family P450 genes in a variety of experimental
models,87 but tend to function in the low to mid micromolar range, compared
to a low nanomolar range for the prototypical AhR agonist TCDD.87,88
Many other phytochemicals will bind to the AhR and induce CYP1A expres-
sion, including phenethyl isothiocyanate (PEITC) and sulforaphane (brassica),
curcumin (turmeric), isoxanthohumol and 8-prenylnaringenin (flavonoids in beer
hops), chrysin (various plant sources), phloretin (strawberries, apple skin),
kaempferol (onions, grapes, apples, citrus family), galangin (tree bark, bee pro-
polis), naringenin (citrus fruits and tomatoes), daidzein and genistein (soy), quer-
cetin (onions, apples, tea, cranberries), myricetin (berries, grapes), luteolin
(parsely, celery), baicalein (various herbals), apigenin (parsley, celery), and
diosmin (citrus), cantharidin (beetle toxin), resveratrol (grapes, wine), green tea
extracts, and emodin (aloe).89–91
It is interesting to consider the etiology of AhR activation by phytochemicals
and the implications for cancer risk. It is a clear advantage to be able to induce
forms of P450 capable of metabolizing dietary xenobiotics. Lipophilic xenobiotics
will accumulate and tend to impair fetal brain development (like methyl-mercury,
PCBs, dioxins), which would represent a selective disadvantage. Conversely,
rapid bioactivation of xenobiotics may enhance cancer risk, but given the expected
lifespan throughout the majority of our history and the fact that reproduction
occurs at a younger age, avoiding cancer beyond the age of 30 or 40 years may
not have created any selective pressure.
Of course, the epidemiological picture is that a high intake of fruit and
vegetable phytochemicals is associated with cancer protection rather than
enhanced risk. However, the induction of AhR by phytochemicals can be linked
to increased cancer risk in animal models, most of which are unphysiological in
nature. At high doses, I3C, an AhR agonist, is well recognized to promote
carcinogenesis in several animal models. By inducing CYP1A genes, it increases

the potency of PAHs and HCAs secondary carcinogens. Other genes induced by
AhR activation are oncogenic in nature.45 These levels of I3C are unphysiological
with respect to a food-based diet, but may now be approached in human diets
through the availability of potent supplements. In addition to excessively high
exposures to I3C, some models divide the timing of exposure to I3C and carcin-
ogen, where I3C exposure may induce P450 expression and then be removed
from the diet before carcinogen exposure. Conversely, in other models, carcinogen
exposure may be completed before I3C is introduced into the diet. In both of
these cases, I3C may enhance carcinogenesis by activating the AhR, without the
potential benefits that occur when I3C and secondary carcinogens are being
metabolized simultaneously. Concurrent exposure is a more physiological model
for human cancer risk, and the following sections describe further mechanisms
that will function under these circumstances.
3.9 PHYTOCHEMICAL ANTAGONISM OF THE AHR

In real conditions that determine human health, people are usually exposed to
small amounts of numerous chemicals for up to decades in time. Under these
conditions, small changes in the balance of metabolism between Phase I and II
pathways, or the routes of Phase I that handle certain xenobiotics, can lead to
dramatic differences in life-long cancer risk. These conditions are poorly repli-
cated by the high single-dose rodent models for carcinogen testing. Even when
chemopreventative dietary components are introduced in high-dose rodent mod-
els, the routes of metabolism and role of DNA repair processes are responding
to unphysiologically high metabolic burdens over a very short time frame, and
the results may be a poor model of human diet–cancer relationships.
The induction of P450 genes, through activation of the AhR and other nuclear
receptors, generally reflects a procarcinogenic effect, but this depends on the level
of activation and whether it is accompanied by Phase II changes. While the
previous section described unphysiological conditions under which dietary phy-
tochemicals could be procarcinogenic, they tend to act differently when present
for a long duration at low levels in the presence of other classes of xenobiotics.
Under these conditions, phytochemicals and secondary carcinogens are both
present inside the cell, with the ability to compete for binding and activation or
inhibition of nuclear receptor function. This has been studied most extensively
for the AhR. In general, when phytochemicals and planar carcinogens (PAHs and
HCAs) are present concurrently, phytochemicals will antagonize the strong AhR
activation of planar carcinogens. Some specific examples are provided below.
The experimental models vary from in vitro transcription assays through cell
culture work to in vivo animal data.
Several screening models have been developed based on AhR complex detec-
tion or the activity of reporter genes attached to CYP1A promoters in cultured
cells.91,92 These have been used to identify dietary components that agonize and
antagonize the AhR. Allen et al. found that resveratrol, apigenin, curcumin,
kaempferol, and green tea extract weakly enhanced the AhR function when used
alone (two- to sixfold at 10 µM), and green tea extract, naringenin, and apigenin
significantly antagonized the strong TCDD-induced activation of AhR (10-fold
at 1 nM).91 Curcumin and TCDD produced an additive effect on AhR activation.
Amakura et al. found that green tea catechins and extracts of sage, spinach, and
citrus fruits produced antagonism of TCDD-induced AhR activation.92 In other
studies, I3C and DIM, kaemferol, quercitin, myricetin, and luteolin antagonized
TCDD-induce AhR activation.50,89
Green tea components are the best characterized phytochemicals with respect
to AhR interactions. Individual catechins have shown lower antagonism than
green tea extracts.91 Williams et al. found that a mixture of the four major
catechins displayed a strong antagonism similar to whole tea extract,93 while
Fukuda et al. showed that tea-derived lutein and chlorophyll a and b are also
antagonists of AhR activation.94 Inconsistent responses to epigallocatechin-3-
gallate (EGCG), a major tea catechin, may be explained by the finding that this
phytochemical does not compete at the binding site of the AhR, but appears to
inhibit the conformation changes subsequent to binding, through interactions with
accessory proteins.95 Black tea theaflavins also suppress the transformation of
TCDD-bound AhR to a transcriptionally active form.96
The importance of these observations is that, while many phytochemicals
alone are weak agonists of the AhR, they are also antagonists of the strong
activation of the AhR by compounds like TCDD and planar carcinogens. In a
typical human diet, these compounds will be presented together and the effect of
phytochemicals may be to modulate the AhR activation to a safer level, limiting
the degree of P450 induction and encouraging a safe balance between Phase I
and II metabolism. This has been verified by an elegant series of experiments in
a physiologically relevant rat model by Thapliyal et al.79,97,98 This model is based
on the prevention of BaP and DMN-induced carcinogenesis by the root spice
turmeric, in which the active ingredient is curcumin (diferuloylmethane). In this
model, dietary turmeric provided dramatic protection against the formation of
BaP adducts in liver, lung, and stomach tissues when consumed for 2 weeks
preceding the dose of BaP, but not when the turmeric was introduced after the
BaP dose.98 DMN-induced cancer of the liver was also decreased by ongoing
turmeric consumption, and not by turmeric following DMN treatment.97 Thus,
the effects in this model are mainly in the bioactivation and initiation period as
has been discussed in this chapter. Further experiments showed that dietary
turmeric alone had variable effects on P450 enzymes in the liver, lung, and GI
tract, from small depressions to modest inductions,79 as described in Section 3.8.
When present in the diet during exposure to BaP, turmeric decreased the large
induction of CYP1A1, 1A2, and 2B1 by 10 to 80%.79 These studies also dem-
onstrated direct inhibition of the catalytic activity of the P450 enzymes and the
induction of Phase II enzymes, as will be discussed below.
3.10 PHYTOCHEMICAL INHIBITION OF THE

CATALYTIC ACTIVITY OF P450
While various phytochemicals may cause some level of induction of P450 expres-
sion, many of the resulting enzymes will be rendered less active through different
forms of inhibition if the phytochemicals continue to be present in the diet.
Inhibition of the P450 catalytic cycle has been convincingly demonstrated for a
long list of phytochemicals.
Thapliyal et al. have demonstrated this inhibition in vitro, showing that
curcumins and isothiocyanates inhibit CYP1A1, 1A2, and 2B1 in rat microsomes,
with IC50 values between 1 and 20 µM.79 Diosmin, naringin, naringenin, rutin,
and quercetin from citrus fruits inhibited CYP1A family enzymes and prevented
the first step in the in vitro bioactivation of dietary HCAs and the tobacco
nitrosamine NNK.99,100 Phytochemicals from hops, used in the brewing of beer,
including xanthohumol, isoxanthohumol, and 8-prenylnaringenin, largely elimi-
nate CYP1A1/2 and 2B1 activity at 10 µM concentrations.101 This prevented the
in vitro bioactivation of aflatoxin, with very little impact on CYP2E1 or 3A4.
Quercetin, ethoxyquin, tannic acid, theaflavins, and tea polyphenols inhibit the
N-hydroxylation of HCAs by rat and human liver microsomes.102 Benzyl and
phenyl isothiocyanates and resveratrol, at levels below 1 µM, dramatically inhib-
ited several forms of P450 in hamster liver microsomes.103 Capsaicin (hot peppers)
inhibited PhIP bioactivation by rat, hamster, and human liver S9 fractions.104 PhIP
mutagenicity (Salmonella reversion) was also inhibited by lycopene, caffeine,
daidzein, and genistein.105 Allyl sulfides (garlic) inhibit the bioactivation of nit-
rosamines by rat and human CYP2E1.106
Different flavanoid structures show selectivity for the inhibition CYP1A1 vs.
1A2,107 which could change the route of metabolism of some secondary carcin-
ogens or change the tissue in which metabolism takes place (CYP1A1 in the GIT
and lung, CYP1A2 in the liver). Tea flavanols have demonstrated many protective
effects in vitro and in animal models of carcinogenesis,108,109 while the human
epidemiology is favorable for the most part, but less clear.109 Some of this
uncertainty may be resolved by recent work showing that EGC and EGCG cause
activation of EROD activity in rat liver microsomes at nM levels, while dramat-
ically inhibiting the same activity at µM levels.110 This assay measures CYP1A1
activity in rats and, if this effect also occurs in human CYP1A1 in extrahepatic
tissues, it could explain some inconsistencies in the chemopreventative effects of
green tea.
Soy, mushroom, and grape phytochemicals have been found to inhibit aro-
matase (CYP19), which plays a role in the formation of estrogens and may impact
on hormonally sensitive cancers.111
Brassica isothiocyanates, such as PEITC and sulforaphane, have demon-
strated a rather selective inhibition of rat CYP2E1 in vitro,112 and have been
shown to protect against CYP2E1-mediated in vivo endpoints like DMN-induced
cancer,113 and acetaminophen and ethanol-induced hepatotoxicity.114 In a careful
study of PEITC inhibition of human P450 enzymes, Nakajima et al. found
competitive inhibition of CYP1A2 and 2A6, noncompetitive inhibition of

CYP2B6, 2C9/19, 2D6, and 2E1, and mechanism-based inactivation of
CYP2E1.84 A similar inactivation process occurs between PEITC or BITC and
CYP2B1, and it appears to involve a covalent modification of one amino acid on
the enzyme protein by a reactive intermediate of the isothiocyanate molecule.115
This type of inhibition can be potent and long-lived in an in vivo situation, as the
protein is permanently inactivated by a short exposure to dietary phytochemicals,
and activity will only recover as new protein is synthesized.
A number of studies have verified that phytochemicals can inhibit P450
activity in vivo, using both rodent116 and human models. Human studies are
usually limited to the use of probe drugs, which may not provide accurate data
on the tissue site or species of P450 involved. The herbal products kava and
goldenseal caused inhibition of CYP2E1 and CYP3A4/5, respectively.117 Elderly
patients experienced an induction of CYP3A4 with St. John’s wort and an inhi-
bition of CYP2E1 following garlic oil.118 In these experiments, the treatment
period was 4 weeks long, and changes may have been due to direct effects on
the enzymes or through gene expression. In a more focused design, a single
ingestion of watercress, a source of brassica isothiocyanates, decreased the clear-
ance of a CYP2E1 probe.119
Direct inhibition or inactivation of P450 enzymes by acute exposure to phy-
tochemicals has the potential to overcome any inducing effects that chronic
exposure to phytochemicals or carcinogens may have caused. This effect may
create a healthier balance between Phase I and II metabolism or force carcinogen
metabolism through safer forms of P450.
3.11 PHYTOCHEMICAL INDUCTION OF PHASE II

ENZYMES
As discussed earlier in this chapter, the induction of Phase II enzymes is respon-
sive to the same nuclear receptors that control P450 gene expression, but is also
controlled via cellular redox status, the Nrf2 transcription factor, and antioxidant
response elements in gene promoters. Thus, the pattern of Phase II induction
from a given xenobiotic exposure is generally different from the Phase I induction.
Planar carcinogens, like PAHs and HCAs, are very active in the induction of
CYP1A family enzymes, but are generally mild inducers of Phase II enzyme
expression.
Many brassica isothiocyanates and metabolites are active in the induction of
glutathione-S-transferase (GST), UDP-glucuronyltransferase (UDPGT), quinone
reductase (QR), and gamma-glutamylcysteine synthase (GGCS) enzymes.120
While PEITC, I3C, and DIM are “bifunctional inducers” that also enhance P450
expression, some brassica phytochemicals, like sulforaphane (4-methylsulfinyl-
butyl isothiocyanate), are very specific in the induction of Phase II enzymes.
Sulforaphane is found at high levels in sprouting broccoli seeds, and extracts of
these sprouts are effective chemopreventative agents.121 The involvement of
Phase II enzymes is demonstrated by the enhanced sensitivity of Nrf2-null mice

to BaP-induced cancers, and the fact that Nrf2-null mice are no longer protected
by Phase II-inducing agents.120
Aliphatic sulfides from garlic and onions, including compounds, such as
diallyl disulfide and trisulfide, are also well-known inducers of Phase II
enzymes.122 These have been associated with decreased risk of GIT cancers in
humans and have proved effective in prevention of GIT and lung cancers in animal
models.123 Feeding of soy to rats also induces GST, UDPGT, and QR activities
in various tissues.124
Phase II reactions are not always beneficial. The final step in the bioactivation
of HCAs involves the acetylation or sulfation of an N-hydroxylated HCA metab-
olite. Fast acetylating phenotypes are associated with increased risk of certain
cancers, and rapid sulfation could create similar risks. Garlic sulfides are inhib-
itors of N-acetyltransferase activity and may also decrease the risk of HCA-
induced cancer by this mechanism.125 Similar to the garlic sulfides, kahweol and
cafestol palmitates from unfiltered coffee are inducers of GST and inhibitors of
N-acetyltransferase activities, and feeding of these phytochemicals decreases the
level of PhIP–DNA adducts in rat colon.126 Compounds that inhibit N-acetyl-
transferase activity may be particularly useful for individuals with high activity
polymorphisms for these genes, to avoid the enhanced cancer risk that may be
associated with this genotype.43 Interestingly, soy phytoestrogens are potent inhib-
itors of sulfotransferase activity, which could protect against HCA-induced can-
cers, but enhance the levels of active estrogens and increase breast cancer risk.127
The phytochemicals described above appear to have an ideal mixture of
properties to act as chemopreventative agents. They appear to inhibit enzymes
involved in carcinogen bioactivation (P450, N-acetyl transferase, sulfotransferase)
and enhance the activities of those that are protective (GST, UDPGT, QR, etc.).
While most of these chemicals appear to act as defenses against various stresses
to the plant, some may have been selected over time in cultivated crops as
beneficial to human health. It is important to remember that they are being handled
by xenobiotic metabolism pathways and exerting many of their health benefits
via interactions with potent carcinogens within these pathways. As we move
towards more functional foods and nutraceutical products with enhanced levels
of these products, we need to remember that they have toxic properties at higher
levels of intake, perhaps to a greater extent than we now appreciate.
3.12 EXPERIMENTAL MODELS FOR FUTURE WORK

AND OPPORTUNITIES FOR IMPROVING
PUBLIC HEALTH
Because of the complex interactions between vegetable and meat consumption,
meat doneness, preservative levels, and associated lifestyle variables, the role of
cooked/preserved meats in carcinogenesis has remained controversial. Basic
research has shown that most members of the PAH, HCA, and NA families are
mutagenic in vitro and carcinogenic to rodents in large single dose models, but
these lack physiological relevance to human exposures. The literature also lacks
data on the effects of chronic exposure to low levels of complex mixtures of
PAHs, HCAs, and NAs. Once a model for this is established, it may be used to
examine cancer incidence, short-term biomarkers of exposure and injury, and the
protective impact of plant products and phytochemical fractions.
In 2003, American consumers spent $1.8 billion on hot dogs in supermarkets
alone128 and much more at locations outside the home ($24.2 million in major
league baseball parks128). Hot dogs are representative of a larger group of cured
meats, including luncheon meats, bacon, and sausages. Norat et al. found that
the 70th percentile of processed meat intake was around 40 g/d, while the 90th
percentile was over 80 g/d,5 representing 1 to 2 meals per day. While processed
meats are often viewed as sources of NA exposure, these products are also smoked
(causing PAH formation) and many are slow cooked to a low-moisture content
(causing HCA formation). Children represent a high-risk group that consumes
even greater levels of these products on a body weight basis, and at a young age
are more susceptible to carcinogen exposure.
Consumers remain poorly informed about meat intake and cancer. Red and
processed meat intake is not diminishing and barbequing appears to be increasing
in popularity.129 Interactions between phytochemicals and secondary carcinogens
represent a potent approach to decrease the cancer risks associated with cooked
meats. These phytochemicals can be introduced into processed meats, creating
functional foods that play a role in disease prevention. Green tea is a promising
source of active phytochemicals for this project. Green tea consumption is asso-
ciated with a decreased risk of several human cancers,130 and numerous animal-
based experiments have shown that green tea phytochemical fractions and purified
catechins inhibit high-dose PAH, HCA, and NA-induced cancers.72 As mentioned
earlier, PAHs, HCAs, and NAs are all secondary carcinogens, requiring bioactiva-
tion by P450 enzymes to cause DNA damage. There are about 60 different P450
gene products, with complex regulation in response to exposure to dietary car-
cinogens and phytochemicals.16 This regulation occurs through receptors, like the
AhR, which bind foreign compounds and become transcription factors that lead
to increased expression of P450 proteins. The AhR is very active in the induction
of CYP1A1/2 genes, which form P450 enzymes that bioactivate PAHs25 and
HCAs.75 In human experiments, regular consumption of well-done meat increases
intestinal CYP1A activity,46 an effect that will increase cancer risk from a given
carcinogen exposure. Basic research has demonstrated that green tea phytochem-
icals compete with carcinogens for binding to the AhR, and lessen the degree of
P450 induction caused by carcinogens alone.78 In addition, these phytochemicals
compete directly at the active site of the P450 enzyme, decreasing the rate of
production of reactive intermediates from carcinogens.82 Lastly, green tea phy-
tochemicals induce the expression of Phase II enzymes and conjugating agents.48
The balance of these phytochemical effects is slower production and more rapid
conjugation of reactive intermediates, decreasing the accumulation of harmful
intermediates, DNA adducts, mutations, and cancer.
In addition, phytochemical fractions influence formation of PAHs, HCAs,

and NAs in foods during storage and cooking. The formation of NAs has been
shown to be inhibited by reducing agents, including green tea phytochemicals,131
which encourage the conversion of nitrite to nitric oxide, preventing the formation
of NAs.22 Similarly, green tea phytochemicals have been shown to inhibit the
formation of HCAs in vitro from purified components131 and in cooked meats,19
and appear to decrease nitrosamine formation in the GIT of humans consuming
a nitrate-rich diet.22
3.13 PUBLIC PERCEPTION OF RISK AND RESULTING

BEHAVIOR
The public has a very biased form of risk perception in relation to diet and health.
Artificial “contaminants” of the food supply, including food additives and pesti-
cide residues, are viewed as dangerous and carcinogenic, while natural products
are seen as beneficial to health. Figure 3.4 shows several dose response curves,
with increasing exposure on the horizontal axis, and health problems (cancer?)
on the vertical axis. For the most part, these curves are theoretical extrapolations
from very few actual data points. For example, pesticide X may be tested at very
high doses in rats and found to cause or promote cancer. It is not possible to test
X at lower doses, given the number of animals that would be required and the
length of the experiment. Therefore, the low-dose area of the curve is modeled
mathematically, which can be done using widely varying models. The simplest
model is the linear response, which states that the incidence of disease will
decrease in direct relationship with the exposure to X and will never completely
disappear as long as X is present in the food supply. A second model assumes
that there is a threshold level at which X may be safely metabolized and the risk
of low exposures disappears at some point. The threshold point is very hard to
determine and is obviously critical to regulation policy. There are two more
dramatic curves to consider, the first of which states that low levels of X actually
start to be beneficial, for example, leading to levels of cancer below that observed
spontaneously (with an absence of X in the food supply). This response is called
“hormesis” and it is thought to be very common, but difficult to study experi-
mentally. Some dangerous exposures, like radiation, have been thought to present
a linear form of risk because radiation produces DNA damage at very low levels
of exposure. Surprisingly, radiation exposure produces one of the clearest and
most studied forms of hormesis. Low levels of radiation extend lifespan and
decrease cancer incidence in experimental animals. These effects are even
reflected by non-thyroid cancer incidence data from Japan and Chernobyl that
are well below predictive models.
Another established model of hormesis involves exposure to dioxin, which
demonstrates health benefits in rodents at low doses. It seems likely that some
of the pesticide residues that cause cancer at high doses in rodents may be
harmless or “beneficial” at the levels found in the human diet. Calabrese and
FIGURE 3.4 Risk models for extrapolation of cancer data. The top panel shows risk
models used by toxicologists and public health policy makers. The bottom panel shows
risk models perceived by the public, in which much higher risk is attached to artificial
components in the food supply, while natural products are perceived as increasingly
beneficial at high exposures, in the form of natural health products.
Baldwin have examined published data for evidence of hormesis in various areas
of toxicology and found that close to 10% of toxicological studies demonstrate
hormesis, although few are properly designed to do so. Among the different
categories of compounds, metals are very likely to produce hormetic dose
response curves, with herbicides, pesticides, insecticides, and hydrocarbons aver-
aging around 5% of studies. Hormesis is poorly dealt with by health policy and
basically incomprehensible in the forum of public risk perception.
The last curve to consider is the public perception curve, where a food additive
or residue shown to cause cancer in rodents at high doses is considered to be as
dangerous, or more so, in trace amounts in human populations. This is the top
curve in the lower panel, and represents the illogical public perception of pesticide
residues and additives like aspartame. To gain an accurate working knowledge
of diet and cancer, the public has to stop considering natural products as healthy
and artificial products as dangerous. They share common pathways of
metabolism, and many of the benefits of phytochemicals are due to the fact that
they are xenobiotics and they interact directly with more potent carcinogens in
Phase I and II metabolism. Some of the problems resulting from public perception
of risk are summarized below.
One of the strongest public fears is of pesticide residues in food, which is
often focused on fruits and vegetables. This may lead to decreased intake through
avoidance or due to the higher cost of organic products. Insufficient intake of
fruits and vegetables is the major nutritional problem in developed countries. A
low intake of fruits and vegetables may lead to increased meat intake, with
associated exposure to carcinogens produced by cooking and preservation.
Producers of natural health products are taking advantage of the public desire
to attain the benefits of fruit and vegetable consumption without incurring the
risk of pesticide exposure or the basic lack of desire to eat these foods. This leads
us to the final curve shown in Figure 3.4. Indole-3-carbinol is a breakdown product
of brassica glucosinolates, known to be associated with decreased cancer risk in
human populations (through consumption of brassica vegetables) and shown to
prevent cancer, at low levels, in rodent models. An unbiased observer might place
this data point in the lowest region of a hormetic response curve and expect that
at high levels the health benefits would diminish and the compound would
eventually become toxic. This follows the basic assumptions of toxicology that
all substances become toxic if the dose is increased sufficiently (Paracelsus, 1493-
1541). This has been established for indole-3-carbinol, which is shown to promote
cancer development at high doses in rodent models. The public views this natural
health product in a different light, however, and assumes that the benefits will
increase with increasing intake, as shown in the bottom curve in the lower panel
of Figure 3.4. Indole-3-carbinol is currently marketed in 200 mg capsules, with
recommended intakes of 2 per day, equivalent to 30 pounds of cabbage per day.
The justification for this extremely high intake by marketers of these compounds
uses data from the regression of cervical dysplasia, which is actually an example
of chemotherapy-like treatment of a dividing cell population, and in themselves
could be interpreted as a toxic end point. This chemotherapy-like dose of I3C is
being recommended and marketed to an unsuspecting public.
3.14 CONCLUSIONS
Current dietary patterns in developed countries favor exposure to carcinogens
derived from meat processing and cooking techniques, and dietary habits have
decreased the intake of chemopreventative phytochemicals. The growth of
research, business, and public interest in functional foods and nutraceuticals
provides opportunities to bring some of the beneficial metabolic effects back to
the modern, highly processed diet. This movement also increases the risk of
toxicity due to unnaturally high exposures to natural xenobiotics. Further research
will help to maximize the benefits and minimize the risks associated with the
novel use of natural products in cancer prevention.
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4 Nutrient and
Phytochemical
Modulation of Cancer
Treatment
Kelly Anne Meckling
CONTENTS
4.1 Introduction................................................................................................95
4.2 Magnitude of the Problem ........................................................................96
4.3 Effects of Overall Nutritional Status on Treatment Outcomes ................97
4.3.1 Changes in Nutritional Status of the Cancer Patient....................97
4.3.2 The Antioxidant Conundrum.......................................................101
4.4 Specific Molecular Targets ......................................................................102
4.4.1 P-Glycoprotein and Other Drug Efflux Pumps...........................102
4.4.2 NFκB, AP-1, and COX-2 ............................................................104
4.4.3 The Mevalonate Pathway ............................................................106
4.4.4 Immunomodulators......................................................................107
4.5 Polyunsaturated Fatty Acids as Therapeutic and Adjuvant
Chemotherapy Agents .............................................................................108
4.6 Other Foods, Phytochemicals, and Supplements Useful in
Adjuvant Chemotherapy..........................................................................113
4.7 Conclusions..............................................................................................115
References .........................................................................................................115
4.1 INTRODUCTION
Epidemiological data have consistently demonstrated that fruit and vegetable
consumption are associated with decreased risk of cancer at a variety of tissue
sites. A number of nutrients and phytochemical components of these foods have
been identified as potential cancer chemopreventive agents acting through a
variety of mechanisms (as described in Chapter 3). Many of these same com-
pounds or additional ones may also have activity in the later stages of cancer
95
development, including metastasis and the response of the patient to surgery,

chemotherapy, and radiation therapy. In some cases, the effects may be opposite
to those found in the early initiating events of carcinogenesis. Good examples
here are the results from clinical intervention trials using β-carotene in smokers
and finding an increased risk of lung tumor development, despite the fact that
high carotenoid intake is frequently associated with decreased risk of developing
lung cancer.1,2
Additional examples include phytochemicals that modify expression of P-
glycoprotein that may be useful as chemopreventive agents since they can promote
the efflux of certain carcinogens including 7,12-dimethylbenz(a)-anthracene. On
the other hand, researchers are looking for ways to antagonize P-glycoprotein
activity during chemotherapy since this activity in tumor cells can promote drug
resistance and treatment failure. It is clear then that treatment of the cancer patient
is a separate case from the prevention of cancer itself. Some strategies may be
specifically useful in the treatment phase, and it can be appreciated that potential
risks may be more acceptable at this point in the disease than they might be when
considered for long-term use in otherwise healthy populations.
4.2 MAGNITUDE OF THE PROBLEM

Based on the best available current data, a comprehensive study estimated that
in 2002 there were 10.9 million new cases of cancer worldwide, 6.7 million
deaths, and 24.6 million people who had been diagnosed with cancer in the last
5 years.3 The risk of developing cancer is highest in North American men and
women, but the risk of dying from cancer is highest in Eastern European men
and Northern European women.4 Lung cancer has been the most common cancer
in the world since 1985. However, because of differences in survival statistics,
four times as many women are breast cancer survivors (5 years since diagnosis)
as men and women who survive lung cancer. While the impression has been that
cancer is largely a problem of the developed world, this is rapidly changing with
cancer rates at many sites steadily rising in the developing world and only barely
lagging behind rates in North America and Europe.
One estimate puts the number of new cancer cases in 2020 at 15 million, a
50% increase over the rate in the year 2000. The increase in the average age of
the population as a whole is one contributor, but others include lifestyle factors,
such as smoking, decreased physical activity, and increased consumption of
“unhealthy diets,” as well as communicable diseases. Given that we can expect
an increase in the burden of cancer worldwide, at least for the next few decades,
improved treatment strategies to effect cure and/or improve the quality of life for
those suffering, are international objectives.
Given that a large number of cancer cases are likely to be attributed to dietary
and lifestyle factors, it is perhaps not surprising that a significant proportion of
the population is using supplements to “ward off disease” and may increase their
use following a diagnosis of cancer.5 Depending on the population studied, as
many as 84% of patients may report using complementary and alternative
Nutrient and Phytochemical Modulation of Cancer Treatment 97
medicine (CAM) and half of these report this use to their treating physician. One-
third of Australian and Finnish women with breast cancer reported substantial
changes to their dietary and lifestyle habits in the months following diagnosis.6
These included smoking cessation, changes in macronutrient composition,
increasing physical activity, and the use of vitamin and herbal supplements as
well as visits to CAM practitioners, including naturopaths. The highest users of
CAM appear to be pediatric oncology patients (84%)7 followed by breast cancer
survivors with more than 66% of Canadian women, living in Ontario, reporting
use of CAM.5 Sixty-one percent of U.S. armed forces veterans diagnosed with
cancer admitted using dietary supplements8 and about 30% of Canadian men with
prostate cancer report using some form of alternative medicine, although only
one-third of these were seeing a CAM practitioner.9
While the move toward informing clinicians about the use of CAM therapies
may be increasing, there are still 30% or more of certain patient groups that fail
to report. In order to provide useful resources for patients and clinicians on
specific therapies, prevalence data and outcome measures need to be available.
Both positive and negative interactions of CAMs with other therapies, including
chemotherapy, need to be identified and reported by both traditional and alterna-
tive healthcare practitioners. If properly applied, diet, nutritional, and herbal
supplements could increase drug efficacy in cancer treatment and potentially
lessen drug toxicity, leading to increased cure and better quality of life for those
suffering from cancer.
4.3 EFFECTS OF OVERALL NUTRITIONAL STATUS

ON TREATMENT OUTCOMES
There is marked individual variation in drug pharmacokinetics based on genetic
polymorphisms, gene-environment interactions, presence or absence of disease,
the type and pattern of nutrient intake, and the scheduling/dosage of drug admin-
istration. In the area of food–drug interactions the main focus has been on the
effects of food on drug absorption in the small intestine. Some adverse reactions
to particular treatment protocols have been attributed to food–drug interactions
without ever identifying the mechanisms or factors involved. Even when the
complicating food/nutrient is identified, the typical response by treating physi-
cians and pharmacists is to put the “food” on the “do not consume” list. While
this may seem like a reasonable approach, the possible nutritional deficits that
may result from eliminating large numbers of foods from one’s diet, combined
with the possible benefits that a mechanistic understanding of the nature of the
interaction may have on drug efficacy, side effects, and normal tissue toxicity,
speaks to the need for more rational treatment design (see Table 4.1).
4.3.1 CHANGES IN NUTRITIONAL STATUS OF THE CANCER PATIENT

Substantial weight loss is often observed in cancer patients, particularly those
with advanced disease. This may come about as a result of the disease process
TABLE 4.1
Nutritional Components with Potential Activity in Adjuvant Cancer
Therapy
Nutrient/
Phytochemical Molecular Target(s) or Activity Confounding Factors
Green tea Multidrug resistance proteins

polyphenols NFB signaling
AP-1 transcription factor
Cell cycle inhibitory proteins p21, p27,
cyclins D1, E
IGF-II
EGFR
Increase ovarian cancer survival
Sulforaphane NFκB signaling
Resveratrol NFκB signaling
Soy isoflavones Decrease breast cancer risk May stimulate growth of ER+ breast
(genistein) Decrease prostate cancer in TRAMP tumors
model Antagonizes benefit of Tamoxifen
Curcumin Antiangiogenic Possible activation of multidrug
Cardioprotectant resistance proteins
Lung protection from bleomycin Inhibition of camptothecin and
doxorubicin killing of breast cancer
cells
Ellagic acid VEGFR-2
PDGFR
Antiangiogenic
Lutein Bax:Bcl-2 ratio May increase ACF formation at high
Angiogenesis concentrations
Induce apoptosis
Grape seed extract Angiogenesis (VEGF)
(procyanidins) IGFBP-3
Apoptosis induction
Mushroom Immune enhancement
polysaccharides Th1/Th2 balance
(Krestin) Dendritic cell differentiation
Flaxseed lignans HER-2/neu
Apoptosis cascade
Estrogen and progesterone receptors

Nutritional Components with Potential Activity in Adjuvant Cancer
Therapy
Nutrient/
Phytochemical Molecular Target(s) or Activity Confounding Factors
Omega-3 fatty Cachexia reversal agent Low levels inhibit breast

acids 20 S proteosome carcinogenesis, but high levels
EPA/DHA Lipoxygenase increase risk if exposed during
Cyclooxygenase prepubescent period
NFB signaling
Improve cytokine profile
Decrease iNOS
Decrease metastatic spread
Improved liver and pancreatic function
Antioxidant Enhance chemotherapy in some adult Interfere with chemotherapy in some
vitamins populations adult and childhood cancers
(E, C, β-carotene) Low dose -carotene inhibits formation of May promote tumor progression in
colonic aberrant crypts smokers
High carotenoid levels associated with High dose β-carotene stimulates
recurrence-free survival in breast cancer ACF formation
patients
itself or from therapies that affect tissue metabolism or the patient’s ability to
assimilate nutrients.10 As well as there being different incidence between cancer
types, there are also differences in the presentation and pattern of body com-
position changes between men and women.11 Regardless of treatment modality,
the incidence of deficiency signs and symptoms frequently increases as treat-
ment progresses.12 These can include the appearance of stomatitis, edema, and
changes in body composition. The anorexia-cachexia syndrome is typical of
patients suffering from many solid tumors and occurs in greater than 50% of
all cancer patients and more than 85% of pancreatic and lung cancer patients.13
Presence of cachexia is associated with increased morbidity and mortality, and
research efforts are focused on improving quality of life and functional capacity
for those suffering.14 Both fat stores (predominantly triglyceride) and skeletal
muscle protein are targets of the hypermetabolic state resulting from a number
of inflammatory mediator and cytokine changes, including release of lipid
mobilizing factor (LIF) and proteolysis inducing factor (PIF) from tumor cells
and other tissues.
Recent studies using oral nutritional supplements containing eicosapen-
taenoic acid (EPA, 20:5n-3) in cachectic patients have demonstrated improve-
ments in weight or decreases in the rate of weight loss, improvements in body
composition, functional status, and quality of life.15–18 Many of the early studies
focused on untreated pancreatic patients where effective therapy and survival are
limited. However, a recent pilot study demonstrated improved outcomes in lung
cancer and pancreatic cancer patients with cachexia undergoing gemcitabine

chemotherapy.19 These studies have demonstrated that an energy- and protein-
rich supplemental drink, providing at least 1 g of EPA per day, can improve
weight gain, physical activity, and quality of life.18,19
EPA has been shown to inhibit the muscle catabolism induced by PIF through
antagonism of the ubiquitin proteosomal pathway.20 EPA attenuates the release
of arachidonic acid (AA, 20:4n-6) from membrane phospholipids and its subse-
quent metabolism to various eicosanoid products, including 15-hydroxyeicosatet-
raenoic acid (15-HETE).21 Both PIF and 15-HETE can directly modify the pro-
teosomal pathway by promoting NFB translocation to the nucleus. β-hydroxy-β-
methybutyrate (HMB), a metabolite of the amino acid leucine, has similar antag-
onistic activity to EPA; however, it appears to act at a step downstream from AA
release by inhibiting phosphorylation of the p42/44 mitogen-activated protein
kinase, key to the upregulation of the 20 S proteosome.22
Even when patients with advanced cancer are in seemingly good medical
condition, there is considerable interindividual response to therapy. The ability
to identify those factors that predict a patient’s ability to tolerate a specific
therapeutic regimen is highly desirable. Severe clinical hematopoietic toxicity is
a frequent and often life-threatening complication of chemotherapy. Although the
incidence does correlate with dose, complications can follow or copresent with
any cytotoxic therapy. It is conceivable that chemotherapy-induced DNA and
cellular damage may be more toxic to normal tissues, promoting hematopoietic
toxicity, in the presence of inflammatory cytokines and metabolic factors asso-
ciated with the anorexia-cachexia syndrome. Alexandre and coworkers recently
reported on the frequency of neutropenic fever or severe thrombocytopenia in
patients undergoing chemotherapy for advanced breast, gynecological, or gas-
trointestinal neoplasias.23 Drugs that were included in the various regimens
included anthracyclines, topoisomerase I or II inhibitors, platinum compounds,
and alkylating agents. Using an NIS index, calculated as the ratio (C-reative
protein X alpha-1 glycoprotein)/(albumin X prealbumin),24 the authors were able
to predict severe hematopoietic toxicity with a sensitivity of 89% and specificity
of 66%. Combined with the WHO measure of performance status, these two
measurements together should be able to identify those most likely to experience
severe chemotherapeutic toxicity. It remains to be seen whether nutritional sup-
plementation or antiinflammatory drug treatment can reverse this sensitivity in
such patients.
In addition to hematopoietic toxicity, a number of drug and radiation regimens
produce substantial dose-limiting gastrointestinal toxicity, including mucositis.25
This cannot only impair the patient’s desire to eat, but can also make them
vulnerable to infection at the site of injury. In a model of intestinal injury repair,
we showed that long-chain omega-3 fatty acids, docosahexaenoic acid (DHA,
22:6n-3) and eicosapentaenoic acid (EPA, 20:5n-3) promoted faster wound heal-
ing than short-chain fatty acids or omega-6 fatty acids 26 and decreased mucosal
damage in the small intestine of drug-treated rats.27 Compared to other areas of
nutrition and cancer research, there have been relatively few studies examining
effective therapies for mucositis. Most efforts have been on palliation and pain
control (for review, see Reference 25). One clinical trial demonstrated that sup-
plementation of parenteral nutrition formula with glutamine reduced the severity
of mucositis in patients undergoing high dose chemotherapy followed by autol-
ogous bone marrow transplantation.28 Another found that treatment with human
recombinant keratinocyte growth factor substantially improved oral mucositis
symptoms in patients about to undergo myeloablation and autologous stem cell
transplantation for hematologic malignancies.29 Though not specifically tested in
cancer patients, a traditional herbal medicine, Throat Coat®, containing elm bark,
licorice root, and marshmallow root, substantially improved symptoms of phar-
yngitis in clinical patients.30
4.3.2 THE ANTIOXIDANT CONUNDRUM

The association between consumption of foods rich in antioxidants and reduced
risk of several types of cancer has often been interpreted as indicating that high
antioxidant intake will inevitably reduce cancer risk and improve cancer therapy.
As noted in Chapter 3, antioxidants can also act as prooxidants depending on
dose and interaction with other biologicals, including other antioxidant vitamins,
minerals, and phytochemicals. While some trials have demonstrated that dietary
supplementation with antioxidants may enhance the response to chemotherapy,
improve the quality of life of patients by reducing toxic side effects, or recovery
of normal tissues to therapeutic insult,31–34 others have shown that supplementa-
tion of antioxidants during chemotherapy may actually interfere with the activity
of the drug.35–38 This is particularly the case when oxidant stress and generation
of free radical damage are part of the mechanism by which the drugs kill tumor
cells. For example, the enhanced therapeutic effect of anthracyclines in the back-
ground of an omega-3 fatty acid supplementation on mammary cancer treatment
is suppressed in the presence of α-tocopherol (vitamin E).38
Saintot and coworkers also found a negative correlation between α-tocopherol
levels and subsequent survival in women with breast cancer.39 In other models,
however, vitamin E has been shown to reduce cardiotoxicity of doxorubicin,40
healing of chemotherapy-induced stomatitis41 and increase efficacy of 5-fluorou-
racil treatment of colon cancer.42 Similar inconsistencies exist for other antioxi-
dant vitamins including vitamin C and beta-carotene (reviewed in Reference 42).
In a recent trial, patients with low vitamin C plasma concentrations had shorter
survival times than those with higher levels.44 While vitamin C levels did correlate
with high C-reactive protein levels, none of these patients were undergoing
chemotherapy, as treatment was strictly palliative.
In contrast to the results in adult cancer patients, children suffering from
acute lymphoblastic leukemia (the most common cancer in children) seem par-
ticularly vulnerable to low antioxidant vitamin status. In this population, higher
intake of vitamins A, C, and E were associated with fewer therapy delays, less
chemotherapeutic drug toxicity, fewer days spent in hospital, and decreased
incidence of infection.45 The authors of this work suggested that the additional
burden of chemotherapy combined with the disease process itself would lead to
an increased free radical load and, therefore, tax the antioxidant defenses even
in the presence of apparent dietary adequacy. Given the inconsistencies between
reports and the large variation between specific patient populations, generaliza-
tions about the benefits and/or risks of antioxidant supplementation during che-
motherapy cannot be made. Rather, recommendations for their use must be based
on the specific drug regimen and tumor type being treated and a molecular
understanding of the relevant targets (see Table 4.1).
4.4 SPECIFIC MOLECULAR TARGETS

4.4.1 P-GLYCOPROTEIN AND OTHER DRUG EFFLUX PUMPS
P-glycoprotein (Pgp) is a 170 kDa phosphorylated glycoprotein encoded by the
human MDR1 gene. It belongs to a family of adenosine triphosphate (ATP)-
dependent membrane ABC cassette transporters normally responsible for trans-
port of structurally unrelated chemicals, including xenobiotics, lipophilic, and
amphipathic natural compounds, hormones, and pharmacological agents. Other
members include ABCG2 (breast cancer resistance protein) and MRPs 1 to 8
(multidrug resistance proteins, reviewed by Choi,46). MRP1 is a 190 kDa protein
with several conserved regions common to ABC members, but with a distinct
transport mechanism and substrate specificity from Pgp. Substrates for MRP
include leukotriene C4 and glucuronide conjugates of 17-β-estradiol,47 and for
drug transport, it is frequently dependent on the presence of reduced glutathione.48
Pgps, along with several other members of this family, are ubiquitously expressed,
but tend to be concentrated on the apical surfaces of epithelial cells, particularly
those in the gastrointestinal tract tissues,49 the blood–brain barrier,50 and in bone
marrow progenitors.51
One of the major reasons for treatment failure in chemotherapy is the devel-
opment of drug resistance. This can occur as a result of over-expression of one
or more of the drug resistance transporters (MDR) listed above, in tumor targets.
Drug therapy becomes ineffective, even at high dose levels, because the activity
of these transporters to extrude drug from tumor cells becomes incredibly effi-
cient. A variety of strategies including the development of chemosensitizers has
been attempted to overcome the resistant phenotype. Some chemosensitizers are
active against more than one MDR product while others are highly selective.
Verapamil, a calcium channel blocker, was shown to be an effective sensitizer in
both models and in a recent clinical trial of metastatic breast cancer;52 however,
at effective doses that bring about sensitization, verapamil causes cardiotoxicity.
Cyclosporin A and related derivatives have also shown some promise clinically,
but again the side effects, in this case immunosuppression, are major clinical
issues. A variety of second-generation chemosensitizing agents are currently
under development by a number of companies with the emphasis being on
reducing toxicity while maximizing drug reversal in tumor cells (reviewed by
Choi46). It is interesting to note that a variety of flavonoid compounds are being
examined as potential chemosensitizers, as it has been proposed that these would

have fewer side effects than pharmacologic agents. Whether this is the case
remains to be determined.
As are the cases with many of the P450 enzymes, Pgp and MRP1 expression
and activity can be regulated by nutrients and phytochemicals, including many
herbal constituents. Identifying the chemical components and target specificity
are keys to developing these “natural” therapeutic agents that may be active alone
or in combination with conventional pharmacologic agents. Dr. Susan Cole’s
group was the first to report that a wide range of flavonoids found in commonly
consumed foodstuffs were inhibitors of conjugated organic anion transport via
MRP1.53 Apigenin, naringenin, genistein, and quercetin were all shown to inhibit
the transport of LTC4 and 17-β-estradiol 17-(β-D-glucoronide), while at the same
time promoting GSH transport by MRP1 without acting as substrates them-
selves.53,54
An important consideration for their utility as cancer therapeutic agents is
their bioavailability in vivo. While many polyphenols have been shown to be
active in vitro, the real question is whether they are effective, particularly when
given orally to patients. For example, curcumin, a polyphenol found in Curcuma
longa (the spice tumeric), has been shown to have antioxidant, antiinflammatory,
hypolipidemic, antimutagenic, anticancer, and antimetastatic properties, presum-
ably through regulation of key signaling cascades involving NFB, c-myc, p21,
and bcl-XL.55 Even though curcumin has been shown to be nontoxic at doses up
to 10 g/d in humans, serum metabolites are virtually undetectable (or at most in
the nM range) at this level largely because liver and intestinal conjugation are so
efficient.56 Curcumin at doses of 50 to 150 µM is cytotoxic to freshly isolated
hepatocytes, but if added to progressively older cultures is nontoxic as a result
of upregulation of Pgp.57 In cervical carcinoma cells, curcumin down regulates
MDR1 activity and increases sensitivity to vinblastine, possibly by directly bind-
ing Pgp and antagonizing its activity.58 However, once again these results were
only achieved at µM concentrations of curcumin. Thus, although curcumin
appears to be a good drug reversal agent in vitro, the key to its clinical utility
will be in improving its bioavailability.
Similarly, various ginseng components including ginsenosides have been
shown to have activity in reversing drug resistant in several tumor cell lines by
inhibiting Pgp.59 Several were active in vivo in prolonging the life of leukemia
P388 transplanted mice treated with doxorubicin60 and in reversing drug resistance
in vitro in several daunorubicin and doxorubicin acute myelogenous leukemia
cell lines with activity greater than observed with verapamil.61 While there are,
as yet, no clinical reports of specific drug interactions with Pgp substrates, one
could predict that Panax ginseng could be used as a complementary therapy in
human cancer chemotherapy.
A variety of catechins in green tea have been associated with decreased cancer
risk at various sites. While some of these same components (EGCG, epicatechin
gallate, and catechin gallate) have been shown to be active Pgp inhibitors
and, therefore, may be useful in adjuvant chemotherapy settings, others (i.e,
epicatechin) have shown the opposite activity.62,63 Whether these heterologous

activities have importance clinically for drug interactions in cancer or any other
disease, remains to be determined. Similarly, while St. John’s wort has been
shown to modify drug activity modulated by CYP3A4 and has a particularly
critical effect when used in combination with psychiatric drugs (see Chapter 6,
Nutrients and Herbals in the Pharmacotherapy of Unipolar/Major Depression), it
has also been shown to stimulate Pgp expression in a number of cell lines and
peripheral blood lymphocytes.64–66 The potential impact of hyperforin and other
St. John’s wort constituents on cancer chemotherapeutics have not been examined
clinically and, thus, the potential for significant worsening of the drug-resistant
phenotype during therapy is entirely possible.
4.4.2 NFκB, AP-1, AND COX-2

NFκB (nuclear transcription factor κB) describes a family of dimeric transcription
factors involved in cell cycle control, cell differentiation, apoptosis, cell adhesion
and migration, and angiogenesis. NFκB is maintained in the cytoplasm in an
inactive form by binding to one of the inhibitory κIBs (inhibitory kappa binding
protein). Phosphorylation of κIB by an Iκkinase (IKK) promotes its ubiquitination
and proteosomal degradation, thereby releasing NFκB for translocation to the
nucleus to target specific genes. While controlled expression of NFκB is critical
to normal cell fate determination, abnormal expression has been associated with
the transformed phenotype (reviewed by Dorai and Aggarwal67 and Nakanishi
and Toi68).
Activated protein-1 (AP-1) is also a hetero- or homodimeric transcription
factor composed of various combinations of fos and jun proteins. Targets of AP-
1 activity include genes such as urokinase-type plasminogen activator (uPA) and
matrix metalloproteinases (MMPs), two types of proteins critical to acquisition
of the metastatic phenotype and the androgen receptor in prostate cancer.69 Several
phytochemicals, including green tea catechins, curcumin, resveratrol, and capsa-
icin, have been shown to suppress AP-1 activation (possibly NFκB as well70) and
may specifically down-regulate Bcl-2 and Bcl-XL, two antiapoptotic proteins. In
addition, some of these same compounds and additional ones from grape seed
extract (procyanidins) have been shown to inhibit growth factor receptors includ-
ing those for insulin-like growth factor (IGF-1), epidermal growth factor (EGF),
fibroblast growth factor-2 (FGF2), and vascular endothelial growth factor
(VEGF), which have roles in promoting tumor growth and angiogenesis.71–78 The
activity and expression of HER-2/neu, a receptor that is over-expressed in many
breast cancers and associated with poor prognosis, is inhibited by curcumin79 and
flaxseed lignans.80
Curcumin has additional targets, such as the Src-homology domain tyrosine
phosphatase (SHP-2), where it activates the protein’s activity, thereby interfering
with the JAK-STAT signaling pathway and suppressing proliferation in tumors,
such as multiple myeloma.81 The tetracyclic phenolic, ellagic acid, has similar
molecular targets to its cousin curcumin. Ellagic acid is richly produced in
raspberries, strawberries, cranberries, walnuts, and pecans, where it is concen-

trated in the seeds/nuts. As well as inhibiting VEGFR-2 in endothelial cells,
ellagic acid also inhibits PDGFR activity in smooth muscle cells.82 These dual
effects result in profound inhibition of angiogenesis and point to ellagic acid’s
potential as a useful adjuvant in human cancer therapy.
In addition to their activities on NFκB and growth factor signaling, directly
or indirectly, a number of food components also modulate the levels of
eicosanoids. Depending on the initial polyunsaturated fatty acid substrate, and
relative levels of the various cyclooxygenase and lipoxygenase activities, a wide
variety of pro- and antiinflammatory eicosanoid metabolites can be produced.
Typical substrates, such as arachidonic acid (AA), can be metabolized to the
proinflammatory hormone-like compound, prostaglandin E2, via cyclooxygenase
enzymes (COX-1 or COX-2) (Figure 4.2). COX-1 is ubiquitously expressed and
tends to be constitutively expressed in most cells, suggesting a generalized house-
keeping role for this form of the enzyme. On the other hand, COX-2, the inducible
form of the enzyme, tends to be stimulated in response to inflammatory signals.
A large body of literature now exists demonstrating an important role for COX-
2 products in tumorigenesis (for review, see Wallace83).
Chemoprevention studies using nonsteroidal, antiinflammatory agents
(NSAIDS), such as celecoxib and aspirin, showed that colon carcinogenesis could
be profoundly inhibited in Min mice and in controlled trials of patients with
familial adenomatous polyposis (FAP). There are a whole host of side effects of
long-term NSAID treatment and, thus, this approach is of limited utility in
populations other than those with FAP. However, given that COX-2 over expres-
sion is an extremely common occurrence in malignancy and interferes with tumor
cell killing and promotes angiogenesis, targeting this enzyme in adjuvant cancer
therapy may be a realistic option. In this respect, it is interesting that certain
phytochemicals, such as curcumin, can act similarly to NSAIDs in that they
inhibit COX-2 activity.
COX-2 is overexpressed in more than 80% of human colorectal adenomas
and carcinomas, and is associated with constitutively higher levels of PGE2 than
normal surrounding mucosa.84,85 Colon cells expressing high levels of COX-2 are
also resistant to the differentiation effects of the bacterial fiber fermentation
metabolite, butyrate. In general, high level COX-2 expression is associated with
more advanced tumor grade, increased frequency of metastasis, and shorter sur-
vival times in colon cancer patients86,87 Glioblastoma multiforme also seems to
be a cancer where COX-2 over expression is predictive of poor survival, and
there is some suggestion that inhibitors may be helpful as adjuvant therapy for
brain tumors. In breast cancer, an association has been made between overactive
HER-2/neu and elevated COX-2 expression. Activation of HER-2/neu in cultured
cells leads to upregulation of COX-2 and increased production of PGE2 in the
medium. Conversely, treatment with the antibody Herceptin reduced COX-2 and
PGE2 expression. Additionally important in the breast, COX-2-derived prostag-
landins stimulate aromatase activity in the breast providing local production of
active estrogens for breast cancer promotion.88,89 Other cancers where a role for
COX-2 in tumor progression have been suggested include cervical carcinoma,

endometrial carcinoma, squamous cell carcinoma of the head and neck, adeno-
carcinoma of the lung, pancreatic cancer, prostate cancer, and bladder cancer
(reviewed by Wallace83).
COX-2 products including PGE2, PGI2 , and thromboxane A2 (TXA2) reduce
endothelial apoptosis and directly stimulate endothelial migration90,91 by stimu-
lating production of basic fibroblast growth factor (BFGF), platelet-derived
growth factor (PDGF), and VEGF. VEGF, in particular, seems to be critical in
establishing the vasculature necessary for tumor angiogenesis and progression.
Furthermore, these products also stimulate tumor production of MMPs facilitating
extravasation and metastatic spread.92 In addition, COX-2 products have been
reported to upset the balance of interleukins and cytokines necessary for normal
immunocompetence (see below), thereby impairing host attempts at tumor cell
killing. There are a wide variety of natural inhibitors of COX-2 that have been
described, many of which have been tested in vitro. Many of these are flavonoids
or related in structure to these. Scutellaria baicalensis and several other Chinese
medicinal herbs demonstrate breast and prostate anticancer activity through sup-
pression of PGE2 via COX-2.93
In addition to being active antitumor agents on their own, curcumin and
resveratrol may also be important in adjuvant therapy settings. They can make
cells more sensitive to radiation and chemotherapy by inducing p21Cip1/waf-1, which
promotes cell cycle arrest, decreasing drug resistance through depressed glu-
tathione levels through GST-Pi inhibition, and down-regulation of the survival
protein surviving (reviewed by Dorai and Aggarwal67). As well, several chemo-
therapy agents may inadvertently transactivate NFκB signaling as they simulta-
neously trigger apoptotic cascades through activation of p53 (reviewed by Nakan-
ishi and Toi68). Taxanes, Vinca alkaloids, and topoisomerase I and II inhibitors
are all examples of drugs that appear to simultaneously induce NFκB nuclear
translocation. This, in itself, could contribute to drug resistance and may be
overcome by simultaneous administration of NFκB targeting phytochemicals. The
exploitation of green tea polyphenols, resveratrol, curcumin, and related phy-
tochemicals in adjuvant therapy needs to be examined in clinical trials of human
cancer patients. Furthermore, combining some of these compounds into “nutri-
tional cocktails” may be particularly useful. A mixture of lysine, proline, arginine,
vitamin C, and epigallocatechin gallate was recently shown to inhibit osteosar-
coma cell production of MMPs, and VEGF.94 Given that osteosarcoma is associ-
ated with poor prognosis and increased frequency of second site tumors as a result
of chemotherapy, new therapies like this need to be tested in clinical trials.
4.4.3 THE MEVALONATE PATHWAY

Statins are probably the most safe and effective hypolipidemic medicines cur-
rently available. Specifically, they are competitive inhibitors of the rate-limiting
enzyme in cholesterol biosynthesis, 3-hydroxy-3-methylglutaryl-coenzyme
A reductase (HMG-CoAR). Mevalonate, in addition to being a precursor of
cholesterol, is also a key intermediate in the formation of isoprenoid groups that

localize and maintain the activity of small GTP-binding proteins, including ras,
rac, and rho. Activating ras mutations or defects in ras signaling are typical of
many cancers. A number of studies have now shown that treatment with statins
has antitumor activity both in vitro and in vivo (reviewed by Katz95). Recently,
Duncan and coworkers showed that EPA and DHA attenuated the up-regulation
of HMG-CoAR in MCF-7 breast cancer cells in response to mevastatin and
suggested that these may be useful in adjuvant breast cancer therapy.96
4.4.4 IMMUNOMODULATORS
Patients undergoing surgery, radiation, or chemotherapy have a high risk of
complications, including wound infections and septicemia. Alterations in host
defense associated with disease or treatment may make the cancer patient par-
ticularly vulnerable. Although there is no single contributor to immune dysfunc-
tion, there are indicators that specific nutritional substrates may be effective in
rebalancing the immune system in cancer patients. As already mentioned, the n-
3 fatty acid, EPA, has anticachectic properties that lead to decreased weight loss
in treated patients. Recent studies have shown that EPA and the amino acids,
arginine or glutamine, when supplemented orally, improved postoperative infec-
tious and wound complications in head and neck cancer97,98 pancreatic cancer,99
colon cancer100 patients, and during bone marrow transplantation.101
Recently, green tea polyphenols were also shown to promote cytotoxic CD8+
T cell invasion of UV-induced tumors and promoted apoptosis via activation of
caspase-3 in animals.102,103 As well, mistletoe lectins (ML-1) have considerable
immunomodulatory activity in human beings. Healthy human subjects adminis-
tered mistletoe lectins responded with an initial proliferation of peripheral blood
mononuclear cells, accompanied by increased production of TNFα and IL-6 and
less pronounced elevation of INF-γ and IL-4 (reviewed by Pryme et al.104).
Expansion in the CD8+ cytotoxic T cell population and NK activity have also
been reported in response to ML-1.105
Additional compounds that have interesting immunomodulatory activity
include a variety of polysaccharides from herbs used in traditional Chinese
medicine (TCM). There are important structure–function relationships that have
not been completely delineated but that seem critical to their activity (reviewed
by Chang106). The most important bioactives appear to have a β1,3 1,4 or 1,6
branching pattern, and the more highly branched and higher molecular weight
species seem to be the most potent.107 More than 200 species have been identified,
mostly fungal or botanicals. One representative example is the polysaccharide,
Lentinan, from Shiitake mushrooms, Lentinus edodes; it inhibits the growth of
transplanted tumors in rodents108 and, in a controlled clinical trial, it increased
survival in recurrent and metastatic gastric and colorectal cancer patients when
given in combination with chemotherapy.109 This latter study was carried out in
Japan, published in Japanese, and, as far as one can tell, has not been reproduced
in larger clinical trials. Thus, the utility of this agent remains to be examined.
However, another agent, Krestin, has been shown to extend survival in

several clinical trials, including patients with gastrointestinal tumors at various
tissue sites, nonsmall-cell lung carcinoma, and some breast cancers.110-115 This
protein-bound polysaccharide has recently shown great promise as an adjuvant
with 5-fluorouracil in colon cancer patients after surgical resection.111,116,117 In
an animal model of bone metastatic breast cancer, Krestin also substantially
affected the rates of metastases and long-term survival when combined with
Tamoxifen118,119 and, in one clinical trial, improved response in combination
chemotherapy for operable breast cancer.120 It has also been shown to restore
the cytotoxic T-cell profile toward Th1 dominance, critical to successful vaccine
therapy, and promoted the differentiation of dendritic cells in similar patient
populations.121 Jimenez et al. also suggested that this compound may have
important NK stimulatory activity as it was potent in NK-derived cell lines in
vitro.122 Polysaccharide preparations from the mushroom Coriolus versicolor
have antileukemic activity by lowering IL-8 and increasing IL-1β and IL-6
production.123 IL-8 excess is thought to be important in cancer progression by
promoting mitotic, angiogenic, and migrational activity of tumor cells. Inter-
estingly, glutamine restriction or deprivation promotes the expression of IL-8
and several other genes in breast cancer and other cell types124 through NFκB.
An oral supplement mixture containing arginine, n-3 fatty acids, and RNA as
a source of nucleosides improved the IL-8 and cytokine profile, markers of
inflammation (including prostaglandins and leukotrienes) in preoperative
patients about to undergo major surgery for cancer.125
4.5 POLYUNSATURATED FATTY ACIDS AS

THERAPEUTIC AND ADJUVANT
CHEMOTHERAPY AGENTS
Long-chain polyunsaturated fatty acids (PUFA) found in the human body are
derived either from foods and/or metabolism of shorter versions consumed in the
diet. Omega-3 (ω-3 or n-3) and omega-6 (ω-6 or n-6) represent two families of
fatty acids required for normal human growth and development. Human beings
do not have the ability to desaturate fatty acids at the 3 or 6 positions from the
methyl end and, thus, these must be provided in the diet as essential fatty acids.
Two examples of essential fatty acids are linoleic acid (LA, 18:2n-6) and alpha-
linolenic acid (LNA, 18:3n-3), both of which can be elongated and desaturated
to form longer versions, such as arachidonic acid (20:4n-6), eicosapentaenoic
acid (EPA, 20:5n-3), and docosahexaenoic acid (DHA, 22:6n-3) (Figure 4.1).
Rich natural sources of LA include sunflower and corn oil, LNA includes flaxseed
and canola oils, and EPA and DHA are found in some algae and cold water fishes.
These fatty acids can in turn be esterified into mono-, di- and triglycerides and
into various membrane phospholipids. Following their release through the action
of phopholipases (including phospholipase A2 [PLA2], which releases fatty acids
from the sn-2 position), 20-carbon (and possibly 22 carbon) fatty acids can either
6-series 3-series
18:2n-6 18:3n-6
(linoleic acid, LA) alpha linolenic acid, LNA)
Δ6-desaturase
18:3n-6 18:4n-3
(gamma linolenic acid, GLA) (steariodonic acid)
elongase
20:3n-6 20:4n-3
(dihomo-gamma-linolenic acid, DGLA) eicosatetraenoic acid)
Δ5-desaturase
20:4n-6 20:5n-3
(arachidonic acid, AA) eicosapentaenoic acid, EPA)
elongase
22:5n-3
(docosapentaenoic acid, DPA)
elongase
24:5n-3
(tetracosapentaenoic acid, TPA)
Δ6-desaturase
24:6n-3
(tetracosahexaenoic acid, THA)
peroxisomal
β-oxidation 22:6n-3
(docosahexaenoic acid, DHA)
FIGURE 4.1 Metabolism of essential fatty acids in man
be reesterified or metabolized into one of several eicosanoid products including

prostaglandins, leukotrienes, and thromboxanes (Figure 4.2).
The balance of eicosanoid products is dependent on the relative concentra-
tions of n-6 and n-3 PUFAs in membrane phospholipids (which is largely depen-
dent on dietary intake), the activity of PLA2 (and related enzymes), and the activity
of the various isoforms of COX, LOX, and cytochrome P450 enzymes. In addition
to enzymatic catalysis, AA is particularly vulnerable to free radical attack pro-
moting the formation of longer-lived isoprostanes, which are potent inflammatory
molecules. Compared to AA metabolites, DGLA and EPA eicosanoids tend to
be less inflammatory or may even be antiinflammatory depending on the specific
series created. Many of the benefits of fish consumption on cardiovascular disease
risk are related to the antiinflammatory properties of EPA metabolites and their
roles in inhibiting platelet aggregation, promoting vasodilation, and lowering
arterial blood pressure.
Most eicosanoid products are relatively short-lived, surviving only seconds
or minutes after release and only able to act locally. In contrast, some LOX by-
products, such as 5-HETE and 12-HETE, may survive up to hours in the circu-
lation, giving them ample time to act and reach more targets. As well, several
eicosanoid products have the ability to bind to fatty acid binding proteins in blood,
20:3n-6 20:5n-3
(DGLA) (EPA)
COX LOX COX LOX
PGE1 and LTB3 and PGE3 and LTB5 and

series-1 series-3 series-3 series-5
eicosanoids leukotrienes eicosanoids leukotrienes
20:4n-6
COX (AA) Cytochrome
P450
PGG2 and
series-2 12-, 16-HETEs
prostaglandins LOX
and thromboxanes
5-, 12-, 15- HPETEs
LOX
5-, 12-, 15- HETEs
and 4-series
leukotrienes
FIGURE 4.2 Eicosanoid metabolism of n-6 and n-3 fatty acids.
which can substantially stabilize them and extend their half-lives further.126 5-
HETE and 12-HETE tend to be associated with increased cancer cell growth and
may act through specific receptors or receptor-independent pathways.127,128 Some
tumor types, including gliomas, breast cancer, and leukemia cells, both produce
and respond to HETEs in an autocrine fashion; LOX inhibitors have been dem-
onstrated to have antitumor activity in these cell lines in vitro.129 5-HETE is a
particularly important survival factor for prostate cancer cells. When its synthesis
is blocked, both hormone responsive and nonresponsive tumor cells undergo rapid
apoptosis.130,131 12-HETE promotes proliferation and the metastatic phenotype of
colon, pancreatic, breast, melanoma, and prostate tumor cell lines.83,131 Elevated
12-LOX activity is typical of high-grade tumors, which release 12-HETE pro-
moting angiogenesis in neighboring endothelial cells, changes in extracellular
matrix proteins, and invasive properties in the primary tumor.132,133 Molecular
targets of 12-HETE include protein kinase C, activation of Erk1/2, src, and
activation of NFκB.134 Often times, both COX and LOX products have cancer
promoting and antiapoptotic activity in the same tumor. MCF-7, ER+ breast
cancer cells are stimulated by both LOX and COX products of AA; inhibition of
only one or the other produces much less growth inhibition than inhibiting both
routes simultaneously.135
From a therapeutic view, it seems reasonable to use COX and LOX inhibitors
in chemopreventive or in adjuvant cancer therapies. Despite the evidence from
epidemiological data demonstrating clear chemopreventive effects of NSAIDs,
the long-term use of such pharmacologic agents is associated with substantial
side-effects including gastric ulceration, perforation, and obstruction and have
been linked to thousands of deaths per year.136 This is where dietary modification
and nutritional supplementation come into play. The goal here is to reset the
balance of eicosanoids so as to inhibit tumorigenesis while leaving normal cells

and developmental programs intact. The key here is to use inherent differences
in metabolism of lipids and fatty acids between tumor and normal cells to achieve
selective targeting.
With the exception of Inuit or Japanese following traditional diet and lifestyles
patterns, most populations around the world consume ratios of n-6:n-3 fatty acids
on the order of 10 to 20:1. While there is no definitive proof of the ideal ratio,
the suggested ratio for optimal health is about 2 to 4:1. This is the ratio that
would be consumed in a traditional Japanese diet (where total fat is low and total
carbohydrate high) or Greenland Inuit diet (high in total fat, low in carbohydrate).
Thus, most North Americans and many other populations around the world
consume diets rich in n-6 fatty acids that will inevitably lead to higher membrane
and tissue levels of LA, DGLA, and AA, relative to the levels of LNA, EPA, and
DHA. In order to achieve this “ideal ratio,” one would need to consume at least
2 to 3 fatty fish (herring, mackerel, salmon, trout, tuna) meals per week. While
plant sources, such as canola and flax, do provide short-chain n-3 fatty acids,
predominantly LNA, these are only very poorly converted to the longer chain
PUFAs, EPA, and DHA in humans. The efficiency of conversion has been esti-
mated to be between 1 and 5% in healthy tissues, and cancer cells are notoriously
deficient in ∆6-desaturase compared to normal cells from the same tissue.137–140
As a result, fish oil supplements appear to be the most effective way of getting
additional long-chain PUFAs into the human diet. A variety of studies both in
vitro and in vivo using fish oil supplements and purified EPA and DHA have
demonstrated selective antitumor action. Supplementation with n-3 fatty acids
has been shown to inhibit proliferation and invasion of various tumor cell lines,
induce cell cycle arrest and/or apoptosis, and enhance response to cell differen-
tiation agents.141–147
EPA and DHA compete with AA and DGLA for incorporation into membrane
phospholipids (PLs). As one might expect, providing additional EPA and DHA
in culture medium through dietary supplementation increases the relative con-
centrations of these fatty acids in specific PL classes. This occurs via direct
competition for esterification, particularly at the sn-2 position of PLs, where
highly unsaturated fatty acids are the preferred substrates. As well, because EPA
and DHA are more highly unsaturated than AA precursors, they bind with higher
affinity to desaturase enzymes, thereby inhibiting de novo AA synthesis. Thus,
upon appropriate stimulation of PLA2, relative levels of EPA will be higher than
AA and, thus, fewer AA fatty acids will be available for eicosanoid production.
Not only are PGE2, 5-HETE, and LTB4 produced in lower amounts, but higher
levels of EPA metabolites, which do not have cancer promoting activity, are
created through LOX and COX activity. Interestingly, EPA and DHA seem to
selectively inhibit COX-2 with little or no effect on COX-1.148
In addition to providing a reduction in inflammatory AA eicosanoids, sup-
plementation with EPA and DHA also affect a number of other activities in cells
in which they are incorporated. Human trials have shown that TNF-α and IL-1
synthesis are inhibited.149 Activity of membrane receptors including those for
insulin, 150 RANKL, 151 HER-2/neu, 152 nuclear PPARs, and mitochondrial
activity153 have all been shown to be influenced by EPA and/or DHA. As well,
DHA appears to down-regulate the expression of inducible nitric oxide synthase
in colon cancer cells, which indirectly leads to decreased COX-2 production and
associated decreases in NFκB, interferons, cyclic GMP, and up-regulation of a
number of cyclin-dependent kinase inhibitors, including p21 and p27.154 All of
these activities have been associated with reduced tumorigenesis, tumor cell arrest
or apoptosis, and decreased tumor progression. As well, recent studies have
suggested that liver and pancreatic function in postoperative cancer patients can
be improved with n-3 supplementation.155 The benefits of n-3 supplementation
alone seem to be relatively limited, though. No “cures” have been demonstrated
with this approach by itself and the levels of fatty acid needed to achieve many
of the effects in vitro were in the high µM range, which is difficult to achieve
through oral supplementation of patients. Where the n-3s are showing the greatest
promise is in combination or adjuvant therapy settings, with surgery, radiation,
and chemotherapy.
More than a decade ago, it was shown that supplementation of fibrosarcoma
and murine leukemia cells with EPA and DHA increased the toxicity of nucleoside
drugs and doxorubicin selectively in the tumor cells while leaving the normal
cells unaffected or even protected.141,142 This resulted in a substantial theoretical
improvement in therapeutic index for arabinosylcytosine (araC) and doxorubicin.
Later, we demonstrated that in vivo supplementation of rat or mouse diets with
purified DHA decreased tumor burden (fibrosarcoma and leukemia) and enhanced
the therapeutic activity of araC in these animals while simultaneously protecting
bone marrow progenitors and gastrointestinal mucosa, two key sites of potential
dose-limiting injury.27,156,157 While we have been able to demonstrate altered
nucleoside transport activity in a variety of cell types incubated with DHA and
EPA,158,159 we showed that at least part of the chemopotentiation by DHA was
due to altered activity of enzymes involved in araC activation and degradation
that was tumor-specific.160 In our models, both in vitro and in vivo, adding vitamin
E did not attenuate the beneficial effects of DHA on drug toxicity, suggesting
that oxidant-stress per se was not critical; however, a study by Colas and
coworkers38 showed that α-tocopherol did suppress mammary tumor sensitivity
to anthracyclines, so this possibility remains open.
In experimental breast cancer, long-chain n-3 PUFAs themselves have anti-
angiogenic and antmetastatic activity, but tumor kill can synergistically be
improved with several drugs, including mitomycin C, doxorubicin, and cyclo-
phosphamide.161,162 Importantly Colas showed that the effectiveness of the n-3
supplementation with doxorubicin in a breast cancer model was attenuated with
α-tocopherol, suggesting that oxidant stress was part of the killing synergism.38
Their studies had previously indicated that women with higher n-3 levels in breast
adipose tissue fared better following chemotherapy to a variety of regimens than
those with low n-3 status in breast adipose.163 It is not clear from the human
studies to date whether the antioxidant status of the patient is important during
the adjuvant treatment with omega-3 supplements and any of the other drugs.
Colon cancer, despite its prevalence and the availability of tools that have
generally aided in early detection and removal of preneoplastic polyps resulting
in increased cure, remains one of the most difficult cancers to treat. Less than
25% of colon cancer patients treated with chemotherapy are responsive. The most
efficacious therapy utilizes a combination of 5-fluorouracil with leucovorin res-
cue. Some recent successes have also been demonstrated with new therapeutic
regimens utilizing two other classes of drugs, irinotecan/camptothecin, and oxali-
platin.164,165 While not typically used in colon cancer therapy (Grem et al., 1995),
araC and gemcitabine have recently been explored as antitumor agents in colon
cancer cells.166 We showed that the therapeutic index of araC could be improved
40-fold with supplementation of rat colon tumor cells while protecting normal
colonic cells from toxicity.167 Jordan and Stein demonstrated that a fish oil-based
lipid emulsion, combined with 5-fluorouracil, promoted apoptosis and cell cycle
arrest in Caco-2 human colon cancer cells as well.168 Wynter et al. showed
increased sensitivity of the MAC16 colon tumor in mice when EPA/DHA were
given in combination with epothiline, gemcitabine, 5-fluorouracil, and cyclophos-
phamide.169 As well as sarcomas, breast cancers, colon cancers, and some leuke-
mias, DHA and EPA have also been tested and found effective as adjuvant agents
in prostate cancer with celecoxib170,171 and in cervical carcinoma.172 A recent study
using more than 30 different colon tumor cell lines identified a series of genes
that identified tumor cells that were, or were not, responsive to 5-fluorouracil
and/or camptothecin.165 Signatures such as these may be useful in the future to
predict which adjuvant therapy setting may provide the best outcome in specific
clinical patients (see Chapters 2, 8, and 9).
4.6 OTHER FOODS, PHYTOCHEMICALS, AND

SUPPLEMENTS USEFUL IN ADJUVANT
CHEMOTHERAPY
Flaxseed, in addition to being a great source of LNA, is also one of the richest
sources of the phytoestrogen secoisolariciresinol diglycoside (SDG), a plant lig-
nan that can be metabolized by colonic bacteria to the mammalian lignans
enterodiol and enterolactone. A related lignan to the one identified in flaxseed,
7-hydroxymatairesinol, is richly expressed in Norway spruce (Picea abies) and
can also be metabolically converted to the mammalian lignan enterolactone. SDG
itself antagonizes mammary tumor growth in nude mice, both for ER+ and ER-
tumor types.173 7-hydroxymatairesinol also has activity against breast tumors in
animal models, but has additional activity against prostate cancer as well.174 When
used in combination with Tamoxifen, even in the presence of high circulating
17-β-estradiol levels, flaxseed enhanced the antitumorigenic response by inhib-
iting tumor cell growth and promoting apoptosis.175 This could have important
implications for human breast cancer therapy where Tamoxifen is being used in
treatment or chemoprevention. A recent placebo-controlled trial using 25 g/d
flaxseed in patients with newly diagnosed breast cancer demonstrated that 4 weeks
of treatment with the flaxseed muffin (between the confirmation biopsy and
surgical resection of the tumor) significantly reduced Ki-67 labeling index, HER-
2/neu expression, progesterone, and estrogen receptor levels, and increased apo-
ptosis in tumor tissue.80 It will be interesting to see if this approach results in
long-term successful treatment outcomes and/or benefits those who go on for
radiation or chemotherapy treatments.
Indole-3-carbinol has been shown to inhibit the hepatotoxicity of the tetrahy-
droisoquinoline alkyloid, trabectidin, without compromising the drug’s mammary
carcinoma killing ability in rats.176 Maitake mushroom β-glucan was recently
shown to enhance the in vitro activity of carmustine in drug-resistant prostate
cancer cells;177 human clinical trials using this preparation in hormone refractory
prostate cancer are needed. Several studies have suggested utility of other natural
medicines, including Rasayanas, which is a complex group of herbals used in
Ayurveda traditional Indian medicine.178 This complex formulation showed
reduced leukopenia and enhanced bone marrow cellularity in cyclophosphamide-
treated mice and enhanced NK activity following radiation therapy179 and inhib-
ited tumor growth and metastasis in melanoma and prostate cancer models.180,181
There is a single report of reduced myelosuppression in a pilot study using human
cancer patients receiving radiation or chemotherapy.182
In addition to certain supplementation conditions, there is evidence that
dietary deficiency of some specific nutrients may actually improve therapy out-
come. While this is generally not the case, a specific example where limiting
nutrient availability may be helpful for cancer treatment is methionine restriction.
A number of studies suggest that tumor cells are dependent on methionine, while
normal cells can use homocysteine instead if provided (reviewed by Cellarier et
al.183). Many tumor types including gliomas and melanomas appear to have low
methionine synthase activity compared to normal surrounding tissue. This could
result from enzymatic mutations or defects in the generation of cofactors B12 and
folate for activity. Alternatively, methionine salvage for polyamine biosynthesis
may be limiting because of deficiencies in methylthioadenosine phosphorylase,
which seems typical of nonsmall-cell lung cancer, leukemia, glioma, rectal ade-
nocarcinoma, and melanoma. Additionally, there is evidence that the requirement
for methionine in tumor cells may simply be much higher than in normal cells
because of increased rates of transmethylation. Capitalizing on these metabolic
differences, some success has occurred in animal models combining methionine-
deficient diets with methionine analogs, such as ethionine in prostate, glioma,
and sarcoma.183 Furthermore, because of the link between folate metabolism and
methionine, deficiency combined with 5-fluorouracil treatment has synergistic
antitumor activity in these same models. Benefits have also been observed with
methionine deficiency in combination with doxorubicin, antimitotics (such as
vincristine), with the alkylating agent carmustine and with cisplatin in human
colon and breast cancer models.184–188 Large-scale clinical trials are still lacking
for these approaches, but are certainly worthy of additional study.
4.7 CONCLUSIONS
As the population continues to age and the numbers of individuals afflicted with
cancer continues to climb over the next few decades, substantial new treatments
will need to be available. While there have been considerable strides in the
detection and treatment of some cancers, there continues to be a substantial
problem of drug resistance and failure of therapies for a large portion of those
with disease. Furthermore, incidence of secondary or recurrent malignancies
continues to be a problem for those who survive their first diagnosis and go on
to surgery, radiation, and/or chemotherapy. A large number of children and adults
are currently using nutritional strategies to either decrease their risk of developing
cancer or to improve their likelihood of cure. While there are still numerous
unanswered questions regarding mechanisms, interactions with conventional
drugs, and very few human clinical intervention trials, this “adjuvant” strategy is
likely to be adopted and expanded over the next several years. Understanding the
molecular targets of drugs and nutrients/phytochemicals should lead to improve-
ments in cancer therapies, increased cure rates, and much more immediately,
improved quality of life for cancer patients. The findings presented here are
encouraging and give hope that together conventional practitioners and those that
practice complementary and alternative medicine can cooperate to provide the
best possible treatment options for their patients.
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5 Role of Nutritional
Antioxidants in the
Prevention and
Treatment of
Neurodegenerative
Disorders
Ennio Esposito
CONTENTS
5.1 Introduction..............................................................................................130
5.1.1 Major Neurodegenerative Disorders ...........................................130
5.1.2 Oxidative Stress and Antioxidants ..............................................131
5.1.3 Epidemiological Studies..............................................................133
5.2 Natural Dietary Antioxidants ..................................................................134
5.2.1 Presence of Flavonoids in Foods ................................................138
5.2.2 Absorption and Metabolism of Polyphenols ..............................140
5.2.3 Biochemical Actions of Polyphenols ..........................................141
5.2.4 Putative Health Benefits of Flavonoids.......................................142
5.3 ROS, NFκB, and Neurodegenerative Disorders .....................................143
5.4 Alzheimer’s Disease, Oxidative Stress, NFκB, and Antioxidants .........146
5.5 Parkinson’s Disease, Oxidative Stress, NFκB, and Antioxidants ..........149
5.6 Amyotrophic Lateral Sclerosis, Oxidative Stress,
NFκB, and Antioxidants..........................................................................154
5.7 Conclusions..............................................................................................156
5.8 Acknowledgments ...................................................................................157
References .........................................................................................................157
129
Keywords: Neurodegeneration, Alzheimer’s disease, Parkinson’s disease, amyo-

trophic lateral sclerosis, oxidative stress, nuclear factor κB, antioxidants, polyphe-
nols, neuroprotection, nutritional.
5.1 INTRODUCTION
There is an increasing attention toward the role played by certain nutritional
components found in foods, including dietary flavonoids, in fruit, vegetables and
beverages in the prevention of age-related decreases in cognitive, memory and
learning tasks. Thus, aging is a major risk factor for neurodegenerative diseases
including Alzheimer’s disease (AD), Parkinson’s disease (PD), and amyotrophic
lateral sclerosis (ALS). An unbalanced overproduction of reactive oxygen species
(ROS) may give rise to oxidative stress, which can induce neuronal damage,
ultimately leading to neuronal death by apoptosis or necrosis. A large body of
evidence indicates that oxidative stress is involved in the pathogenesis of AD,
PD and ALS. An increasing number of studies show that nutritional antioxidants
(especially vitamin E and polyphenols) can block neuronal death in vitro, and
may have therapeutic properties in animal models of neurodegenerative diseases
including AD, PD and ALS. Moreover, clinical data suggest that nutritional
antioxidants might exert some protective effect against AD, PD and ALS. In this
chapter, the biochemical mechanisms by which nutritional antioxidants can
reduce or block neuronal death occurring in neurodegenerative disorders are
reviewed. Particular emphasis will be given to the role played by the nuclear
transcription factor-κB (NFκB) in apoptosis, and in the pathogenesis of neuro-
degenerative disorders, such as AD, PD and ALS. The effects of ROS and
antioxidants on NFκB function and their relevance in the pathophysiology of
neurodegenerative diseases will also be examined.
5.1.1 MAJOR NEURODEGENERATIVE DISORDERS

Neurodegenerative disorders, such as AD, PD and ALS, are among the most
common neurological diseases. As the population of elderly increases, the prev-
alence of these age-related diseases is likely to increase. Thus, of the few risk
factors that have been identified for these diseases, increasing age is the only one
that is common to AD, PD and ALS. For AD, the incidence and prevalence of
the disease increase dramatically after age 60; one study showed a 47% prevalence
for patients over age 85.1 Alzheimer’s disease currently affects almost 2% of the
population in industrialized countries,2 and it is predicted that the incidence of
AD will increase three-fold within the next 5 decades.2 The causes of AD, PD
and ALS are not known and, with the possible exception of PD, there is no
treatment that alters significantly the progression of any of these disorders.
Although the vast majority of these neurodegenerative disorders are sporadic,
genetic and environmental factors can determine the individual risk for them.2
For example, a low calorie diet decreases the risk of major neurodegenerative
disorders including AD and PD,3 and there is evidence that suppression of
Role of Nutritional Antioxidants 131
oxidative stress is one mechanism by which dietary restriction protects neurons.4

However, aging appears to be the single most important factor in the occurrence
of AD, PD and ALS.1 In addition to their possible involvement in aging, mito-
chondrial dysfunction and oxidative damage may play important roles in the
slowly progressive neuronal death that is characteristic of several different neu-
rodegenerative disorders including AD, PD and ALS.5–9
5.1.2 OXIDATIVE STRESS AND ANTIOXIDANTS

There is substantial evidence that the brain, which consumes large amounts of
oxygen, is particularly vulnerable to oxidative damage. Free radicals are normal
products of cellular metabolism.10 The predominant cellular free radicals are the
superoxide (O2–.) and hydroxyl (OH.) species.6,11 Other molecules, such as hydro-
gen peroxide (H2O2) and peroxynitrite (ONOO–), although not themselves free
radicals, can lead to the generation of free radicals through various chemical
reactions. Thus, H2O2, in the presence of reduced metal, forms the highly reactive
OH. via the Fenton reaction.11 Peroxynitrite (ONOO–), formed by the reaction of
nitric oxide (NO.) with O2–., is a highly reactive molecule that also breaks down
to form OH.. Together, these molecules are referred to as reactive oxygen species
(ROS) to signify their ability to lead to oxidative changes within the cell.11,12
Problems occur when the production of ROS exceeds the ability of cells to defend
themselves against these substances. This imbalance between cellular production
of ROS and the ability of cells to defend themselves against ROS is referred to
as oxidative stress.11 Oxidative stress can cause cellular damage and ROS oxidize
critical cellular components such as membrane lipids, proteins and DNA, thereby
inducing apoptosis or necrosis.13–17 Necrosis is characterized by a loss of plasma
membrane integrity, the formation of large vacuoles and cell swelling, whereas
typical features of apoptotic cells are nuclear changes that include chromatin
margination and condensation, DNA fragmentation, membrane blebbing and cell
shrinkage.18 There is large scientific literature regarding the relation between ROS
production, the induction of apoptosis (or necrosis) and the pathogenesis of
neurodegenerative disorders.12,18–26 Although this subject is still a matter of debate,
increasing evidence supports the hypothesis that neuronal death may occur pri-
marily by apoptotic mechanisms in AD, PD and ALS.27–31 Therefore, clinical
evidence shows signs of apoptosis in patients with AD, PD and ALS.30–33
Cells normally have a number of mechanisms to resist against damage
induced by free radicals.10 The major antioxidant defenses consist of antioxidant
scavengers, such as glutathione (GSH), vitamin C (ascorbic acid), vitamin E (α-
tocopherol), carotenoids, polyphenols, flavonoids and antioxidant enzymes.
Severe depletion of GSH in mice by administration of buthionine sulphoximine,
which inhibits GSH synthesis, causes neuronal damage and mitochondrial degen-
eration.13 There is a high concentration of ascorbic acid in the gray and white
matter of the central nervous system in all species that have been examined.34
Indeed, the brain, spinal cord and adrenal glands have the highest ascorbate
concentrations of all the tissues in the body.34 Ascorbate is a broad-spectrum
radical scavenger that is effective against peroxyl and hydroxyl radicals, super-
oxide, singlet oxygen and peroxynitrite.34 Also, the lipid-soluble, chain-breaking
antioxidant vitamin E exerts a very important protective function against oxidative
stress in the brain10 and interacts with ascorbate enhancing its antioxidant activ-
ity.35 Little information is available on the levels of carotenoids and flavonoids
in the human brain. The antioxidant enzymes in the brain include Cu/Zn super-
oxide dismutase (SOD-1) and Mn superoxide dismutase (SOD-2), which catalyze
the conversion of O2–. to H2O2.36 H2O2 is then converted to H2O by either catalase
or glutathione peroxidase (GSH-Px). Antioxidant defense mechanisms can be
upregulated in response to increased ROS or peroxide production.37 Although
upregulating antioxidant defense systems may confer protection against ROS,
they are not completely effective in preventing oxidative damage. Moreover, the
efficiency of gene expression may decline with age or become defective as
oxidative damage to the genome increases.
As already mentioned, the brain is especially vulnerable to ROS damage
because of its high oxygen consumption rate, abundant lipid content and relative
paucity of antioxidant enzymes compared with other organs.38 If the increased
demand on the cell’s capacity to detoxify ROS is not met, alterations, such as
aldehydes or isoprostanes from lipid peroxidation, protein carbonyls from protein
oxidation, and oxidized base adducts from DNA oxidation may accumulate.10
Oxidation of polyunsaturated fatty acids (PUFA) results in the production of
multiple aldehydes with different carbon chain lengths including propanal, buta-
nal, pentanal, hexanal and 4-hydroxy-2-trans-nonenal (4-HNE). There is evidence
that 4-HNE is capable of inducing apoptosis in PC12 cells and cultured rat
hippocampal neurons, suggesting that it is a mediator of oxidative stress-induced
apoptosis.39 These findings suggest that in addition to direct ROS damage to
phospholipid membranes, there is an indirect mechanism involving 4-HNE, which
may also be involved in neuronal death. In this regard, it noteworthy that 4-HNE
has been suggested to be involved in the pathogenesis of PD.40,41 Oxidative
damage to proteins can be revealed by measuring protein carbonyl content,42
which was found to be elevated in AD and ALS patients.43 Another indication of
protein oxidation is the formation of nitrotyrosine by peroxynitrite. This might
represent a useful clinical parameter of the occurrence of oxidative stress in
neurodegenerative diseases, inasmuch as increased levels of nitrotyrosine have
been found in AD, PD and ALS.21,44–49 The most useful marker of DNA oxidation
is 8-hydroxy-2′-deoxyguanosine (8-OHdG), which is elevated in patients with
AD, PD and ALS.50–54
Another index of oxidative is the activation of the transcription factor NFκB
(nuclear factor kappa B). Thus, a large body of evidence indicates that ROS can
act as second messengers mediating intracellular responses, including NFκB
activation.55–59 In turn, activated NFκB can influence the expression of a large
number of genes, including SOD-2.55,58,60 Hence, NFκB activation can be con-
sidered as the executive branch of a feed-back mechanism that operates to regulate
the intracellular concentration of ROS, trying to dampen an excessive accumu-
lation of ROS, which can be dangerous for the cell. Moreover, NFκB induces
the expression of the so-called IAPs (inhibitor of apoptosis proteins), Bcl-2, and
calbindins.60,61 All these biochemical actions of NFκB indicate that this transcrip-
tion factor can exert an antiapoptotic effect, thereby protecting neurons against
degeneration.58,60 As we will discuss below, these data are consistent with clinical
findings showing increased levels of NFκB in vulnerable regions of the central
nervous system of AD, PD and ALS.62–64
5.1.3 EPIDEMIOLOGICAL STUDIES

Although the available data are still limited, epidemiological studies indicate that
dietary habits can influence the incidence of neurodegenerative disorders, such
as dementia (including AD) and PD.65–69 For example, incidence data from the
so-called PAQUID (Personnes Agees Quid) study showed that people drinking
three to four glasses of wine per day had an 80% decreased incidence of dementia
and AD 3 years later, compared to those who drank less or did not drink at
all.67,68,70 This protective effect was still highly significant after adjusting the data
for potential confounding factors such as age, sex, education, occupation and
baseline MMSE (Mini-Mental State Examination). However, in another study,
moderate wine consumption was found to be associated with a fourfold reduction
of the risk for AD, but this effect disappeared when institutionalization was taken
into account.71 These protective effects are most likely due to the presence of
antioxidants in food and beverages,65,68 inasmuch as it has been found that wine
drinking and the consumption of other foods and drinks, which are rich in
polyphenols, can increase the antioxidant activity in serum.72–74 More recently,
investigators in the Rotterdam Study75 reported that any form of moderate alcohol
would have the same beneficial effects. The risk reduction associated with alcohol
is possibly related to its antioxidant properties or its effects on lipid metabolism.
Some observational studies have found a beneficial effect on the risk of
dementia associated with vitamin supplements intake, although this effect has
been observed with vitamin C alone in two studies76,77 and only with combination
of vitamin E and C in another.78 Subsequently, other researchers failed to find
such an association between vitamin supplement intake and incident AD, but they
have demonstrated a decreased risk of incident AD among subjects with high
dietary intake of vitamins, particularly vitamin E.79,80 However, Varner81 suggested
a different interpretation for the inverse relationship between intake of vitamin
E from food (not supplements) with the risk of AD; the predominant form of
vitamin E, γ-tocopherol, but not the form found in supplements, α-tocopherol,
has been shown to inhibit cyclooxygenase 2 (COX-2) production of prostaglandin
E2 in macrophage and epithelial cells.82 Indeed, long-term use of nonsteroidal
antiinflammatory drugs and COX-2 inhibition may protect against AD.83 More-
over, commenting on a paper of Engelhart et al.,79 Brenner84 proposed that silicon
might be the dietary element responsible for the positive effect of vitamin E-rich
foods on the risk for AD. Thus, plant-based foods (e.g., grains and cereal products,
which have high levels of vitamin E) are also the major dietary sources of silicon,85
which could be associated with reduction in development of AD. Asian Indians
have much higher silicon intakes than do Western populations because of higher
intakes of plant-based foods86 and it may be significant that Indians have among
the lowest rates of AD.87 More recently, it was found that subjects with low plasma
vitamin E concentrations are at higher risk of developing a dementia in subsequent
years.88 However, this is a very controversial issue in that data obtained from the
Honolulu–Asia Aging Study, a prospective community-based study of Japanese-
American men who were age 45 to 68 in 1965 to1968, show that midlife dietary
intake of antioxidants does not modify the risk of late-life dementia or its most
prevalent subtypes.89 Thus, intake of β-carotene, vitamin C and flavonoids was
not associated with the risk of dementia and its subtypes either at 6 years or at
26 years of follow-up.89,90
Epidemiological studies have also found an inverse association between high
intake of dietary vitamin E (but not flavonoids or vitamin C) and the occurrence
of PD.69,91 However, these data were not confirmed by other studies,66,92 although
Hellenbrand et al.66 reported a significant statistical trend toward a protective
effect of vitamin C in PD. Individuals over the age of 65 that had higher levels
of β-carotene performed better on learning and memory tests compared with
individuals with low β-carotene levels.93 Lycopene is a carotenoid that has been
suggested to protect against heart disease, stroke and certain cancers.94,95 Lyco-
pene can protect cultured hippocampal neurons against amyloid-beta (Aβ) and
glutamate toxicity.3 Uric acid is markedly effective in protecting cultured neurons
against insults relevant to AD and PD, including exposure to Aβ and iron.96,97
The clinical findings indicating a protective effect of dietary flavonoids against
neurodegenerative disease are supported by data obtained in laboratory animals
showing that a diet supplemented with fruits and vegetables rich in antioxidants
(blueberries, strawberries and spinach) can have beneficial effects on age-related
decline of neuronal and cognitive function in old rats.98
This chapter will focus on the actions of in vitro application of natural
nutritional antioxidants in experimental models of neurodegenerative disorders.
The capability of these compounds to counteract the damaging effects of ROS,
and the relevance of this biochemical effect in their putative neuroprotective action
will be examined. Among the numerous biochemical effects of ROS and antiox-
idants, particular emphasis will be given to their interference with NFκB function,
whose role in the pathophysiology of neurodegenerative disorders is gaining
increasing attention. Moreover, the effects of the administration of “pharmaco-
logical” doses of nutritional antioxidants in animal models and in patients with
AD, PD and ALS will be reviewed. Finally, a detailed analysis on the role of
dietary intake of polyphenols and other antioxidant vitamins in the prevention of
AD and PD will be carried out.
5.2 NATURAL DIETARY ANTIOXIDANTS

Natural dietary antioxidants include vitamins A, C and E, carotenoids, polyphe-
nols and flavonoids. Vitamin C (ascorbate) and vitamin E (α-tocopherol) are
absorbed from the gut. Ascorbate is rapidly distributed to all tissues, whereas
α-tocopherol is incorporated into lipoproteins in the liver and is then secreted

together into plasma.13 Ascorbate can scavenge many reactive species including
O2-., OH. and lipid hydroperoxides,34 and may stabilize catecholamines from
forming ROS. α-tocopherol is a powerful chain-breaking antioxidant that inhibits
lipid peroxidation.35 Carotenoids can scavenge singlet oxygen and a range of
other ROS in vitro, but there is still little evidence that they contribute significantly
to the antioxidant defense system in the central nervous system.13 Several thou-
sand molecules having a polyphenolic structure (i.e., several hydroxyl groups on
aromatic rings) have been identified in higher plants and are generally involved
in defense against ultraviolet radiation or aggression by pathogens.99 These com-
pounds may be classified into different groups as a function of the number of
phenolic rings that they contain and of the structural elements that bind these
rings to one another (Figure 5.1). Distinctions, thus, are made between the
phenolic acids, flavonoids, stilbenes and lignans (Figure 5.1). The flavonoids,
which share a common structure consisting of two aromatic rings (A and B), are
bound together by three carbon atoms that form an oxygenated heterocycle (ring
C) (Figure 5.2). Thus, the flavonoids belong to a group of natural substances with
variable phenolic structures and are found in fruit, vegetables, grains, flowers,
tea and wine.100 More than 4000 varieties of flavonoids have been identified,
many of which are responsible for the attractive colors of flowers, fruits and
leaves.70 Flavonoids represent the single most widely occurring group of phenolic
phytochemicals.101 They can be divided into various classes on the basis of their
molecular structure. The six main groups of flavonoids are:
1. Flavones
2. Flavanones
3. Isoflavones
4. Flavonols
5. Catechins
6. Anthocyanins
The flavones are characterized by a planar structure because of a double bond

in the central ring. One of the best described flavonoids, quercetin, is a member
of this group. Quercetin is found in abundance in onions, apples, broccoli and
berries. The second group is the flavanones, mainly found in citrus fruit. Fla-
vonoids belonging to the catechins are mainly found in green and black tea and
in red wine, whereas anthocyanins are found in strawberries and other berries,
grapes, wine and tea.70 Another phenolic antioxidant is curcumin, a yellow curry
spice derived from turmeric, which is used as a food preservative and herbal
medicine in India. Most flavonoids are glycosylated in their natural dietary forms
with the exception of the catechins.101 Generally, flavonoids may undergo three
forms of intracellular metabolism: (1) conjugation with thiols, particularly GSH;
(2) oxidative metabolism; and (3) P450-related metabolism.102 Metabolic modi-
fications of flavonoids will alter their “classical” antioxidant nature, which is
defined mainly by the presence of a B-ring catechol group (dihydroxylated B-ring)
Hydroxybenzoic acid Hydroxycinnamic acids
R1
R1
O
O R2
R2
OH
OH R3
R3
R1 = R2 = OH, R3 = H : Protocatechuic acid R1 = OH : Coumaric acid
R1 = R2 = R3 = OH : Gallic acid R1 = R2 = OH : Caffeic acid
R1 = OCH3 = OH : Ferulic acid
O
HO OH
O
OH
H O
HO H
H OH OH
Chlorogenic acid
Flavonoids
Stilbenes Lignans
CH3O CH2OH
HO
HO CH2OH
OH
OCH3
HO OH
Resveratrol Secoisolariciresinol
FIGURE 5.1 Chemical structure of polyphenols.
capable of readily donating hydrogen (electron) to stabilize a radical species.102

Other structural features for antioxidant nature include the presence of 2,3 unsat-
uration in conjunction with 4-oxo- function in the C-ring and the presence of
functional groups capable of binding transition metal ions, such as iron and
copper. Circulating metabolites of flavonoids, such as glucuronides and O-meth-
ylated forms, and intracellular metabolites (e.g., flavonoid–GSH adducts) have
reduced ability to donate hydrogen and are less effective scavengers of ROS and
nitrogen species relative to their parent aglycone forms.102
Flavonols Flavones
R1
R1
R2
R2
HO O
R3 HO O
R3
OH
OH O
OH O
R2 = OH; R1 = R2 = H : Kaempferol R1 = H ; R2 = OH : Apigenin
R1 = R2 = OH ; R3 = H : Quercetin R1 = R2 = OH : Luteolin
R1 = R2 = R3 = OH : Myricetin
Flavanones
Isoflavones R1
R2
HO O
HO O
R3
R1 O OH O
OH
R1 = H; R2 = OH : Narigenin
R1 = H : Daidzein R1 = R2 = OH : Eriodictyol
R1 = OH : Genistein R1 = OH; R2 = OCH3 : Hesperetin
Flavanols
Anthocyanidins R1
R1
R2
OH
HO O
+
HO O R3
R2
OH
OH
OH
OH R1 = R2 = OH; R3 = H : Catechins
R1 = R2 = R3 = OH : Gallocatechin
R1 = R2 = H : Pelargonidin
R1 = OH; R2 = H : Cyanidin OH
R1 = R2 = OH : Delphinidin
R1 = OCH3; R2 = OH : Petunidin HO O
OH
R1 = R2 = OCH3 : Malvidin
OH
OH
OH
HO O
OH
OH OH
OH
HO O
OH
OH
OH
Trimeric procyanidin
FIGURE 5.2 Chemical structure of flavonoids.

5.2.1 PRESENCE OF FLAVONOIDS IN FOODS

Flavonols are the most ubiquitous flavonoids in foods; the main representatives
are quercetin and kaempferol. They are generally present at relatively low con-
centrations of 15 to 30 mg/kg fresh weight. The richest sources are onions (up
to 1.2 g/kg fresh weight), curly kale, leeks, broccoli and blueberries. Red wine
and tea also contain up to 45 mg flavonols/l. These compounds are present in
glycosylated forms. The associated sugar moiety is very often glucose or rham-
nose, but other sugars may also be included (e.g., galactose, arabinose, xylose,
glucuronic acid). Flavones are much less common than flavonols in fruit and
vegetables. Flavones consist mainly of glycosides of luteolin and apigenin. The
only important edible sources of flavones identified to date are parsley and
celery.102 In human foods, flavanones are found in tomatoes and certain aromatic
plants, such as mint, but they are present in high concentrations only in citrus
fruit.102 The main aglycones are naringenin in grapefruit, hesperetin in oranges,
and eriodictyol in lemons. Flavanones are generally glycosylated by a disaccha-
ride at position 7: either a neohesperidose, which imparts a bitter taste (such as
naringenin in grapefruit) or a rutinose, which is flavorless. Orange juice contains
between 200 and 600 mg hesperidin/l and 15 to 85 mg narirutin/l, and a single
glass of orange juice may contain between 40 and 140 mg flavanone glycosides.99
Because the solid parts of citrus fruit, particularly the albedo (the white spongy
portion) and the membrane separating the segments, have a very high flavanone
content, the whole fruit may contain up to five times as much as a glass of orange
juice.99
Isoflavones are flavonoids with structural similarities to estrogens. Although
they are not steroids, they have hydroxyl groups in positions 7 and 4′ in a
configuration analogous to that of the hydroxyls in the estradiol molecule. This
confers pseudohormonal properties on them, including the ability to bind to
estrogen receptors, and they are consequently classified as phytoestrogens. Isofla-
vones are found almost exclusively in leguminous plants.99 Soy and its processed
products are the main source of isoflavones in the human diet. The isoflavone
content of soy and its manufactured products varies greatly as a function of
geographic zone, growing conditions and processing. Soybeans contain 580 to
3800 mg isoflavones/kg fresh weight, and soymilk contains between 30 and 175
mg/l. Flavanols exist in both the monomeric form (catechins) and the polymeric
form (proanthocyanidines). Catechins are found in many types of fruits; apricots,
which contain 250 mg/kg fresh weight, are the richest fruit source. They are also
present in red wine (up to 300 mg/l), but green tea and chocolate are by far the
richest sources.99 An infusion of green tea contains up to 200 mg catechins. Black
tea contains fewer monomeric flavanols, which are oxidized during “fermenta-
tion” (heating) of tea leaves to more complex condensed polyphenols known as
theaflavins (dimers) and thearubigins (polymers). Catechin and epicatechin are
the main flavanols in fruit, whereas gallocatechin, epigallocatechin and epigallo-
catechin gallate are found in certain seeds of leguminous plants, in grapes and,
more importantly, in tea.99 In contrast to other classes of flavonoids, flavanols are
not glycosylated in foods. The tea epicatechin is remarkably stable when exposed
to heat as long as the pH is acidic; only 15% of this substance is degraded after
7 h in boiling water at pH 5.99
Proanthocyanidins, which are also known as condensed tannins, are dimers,
oligomers and polymers of catechins that are bound together by links between
C4 and C8 (or C6). Through the formation of complexes with salivary proteins,
condensed tannins are responsible for the astringent character of fruit (grapes,
peaches, kakis, apples, pears, berries, etc.) and beverages (wine, cider, tea, beer,
etc.) and for the bitterness of chocolate.99 This astringency changes over the course
of maturation and often disappears when the fruit reaches ripeness; this change
has been well explained in the kaki fruit by polymerization reactions with ace-
taldehyde. Such polymerization of tannins probably accounts for the apparent
reduction in tannin content that is commonly seen during the ripening of many
types of fruit. It is difficult to estimate the proanthocyanidin content of foods
because they have a wide range of structures and weights. Anthocyanins are
pigments dissolved in vacuolar sap of the epidermal tissues of flowers and fruit,
to which they impart a pink, red, blue or purple color.99 They exist in different
chemical forms, both colored and uncolored, according to pH. Although they are
highly unstable in the aglycone form (anthocyanidins) while they are in plants,
they are resistant to light, pH and oxidation conditions, which are likely to degrade
them. In the human diet, anthocyanins are found in red wine, certain varieties of
cereals and certain leafy and root vegetables (aubergines [eggplant], cabbage,
beans, onions, radishes), but they are most abundant in fruit.
Cyanidin is the most common anthocyanidin in foods. Food contents are
generally proportional to color intensity and reach values up to 2 to 4 g/kg fresh
weight in blackcurrants or blackberries. These values increase as the fruit ripens.
Anthocyanins are found in the skin of certain types of red fruit and may occur
in the flesh (cherries and strawberries) as well. Wine contains 200 to 350 mg
anthocyanins/l, and these anthocyanins are transformed into various complex
structures as the wine ages.99 Stilbenes are found in only low quantities in the
human diet. One of these, resveratrol, for which anticarcinogenic effects have
appeared during screening of medical plants and which has been extensively
studied, is found in low quantities in wine (0.3 to 7 mg aglycone/l and 15 mg
glycosides/l in red wine). However, because resveratrol is found in such small
quantities in the diet, any protective effect of this molecule is unlikely at normal
nutritional intake.99
In most cases, foods contain complex mixtures of polyphenols, which are
often poorly characterized. Apples, for example, contain flavanol monomers
(mainly epicatechin) or oligomers (procyanidin B2 mainly), chlorogenic acid
and small quantities of other hydroxycinnamic acids, two glycosides of phlo-
retin, several quercetin glycosides and anthocyanins, such as cyanidin 3-galac-
toside in the skin of certain red varieties. Apples are one of the rare types of
food for which fairly precise data on polyphenol composition between varieties
of apples have notably been studied. The polyphenol profiles of all varieties of
apples are practically identical, but concentrations may range from 0.1 to 5 g
total polyphenols/kg fresh weight and may be as high as 10 g/kg in certain

varieties of cider apples.99
Methods of culinary preparation also have a marked effect on the polyphenol
content of foods. For example, simple peeling of fruit and vegetables can eliminate
a significant portion of polyphenols because these substances are often present
in higher concentrations in the outer parts than the inner parts. Cooking may also
have a major effect. Onions and tomatoes lose between 75 and 80% of their initial
quercetin content after boiling for 15 min, 65% after cooking in a microwave
oven, and 30% after frying. Steam cooking of vegetables, which avoids leaching,
is preferable. Potatoes contain up to 190 mg chlorogenic acid/kg, mainly in the
skin. Extensive loss occurs during cooking and no remaining phenolic acids were
found in french fries or freeze-dried mashed potatoes.99
Only partial information is available on the quantities of polyphenols that are
consumed daily throughout the world. These data have been obtained through
analysis of the main aglycones (after hydrolysis of their glycosides and esters)
in the foods most widely consumed by humans. In 1976, Kuhnau103 calculated
that dietary flavonoid intake in the U.S. was 1 g/day and consisted of the follow-
ing: 16% flavonols, flavones and flavanones; 17% anthocyanins; 20% catechins;
and 45% “biflavones.” Although these figures were obtained under poorly detailed
conditions, they continue to serve as reference data. Certain studies have subse-
quently provided more precise individual data concerning the intake of various
classes of polyphenols. Flavonols have been more extensively studied. Consump-
tion of these substances has been estimated at 20 to 25 mg/day in the U.S.,
Denmark and Holland.99 In Italy, consumption ranged from 5 to 135 mg/day, and
the mean value was 35 mg/day.99 The intake of flavanones is similar or possibly
higher than that of flavonols, with a mean consumption of 28.3 mg hesperetin/day
in Finland.99
5.2.2 ABSORPTION AND METABOLISM OF POLYPHENOLS

Metabolism of polyphenols occurs via a common pathway. The aglycones can
be absorbed from the small intestine. However, most polyphenols are present in
food in the form of esters, glycosides or polymers that cannot be absorbed in
their native form. These substances must be hydrolyzed by intestinal enzymes or
by the colonic microflora before they can be absorbed. During the course of
absorption, polyphenols are conjugated in the small intestine and later in the liver.
This process mainly includes methylation, sulfanation and glucuronidation. The
conjugation mechanisms are highly efficient and aglycones are generally absent
in blood or present in low concentrations after consumption of nutritional doses.
Circulating polyphenols are conjugated and extensively bound to albumin and
both polyphenols and their derivatives are eliminated chiefly in the urine and bile.
Polyphenols are secreted via the biliary route into the duodenum, where they are
subjected to the action of bacterial enzymes, especially β-glucuronidase, in the
distal segments of the intestine, after which they may be reabsorbed. The entero-
hepatic recycling may lead to a longer half-life of polyphenols within the body.
The partitioning of polyphenols and their metabolites between aqueous and

lipid phases is largely in favor of the aqueous phase because of their hydrophilicity
and binding to albumin. However, in some lipophilic membrane models, some
polyphenols penetrate the membrane to various extents. Quercetin showed the
deepest interaction, probably because of its ability to assume a planar conforma-
tion. At physiologic pH, most polyphenols interact with the polar head groups of
phospholipids at the membrane surface via the formation of hydrogen bonds that
involve the hydroxyl groups of the polyphenols99
5.2.3 BIOCHEMICAL ACTIONS OF POLYPHENOLS

Flavonoids can prevent injury caused by ROS in various ways.104 One way is the
direct scavenging of free radicals.105-107 Structurally important features defining
the reduction potential of flavonoids are believed to be the hydroxylation pattern,
especially a 3′,4′-dihydroxy catechol structure in the B-ring, the planarity of the
molecule and the presence of 2,3 unsaturation in conjunction with a 4-oxo-
function in the C-ring (see Figure 5.1). Thus, flavonoids with an O-dihydroxy
catechol group in the B-ring (quercetin, epicatechin, etc.) are more powerful
reductants/antioxidants and scavengers of ROS than those having a monohydrox-
yphenolic structure. Flavonoids are oxidized by radicals resulting in a more stable,
less reactive radical. In other words, flavonoids stabilize ROS by reacting with
the compound of the radical. Because of the high reactivity of the hydroxyl group
of the flavonoids, radicals are made inactive, according to the following equation:
Flavonoid (OH) + R. → flavonoid(O.) + RH
where R. is a free radical and O. is an oxygen free radical. Selected flavonoids

can directly scavenge superoxide, whereas other flavonoids can scavenge the
highly reactive oxygen-derived radical peroxynitrite.108,109 For example, flavanols
are scavengers of superoxide anions,110 singlet oxygen111 and lipid peroxy radi-
cals,112 and they can sequester metal ions by chelation.113
It has recently been shown that the flavonoid compounds caffeic acid and
(+)-catechin can inhibit peroxynitrite-mediated oxidation of dopamine.114 More-
over, it has been demonstrated that (—)-epicatechin, (—)-epicatechin gallate and
quercetin serve as powerful antioxidants against lipid peroxidation when phos-
pholipid bilayers are exposed to ROS in vitro.107,115 There is also evidence that
flavonoids can inhibit the activities of several enzymes, including lipoxygen-
ase,116-118 cyclooxygenase,116,117 xanthine oxidase,119 phospholipase A2120 and pro-
tein kinases.121 These biological effects are believed to derive from the antioxidant
properties of the related flavonoids.119 However, increasing evidence suggests that
flavonoids might exert modulatory effects in cells independently from classical
antioxidant activity through selective actions at different components of a number
of protein kinase and lipid kinase signaling cascades, such as phosphoinositide
3-kinase (PI 3-kinase), protein kinase B (Akt/PKB), tyrosine kinases,
protein kinase C (PKC), and mitogen-activated protein kinase (MAP kinase).102
Flavonoids have the potential to bind to the adenosine triphosphate (ATP)-binding

sites of a large number of proteins, including mitochondrial ATPase, calcium
plasma membrane ATPase, protein kinase A, PKC and topoisomerase.102 In addi-
tion, interactions with benzodiazepine-binding sites of GABA-A receptors and
with adenosine receptors102 have been shown.
Resveratrol and the citrus flavanones hesperetin and naringenin have been
reported to exert inhibitory activity at a number of protein kinases102 This inhi-
bition is mediated via the binding of the polyphenols to the ATP binding site,
presumably causing three-dimensional structural changes in the kinase leading
to its inactivity. Flavonoids may also interact with mitochondria, interfere with
pathways of intermediary metabolism, and/or down regulate the expression of
adhesion molecules.102 There are a number of additional potential sites where
flavonoids may interact with key signaling pathways. For instance, flavonoid-
mediated inhibition of oxidative stress-induced apoptosis may occur by prevent-
ing the activation of JNK (c-Jun amino-terminal kinase). There is strong evidence
linking the activation of JNK to neuronal loss in response to a wide array of
proapoptotic stimuli in both developmental and degenerative death signaling.121,122
A number of flavonoids have been reported to inhibit the activation of JNK,
although it is not clear if this is mediated by antioxidant activity or is due to
inhibitory actions at signaling molecules.
Much interest has focused recently on the beneficial effects of flavanols, such
as epicatechin, epigallocate chin (EGC), and epigallocate chin gallate (EGCG), and
there is growing interest that the cytoprotective nature of these polyphenols is based
on their interaction within signaling pathways. For example, epicatechin and one of
its major in vivo metabolites, 3′-O-methyl epicatechin, have been shown to elicit
strong cytoprotective effects against oxidative stress in fibroblasts and neurons.102 In
another study, the neuroprotective mechanism of another flavanol, EGCG, against
oxidative stress-induced cell death was also found to involve modulation of signaling
proteins. Thus, EGCG caused a stimulation of PKC and a modulation of cell sur-
vival/cell cycle genes, such as Bax, Bad, Mdm2, Bcl-2, Bcl-w, and Bcl-x.123,124
Together, these findings suggest that protection is likely to be partly mediated through
specific action within signaling pathways, although at this time it remains unclear
exactly where such interactions occur within the pathway.
5.2.4 PUTATIVE HEALTH BENEFITS OF FLAVONOIDS

In recent years, there has been an increasing interest in investigating the many
positive pharmacological properties of flavonoids. Much of this interest has been
spurred by the dietary anomaly referred to as the “French paradox,” the apparent
compatibility of a high saturated fat diet with a low incidence of coronary ath-
erosclerosis.125 It was suggested that the polyphenolic substances, such as fla-
vonoids in red wine, can provide protection against coronary heart disease. The
natural phytoalexin resveratrol and the flavonoids quercetin and (+)-catechin have
been invoked in order to explain the beneficial effects of moderate red wine
consumption against coronary heart diseases.126,127 In addition, epidemiological
studies have shown that moderate wine consumption can be protective against
neurological disorders, such as age-related macular degeneration127,128 and AD.68
Moreover, in vitro and in vivo preclinical studies have shown the neuroprotective
effect of lyophilized red wine,129 grape polyphenols,130 quercetin,131 trans-
resveratrol132–134 and (+)-catechin.135 Taken together, these findings raise the pos-
sibility that red wine constituents may be beneficial in the prevention of age-
related neurodegenerative disorders. There is also increasing interest for the role
of tea (Camellia sinensis) in maintaining health and in treating disease. Although
tea consists of several components, research has focused on polyphenols, espe-
cially those found in green tea. The green tea polyphenols include (—)-epicatechin
(EC), (—)-epigallocatechin (EGC), (—)-epicatechin-3-gallate (ECG), (—)-epi-
gallocatechin-3-gallate (EGCG). Of these, EGCG generally accounts for greater
than 40% of the total.136 Green tea polyphenols are potent antioxidants.106 EGCG
usually has the greatest antioxidant activity and is the most widely studied
polyphenol for disease prevention.136–138 Many of the putative health benefits of
tea are presumed to stem from its antioxidant effects.
The epidemiological evidence indicating the putative role of nutritional anti-
oxidants in the prevention and attenuation of neurodegenerative disorders is
receiving experimental confirmation in a number of laboratory studies. Thus, the
polyphenol epicatechin was shown to attenuate neurotoxicity induced by oxidized
low-density lipoprotein in mouse-derived striated neurons.138 Tea extracts and
EgCG attenuated the neurotoxic action of 6-OHDA in rat PC12 cells, human
neuroblastoma SH-SY5Y cells,137 and was shown to be neuroprotective in a
mouse model of PD.138 Moreover, recent reports have revealed that flavonoids
may be neuroprotective in neuronal primary cell cultures. For example, the ginkgo
biloba extract, enriched with flavonoids, has been shown to protect hippocampal
neurons from nitric oxide or β-amyloid derived, peptide-induced neurotoxic-
ity.139–141 In addition, the extract of ginkgo biloba, referred to as Egb 761, is one
of the most popular plant extracts used in Europe to alleviate symptoms associated
with a range of cognitive disorders.142,143 The mechanism of action of Egb 761
in the central nervous system is only partially understood, but the main effects
seem to be related to its antioxidant properties, which require the synergistic
action of the flavonoids, the terpenoids (ginkgolides, bilobalide), and the organic
acids, principal constituents of Egb 761.144 These compounds, to varying degrees,
act as scavengers of ROS, which have been considered the mediators of the
excessive lipid peroxidation and cell damage observed in AD.145–147
5.3 ROS, NFκB, AND NEURODEGENERATIVE

DISORDERS
The transcription factor NFκB, originally studied in cells of the immune system
wherein it regulates cell survival,148–150 is widely expressed in the nervous system
and exists in neurons in both an inducible and a constitutively active form.151–154
NFκB resides in the cytoplasm in an inactive form consisting of three subunits:
p65, p50 and an inhibitory subunit called IκB.148,149,154 When IκB is bound to
p50/p65, it is inactive; signals that activate NFκB cause dissociation of IκB
releasing p50/p65, which then translocates to the nucleus and binds to specific
κB DNA consensus sequences in the enhancer region of a variety of κB-respon-
sive genes.58,60,148,149,154,155
In neurons, NFκB is activated by various intracellular signals, including
cytokines, neurotrophic factors, and neurotransmitters.58,154,156 Activation of
glutamate receptors and membrane depolarization lead to activation of NFκB in
hippocampal pyramidal neurons and cerebellar granule neurons in culture.152,157
The mechanism whereby diverse stimulants lead to the activation of NFκB has
been a subject of intense research. Most work has focused on the p50/p65 dimer,
the predominant form of NFκB activated in many cells including neurons,58,60,148
and its association with IB. It is now known that upon stimulation with many
NFκB inducers, IκBα is rapidly phosphorylated on two serine residues (S32 and
S36), which target the inhibitor protein for ubiquitination and subsequent degra-
dation by the 26S proteasome.155 Released NFκB dimer can then translocate to
the nucleus and activate target genes by binding with high affinity to κB elements
in their promoters. The phosphorylation and degradation of IκBα are tightly
coupled events, so it is likely that agents that activate NFκB do so by stimulating
a specific IκB kinases, or alternatively by inactivating a particular phosphatase.
Two IκB kinases (IKKs) termed IKKα and IKKβ have been described in
research.155 IKKα and β have been shown to be activated by important inducers
of NFκB, such as IL-1 and TNF, to specifically phosphorylate S32 and S36 of
IκBα, and to be crucial for NFκB activation by these cytokines.155 The IKKs are
part of a larger multiprotein complex called the IKK signalsome. It appears that
multiple pathways can regulate NFκB, most of which lead to IκB phosphorylation
via the IKK-containing signalsome155 One model has been proposed whereby
diverse agents all activate NFκB by causing oxidative stress.57,149,158 This hypoth-
esis is based on four main lines of evidence:
1. Direct application of H2O2 to culture medium activates NFκB in some

cell lines.159–162
2. In some cell types, ROS have been shown to be increased in response
to agents that also activate NFκB.149,159–163
3. Virtually all stimuli known to activate NFκB can be blocked by anti-
oxidants, including L-cysteine (a precursor of glutathione), N-acetyl-
L-cysteine (NAC), caffeic acid phenethyl ester (CAPE), (–)-epigallo-
catechin-3-gallate, resveratrol, thiols, dithiocarbamates, vitamin E and
its derivatives, and thioredoxin (an important cellular protein oxi-
doreductase with antioxidant activity).55,56,127,149,159,164–168
4. Inhibition or over expression of enzymes that affect the level of intra-
cellular ROS has been shown to modulate the activation of NFκB by
some agents.163,169 Ultimately, this theory led to the proposal of H2O2
as the central second messenger in NFκB activation.163
A large body of evidence indicates that NFκB is involved in the control of

cell survival. The great majority of the available data shows that NFκB exerts an
antiapoptotic action. Thus, activation of NFκB can prevent cell death in various
culture paradigms.58,170 Moreover, increasing data suggest that NFκB activation
may transduce anti-cell death signals in neurons.58 For example, TNFα protected
cultured hippocampal neurons against death induced by metabolic, excitotoxic
and oxidative insults.58,171 The involvement of NFκB in such neuronal cell death
paradigms is suggested by data showing that TNFα induces activation of NFκB
in cultured hippocampal neurons against excitotoxic and oxidative insults.171–173
Moreover, in the PC12 neuronal cell line174 and in primary sympathetic neurons,175
activated NFκB has been found to mediate the antiapoptotic effect of nerve
growthh factor (NGF). It has also been shown that the resistance of selected
clones of PC12 cells to oxidative cell death induced by Aβ and H2O2 is mediated
by NFκB.176 An inhibition of NFκB potentiated Aβ peptide-mediated apoptotic
damage in primary cultures of cerebellar granule cells,177 and increased the
apoptotic death of PC12 cells induced by autooxidation of dopamine.178 Similarly,
a lack of p50 subunit increased the vulnerability of hippocampal neurons to
excitotoxic injury.179
Recent studies have shown that NFκB is activated and may play a protective
role in neurodegenerative disorders, such as AD,180 PD62 and ALS,64 and severe
epileptic seizures.179 There is also evidence that NFκB plays a pivotal role in the
cell survival-promoting action of ADNF9, a nine amino acid activity-dependent
neurotrophic factor (ADNF) peptide.181 In addition, it has recently been reported
that NFκB is involved in the neuroprotective effect exerted by subtoxic concen-
tration of N-methyl-D-aspartic acid (NMDA) and can counteract low potassium-
induced apoptosis in cultured cerebellar granule neurons.182,183 Also precondition-
ing-induced neuroprotection in cultured hippocampal neurons seems to be medi-
ated by activation of NFκB.184 The mechanism by which NFκB can exert its
antiapoptotic effect is still unclear. One possible mechanism would be the tran-
scription of genes encoding trophic factors, antioxidant enzymes and calcium-
regulating proteins. One of the first genes shown to be responsive to NFκB was
SOD-2, a mitochondrial antioxidant enzyme that protects cells against apopto-
sis.180 Other genes induced by NFκB include the cell adhesion molecules, such
as ICAM-1,185 the inducible form of nitric oxide synthase,186 Bcl-2, Bcl-x, and
the Bcl-2 homologue Bfl-1/A1.61,187,188
However, in some cases NFκB can promote neuronal death.189–191 Thus, the
neuroprotective effect of acetylsalicylic acid is apparently mediated by inhibition
of NFκB.190 More recently, it was found that NFκB is essential for dopamine-
induced apoptosis in PC12 cells.192 Whether NFκB inhibits or promotes apoptosis
might depend on the cell type and the nature of the apoptosis-inducing stimulus.191
However, the explanation for the conflicting results concerning an antiapoptotic
vs. proapoptotic role of NFκB activation still is not clear and has been described
as the “Janus faces” of NFκB.191
5.4 ALZHEIMER’S DISEASE, OXIDATIVE STRESS,

NFκB, AND ANTIOXIDANTS
The incidence rate of Alzheimer’s disease (AD), or the number of new cases
developing among unaffected individuals over a specified time, increases from
approximately 1% annually among people aged 65 to 70 years to approximately
6 to 8% for people over age 85.193 The rates of disease are slightly higher for
women and for African Americans and Caribbean Hispanics.193 The duration of
illness varies considerably from 2 to 20 years. Two population-based studies
found that the median survival time for patients with AD was 3 to 4 years.194,195
The prevalence, or proportion, of individuals surviving with clinically diagnosed
AD also varies dramatically with age. Thus, the estimated prevalence of senile
dementia in Europe increases with age from 1% in men and women of 60 years
of age to 44.7% in the population 90 to 95 years of age.196 AD is the most common
form of dementia, with a prevalence of 0.4% in women and 0.3% in men aged
60 to 69 years.197 A community-based study has suggested that approximately 4
million persons in the U.S. have contracted AD.1 AD is a progressive dementing
disorder characterized by selective neuronal loss in several areas of the central
nervous system. In AD, the progressive memory deficits, cognitive impairments
and personality changes are due to progressive dysfunction and death of the
neocortex, limbic system, hippocampus and several of the subcortical regions of
the brain.
The majority of cases of AD are age-related and, indeed, age is the only
reliable risk factor for the nongenetic sporadic forms (85% of all cases) and,
therefore, for the majority of cases of this disorder.2,198 However, molecular
genetic analyses suggest that there might be many genes that influence individual
susceptibility to AD. The first such susceptibility gene identified was apolipopro-
tein E for which there are three alleles that encode three different isoforms of
apolipoprotein E (E2, E3 and E4). Subjects that produce the E4 isoform are at
increased risk of AD.199 The mechanism by which E4 promotes AD is not well
understood, but there is evidence that E4 enhances Aβ aggregation and reduces
amyloid precursor protein 42 (APP). In addition, data suggest that E4 might
increase the risk of AD by enhancing amyloidogenic processing of certain iso-
forms of APP, increasing oxidative stress and impairing neuronal plasticity. The
characteristic histopathologic alterations in AD are neuritic or senile plaques (SPs)
composed largely of amyloid β-peptides (Aβ) and neuronal aggregates of abnor-
mally phosphorylated cytoskeletal proteins (neurofibrillary tangles [NFTs]). A
number of data indicate that Aβ is responsible for the neuronal death in AD.
Thus, aggregates of Aβ peptides are toxic to neurons in cultures198,200–202 and can
cause cell death by apoptosis18,30,31,202,203; however, the exact mechanisms of Aβ-
induced neurotoxicity are still unknown.
Several lines of evidence suggest that the overproduction of ROS is implicated
in Aβ neurotoxicity:
• Exposure of cultured neurons or neuronal cell lines to Aβ increases

the intracellular levels of ROS2,3,15,204–208 leading to the activation of
NFκB.63
• Markers of oxidative stress are found to increase in a transgenic mouse
model of AD.147,209
• Neurotoxicity of Aβ is attenuated by antioxidants, such as vitamin E,
the spin-trap compound PBN (α-phenyl-tert-butyl nitrone) and laz-
aroids,15,198,200,201,210–212 and/or free radical scavengers.213
Thus, in 1992 the protective effect of vitamin E was first described on neurons
in culture against Aβ–induced cell death.201 Following these initial findings, a
number of subsequent studies confirmed the role of oxidative stress in the neu-
rotoxic effect of Aβ peptide. For example, Behl et al.214 found that Aβ can induce
the formation of H2O2 in hippocampal neurons, which causes peroxidation of cell
membranes and ultimately leads to neuronal death. Consistent with these findings,
exposure of cultured hippocampal neurons to Aβ induced a significant increase
in 4-HNE.215 Moreover, it has recently been found that the phenolic antioxidant
curcumin, which is largely used as a food preservative and herbal medicine in
India, reduces oxidative damage and amyloid pathology in a transgenic mouse
model of AD.216 However, in another study, Aβ-induced neurotoxicity in rat
hippocampal neurons in culture was not affected by several antioxidants;217 nev-
ertheless, pretreatment of cultures with Aβ significantly increased the sensitivity
of neurons to H2O2, suggesting that Aβ can render neurons more susceptible to
ROS damage.217 Some of the proteins oxidatively modified by Aβ−induced oxi-
dative stress include membrane transporters, receptors, GTP-binding proteins (G
proteins) and ion channels. Oxidative modifications of tau by 4HNE and other
ROS can promote its aggregation and, thereby, may induce the formation of
neurofibrillary tangles. Aβ also causes mitochondrial oxidative stress and dysreg-
ulation of Ca2+ homeostasis, resulting in impairment of the electron transport
chain, increased production of superoxide anion radicals and decreased produc-
tion of ATP.
In agreement with data obtained in experimental models, clinical findings
indicate that oxidative stress occurs in AD, as indicated by the finding that higher
than normal levels of lipid, protein and DNA oxidation are found in the brains
of AD patients.46,48,145,146,218 Thus, lipid peroxidation, measured as thiobarbituric
acid reactive substances (TBARS), was found to be increased in various brain
regions of AD patients.219–221 Moreover, Mecocci et al.53 found a significant three-
fold increase in mitochondrial DNA oxidation in the parietal cortex of AD
patients. In addition, immunohistochemical analysis of brain sections from AD
patients using an antibody with selectivity for the activated nuclear form of p65
revealed that NFκB was activated in neurons and astrocytes.63 Cells with activated
NFκB were restricted to the close proximity of early plaque stages.63 Thus, it is
possible that Aβ-induced NFκB activation contributes to the pathological changes
observed in AD via the induction of proinflammatory and cytotoxic genes or,
more likely, that Aβ-induced NFκB activation is part of a cellular defense pro-
gram.
Based on the preclinical and clinical data indicating the presence of oxidative
stress in AD patients, clinical trials were carried out to test the effect of antiox-
idants in this pathological condition. However, as already indicated above, incon-
sistent findings were reported in the trials investigating the effects of antioxidant
vitamins on cognitive function and dementia. Thus, a controlled clinical trial with
dl-α-tocopherol (synthetic form: 2000 IU/d) in patients with moderately severe
impairment from AD showed some beneficial effects with respect to rate of
deterioration of cognitive function.222 In the same dl-α-tocopherol clinical trial,
selegiline (10 mg/d), a monoamine oxidase inhibitor, produced beneficial effects
similar to that produced by dl-α-tocopherol.222 It is interesting to note that there
was no significant difference in effect between the groups receiving a combination
of dl-α-tocopherol and selegiline and those receiving treatment with the individual
agents.146,222 Several possibilities were proposed to explain the lack of additive
effect. One was that selegiline and vitamin E can act by the same mechanism.
Indeed, both reduce the levels of free radicals, although by different molecular
pathways. Vitamin E protects neurons by destroying formed ROS (“quenching”),
whereas selegiline protects neurons by preventing the formation of ROS and by
inhibiting oxidative metabolism of catecholamines. Therefore, clinical studies
involving vitamin E and selegiline support the concept that ROS are one of the
intermediary risk factors for the progression of neurodegeneration in AD.198
However, in the MRC/BHF Hearth Protection Study, which included 20,536
persons allocated to receive either antioxidant vitamin supplementation (vitamins
E and C and β-carotene) or a placebo, no treatment differences were found in
the percentage of persons defined as cognitively impaired or in mean cognitive
scores after 5 years of treatment.223 In addition, no difference was found in the
number of persons who developed dementia during follow-up.
Another clinical study was conducted among 1059 rural, noninstitutionalized
elderly residents of southwestern Pennsylvania, who were participants in the
Monongahela Valley Independent Elders Survey.224 Current use of nutritional
supplements containing vitamins A, C, or E, β-carotene, zinc or selenium was
measured through self-report. After adjustment for age, education and sex, no
significant differences were found in cognitive test performance between antiox-
idant users and nonusers.224 However, in the Rotterdam Study, Jama et al.225
studied 5182 elderly persons and found that dietary and nutritional supplement
intake of β-carotene was inversely associated with cognitive impairment, even
after adjustment for age, sex, education, smoking, total caloric intake and con-
sumption of other antioxidants. The discrepancy between the results of the study
of Mendelsohn et al.224 and that of Jama et al.225 could be due to differences in
the study populations, such as difference in age distribution or socioeconomic
status or the exclusion of demented persons in the Rotterdam study. Although
Jama et al.225 found an association between β-carotene and cognition, they did
not find similar results with vitamins E or C. Nevertheless, the data of a protective
effect of β-carotene against age-related cognitive impairment were not confirmed
in a more recent study.226 Thus, from the analysis of the data from the Washington
Heights-Inwood Columbia Aging Project (WHICAP), results indicated that the
risk of AD was not associated with supplement, dietary or total intake of car-
otenes, vitamin C or vitamin E.226 In contrast, a recent prospective study has
shown a reduced prevalence and incidence of AD in individuals taking vitamins
E and C in combination.227 However, there was no significant reduction in risk
of incident AD with vitamin E or vitamin C alone or with a multivitamin. There
was also no association between AD risk and use of B-complex vitamins.227 In
two other investigations, an association of high folic acid levels and decreased
homocysteine levels with reduced AD risk was found.228,229 Given the conflicting
data reported in many of the trials to date, it is clear that more effort is necessary
in the future to try to confirm whether or not there is a relationship between
dietary habits (in particular, the amount of polyphenols and other antioxidants
intake) and the risk of dementia.
Increasing evidence suggests that diets high in saturated fats may increase
the risk of AD, whereas diets rich in mono- and polyunsaturated fatty acids may
decrease the risk. Several studies indicate that diets rich in specific long-chain,
polyunsaturated fatty acids of the omega-3 series, such as those found in fish,
can reduce the risk of AD.230 Recent studies have extended and confirmed data
showing the protective effect of omega-3 fatty acids against AD.231 Moreover,
there is epidemiological data suggesting an association between an inadequate
intake of fish oils and a greater than expected incidence of late onset dementia.232
Interestingly, experimental animal studies support these epidemiological data in
that there is evidence that a diet enriched with docosahexaenoic acid (DHA,
22:6n-3) reduces the burden of β-amyloid peptide in a mouse model of AD.233
Although the emerging data linking fatty acids to AD are encouraging, the
potential of dietary modifications of fat intake to affect disease risk remains to
be established. There is also emerging evidence suggesting that cognitively stim-
ulating environments, physical exercise and diets low in energy and fats (choles-
terol and saturated fatty acids) may reduce the risk of AD.193,228,234 Exercise,
cognitive stimulation and dietary restriction may each exert a beneficial effect
through a similar mechanism involving increased production of brain-derived
neurotrophic factor (BDNF).228,235,236 The possibility that the risk of AD can be
reduced by modifications of diet and lifestyle is of considerable interest and
suggests the potential for reducing the incidence of AD by preventive strategies
similar to those that reduce the risk of cardiovascular disease.
5.5 PARKINSON’S DISEASE, OXIDATIVE STRESS,

NFκB, AND ANTIOXIDANTS
Parkinson’s disease (PD) is a neurological syndrome manifested by any combi-
nation of tremor at rest, rigidity, bradykinesia and loss of postural reflexes. The
neuropathological hallmark of PD is the selective degeneration of dopamine (DA)
neurons in the nigrostriatal system.237,238 These neurons synthesize and release
DA, and the loss of dopaminergic influence on other structures in the basal ganglia
leads to the classic Parkinsonian symptoms. Moreover, PD is characterized by
degeneration of monoamine-containing neurons in the brain stem nuclei (pre-
dominantly the locus coeruleus) and is variably associated with pathology in
nonnigral systems causing multiple neurotransmitter dysfunctions.239
Parkinson’s disease develops much less frequently than Alzheimer’s disease,
ranging from 0.1 to 0.5% annually. Depending on the study, the annual incidence
for PD ranges from 0.1 to 0.3% for those over 50 years, to 0.4% for those over
80 years and as high as 2.6% for those 85 to 98 years of age.240–242 Incidence
rates for PD increase with age both in men and women, but the rate in men
exceeds that for women by two-fold.241 The average duration of PD is 9 years.
From birth, the lifetime risk of developing PD is about 2% for men and 1.3% for
women.243 Although idiopathic PD is usually sporadic, it is now well established
that there is a genetic component to the disease.244,245 Approximately 5 to 10%
of PD patients have a familial form of Parkinsonism with an autosomal-dominant
pattern of inheritance.245 Case control studies have typically indicated a 2- to 14-
fold increase in incidence in close relatives of PD patients,246 and although
concordance rates between identical twins are low for overt expression of the
disease, they are much higher when subclinical decline in striatal dopaminergic
dysfunction is measured by positron emission tomography (PET) imaging (53%
in monozygotic twins of PD patients, compared with 13% in dizygotic cases).247
A specific mutation in exon 4 of the gene encoding α-synuclein has been identified
as a causative factor of Parkinsonism in a family from southern Italy.248 Subse-
quently, this same mutation was found in other Greek and Brazilian families.246,249
Kruger et al.250 reported a second mutation in a German family. The parkin
gene mutation was first described in a Japanese family with autosomal recessive,
levodopa-responsive disease characterized by degeneration of the substantia nigra
and the absence of Lewy bodies.251 Over 20 different mutations have since been
identified252 and these mutations are now considered to be the most common
cause of familial PD. Nevertheless, in sporadic PD, environmental factors have
been emphasized.253 Epidemiological studies have correlated a number of poten-
tial factors as those that may increase the risk of developing PD.254 These include
exposure to well water, herbicides, industrial chemicals, wood pulp mills, farming
and living in a rural environment. A number of exogenous toxins have been
associated with the development of Parkinsonism, including trace metals, cyanide,
lacquer thinner, organic solvents, carbon monoxide, and carbon disulfide.245 There
has also been interest in the possible role of endogenous toxins, such as tetrahy-
droisoquinolines and β-carbolines. However, no specific toxin has been found in
the brain of PD patients. The most compelling evidence for an environmental
factor in PD relates to the toxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine
(MPTP). MPTP is a biproduct of the illicit manufacture of a synthetic meperidine
derivative. Some drug addicts who took MPTP developed a syndrome that strik-
ingly resembled PD, both clinically and pathologically.255,256 MPTP induces tox-
icity through its conversion in astrocytes to the pyridinium ion (MPP+) in a
reaction catalyzed by monoamine oxidase B (MAO-B).11 MPP+ is then taken up
by DA neurons and causes a mitochondrial complex I defect similar to that found

in PD.257 This observation supports the possibility that an environmental factor
might cause PD; however, no MPTP-like factor has been identified in PD patients
to date.
The principal cytoskeletal pathology of PD is the Lewy body, which in 85
to 100% of cases occurs in many monoaminergic and other subcortical nuclei,
spinal cord, sympathetic ganglia, and less frequently in cerebral cortex, myenteric
plexuses and adrenal medulla.237,258–261 Lewy bodies are abnormal intracytoplas-
mic neuronal inclusions that are considered to be a major anatomic hallmark of
PD, although they are seen in pigmented nuclei in various disorders and in aging
brains. In the majority of cases, the mechanisms involved in nigral degeneration
in PD are unknown, but evidence from studies of postmortem brain tissue suggests
the involvement of ROS and oxidative stress.11,239,262 Oxidative stress may arise
from the metabolism of DA with the production of potentially harmful free radical
species.262,263 This may be important as surviving neurons increase DA turnover
to compensate for diminishing synaptic transmission.
Circumstantial evidence exists that defects in mitochondrial energy metab-
olism may cause nigral neuronal degeneration in PD. Thus, MPTP produces
dopaminergic neuronal degeneration and Parkinsonian symptoms in humans and
nonhuman primates.264 1-methyl-4-phenylpyridinium (MPP+), produced by the
catabolism of MPTP by monoamine oxidase B (MAO-B) in glia, is selectively
taken up into dopaminergic neurons by the DA transporter. Within dopaminergic
neurons, MPP+ is concentrated by the electrochemical gradient into mitochondria.
MPP+ selectively inhibits NADH CoQ reductase (complex I) of the mitochondrial
electron transport chain and induces neuronal degeneration. Evidence exists that
similar mitochondrial dysfunction may occur in idiopathic PD; a defect in com-
plex I has been reported in the striatum of patients with PD.265–268 Similar defects
have been found in the platelets,269 but not skeletal muscle270 of patients with PD.
Reductions have been found in the substantia nigra, but not in other regions of
the brain, such as the globus pallidus or cerebral cortex.271 Therefore, the speci-
ficity of mitochondrial impairment may play a role in the degeneration of nigros-
triatal dopaminergic neurons. Interestingly, recent evidence indicates that expo-
sure to the complex I inhibitor, rotenone, can cause nigrostriatal dopaminergic
degeneration associated with Parkinsonian-like symptoms and accumulation of
protein aggregates containing ubiquitin and α-synuclein.272
Alterations in pro- and antioxidant molecules have been reported in postmor-
tem tissue from individuals with PD. Increased total iron has been found in the
substantia nigra in PD.263,273–275 Iron could increase oxidative stress by promoting
the formation of OH. from H2O2 via the Fenton reaction. Reductions in GSH
levels in the substantia nigra have also been reported.186,276–280 These reductions
were not detected in other neurodegenerative diseases in which nigral cell loss
occurs, suggesting they are specific to PD and not secondary to cell loss alone.
Decreases in GSH have also been found in the substantia nigra in individuals
with incidental Lewy bodies at postmortem, a potential marker of preclinical PD,
suggesting that alterations in GSH are an early event.281 Reductions in GSH levels
could promote and/or be a consequence of oxidative stress. Because GSH is

involved in the detoxification of H2O2, reductions in GSH could result from
increased concentrations of H2O2 and in the presence of metals, the highly reactive
OH.. The presence of lipid peroxidation and oxidative DNA damage further
supports the existence of oxidative stress in PD.262,282–284
As already mentioned, the hallmark of PD is a severe reduction of DA in all
components of the basal ganglia. DA and its metabolites are depleted in the
caudate nucleus, putamen, globus pallidus, nucleus accumbens, the ventral seg-
mental area, and the substantia nigra pars compacta and reticulata. Moderate
losses of DA are found in the lateral hypothalamus, medial olfactory region and
amygdaloid nucleus.285 In early Parkinsonism, there appears to be a compensatory
increase in DA receptors to accommodate the initial loss of DA neurons.286,287 As
the disease progresses, the number of DA receptors decreases, apparently due to
the concomitant degeneration of DA target sites on striated neurons. In the
remaining neurons in patients with PD, DA turnover seems greatly increased,
judging from the concentrations of homovannilic acid (HVA) in the nerve termi-
nals in the striatum and the cell bodies and dendrites in the substantia nigra,288
and the ROS production may very well increase as a consequence.
This hypothesis is strengthened by a study showing that the concentrations
of GSH decrease when DA turnover increases after reserpine treatment in rats,
indicating increased activity of the peroxide scavenging enzyme GSH-Px.289 If
the increase in ROS production (due to increased DA turnover) is not buffered
by the scavenging enzymes (SOD, catalase and GSH-Px), the compensatory
hyperactivity of the dopaminergic neurons may become self-destructive. Chronic
administration of L-DOPA would then only exacerbate the production of destruc-
tive ROS.290,291 The administration of L-DOPA itself has been postulated to
enhance the accumulation of ROS.292,293 Hiramatsu et al.294 using electron spin
resonance spectrometry have shown that 10 mM L-DOPA by itself was inactive,
whereas it produced ROS in the presence of 10 mM Fe-diethylenetriaminepan-
taacetic acid. This effect was blocked by deprenyl, an inhibitor of monoamine
oxidase B (MAO-B), which has been advocated as a symptomatic and protective
therapy in PD,295 as well as MPTP-induced Parkinsonism.296 Another index of
oxidative stress in PD might be the evidence of a robust increase of NFκB in the
nuclei of dopaminergic neurons in the substantia nigra of PD patients.62 This
clinical finding is consistent with in vitro data showing that oxidative stress
induced by C2-ceramide treatment causes nuclear translocation of NFκB in cul-
tured mesencephalic neurons.62 More recently, it has been shown that the neuro-
toxin, 6-OHDA, activates NFκB in PC12 cells by enhancing intracellular ROS
levels.297 Interestingly, in this experimental model, NFκB seems to sustain cell
survival by stimulating the expression of the antiapoptotic proteins bcl-2 and bfl-
1.297 Moreover, as already mentioned, the potent green tea polyphenol antioxidant
EGCG exerts a neuroprotective effect in a MPTP mouse model of PD.137
When induced by the toxins 6-OHDA or MPTP in animal models of PD,
nigral cell death seems to involve both necrotic and apoptotic processes. In human
PD, there has been some debate about whether key features of apoptosis could
be demonstrated, at least when based, alone, on morphological features or ter-

minal deoxynucleotidyl transferase-mediated dUTP-fluorescein nick end labeling
(TUNEL).298,299 However, the recent development of techniques involving double
labeling with TUNEL to demonstrate DNA fragmentation in conjunction with
cyanine dye that binds to DNA to provide additional structural details. Using this
technique, it has been demonstrated that chromatin condensation and DNA frag-
mentation within the same nuclei in the substantia nigra of PD patients is greater
than that seen in normal aging, consistent with the 10-fold higher rate of cell loss
seen in patients with the disease.245,300
The progressive nature of PD and the fact that neuronal degeneration in the
substantia nigra is slow and protracted301 present opportunities for therapeutic
intervention aimed at blocking or slowing down the degenerative process. Recent
neuroimaging and autopsy data indicate that there is a preclinical period of 4 to
5 years before symptoms appear. The rate of cell loss and decline of dopaminergic
function in the striatum is likely to be on the order of 10% per year, with the
disease progressing more rapidly during the early phases than the more advanced
stages of the disease.247,301 Both PET and SPECT (single-photon emission com-
puted tomography) imaging seem to be able to detect a decline in striatal dopa-
minergic function before clinical symptoms appear,247 which may make it possible
to begin neuroprotective intervention during the preclinical phase.
The largest neuroprotective trial conducted to date, the DATATOP (Deprenyl
and Tocopherol Antioxidant Therapy of Parkinsonism) study,302 involved two
putative antioxidant agents, vitamin E and deprenyl.303–305 Vitamin E had no
significant effect at the doses used, but deprenyl slowed the early progression of
symptoms and delayed the emergence of disability by an average of 9 months.
However, being an MAO-B inhibitor, this drug has symptomatic effects of its
own, which has confounded interpretation of the results.302 Interestingly, animal
studies have suggested that the neuroprotective effect is not dependent on MAO-
B inhibition per se, but rather on an antiapoptotic effect of the metabolite desm-
ethyl-deprenyl, possibly acting on protein translation.300,306 Before the completion
of the large DATATOP study (n = 800), an open trial with high dosages of α-
tocopherol and ascorbate, administered to a small group of early PD patients (n
= 15), found that this combination of natural antioxidants delayed by 2.5 years
the time necessary to begin the therapy with L-DOPA.307 There are many alter-
native antioxidative approaches that may be considered in future clinical trials,
including free-radical scavengers, GSH, GSH enhancing agents, ion chelators
and drugs that interfere with oxidative metabolism of DA. Interestingly, the classic
directly acting DA receptor agonists may belong to the last group; by stimulating
DA autoreceptors, these drugs reduce DA synthesis, turnover and release, so that
less L-DOPA is needed.308 In addition, some of these compounds have direct
antioxidant effects.309,310 More recently, the DA receptor agonist, pramipexole,
has been used as a monotherapy for the treatment of PD, and it has been shown
that it may have neuroprotective effects.311
5.6 AMYOTROPHIC LATERAL SCLEROSIS,

OXIDATIVE STRESS, NFκB, AND ANTIOXIDANTS
Amyotrophic lateral sclerosis (ALS) is a fatal paralytic neurodegenerative disor-
der of unknown cause, mainly characterized by a progressive loss of motor
neurons in the cerebral cortex, brainstem and spinal cord. ALS is a progressive
disease that invariably leads to death within approximately 3 to 5 years from the
onset of symptoms.312 The annual worldwide incidence for ALS ranges between
0.4 and 1.8 per 100,000 people and the prevalence is estimated at 4 to 6 per
100,000, with an overall male predominance.313 Although most cases are sporadic,
about 5 to 10% are familial, with inheritance following an autosomal dominant
pattern. About 15 to 20% of patients with familial ALS (FALS), which is clinically
indistinguishable from the more common sporadic ALS, carry mutations in the
gene encoding for the free radical scavenging enzyme SOD-1.314,315 Over 50
different SOD-1 mutations have been documented in FALS patients.316 Transgenic
mice have been generated that express mutant forms of SOD-1 found in FALS
cases, including gly93 → ala (G93A)317–319 and gly37 → arg,320 which develop
motor neuron disease and death within 4 to 6 months if the mutant enzyme is
expressed at sufficient levels. Studies of FALS patients with mutations of SOD-
1 indicate that SOD-1 activity is decreased 20 to 50%.315,321 This suggested
initially that the disease was due to ROS-induced damage resulting from a struc-
turally defective enzyme with reduced activity.315 However, no deletions of the
SOD-1 gene have been found in FALS families, implying that expression of the
mutant protein is required for pathogenesis. Studies in transgenic mice suggest
that, rather than causing a loss of function, the mutations of SOD-1 in FALS
patients cause a gain of function that results in neuronal degeneration.319,322
Because transgenic mice expressing wild-type human SOD-1 with comparable
elevation of brain SOD activity do not develop motor neuron disease319,320 and,
in fact, show enhanced resistance to oxidative stress,323,324 the disease is due to
expression of the mutant protein and not to elevation of SOD activity in the
brain.325–327
Several investigators have found increased levels of ROS in animal models
of ALS.328–330 Consistent with animal data, a number of clinical studies indicate
that oxidative stress may be involved in the pathology of ALS, as suggested by
increased levels of oxidative damage products, such as protein carbonyls, 4-HNE,
8-OHdG and nitrotyrosine.21,44,51,330–333 In addition, fibroblasts from ALS patients
were found to be more sensitive to oxidative stress.334 Moreover, immunohis-
tochemical studies have shown that NFκB is strongly activated in astrocytes of
the spinal cord of ALS patients, probably as a consequence of the oxidative
stress.64 Thus, the occurrence of oxidative stress and activation of NFκB is a
common characteristic of AD, PD and ALS. In this regard, it is noteworthy that
overlapping syndromes with clinical and pathological features of dementia, ALS
and PD have been described.335 It is also important to mention that degeneration
of midbrain DA neurons occurs in a mouse model of ALS.336
Various drugs, which can act by reducing oxidative stress, have been used as
potential therapeutic agents in transgenic mice expressing the mutated human
SOD-1 enzyme. Thus, polyamine- or putrescine-modified catalase, an antioxidant
enzyme that removes hydrogen peroxide and has good permeability at the
blood–brain barrier, increases the survival of transgenic mice bearing the human
mSOD-1G93A.337,338 Moreover, the copper chelator and thiol compound penicil-
lamine, the copper chelator trientine, carboxyfullerenes, vitamin E and N-acetyl-
cysteine have been reported to increase the survival time in this mouse model
and/or delay the onset of the disease to a small extent.339–342 The drug riluzole,
which inhibits glutamate release at presynaptic terminals, also extends lifespan
slightly in human mSOD-1G93A transgenic mice.342 Interestingly, riluzole, which
is used clinically in patients with ALS,343 has been shown to have direct antiox-
idative effect on cultured cortical neurons.344 However, no clear evidence for a
beneficial effect of α-tocopherol, selegiline, N-acetylcysteine or an antioxidant
cocktail has been obtained in humans.345–347
Li et al.348 have recently reported that blockade of caspase-1 and caspase-3
activity by N-benzyloxycarbonyl-Val-Asp-fluoromethylketone (zVAD-fmk) pro-
longs the survival of transgenic mice expressing the human mSOD-1G93A that
begin to develop ALS symptoms at the mean age of 3 months. These findings
open new perspectives for the use of caspase inhibitors as potential therapeutic
agents in the treatment of ALS and other neurodegenerative diseases. However,
because of the low oral bioavailability and limited brain penetrance, zVAD-fmk
was delivered by intracerebral administration. Thus, the physicochemical char-
acteristics of zVAD-fmk might limit its clinical usefulness. Based on these find-
ings and on the hypothesis that in transgenic mice expressing the human mSOD-
1G93A an increased formation of ROS occurs, we decided to treat them with
lyophilized red wine (which is rich in antioxidant compounds) dissolved in the
drinking water that was freely available to the animals. This treatment regimen
caused a significant reduction in the overall mortality of the treated mice as
compared with control animals. Thus, lyophilized wine prolonged by 6% the
survival of mSOD1G93A mice.349
In the first series of experiments, the onset of treatment was variable and
ranged from 43 to 66 days of age for the mice.349 We have recently repeated the
experiments on mSOD1G93A mice that were treated with the same concentration
of lyophilized red wine, but the treatment was started earlier, i.e., 30 to 40 days
from birth. By using this protocol, we have found that administration of lyo-
philized red wine significantly increased the mean survival time by 15%, as
compared with control transgenic mice given drinking water only. The calculated
concentration of polyphenolic compounds, expressed as gallic acid equivalent
(GAE), was 4824 mg/l. Considering that each mouse drank about 4 ml of liquid
daily, it is possible to calculate the daily intake of GAE, which was about 20 mg
per mouse. It is tempting to speculate that the mechanism of neuroprotection
exerted by lyophilized red wine on mSOD1G93A mice might be due to its ability
to inhibit caspase-3 activity. This hypothesis is based on in vitro experiments
showing that lyophilized red wine (5 µg/ml) caused a significant inhibition of
caspase-3 activity on primary cultures of rat cerebellar granule neurons.129 How-

ever, it is presently impossible to establish whether the effect of lyophilized red
wine on caspase-3 is direct or mediated by inhibition of ROS formation. Further-
more, ex vivo experiments aimed at investigating the inhibitory effect of lyo-
philized red wine on activated caspase-3 in mSOD-1G93A mice are necessary to
confirm our hypothesis. Unfortunately, treatment with CAPE, curcumin, trans-
resveratrol, quercetin, hydroxytirosol and EGCG did not cause any significant
effect on the survival of mSOD-1G93A mice either when given orally in the drinking
water or when administered daily intraperitoneally. These findings suggest that
treatment with single antioxidant compounds is not an efficient strategy in the
therapy of this devastating disease, and the use of a mixture of multiple com-
pounds might be preferable.
5.7 CONCLUSIONS
There is growing evidence that oxidative stress may play an important role in the
pathogenesis of AD, PD and ALS. However, in spite of the large body of exper-
imental data showing the protective effect of antioxidants in in vitro models of
neurodegeneration and in some in vivo animal models, there is still limited
evidence for a neuroprotective effect of antioxidants in the treatment of neuro-
degenerative disorders in humans. There may be several reasons for this discrep-
ancy between preclinical and clinical data. Many laboratory studies use models
of oxidative stress and investigate rescue by antioxidant agents. These models
normally use acute high doses of antioxidants that far exceed those usually
ingested via dietary sources.350 These types of studies are, therefore, unlikely to
be comparable to dietary exposure to antioxidants. Moreover, it is conceivable
that the therapeutic regimen used so far (e.g., one or two antioxidants) might not
be sufficient to halt the neuropathologic process. As pointed out by others, a more
efficient strategy would be the use of multiple antioxidants in the treatment of
AD, PD and ALS.146 In this regard, it is important to point out that one possible
advantage of the use of extracts of fruits, vegetables or beverages (such as red
wine, green tea or ginkgo biloba) in the treatment of neurodegenerative disorders,
is that they often contain multiple antioxidant compounds that can potentiate each
other. Consistent with this line of reasoning, it has recently been shown that a
complex antiaging dietary supplement composed of 31 ingredients, most of them
with antioxidant activity, is capable of blocking age-related cognitive decline in
transgenic mice expressing high levels of ROS-mediated processes.351 Particularly
important would be the use of lyophilized red wine,349 which is provided with
strong antioxidant capacity.73,74
One possible limitation of the neuroprotective strategy (including antioxidant
administration) might be consequent to the fact that when overt symptomatology
of AD, PD and ALS occurs, a certain amount of neuronal death has already
happened. Thus, the neuroprotective agents (including antioxidants) can, at best,
only rescue the surviving neurons, an effect which might not be sufficient to
attenuate the neurological symptomatology. Nevertheless, recent advances
suggest that the goal of curing patients with age-related neurodegenerative dis-
orders is worth pursuing. One reason for optimism is that the extent of neuronal
loss in AD and PD patients during the early period of the disease may not be as
great as initially thought because many dysfunctional neurons may be able to
recover.350 It is, therefore, important to start the therapeutic intervention at an
early stage of the disease process. In this regard, it is interesting to note that some
epidemiological studies have shown that dietary habits can influence the incidence
of neurodegenerative disorders. In particular, it was found that a diet rich in
vitamin E can reduce the risk for PD69,91 and that moderate wine consumption
may decrease the risk for AD.67,68 However, there are still few and controversial66,92
epidemiological data on this important point, which might be partly due to the
intrinsic difficulties in performing epidemiological surveys regarding the dietary
habits of large populations. Nevertheless, it is desirable that future studies aimed
at investigating the relationship between dietary antioxidant intake and the relative
risk for neurodegenerative disorders such as AD, PD and ALS will shed more
light on this very important aspect of public health.
5.8 ACKNOWLEDGMENTS
This work was supported by the Italian MIUR (Ministero Istruzione Università
Ricerca) L488/92 project n. s209-p/f. The author is very grateful to Dr. Andreina
Poggi for her critical reading of this manuscript.
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6 Nutrients and Herbals

in the Pharmacotherapy
of Unipolar/Major
Depression
Matthew Chronowic
CONTENTS
6.1 Introduction..............................................................................................179
6.2 Unipolar Depression................................................................................180
6.2.1 Amino Acids ...............................................................................182
6.2.2 Folate, Vitamin B12, and Homocysteine......................................186
6.2.3 S-adenosylmethionine..................................................................190
6.2.4 Zinc ..............................................................................................192
6.2.5 St. John’s Wort (Hypericum perforatum)....................................193
6.2.6 Ginkgo Biloba and Panax Ginseng.............................................194
6.2.7 Vitamin B6 ...................................................................................195
6.2.8 Tyramine ......................................................................................196
6.2.9 Omega-3 Fatty Acids...................................................................197
6.3 Conclusion ...............................................................................................198
References .........................................................................................................203
6.1 INTRODUCTION
Approximately 7% of the North American population is currently affected by
mood disorders.1 The bouts of depression and mania that characterize these
conditions cause a great deal of morbidity and increase the risk of mortality,
ranking these disorders among the top 10 causes of disability.1 Therefore, effective
treatment options are of the utmost importance to increase quality of life for these
individuals, as well as to increase the labor capacity of the work force. Originally
the domain of the psychiatrist, the pharmacotherapy of these disorders is now
often in the hands of the general physician. This requires considerable knowledge
179
of the disease processes and appropriate pharmacotherapy choices by healthcare

professionals.
Understanding the biological basis of these mood disorders has been a diffi-
cult process and a good deal of research continues in this area today. As our
understanding has grown, so too has the wide range of pharmaceuticals available
to treat these disorders. With such a broad spectrum of pharmaceuticals to choose
from, it is not only important to understand how the drugs may interact with one
another, but also to understand how nutrition and diet can affect pharmacotherapy.
Nutritional augmentation of a drug regimen could have the potential to decrease
the dose of medication required, subsequently lessening the adverse side effects
that are common to most pharmaceuticals. Conversely, nutrition also has the
capacity to affect the metabolism of drugs, resulting in a lesser or greater efficacy,
which would require compensatory dosing by the prescribing physician. There-
fore, it is the aim of this chapter to present some of the observed drug–nutrient
interactions that have been noted in the pharmacotherapy of unipolar depression.
6.2 UNIPOLAR DEPRESSION

The most prominent hypothesis of the etiology of unipolar depression, or major
depressive disorder, is the idea that a deficit of monoamine neurotransmission
underlies the disorder; specifically a lack of norepinephrine and serotonin (5-
hydroxytryptamine or 5-HT) neurotransmission. Despite being relatively few in
number (<1 in 200,000), monoaminergic neurons send axonal branches through-
out the brain and likely act through G-protein coupled receptors to alter overall
brain activity.1 Our current understanding of the disease does extend beyond this
simple model, but it is evident that pharmacologically elevating monoamines do
have the capacity to elevate mood in many individuals affected by this disorder.2
Furthermore, it is mainly in the context of this simple model that we understand
the mechanistic basis of the current pharmaceuticals used to treat depression.
The first class of drugs developed to treat unipolar depression was the
monoamine oxidase inhibitors (MAOIs), introduced in the mid 1950s. As the
name suggests, these drugs have the capacity to inhibit the enzyme, monamine
oxidase. This enzyme, found at high concentrations in neurons and liver cells,
degrades monoamine neurotransmitters. By inhibiting this process, the amount
of monoamine neurotransmitters is increased in the synaptic cleft, resulting in a
subsequent increase in monoaminergic neurotransmission. However, this class of
drugs can be quite dangerous if combined with substances that further increase
the amount of serotonin in the synaptic cleft, leading to a condition known as
the serotonin syndrome.2 This syndrome is characterized by altered mental status,
agitation, diaphoresis, hyperthermia, and hypertonicity, putting the individual in
grave danger.2 Clinically, this syndrome causes mental, autonomic, and neuro-
logical disorders that appear very suddenly, often less than 24 hours after the
offending substance is taken.3 Also, there are a number of foods that must be
avoided when taking MAOIs, which, if consumed, could result in hypertensive
crisis. This will be discussed in greater detail below. Due to the significant risks
Nutrients and Herbals 181
associated with these pharmaceuticals, they are often a last line of defense used
in treatment-resistant depression. The current MAOIs in use are phenelzine and
tranylcypromine.2
Also brought onto the market in the 1950s were the tricyclic antidepressants,
which are a major class of drugs used to treat depression. This class of drugs
works by nonselectively blocking the reuptake of monoamines. Reuptake is a
mechanism that prevents over-stimulation of the postsynaptic neuron by bringing
neurotransmitters back into the presynaptic neuron. By nonselectively blocking
monoamine reuptake, the concentrations of serotonin, norepinephrine, epineph-
rine, and dopamine are all increased in the synaptic cleft. Consequently, a number
of different monoamine receptors are activated both in the brain and many other
tissues where receptors are expressed. This lack of selectivity explains why the
tricyclic antidepressants are associated with a plethora of side effects: drowsiness,
dry mouth, urinary retention, constipation, blurred vision, low blood pressure,
weight gain, and cardiac effects.2 The current tricyclic antidepressants in use
include amitriptyline, clomipramine, desipramine, nortriptyline, and imipramine.2
A major development in unipolar depression pharmacotherapy was the intro-
duction of selective serotonin reuptake inhibitors (SSRIs) in the 1980s. This class
of drugs also works by inhibiting the reuptake of neurotransmitters; however, it
differs from tricyclic antidepressants in that it is selective only for serotonin. By
binding only to the serotonin reuptake transporter protein, there is an increase in
serotonin in the synaptic cleft, but the other monoamine concentrations remain
unchanged. This property of the SSRIs results in fewer side effects than the
tricyclic antidepressants, although several are still observed, including nausea,
nervousness, insomnia, sexual dysfunction, and headache.2 Given the increased
selectivity and decreased severity of side effects, SSRIs (fluoxetine, sertraline,
paroxetine, citalopram, and escitalopram) are usually the first-line therapy in
unipolar depression.2
Similar to the SSRIs, another class of drugs named the serotonin and nore-
pinephrine reuptake inhibitors (SNRIs), which block the reuptake of both sero-
tonin and norepinephrine, were developed for the treatment of unipolar depres-
sion. The SNRIs still have fewer side effects than tricyclic antidepressants and
are more efficacious in some individuals than the SSRIs. However, there is some
risk of hypertension as well as other side effects with the SNRIs (venlafaxine
and duloxetine), which requires the doctor’s consideration when prescribing.2
Finally, the last class of drugs used for the treatment of unipolar depression
is the atypical antidepressants. They fall outside the four aforementioned catego-
ries due to differences in both structure and function. Some operate by weakly
inhibiting reuptake of specific neurotransmitters, some by inhibiting neurotrans-
mitter receptors directly, whereas the mechanisms of others still remain to be
clarified. These mechanisms will be elaborated upon as needed in the following
sections. In line with the other classes of drugs, the atypical antidepressants,
mirtazapine, bupropion and trazodone, are also associated with a range of side
effects.2
There are a few problems with the monoamine theory of unipolar depression.
First of all, it cannot account for the fact that antidepressant drugs produce their
biochemical effects within minutes or hours, yet the onset of therapeutic benefit
usually takes weeks. Furthermore, the severity of the depressive state and the
amount of monoamine depletion do not necessarily correlate.4 Our knowledge of
intracellular signaling has drastically improved over the past decade, and accu-
mulating evidence seems to suggest that protein phosphorylation is part of the
long-term mechanism of antidepressants.4 Evidence has shown that various anti-
depressants can alter both the activity and translocation of second-messenger
regulated protein kinases; in particular, protein kinase C (PKC), cyclic AMP-
dependent protein kinase (PKA), and Ca2+/calmodulin-dependent protein kinase
II (CaMKII).4 Increasing activation of protein kinases allows for increased phos-
phorylation of certain subcellular components (of particular interest are micro-
tubule-associated protein 2 [MAP2] and synaptotagmin) that may consequently
affect cytoskeleton remodeling, a process involved in neurotransmitter release.4
This new theory of the mechanistic basis of antidepressant action continues to
develop at a rapid pace and shows great promise at aiding in the development of
more effective antidepressants as well as furthering our understanding of the
etiology of the disease.
Another theory that accounts for the temporal discrepancy between the bio-
chemical and clinical effects of antidepressants is the idea that these compounds
decrease N-methyl-D-aspartate (NMDA) receptor function. This is supported by
the observation that chronic (14 day) administration, but not acute (1 day), of 17
different antidepressants produces adaptive changes in the binding of radioligand
to NMDA receptors in a mouse model.5 This is an interesting finding as it
corresponds quite well with the time course required for the clinical benefit of
antidepressants.
It is in the context of these mechanisms that the interaction of nutritional and
dietary components with unipolar depression drugs will be examined.
6.2.1 AMINO ACIDS

With the monoamine hypothesis of unipolar depression in mind, the most logical
point to begin the discussion of nutrient–drug interactions is with amino acids.
The essential amino acid tryptophan is the dietary precursor to the monoamine—
serotonin. On the other hand, the monoamines norepinephrine, epinephrine, and
dopamine are synthesized from the amino acid tyrosine. The first inclination is
to suspect that more amino acid precursor could give rise to more monoamine
synthesis, and subsequently resolve the deficit in monoaminergic neurotransmis-
sion seen in unipolar depression. Furthermore, considering that most antidepres-
sants inhibit serotonin reuptake as part of their mechanism,6 one would think that
having more amino acid precursors available would provide a more suitable
environment for pharmacotherapy to work. Unfortunately, the relationship is not
so simple.
There have been many studies conducted to determine if baseline amino acid
status is correlated to pharmacotherapy success. In these studies, it is common
to use a ratio of tryptophan over the sum of all other large, neutral amino acids
(Trp/LNAA). The sum of plasma LNAA is typically obtained by adding valine,
isoleucine, leucine, tyrosine, and phenylalanine together. This Trp/LNAA ratio
is used because tryptophan competes with the other LNAA to get across the
blood–brain barrier (BBB). Hence, an excess of other LNAAs will hinder tryp-
tophan’s uptake into the brain, causing a subsequent decrease in serotonin syn-
thesis. A similar ratio is used for tyrosine (Tyr/LNAA) when the study is con-
cerned with noradrenergic neurotransmission, except that tryptophan replaces
tyrosine in the denominator.
The baseline Trp/LNAA ratio has been shown to be inversely proportional
to improvement on the tricyclic antidepressants, amitriptyline and clomipramine,
with a Trp/LNAA ratio below the group mean predicting a better treatment
response.7 In addition, the same study also showed that the plasma concentration
of tryptophan was inversely proportional to the clinical response to the SSRIs,
citalopram, and paroxetine.7 Another study using a mixture of SSRIs (fluvoxam-
ine, fluoxetine, and citalopram), along with SNRIs (amitriptyline and clomi-
pramine), and a tricyclic antidepressants (TCA) (nortriptyline), showed that the
Trp/LNAA ratio during the first week of therapy could be used to predict clinical
efficacy of the group as a whole.8 However, no such relationship was observed
when examining treatment of major depression with the MAOI, moclobemide.9
The Tyr/LNAA has also shown some utility in predicting the response to treatment
with the TCA, nortriptyline, with the baseline ratio similarly being inversely
correlated to subsequent clinical improvement on the medication.10 Unfortunately,
not all results are in agreement with the above trends. In one of the larger studies
conducted in this area (n = 147), baseline Trp/LNAA and Tyr/LNAA ratios were
not correlated with the 6-month treatment outcome of depressed subjects placed
on fluoxetine or nortriptyline.11 It was initially hoped that a clear pattern of plasma
amino acid status could suggest the best choice for pharmacotherapy. However,
the mixed nature of the results has made this exceedingly difficult.
It has been suggested that the ability of the 5-HT postsynaptic receptors to
adapt to serotonin availability is more important than the availability of precursor
amino acids.11 This could also mean that a reduction in receptor plasticity is a
marker of treatment resistance in depression.12 If this were the case, then the
baseline amino acid profile could be a more meaningful predictor of treatment
response in those individuals who have a greater ability to regulate their 5-HT
receptors. On the other hand, those individuals who lack such plasticity would
likely be those that do not respond well to increasing monoamine precursors via
diet, or by increasing monoamines directly using antidepressants.
An interesting observation is that subjects given the TCA antidepressant,
clomipramine, for 6 months had their plasma tryptophan concentration reduced
to 28% of its initial value over the treatment period, but this value was still only
68% of the initial value 3 months after successful pharmacotherapy was com-
pleted.13 This was interpreted as a rebound phenomenon due to the fact that
antidepressants commonly inhibit the liver tryptophan pyrrolase enzyme, which

is the first enzyme in the kynurenine pathway and, therefore, determines how
much tryptophan is degraded.13 Once pharmacotherapy ceases, this enzyme loses
its inhibition and resumes the degradation of tryptophan. Such a process could
possibly contribute to the relapse that is common in the treatment of unipolar
depression, and monitoring of plasma amino acid levels could become increas-
ingly important. More research is needed to look at the possibility of modulating
plasma tryptophan levels in order to deter relapse.
A commonly used experimental methodology in this field is to cause an acute
depletion of either tryptophan or tyrosine by giving an amino acid drink devoid
of either tryptophan or tyrosine and phenylalanine, respectively. A tryptophan-
free mixture will induce protein synthesis in the liver, but all of the required
tryptophan will be taken from the plasma causing a rapid depletion of the circu-
lating levels. The same is true when a drink devoid of tyrosine and phenylalanine
is used. The acute depletion of tyrosine has been shown to have little effect on
a small group of subjects who had responded successfully to pharmacotherapy,
the majority of whom had been on SSRIs.14 This suggested that the status of
dopaminergic neurotransmission was not of critical importance to SSRI pharma-
cotherapy.14 However, acute tryptophan depletion has been shown to cause relapse
in successful responders to the SSRI, fluoxetine, but to a much lesser extent in
responders to the TCA, desipramine.15 Not only does this provide the mechanism
of action of these drugs, but it also suggests that closely moderating plasma levels
of the serotonin precursor, tryptophan, may be more important to preventing
relapse in subjects that have responded successfully to SSRIs than to TCAs. More
studies are needed to confirm such an observation, but knowing that tryptophan
plasma levels are more crucial in responders to a certain type of medication could
suggest a novel method of preventing relapse. In addition, because subjects were
randomized to each treatment group in this study, it suggests that the response
to acute tryptophan depletion is more related to the type of antidepressant used,
and not to patient variables.15
In stark contrast to these results, a small study (n = 14) showed that currently
depressed patients receiving the SSNRI, venlafaxine, improved in clinical out-
come when they were subjected to acute tryptophan depletion.16 The authors
hypothesized that an initial decrease in serotonergic neurotransmission that typ-
ically occurs after administration of SSRIs may be prevented by acute tryptophan
depletion.16 This suggests that the stage of pharmacotherapy that an individual is
in might determine how they respond to acute tryptophan depletion.
There are a few different nutritional supplements that have been co-admin-
istered with conventional antidepressant medications in an effort to improve
monoaminergic neurotransmission in unipolar depression. These supplements
include L-tryptophan, 5-hydroxytryptophan (5-HTP), and α-lactalbumin. Tryp-
tophan is acted upon by the enzyme tryptophan hydroxylase, which converts it
to 5-HTP.6 The 5-HTP is subsequently acted upon by L-amino acid decarboxylase
to form serotonin.6 α-lactalbumin is a whey protein that is high in tryptophan,
which is not a characteristic found in many proteins. Supplementation focuses
on monoamine precursors because serotonin itself lacks the ability to cross the
blood–brain barrier, hence making supplementation ineffective.
One study has shown that co-administration of tryptophan with the SSRI,
fluoxetine, caused a significant decrease in clinical symptoms of unipolar depres-
sion in the first week of pharmacotherapy, but at no later points in time.17 This
is an important observation considering that side effects of antidepressants begin
immediately with clinical response beginning after approximately 2 weeks, which
makes the initial phase of treatment notoriously difficult.17 It should be noted that
the over-the-counter sale of L-tryptophan was banned in the U. S. by the U.S.
Food and Drug Administration (USFDA) in 1990 as a result of a number of deaths
related to a contaminated batch.6 This batch caused at least 38 deaths due to
eosinophilia myalgia syndrome.6 However, α-lactalbumin supplementation is still
available over-the-counter and may be a worthwhile alternative.
There is much interest in using the -lactalbumin protein to increase seroton-
ergic activity in the brain. One study showed that a 2-day experimental diet
supplemented with α-lactalbumin could decrease depressive feelings under stress
in subjects that were stress-vulnerable.18 This is likely due to the fact that, when
combined with a regular diet, α-lactalbumin has the capacity to increase the
Trp/LNAA ratio.19 It still remains to be determined if α-lactalbumin co-admin-
istered with antidepressants has a beneficial effect, but there is some evidence
for the utility of 5-HTP in this regard.
5-HTP may be more efficacious than L-tryptophan, or α-lactalbumin, as it
bypasses the tryptophan hydroxylase enzyme in serotonin synthesis, which is the
rate-limiting step, and, in addition, it cannot be shunted into niacin or protein
production.20 A review of the literature suggesting that 5-HTP is effective in
treating unipolar depression showed that only a few studies were of sufficient
quality to show “statistical superiority to placebo” and they were mostly aug-
mentation studies.6 In light of this observation, it is clear that more studies looking
at the augmentation of classical antidepressant pharmacotherapy with 5-HTP are
warranted. It has been suggested that the advent of the SSRIs in the 1980s caused
a loss of interest in 5-HTP as a treatment, but in light of the augmentation studies
perhaps it should be reconsidered as a treatment option.6
Both clinicians, as well as patients, need to be aware of the remote possibility
of serotonin syndrome in regards to co-administration of more than one com-
pound. The most commonly reported substances involved in causing this syn-
drome are the MAOIs in combination with L-tryptophan or fluoxetine, but there
has been a reported case of an over-the-counter cough medicine causing serotonin
syndrome in a man with vascular disease.21 However, there have been no reports
of serotonin syndrome with the use of 5-HTP as a monotherapy, or in augmen-
tation studies with antidepressants.6 Despite these promising results, patients
should probably only use 5-HTP under the supervision of their clinician.
Ultimately, the efficacy of L-tryptophan, 5-HTP, and α-lactalbumin to aug-
ment traditional pharmacotherapy in unipolar depression likely depends on a
number of factors. First, the type of drug being used may determine whether or
not modulating monoaminergic precursors has any beneficial effect. Second,
individual variation, such as 5-HT receptor plasticity and the relative abundance
of precursor amino acids already available, could also determine whether more
monoamine precursor would be of any clinical benefit. Furthermore, the degree
of inhibition of the liver enzyme tryptophan pyrrolase may very well determine
how much tryptophan is being catabolized and, hence, how much is needed in
the diet. Finally, the involvement of serotonergic deficits, noradrenergic deficits,
or both in unipolar depression etiology could ultimately determine the utility of
the amino acid precursors to monoamines. As our understanding of the biological
and molecular etiology of depression becomes clearer, it is likely that examination
of the above factors in individual patients could suggest which pharmacotherapy
may be the most effective, and whether augmentation with monoamine precursors
would have any therapeutic benefit.
6.2.2 FOLATE, VITAMIN B12, AND HOMOCYSTEINE

There is accumulating evidence that a deficiency in one-carbon metabolism may
be linked to unipolar depression, as well as the clinical response to pharmaco-
therapy. Vitamin B12 and folic acid (folate) are integral to maintaining appropriate
one-carbon metabolism and, therefore, may play a role in the prevention and
treatment of unipolar depression (see Chapters 2, 8, and 9 for detailed figures).
One-carbon metabolism centers on the methylation cycle, which involves
the production of S-adenosylmethionine (SAM) from the amino acid methion-
ine. SAM contains a reactive methyl group making it a crucial methyl donor
in the human body. The number of substrates that SAM is responsible for
methylating is extremely large and includes nucleic acids, proteins, phospho-
lipids, myelin, polysaccharides, choline, catecholamines, and numerous other
small molecules.22 With this list in mind, it is easy to see that a reduced capacity
for one-carbon metabolism could have a plethora of consequences. Once SAM
donates its methyl group it becomes S-adenosylhomocysteine (SAH), which is
reversibly hydrolysed to homocysteine. Homocysteine can then have three
possible fates: either directed towards the transulfuration pathway, converted
back to SAH, or recycled back to methionine via the enzyme methionine
synthase. Conversion of homocysteine back to methionine allows for the for-
mation of more SAM and the subsequent methylation of numerous substrates.
However, methionine synthase requires Vitamin B12 as a cofactor and meth-
yltetrahydrofolate, a metabolite of dietary folate, as a substrate. The production
of methyltetrahydrofolate requires the enzyme methyltetrahydofolate reductase
(MTHFR), giving it an indirect role in the recycling of homocysteine back to
methionine. One polymorphism of the MTHFR gene (677C > T) has been
extensively studied and appears to result in a 25% average increase of homocys-
teine levels, depending on folate status of the individual.22 The reason for this
is that this polymorphism codes for a thermolabile form of the MTHFR protein,
resulting in reduced enzyme activity and, hence, a decreased ability to form
methyltetrahydrofolate.22 The status of a number of the components of this
cycle may ultimately have an influence on the etiology of unipolar depression

as well as the capacity for pharmacotherapy to work.
In an ethnically diverse sample of the U.S. population of approximately 3000
individuals, those who met the criteria for a lifetime diagnosis of unipolar depres-
sion had lower concentrations of folate in serum and red blood cell samples than
subjects who had never been depressed.23 Furthermore, numerous studies have
examined baseline levels of compounds involved in the methylation cycle to see
if they correlate with the subsequent response to pharmacotherapy. The three
main compounds of interest, vitamin B12, folate, and homocysteine, are those
which could potentially be modulated in order to better accommodate treatment
of unipolar depression. One study showed that low folate levels prior to 8 weeks
of treatment with fluoxetine predicted a poor treatment outcome in outpatients
with unipolar depression, but homocysteine and vitamin B12 levels were unre-
lated.24 In 2002, a literature review was conducted to examine the relationship
between homocysteine and neuropsychiatric disorders and, based on the five
studies that were included examining unipolar depression, the authors concluded
that a significant proportion of depressed patients had either elevated total
homocysteine levels or low levels of folate or vitamin B12.25 Unfortunately, no
consistent relationship could be found for any one of the three compounds.
Furthermore, it was concluded that there is preliminary evidence that both folate
and homocysteine levels have some capacity to predict antidepressant treatment
efficacy.25
The following year a prospective study showed that 6-month treatment out-
come in subjects with unipolar depression being treated with a wide variety of
antidepressants was associated with baseline vitamin B12 status, but only weakly,
and likely not independently from baseline folate status.26 Higher levels of vitamin
B12 were associated with a better treatment outcome in this study, but the wide
variety of medications in the study limits the specificity of the results.26 Further-
more, an archival study of inpatients in a geriatric psychiatric unit suggested that
elevated homocysteine and low folate, but not vitamin B12, were associated with
radiological markers of neuropathology.27 Of particular interest was the fact that
none of the patients in the study were clinically deficient in folate.27 This could
suggest that a reevaluation of what defines a functionally significant folate defi-
ciency may be in order, at least in regards to certain organs and particular health-
status groups.27 Overall, it is difficult to be certain that baseline measures of any
one compound associated with one-carbon metabolism can predict treatment
response in major depression, but it is possible that interplay between folate,
homocysteine, and vitamin B12 could predict response.
The status of one-carbon metabolism may also have the ability to predict
treatment resistance and possibility of relapse in subjects with unipolar depres-
sion. One study showed that low folate status predicted further treatment resis-
tance in subjects with unipolar depression who did not respond to initial treatment
with fluoxetine.28 These subjects, who participated in a 4-week, double-blind
augmentation trial with either a fluoxetine dose increase, fluoxetine augmented
with lithium, or fluoxetine augmented with desipramine, showed a strong
relationship between low baseline folate status and further treatment resistance.28
However, elevated homocysteine levels and low vitamin B12 levels were not
associated.28 Furthermore, this group also showed that low baseline serum folate
levels, but not elevated homocysteine or low vitamin B12 levels, were associated
with increased risk of relapse during the continuation phase of fluoxetine phar-
macotherapy.29 This study examined 71 patients, who had previously remitted
from unipolar depression for at least 3 weeks while undergoing fluoxetine treat-
ment and were followed for a subsequent 28 weeks to be monitored for depressive
relapse.29 These studies suggest that low folate status might have the ability to
predict treatment resistance as well as possibility of relapse, in addition to its
possible ability to predict treatment outcome, as noted above. If this is substan-
tiated by further research, folate status could prove to be a valuable clinical
indicator.
There are a few points of detail that should be considered while contemplating
the above relationships. The first of which is the possibility that macrocytic
anemia could be a predictor of poor treatment response in unipolar depression,
as it is often caused by either a folate or vitamin B12 deficiency. However, one
study found that neither macrocytosis nor anemia could predict antidepressant
refractoriness, in addition to not being able to predict low serum folate or vitamin
B12.30 Based on these results, it does not appear that macrocytosis or anemia is
useful in predicting treatment response. Another important finding is that the
relationship between low folate levels and an increased incidence of unipolar
depression may not exist in all populations. For instance, it was shown that 117
newly admitted Chinese inpatients with unipolar depression had normal levels of
folate, and that folate levels could not predict outcome on standard assessments
of depression.31 However, both the subjects and controls were found to have a
high intake of green vegetables.31 Therefore, the utility of serum folate levels to
predict incidence of depression might be limited in populations with a high intake
of green vegetables. Whether or not this relationship extends the likelihood of a
favorable response to pharmacotherapy remains to be determined.
A few studies have been undertaken to investigate the use of supplemental
folate in augmenting the traditional pharmacotherapy of unipolar depression. It
was shown that the addition of 15 mg daily folate to the standard treatments of
123 subjects, having either major depression or schizophrenia, with borderline
or deficient folate status, resulted in significant clinical and social improvements
in both groups over the course of a 6-month period.32 Later, a study was conducted
to see if subjects with unipolar depression, randomly assigned to treatment with
20 mg of fluoxetine plus 500 µg of folic acid or a similar looking placebo,
responded differently.33 Interestingly, only the female subjects receiving folate
showed a significantly greater improvement compared to placebo, as well as a
significant decrease in plasma homocysteine levels.33 It was suggested that the
dose of folic acid needed to enhance pharmacotherapy in males could be higher
than what was given in the study and should be sufficient to decrease plasma
levels of homocysteine.33 It is clear that more research needs be done in order to
determine the optimum doses for different subjects.
Folinic acid, which is metabolized to methylfolate in the body, may have

some capacity to improve treatment in SSRI-refractory individuals with unipolar
depression. In a study of 22 subjects with major depression, who had either partial
response or were nonresponders to an SSRI, the addition of folinic acid at 15 to
30 mg daily was modestly effective at improving response to SSRI treatment.34
Of those who completed the trial, 31% of subjects achieved a 50% or greater
reduction in the Hamilton Depression Rating Scale, and 19% of subjects achieved
clinical remission.34 This study was not placebo-controlled, but certainly shows
the need for more research investigating the use of folate to augment traditional
antidepressant treatment in subjects refractory to SSRI treatment.
A systemic review of the literature investigating the role of folate for the
treatment of unipolar depression found only two randomized, controlled studies
involving a combined total of 151 people that assessed the use of folate to augment
traditional pharmacotherapy.35 However, the combined data from these two stud-
ies showed that the addition of folate to treatment reduced Hamilton Depression
Rating Scores on average by 2.65 points, and there was no evidence of problems
with tolerability.35 These are promising results, yet more studies are needed to
confirm them to determine appropriate dosing for different individuals and to
elucidate what baseline characteristics influence the required dose.
There have been a number of proposed mechanisms suggesting how one-
carbon metabolism may influence brain function, and these mechanisms could
help explain how folate augmentation improves the efficacy of traditional phar-
macotherapy. These mechanisms are reviewed by Coppen and Bolander-
Gouaille.22 The first possible explanation is that a decreased recycling of
homocysteine back to SAM decreases the body’s methylation capacity, which
could impair production of neurotransmitters and membrane phospholipids. Both
of these effects could alter the molecular environment in which antidepressants
work. The second possible explanation is that a disturbed one-carbon metabolism
will result in reduced levels of tetrahydrobiopterin (BH4), a compound that is
dependent on folate for its turnover. BH4 is a cofactor for the enzymes tryptophan
hydroxylase and tyrosine hydroxlyase, which represent the rate-limiting steps in
serotonin, and dopamine/norepinephrine synthesis, respectively. Therefore, a
reduction in BH4 levels could also mean a reduced capacity for synthesis of
neurotransmitters. Finally, it is also possible that reduced levels of BH4, a potent
antioxidant, and increased levels of homocysteine, a molecule that could cause
vascular injury via several oxidative mechanisms, could together result in cellular
injury leading to cerebral dysfunction. At this point it is not clear which of these
mechanisms play a role in the pathogenesis of depression, or in the subsequent
success of pharmacotherapy. However, it could very well be possible that more
than one of these mechanisms is involved.
Currently, it appears that disturbed one-carbon metabolism plays a role in the
pathogenesis and treatment efficacy of depression. However, to what extent it
is involved and the exact mechanism of this interaction still remains to be eluci-
dated. More research is needed to determine the appropriate dosing for augmen-
tation of traditional pharmacotherapy as well as to determine what baseline
characteristics influence this relationship. Furthermore, the individual roles of

folate, vitamin B12, and homocysteine in the disease process and treatment poten-
tial need to be further investigated. Currently, it appears that modulating folate
levels has the greatest capacity to help prevent and treat unipolar depression, but
it may be possible that the modulation of all three compounds could be the most
beneficial. Nevertheless, the modulation of one-carbon metabolism remains a
very promising avenue of research for unipolar depression.
6.2.3 S-ADENOSYLMETHIONINE
The use of S-adenosylmethionine (SAM) in the treatment of unipolar depression
makes perfect sense considering the previous section discussing one-carbon metab-
olism and its possible relationship to this disease. It functions as a cofactor in the
rate-limiting steps of the tryptophan hydroxylase and tyrosine hydroxylase reactions,
which are responsible for the production of serotonin and dopamine/norepinephrine,
respectively.36 Theoretically, by providing exogenous SAM, one should be able to
improve the capacity for methylation reactions that are necessary for neurotransmitter
synthesis, along with a myriad other reactions. However, the function of SAM might
extend beyond its role in methylation reactions.
As a monotherapy, SAM has shown some clinical utility. One open, multicenter
study showed that in 145 patients with unipolar depression, the parenteral adminis-
tration of 400 mg/day SAM for 15 days caused scores of depression to decrease
after both 7 days and 15 days.37 This is an important finding as most traditional
antidepressants have a 2-week lag period, during which there is no clinical benefit.
This rapid onset of action of SAM treatment is a common finding, with improvement
seen anywhere from 2 days to 2 weeks after treatment begins.36 A number of studies
have compared the efficacy of SAM monotherapy to traditional antidepressants. One
small study comparing intravenous administration of SAM at 400 mg/day with oral
administration of the tricyclic antidepressant, imipramine, found that SAM produced
superior results at the end of the first week and, by the end of the study, 66% of the
patients in the SAM group and only 22% of the patients in the imipramine group
had a clinically significant improvement.38 Later, two multicenter studies, each with
approximately 300 subjects with unipolar depression, were conducted simulta-
neously to compare 1600 mg/day SAM given orally and 400 mg/day given intra-
muscularly, both to 150 mg imipramine/day given orally.39 The clinical responses
for the SAM and imipramine arms did not differ, but significantly fewer side effects
were observed in subjects treated with SAM.39 Overall, in six of the eight studies
that have compared SAM with tricyclic antidepressants, SAM was of equivalent
efficacy, and in one study it was superior to imipramine.36 These are promising results
for SAM as a monotherapy, considering that it seems to be well tolerated with few
side effects and has an onset of action that is faster than traditional antidepressants.
In addition to these clinical trials, electroencephalogram/event-related potential map-
ping identified SAM as an antidepressant when given intravenously at 800 mg/day.40
Unfortunately, there are far fewer studies that have looked at augmenting
traditional pharmacotherapy of unipolar depression with SAM. It has been shown
that combining orally administered imipramine with 200 mg/day of intramuscu-

larly administered SAM caused an earlier decrease in depressive symptoms than
subjects who received a placebo along with imipramine.41 When the research
surrounding the use of SAM in the treatment of unipolar depression was reviewed
in 2002, there were only two augmentation studies included and they both showed
that combining SAM with tricyclic antidepressants resulted in an earlier onset of
action than tricyclic antidepressants alone.36 These are promising results, but they
certainly emphasize the need for more placebo-controlled studies investigating
the use of SAM to augment traditional pharmacotherapies. Furthermore, accurate
dosing information, as well as the use of SAM to augment other types of anti-
depressants, such as the SSRIs, still needs to be investigated.
There are a few theories proposed to explain SAM’s ability to lessen clinical
signs of unipolar depression sooner than traditional antidepressants. The first is
that SAM can increase synapsin I, which regulates the number of vesicles that
are available to be exocytosed from the presynaptic neuronal terminal.42 This is
supported by the fact that administration of 100 mg/kg/day of SAM for 12 days
in Sprague-Dawley rats induced changes in calcium/calmodulin-dependent pro-
tein kinase II (CaMKII) activity and increased synapsin I levels in the hippocam-
pus and frontal cortex nerve terminals.42 Synapsin I is a substrate for CaMKII,
so both of these results could be interpreted to mean that SAM has the ability to
modulate neurotransmitter release. This could account for its rapid mode of action
that is commonly observed in human trials. Furthermore, again in the rat model,
SAM treatment prevented the 5-HT1A receptor upregulation that normally accom-
panies acute administration of imipramine.43 This could also help explain the
quick response time seen in patients treated with SAM. More research needs to
be conducted to clarify SAM’s mode of action, but it is likely that it has the
capacity to modify intraneuronal signaling mechanisms.
Overall, it appears that SAM may have some capacity to treat unipolar
depression as well as augment traditional pharmacotherapy; however, more clin-
ical studies are needed to confirm this. The current body of evidence suffers from
the fact that a wide range of doses (200 to 1600 mg/day) and a variety of methods
of administration (oral, intramuscular, and intravenous) have been used.36 Fur-
thermore, having an additional diagnosis of bipolar disorder might contraindicate
the use of SAM, as it has been shown to increase anxiety, mania, and hypomania
in subjects with bipolar depression.36 SAM has also been criticized for the lack
of evidence investigating its long-term side effects and toxicity,44 as well as the
lack of information describing drug interactions. All of these areas require further
study if SAM is to be commonly used for unipolar depression. Furthermore, there
is concern that over-the-counter SAM supplements may have varying amounts
of active compound in them, as it is very unstable at room temperature when
exposed to air.44 Nevertheless, SAM remains a potential treatment strategy for
unipolar depression and augmentation of traditional pharmacotherapy, but needs
further investigation into appropriate dosing, long-term toxicity, drug interactions,
and stability when stored.
6.2.4 ZINC
Some studies have shown that plasma zinc levels in depressed subjects are
significantly lower than levels in healthy controls,45 yet this relationship was not
statistically significant in other studies.46 However, regardless of baseline zinc
levels, both these studies showed that plasma zinc levels significantly increased
after successful treatment with antidepressants.45,46 Furthermore, in a rat model,
treatment with citalopram and imipramine as well as electroconvulsive shock
therapy, all elevated hippocampal zinc concentrations.47 These studies suggest
that zinc may have a role in the pharmacotherapy of unipolar depression.
A number of studies have investigated the use of zinc supplementation to
augment traditional antidepressant efficacy. One study showed that zinc (ZnSO4)
at a dose of 30 mg/kg and/or imipramine at 30 mg/kg reduced the immobility
time in the forced swim test, also known as Porsolt’s test, in both mice and rats.48
Furthermore, a combination of zinc and imipramine at ineffective doses (1 and
5 mg/kg, respectively) was clinically effective in rats.48 Research has suggested
that the state of immobility in the forced swim test reflects lowered mood in rats
and is improved with antidepressant treatment as well as electroconvulsive shock
therapy.49 Therefore, the aforementioned study may suggest that augmentation of
traditional pharmacotherapy with zinc may have the capacity to improve treatment
of unipolar depression. This notion was further supported by the finding that low,
otherwise ineffective doses of imipramine and citalopram, administered together
with low, ineffective doses of zinc were effective in decreasing immobility times
in mice subjected to the forced swim test.50 In addition to these findings, it has
been shown that the use of zinc hydroaspartate alone has antidepressant properties
in both the forced swim test as well as the olfactory bulbectomy animal models
of depression.51 Furthermore, there was a concomitant rise in serum levels of zinc
in association with the antidepressant-like effects, and no tolerance was developed
following chronic treatment.51 These results all support a role for zinc status in
the treatment of unipolar depression.
In addition to the research conducted in animal models of depression, there
is some preliminary evidence showing zinc augmentation of traditional pharma-
cotherapy in humans. One small (n = 14), placebo-controlled, double-blind study
showed that zinc supplementation was able to augment the treatment of unipolar
depression with tricyclic antidepressants as well as SSRIs.52 Subjects in the zinc
supplementation group showed a significant decrease in both Hamilton Depres-
sion Rating Scale and Beck Depression Inventory measures of clinical symptoms
after 6 and 12 weeks of supplementation, as compared with placebo supplemen-
tation.52 These preliminary findings are encouraging and emphasize the need for
additional clinical intervention trials of zinc and conventional antidepressants.
The mechanistic basis of zinc’s action in the treatment of unipolar depression
is an area of great interest. One line of evidence suggests that the chronic
administration of antidepressants ultimately results in a region-specific dampen-
ing of N-methyl-D-aspartate (NMDA) receptor function, and conversely, the
administration of compounds that reduce transmission at NMDA receptors act as
antidepressants.53 In accordance with this theory, it has been shown that zinc has
the capacity to modulate binding of NMDA receptor agonists and antagonists to
NMDA receptors in the forced swim test, resulting in antidepressant-like effects.54
Furthermore, it has been shown that there is a significant decrease in the potency
of zinc to inhibit binding at NMDA receptors in the hippocampal tissue of suicide
victims; a group with a high likelihood of suffering from depression.55 This
interaction of zinc with NMDA receptors requires further research, as animal
models of depression are somewhat controversial, and not all suicide subjects
had a diagnosis of depression.55
It has been suggested that it is possible that zinc may reduce NMDA receptor
function via single or multiple mechanisms, but experiments to reveal the exact
nature of zinc’s influence on the NMDA receptor are currently being conducted.56
The observed actions of zinc include a direct antagonism of the NMDA receptor,
the ability to act on the AMPA receptor, and also to inhibit group I metabotropic
glutamate receptor function, all of which may ultimately affect NMDA receptor
function (reviewed in Nowak56).
In addition to the work on NMDA receptors, it has been shown that zinc has
the ability to potentiate the action of acetylcholine on the nicotinic acetylcholine
receptors (nAChRs), and fluoxetine has the ability to inhibit acetylcholine’s
action.57 Furthermore, when the two substances are combined, fluoxetine is able
to greatly reduce or even abolish the potentiating action of zinc.57 How this action
of zinc is involved in the etiology and treatment of depression, and whether it is
related to the NMDA mechanism in some way, still remains to be elucidated.
Another interesting finding is that zinc was able to enhance 5-HT uptake in
certain areas of adult rat brain, and that zinc reversed the inhibition of 5-HT
uptake exerted by fluoxetine, imipramine, and 6-nitroquipazine.58 This recent
finding suggests that zinc may very well play a role in modulating serotonergic
neurotransmission. Whether this is related to its ability to inhibit NMDA receptor
function, or interact with nAChRs, requires additional study.
Overall, it appears as though zinc has the capacity to augment pharmacother-
apy in animal models of depression as well as in a preliminary trial in human
subjects. Although the mechanistic basis of this still requires much investigation,
zinc has shown some ability to modulate NMDA receptor activity, nAChR activ-
ity, and influences 5-HT uptake. It is possible one or more of these mechanisms
may explain zinc’s capacity to augment antidepressant pharmacotherapy, but
much more work is needed to clarify these relationships.
6.2.5 ST. JOHN’S WORT (HYPERICUM PERFORATUM)
While it does not promote growth and, as such, cannot be classified as a nutrient,
St. John’s wort is included in this chapter as it is commonly taken by individuals
suffering from unipolar depression, either as a monotherapy or in combination
with other antidepressant compounds. However, individuals taking this extract
should be aware of the spectrum of drugs with which it can interact. This chapter
will discuss only the interactions related to antidepressants, but for a more
complete list of drug interactions associated with St. John’s wort, see Zhou.59
Evidence of the efficacy of St. John’s wort in the treatment of major depression
is inconsistent, as reported by a recent meta-analysis.60 Nevertheless, it is still
extremely popular for the treatment of depression, possibly because of the mis-
informed and commonly held notion that all “natural” compounds are safe and
superior to pharmaceuticals.
A review of the literature suggests that co-administration of St. John’s wort
with the tricyclic antidepressant, amitriptyline, results in decreased plasma levels
of the drug as well as two of its metabolites.59 Therefore, co-supplementation of
St. John’s wort with amitriptyline can have drastic effects on the plasma concen-
tration of the drug, reducing its capacity for effective treatment. Cytochrome P450
3A4 (CYP 3A4), which is found in the liver and intestine, plays a role in two
different steps of amitriptyline metabolism.61,62 A literature review showed that
St. John’s wort had the capacity to induce both hepatic and intestinal CYP 3A4
expression,59 which likely results in increased clearance of amitriptyline. Obvi-
ously, healthcare practitioners need to be aware of this important interaction when
treating subjects with unipolar depression.
In addition to its capacity to increase clearance of the tricyclic antidepressant
amitriptyline, St. John’s wort may also contribute to induction of the serotonin
syndrome when taken along with an SSRI. A number of case reports have linked
co-consumption of St. John’s wort and an SSRI with symptoms typical of the
serotonin syndrome.63-65 A number of biologically active components of St. John’s
wort, including hyperforin, adhyperforin, and procyanidins, have the capacity to
inhibit synaptosomal uptake of neurotransmitters.66 St. John’s wort can be con-
sidered a nonselective reuptake inhibitor as it has the capacity to inhibit reuptake
of serotonin, dopamine, and norepinphrine, as well as GABA and glutamate. In
the case of the latter two substances, these are not typically affected by other
antidepressants.67 Therefore, the combination of SSRI treatment, which also
inhibits the reuptake of serotonin, with St. John’s wort could have the capacity
to cause an accumulation of excess serotonin in the synaptic cleft, resulting in
serotonin syndrome. Due to our limited knowledge of the safety of combining
SSRIs with St. John’s wort, it has been suggested that combining these two
substances should be avoided.59
It is crucial that healthcare practitioners, as well as patients being treated with
antidepressants, be aware of the interactions between amitriptyline, SSRIs, and
St. John’s wort. Combining these substances can not only have negative effects
on the outcome of pharmacotherapy, but could also be life threatening. Further-
more, St. John’s wort interacts with a multitude of other drugs, some of which
unipolar depressed patients may be taking for other health problems. Clearly its
use must be carefully monitored by this population.59
6.2.6 GINKGO BILOBA AND PANAX GINSENG

The use of herbal supplements and alternative medicines for various reasons is
becoming increasingly popular. Therefore, it is worthwhile to note a few possible
interactions between certain herbal supplements and particular traditional anti-

depressant drugs.
There has been one case report that described an elderly woman with Alzheimer’s
disease who went into a coma while taking a low dose of trazodone, an atypical
antidepressant, and Ginkgo biloba.68 The underlying mechanism behind this remains
unclear, but it may be related to the ability of Ginkgo biloba to enhance gamma-
aminobutyric acid (GABA)-related neuronal activity in the brain, and/or its capacity
to increase CYP 3A4 activity, which results in increased formation of the active
metabolite of trazodone.69 This is supported by the fact that bilobalide, a constituent
of Ginkgo biloba, has been shown to increase GABA levels, possibly by its ability
to increase levels of glutamic acid decarboxylase in mouse brain.70 Furthermore, it
is supported by the observation that administration of Ginkgo biloba extract to rats
has the ability to enhance the expression of certain cytochrome P450 enzymes.71
More research is needed to investigate the interaction between Ginkgo biloba and
trazodone in order to determine if such a combination is capable of inducing coma
and, if so, what mechanism underlies this.
A review of the literature in this area also found several case reports describing
a possible interaction between Panax ginseng and the MAOI, phenelzine, resulting
in mania.69 The mechanism underlying this potential interaction is not currently
understood, but it has been speculated that it may relate to the psychoactive effects
of ginseng.69 Such psychoactive effects include the ability of a ginseng saponin
to block human nicotinic acetylcholine receptors (nAChRs)72 as well as the ability
of a different saponin to inhibit NMDA receptors in cultured rat hippocampal
neurons.73 A better understanding of Panax ginseng’s psychotropic effects could
help to explain the potential for this herbal product to induce mania when taken
simultaneously with phenelzine.
Due to the increasing popularity of herbal supplements and their likely con-
comitant use with various drugs, it is of the utmost importance that healthcare
practitioners report all potential interactions with medications. As we improve
our understanding of the effects that these herbal compounds have on the central
nervous system and drug-metabolizing enzymes, it will become more clear as to
how their concomitant use with antidepressant medications could result in
unwanted complications.
6.2.7 VITAMIN B6
There is a fairly well-established relationship between treatment with the MAOI,
phenelzine and vitamin B6 deficiency. Over the years, there have been a number
of reports describing vitamin B6 deficiency associated with phenelzine use.74–76
One open treatment study detected no effects of phenelzine on vitamin B6 levels,
but these results may have been confounded by concomitant use of vitamin
supplements by the subjects.77
Despite finding an average 54% reduction of vitamin B6 levels in subjects
treated with phenelzine as compared to a control group, one study did not find
a correlation between the pyridoxal phosphate (active form of vitamin B6 in the
body) level and daily dose.76 This indicates that there could be other factors
involved, yet still suggests that phenelzine use can induce a vitamin B6 deficiency.
However, administration of a pyridoxine supplement along with phenelzine has
alleviated the symptoms of vitamin B6 deficiency in patients.74,75
The suspected reason for the vitamin B6 deficiency associated with phenelzine
use, as discussed in Malcolm et al.,76 is the nonenzymatic formation of a pyri-
doxalhydrazone complex involving the two compounds. This could result in a
rapid decrease in plasma levels of pyridoxal phosphate after taking phenelzine.
As is the case for the other supplements listed above, healthcare practitioners as
well as patients need to be aware of this relationship in order to avoid deficiencies
of vitamin B6 while taking phenelzine.
6.2.8 TYRAMINE
Tyramine is a monoamine derived from the amino acid tyrosine and can be found
in many foods. The enzyme, monoamine oxidase, is responsible for the oxidative
deamination of tyramine and, hence, its metabolism. It has been well established
that individuals being treated with a MAOI can have a hypertensive crisis if they
consume large amounts of tyramine. When an individual is taking a MAOI drug,
MAO-A type in the gut wall and liver is inhibited, allowing tyramine from the diet
to enter the circulation.78 Tyramine has vasopressor activity of its own, but also
promotes the release norepinephrine from nerve terminals, both of which contribute
to an increase in blood pressure.78 The hyperadrenergic state induced by consumption
of large quantities of tyramine while taking a MAOI is commonly referred to as “the
cheese reaction” and can result in a number of symptoms. This condition can take
three different forms as reviewed by Brown et al.78 The most common is characterized
by a sudden, severe headache, along with pallor, chills, and neck stiffness; the second
form relates almost exclusively to the cardiovascular system and is characterized by
sudden palpitations, hypertension, chest pain, apprehension, headache, pallor, and
collapse; the third and most devastating manifestation of the condition is character-
ized by intracerebral hemorrhage and death.78
A diet has been devised in order to assure that individuals taking MAOI drugs
do not consume large amounts of tyramine. This diet has gone through extensive
refining in order to be as unrestrictive as possible, helping to ensure compliance
with medications. Foods that are to be avoided on this diet, due to their high
levels of tyramine, include aged cheese, aged or cured meats, any potentially
spoiled meat/poultry/fish, broad (fava) bean pods, marmite (concentrated yeast
extract), sauerkraut, soy sauce and soy bean condiments, and tap beer.79 However,
pizza bought from large-chain commercial outlets appears to be safe, even with
double-cheese and double-pepperoni.80 Nevertheless, it has been suggested that
caution be exercised if ordering from smaller pizza outlets or if purchasing a
gourmet pizza that could contain aged cheeses.80 By receiving accurate dietary
advice about the MAOI diet, patients taking this class of drug should be able to
prevent potentially lethal hypertensive crises.
6.2.9 OMEGA-3 FATTY ACIDS

In recent years, much attention has been given to the role that polyunsaturated
fatty acids (PUFAs) may play in mood disorders. This stems from the observation
that some epidemiology has shown that frequent fish consumption, which is
indicative of a high intake of the omega-3 fatty acids eicosapentaenoic acid (EPA)
and docosahexaenoic acid (DHA), is associated with a lower incidence of depres-
sive symptoms in the general population.81 Furthermore, some studies have shown
that subjects with unipolar depression have lower serum levels of omega-3 fatty
acids and a compensatory increase in omega-6 fatty acids, when compared to
nondepressed controls.82 Such observations have stimulated investigations into
the potential roles that PUFAs may play in disease etiology, therapy, and antide-
pressant augmentation.
A number of studies have investigated the efficacy of adding omega-3 PUFAs
to the traditional antidepressant treatment of unipolar depression. One study
showed that adding EPA, at a dose of 1 g/day, to the pharmacotherapy of 70
subjects who remained depressed while on traditional antidepressants, was effec-
tive in treating their depression.83 Similarly, adding the ethyl ester of EPA, to the
maintenance antidepressant therapy of 20 subjects who continued to have depres-
sive episodes, caused significant clinical benefit compared to augmentation with
placebo, by the third week of treatment.84
When the literature in this area was recently reviewed, four of seven ran-
domized, double-blind, placebo-controlled trials investigating the use of omega-
3 fatty acids to treat depression (five studies of unipolar depression, one of bipolar,
and one of postpartum depression) showed a significant improvement in depres-
sion with administration of at least 1 g/day of EPA.85 Results for the administration
of DHA were not as promising, and it remains to be determined whether EPA
alone is equally as effective as a combination of DHA and EPA.85 It should be
noted that in the four studies that demonstrated positive findings, subjects were
concurrently treated with traditional antidepressants or mood stabilizing agents,
but in only one of the three null trials were the subjects receiving concurrent
treatment.85 Therefore, it could not be ascertained whether the benefit of the
omega-3 supplementation was independent of the standard antidepressant treat-
ment that subjects were undergoing.85 Furthermore, it was not clear whether
treatment with omega-3 PUFAs benefited all subjects with depression or only
those with a particularly low concentration of these fatty acids.85
One concern about research in this area is the loss of blinding that may occur
due to a fishy aftertaste that is sometimes noted by subjects.85 Should a subject
note such a taste, it is quite possible that this will affect how they perceive their
condition. Hence, this could add an element of confounding to this area of
research if it is not controlled.
There is much speculation on how it is that omega-3 polyunsaturated fatty
acids participate in the etiology and treatment of unipolar depression and control
of mood.85 One theory suggests that mood disorders may be caused by impaired
phospholipid metabolism and impaired fatty-acid related signal transduction.86
Another suggests that omega-3 fatty acids have some capacity to regulate sero-
tonergic neurotransmission.87 Thirdly, it has been noted that unipolar depression
is associated with an acute phase response, with increased secretions of proin-
flammatory eicosanoids, and excessive secretion of proinflammatory cytokines.88
Therefore, by limiting the available omega-3 fatty acids, the relative abundance
of omega-6 PUFAs is increased (including arachidonic acid), which leads to
excess synthesis of proinflammatory eicosanoids. (For further discussion and
metabolic pathway descriptions, see Chapter 4.) Each of these mechanisms rep-
resents a large area of study currently undergoing intensive research. How, and
to what extent, each of these mechanisms is involved in the therapeutic benefit
of omega-3 fatty acids in unipolar depression requires much clarification. How-
ever, it is possible, and even likely, that these mechanisms are integrated or that
more than one mechanism contributes to the beneficial properties of these fatty
acids. Overall, the omega-3 fatty acids show promise in the augmentation of
traditional antidepressant treatments for unipolar depression. Nevertheless, more
research is required
6.3 CONCLUSION
It appears that there are a number of nutrients that may have the capacity to
interact with current antidepressants in a positive manner. A better understanding
of these relationships could theoretically allow for nutritional modifications, or
the use of dietary supplements, which could improve the efficacy of unipolar
depression pharmacotherapy. Furthermore, such dietary alterations could decrease
the required dose of traditional antidepressants resulting in a consequent lessening
of negative side effects. In addition, there are a number of dietary components
that can cause various detrimental effects when combined with antidepressants.
A clear understanding of these relationships is important for healthcare practi-
tioners when prescribing unipolar depression medications in order to avoid any
unnecessary suffering for the patient.
Understanding the mechanistic basis of the beneficial nutrient–drug interac-
tions is of the utmost importance for their utilization in the pharmacotherapy of
individuals suffering from unipolar depression. However, many of these interac-
tions are currently explained by seemingly disparate mechanisms, which is a good
indication that much more research is needed in this area. Conversely, under-
standing the mechanistic basis of the detrimental nutrient–drug interactions could
help prevent adverse side effects from occurring as well as help improve our
understanding of the etiology of the condition.
While the “monoamine theory of unipolar depression” has proved at least
partly useful in developing treatments for unipolar depression, it has also left
much room for improvement. It is likely that pharmacotherapy will advance
greatly as we attain a better understanding of the molecular and cellular basis of
this condition. Furthermore, such an understanding will also allow for enhanced
utilization of dietary modifications that assist in pharmacotherapy, and avoidance
of dietary components that could result in unwanted side effects.
TABLE 6.1
Summary of Nutrients and Herbals in the Pharmacotherapy of Unipolar/Major Depression
Nutrient or
Herbal Molecular Target or Possible Confounders or
Supplement Suspected Activity Possible Negative Interactions
Tryptophan Amino acid precursor to serotonin Confounders:

Nutrients and Herbals
Other large neutral amino acids compete to get across blood–brain barrier
Tryptophan can be shunted into niacin or protein production (5-HTP cannot)
The type of drug being taken can determine efficacy of tryptophan
Subject variation: 5-HT receptor plasticity, the relative abundance of precursor
amino acids already available, stage of pharmacotherapy
Degree of inhibition of the liver enzyme tryptophan pyrrolase

The relative importance of serotonin and norepinephrine deficits in depression
etiology
Negative Interaction:
Theoretically, use of any serotonin precursor along with a drug that increases
serotonin in the synaptic cleft could possibly cause serotonin syndrome
Tyrosine Amino acid precursor to norepinephrine, epinephrine, and Similar to above
dopamine
5-HTP Precursor to serotonin, but bypasses the tryptophan hydroxylase Similar to above
step that tryptophan must go through to become 5-HTP
α-Lactalbumin A whey protein high in tryptophan, which provides precursor for Similar to above
serotonin synthesis
Folate, vitamin B12, Methionine synthase requires B12 as a cofactor and Confounders:
(and methyltetrahydrofolate as a substrate MTHFR polymorphisms can have an indirect effect of homocysteine recycling
homocysteine) Possible consequences of disturbed one-carbon metabolism: The association between low folate status and incidence of depression may
not exist in populations with high intake of green vegetables
199
200

Nutrient or
Folate, vitamin B12, 1. Decreased recycling of homocysteine decreases the body’s Dose of folate needed to augment pharmacotherapy may be higher in males
(and methylation capacity, which could impair production of than females (it may have to be high enough to decrease homocysteine levels)
homocysteine) neurotransmitters and membrane phospholipids Interactions between folate, B12, and homocysteine may make interpretations
(continued) 2. Decreased production of tetrahydrobiopterin (BH4), which is of results difficult
a cofactor for tryptophan hydroxylase and tyrosine hydroxylase
(rate-limiting steps in monoamine synthesis)
3. Decreased BH4 (antioxidant) and increased homocysteine (a

potential prooxidant) could result in cellular injury leading to
cerebral dysfunction
SAM 1. Needed in the tryptophan hydroxylase and tyrosine Confounders:
hydroxylase reactions and, hence, may affect synthesis of A wide range of doses and routes of administration have been used in studies
neurotransmitters Over-the-counter supplements may contain varying amounts of active
2. Might be able to increase synapsin I, which regulates the compound
number of vesicles available to be exocytosed from the Negative Interactions:
presynaptic neuronal terminal May increase anxiety, mania, and hypomania in individuals with bipolar
3. Has the capacity to prevent 5-HT1A receptor upregulation, Not much is known about long-term side effects and toxicity
which is normally caused by certain antidepressants
Nutrient–Drug Interactions
Zinc Has the capacity to modulate binding of receptor agonists and Confounders:
antagonists to NMDA receptors by three potential mechanisms: Animal models of depression are controversial
1. Direct antagonism of NMDA receptor Suicide victims used in certain studies did not all have a diagnosis of unipolar
2. Ability to act on AMPA receptor depression before death
3. Ability to inhibit group I metabotropic glutamate receptor
Appears to have the ability to potentiate the action of
acetylcholine on nAChRs
Shown to enhance 5-HT uptake in certain areas of rat brain and
Nutrients and Herbals
reverse inhibition of uptake caused by certain antidepressants

St. John’s wort Induces hepatic and intestinal levels of CYP3A4, which increases Negative Interactions:
the metabolism of amitriptyline Interacts with a large number of different drugs used for various conditions
Is a nonselective reuptake inhibitor and, therefore, may cause
serotonin syndrome when combined with an SSRI
Ginkgo biloba Bilobalide may increase GABA levels in the brain by increasing Negative Interaction:
glutamic acid decarboxylase levels Suspected to have caused a coma in an elderly woman with Alzheimer’s
May increase CYP 3A4 activity resulting in increased conversion disease, when taken along with trazodone
of trazodone to its active metabolite
Panax ginseng Ginseng sapoins observed to block nAChRs Negative Interaction:
Ability to inhibit NMDA receptors When taken with phenelzine, it may result in mania
Vitamin B6 B6 and phenelzine nonenzymatically form a complex, resulting Negative Interaction:
in a loss of available B6 Phenelzine treatment observed to cause B6 deficiency
Confounders:
Mixed results of the studies might be the result of concomitant use of vitamin
supplements in some studies
201
202

Nutrient or
Tyramine Monoamine oxidase (MAO) is needed for the oxidative Negative Interaction:
deamination of tyramine A hypertensive crisis may result if foods high in tyramine are consumed by
When MAO is blocked, tyramine enters the circulation an individual taking an MAOI drug
Tyramine has vasopressor activity and also releases
norepinephrine from nerve terminals
The end result is an increase in blood pressure
Omega-3 PUFAs Three theories suggest how fatty acids may relate to mood Confounders:
(EPA and DHA) disorders: EPA, DHA, and the combination of EPA + DHA may have differing efficacies
1. Mood disorders may be caused by impaired phospholipids Patient variables, such as baseline fatty acid profiles, may also determine
metabolism and impaired fatty-acid related signal transduction efficacy
2. Omega-3 fatty acids may have the ability to regulate Loss of blinding may occur in supplementation studies due to a fishy aftertaste
serotonergic neurotransmission after consuming the fish oil
3. Limitation of omega-3 PUFAs leads to excess omega-6 PUFAs,
which are the precursors to proinflammatory eicosanoids
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7 Supplements and
Anesthesiology
Alan D. Kaye, Amir Baluch, and
Jason M. Hoover
CONTENTS
7.1 Introduction..............................................................................................210
7.2 Minerals ...................................................................................................210
7.2.1 Calcium........................................................................................210
7.2.2 Chromium ....................................................................................210
7.2.3 Magnesium ..................................................................................211
7.2.4 Iron...............................................................................................212
7.2.5 Selenium ......................................................................................212
7.2.6 Zinc ..............................................................................................212
7.3 Vitamins...................................................................................................213
7.3.1 Vitamin A.....................................................................................213
7.3.2 Vitamin B12 ..................................................................................213
7.3.3 Vitamin C.....................................................................................214
7.3.4 Vitamin D ....................................................................................215
7.3.5 Vitamin E.....................................................................................215
7.3.6 Folate ...........................................................................................215
7.4 Herbals.....................................................................................................216
7.4.1 Saw Palmetto ...............................................................................216
7.4.2 St. John’s Wort ............................................................................216
7.4.3 Echinacea.....................................................................................217
7.4.4 Feverfew ......................................................................................220
7.4.5 Ephedra ........................................................................................220
7.4.6 Ginger ..........................................................................................221
7.4.7 Garlic ...........................................................................................222
7.4.8 Gingko Biloba .............................................................................223
7.4.9 Kava Kava....................................................................................224
7.4.10 Ginseng ........................................................................................225
7.5 Conclusion ...............................................................................................226
References .........................................................................................................226
209
7.1 INTRODUCTION
The use of supplements such as minerals, vitamins, and herbal products has
increased dramatically in recent years. Reasons for such an increase in prevalence
include anecdotal reports on efficacy, impressive advertisement, lower cost of
products compared to prescription medications, and ease of attainment of the
supplements. Regardless of the reasons, it is important that physicians, particu-
larly the anesthesiologist, be cognizant of the effects of these agents, whether
beneficial or harmful.
7.2 MINERALS
7.2.1 CALCIUM
It may be reasonable for patients to supplement their diet with calcium, as calcium
deficiency is a common problem.1 Many women supplement with calcium to
improve symptoms associated with premenstrual syndrome.2
Calcium may interfere with a host of commonly used drugs. The anesthesi-
ologist must be aware of patients with cardiac problems that may be taking
calcium channel blockers or beta-blockers. The effects of calcium channel block-
ers may be affected by calcium supplementation, as calcium has been shown to
antagonize the effects of verapamil.3 In fact, calcium has recently been used in
the successful management of calcium channel blocker overdose.4 Calcium sup-
plementation may also decrease levels of beta-blockers, leading to a greater
chronotropic and inotropic presentation than one would expect.5
Thiazide diuretics have been shown to increase serum calcium concentrations,
possibly leading to hypercalcemia due to increased reabsorption of calcium in
the kidneys. Dysrhythmias may occur in patients taking digitalis and calcium
together. The antibiotic effect of tetracyclines and quinolone, and pharmacological
blood levels of bisphosphonates and levothyroxine may be decreased with calcium
supplementation; these medications should not be taken within 2 hours of calcium
intake.6,7
Calcium supplementation may also affect the choice of anesthesia used in
operative procedures. Elevated levels of calcium may complicate cardiopulmo-
nary bypass procedures by worsening mechanical injury to erythrocytes. Recent
data suggest that the use of propofol may have a protective effect on erythrocytes
in patients with elevated levels of calcium.8 Documenting the use of calcium by
patients preoperatively may prevent many of these drug interactions.
7.2.2 CHROMIUM
Chromium is an essential nutrient involved in metabolism of carbohydrates and
lipids. Recently, chromium has received attention from consumers in the belief
that it may improve glucose tolerance in diabetics, reduce body fat, and reduce
atherosclerotic formation. These purported effects stem from chromium’s effect
Supplements and Anesthesiology 211
on insulin resistance. However, the evidence regarding its use for insulin resis-
tance and mildly impaired glucose tolerance is inconclusive.9–12
A double blind trial with 180 patients concluded that high doses of chromium
supplementation (1000 mg) could have beneficial effects on hemoglobin A1c,
insulin, cholesterol, and overall glucose control in type 2 diabetics.13 The anes-
thesiologist should consider asking any diabetics if they supplement with chro-
mium in an attempt to attain these effects. Because of chromium’s effects on
insulin resistance and impaired glucose control, some patients will supplement
with this mineral to prevent risk of cardiovascular disease. Human studies have
shown decreased levels of cholesterol and triglycerides in elderly patients taking
200 µg twice a day.14
Chromium is generally well tolerated; however, some patients may experience
nervous system symptoms, such as perceptual, cognitive, and motor dysfunction
with doses as low as 200 to 400 µg.15 In addition, toxicity has been reported with
chromium consumption. In one case, a woman developed anemia, thrombocy-
topenia, hemolysis, weight loss, and liver and renal toxicity when attempting
weight loss with 1200 to 2400 µg of chromium picolinate. These problems
resolved after discontinuation of chromium ingestion.16 A lower dose of only 600
µg was demonstrated to have resulted in interstitial nephritis in another female
patient.17
7.2.3 MAGNESIUM
Magnesium plays many important roles in structure, function, and metabolism
and is involved in numerous essential physiologic reactions in the human body.
The supplementing of magnesium has been used extensively by patients with
cardiovascular disease, diabetes, osteoporosis, asthma, and migraines, although
most individuals consume adequate levels in their diet.18 Patients with a history
of these illnesses may be supplementing with magnesium and, therefore, should
be questioned.
The most obvious anesthetic consideration in treating a patient taking mag-
nesium supplementation has to do with its effect on muscle relaxants. The mineral
can potentiate the effects of nondepolarizing skeletal muscle relaxants, such as
tubocurarine. Therefore, it may be advisable to ask patients about their magne-
sium usage preoperatively to avoid complications.6
Anesthesiologists caring for obstetrical patients must be aware of the effects
of magnesium sulfate in the patient undergoing caesarean section. Literature
suggests that the duration of action of relaxant anesthetics, such as mivacuronium,
may be affected by subtherapeutic serum magnesium levels.19 Magnesium may
also interfere with the absorption of antibiotics, such as tetracyclines, flurorqui-
nolones, nitrofurantoins, penicillamine, ACE inhibitors, phenytoin, and H2 block-
ers. Absorption problems can be ameliorated by not taking doses of magnesium
within 2 hours from these other medications.20–23 The mineral may also make
oral hypoglycemics, specifically sulfonylureas, more effective when used, thus
increasing the risk of hypoglycemic episodes.24
7.2.4 IRON
In both developed and underdeveloped countries, iron deficiency is the most
common nutrient deficiency. Worldwide, at least 700 million individuals have
iron-deficiency anemia.25 More than just a constituent of hemoglobin and myo-
globin, iron is a key component in nearly every living organism and, in humans,
is associated with hundreds of enzymes and other proteins structures. People have
supplemented with iron in order to treat iron deficiency anemia, alleviate poor
cognitive function in children, increase athletic performance, and suppress restless
legs syndrome (RLS).
High concentrations of iron in the blood may worsen neuronal injury sec-
ondary to cerebral ischemia.26 Increased iron levels during pregnancy may lead
to preterm delivery and neonatal asphyxia.27 These complications may occur even
with normal iron intake if the patient also takes vitamin C, as high doses of the
vitamin can increase iron absorption.28
Iron may inhibit absorption of many drugs including levodopa, methyldopa,
carbidopa, penicillamine, thyroid hormone, captopril, and antibiotics in the qui-
nolone and tetracycline family.29–33 Some medications may decrease iron absorp-
tion and lead to decreased therapeutic levels of the mineral. This includes antacids,
histamine (H2) receptor antagonists, proton pump inhibitors, and cholestyramine
resin.6,7 Iron should not be given orally within 2 hours of other pharmaceuticals
to avoid alterations in drug or mineral absorption.
7.2.5 SELENIUM
Selenium, an essential trace element, functions in a variety of enzyme-dependent
pathways, especially those utilizing selenoproteins. Much of its supplemental
efficacy is due to its antioxidant properties. Glutathione peroxidase incorporates
this mineral at its active site and, as dietary selenium intake decreases, glutathione
levels drop.34
Patients supplement with selenium for a variety of reasons, most notably a
supposed improvement in immune status. Elderly patients may be inclined to
supplement with selenium for this reason.Toxicity with selenium supplementation
begins at intake greater than 750 µg/day and may manifest as garlic-like breath,
loss of hair and fingernails, gastrointestinal distress, or central nervous system
changes.35,36 Few interactions with other pharmacological agents have been
found.6
7.2.6 ZINC
Zinc deficiency was first described in 1961, when it was found to be associated
with “adolescent nutritional dwarfism” in the Middle East.37 Deficiency is this
mineral is thought to be quite common in infants, adolescents, women, and the
elderly.38–41 The most well-known use for zinc supplementation is in treatment
of the common cold, caused principally by the rhinovirus.
Patients self-medicating with zinc supplements may inadvertently overmed-

icate themselves. Signs of zinc toxicity include anemia, neutropenia, cardiac
abnormalities, unfavorable lipid profiles, impaired immune function, acute pan-
creatitis, and copper deficiency.42,43 Zinc supplements may interfere with the
absorption of antibiotics, such as tetracyclines, flurorquinolones, and penicil-
lamine.42 Zinc should not be ingested within 2 hours of antibiotics.7
7.3 VITAMINS
7.3.1 VITAMIN A
The term “vitamin A” refers to a large number of related compounds: preformed
retinol (an alcohol) and retinal (an aldehyde). Vitamin A deficiency is common
in teenagers, lower socioeconomic groups, and in developing countries.44 Fur-
thermore, some studies indicate that diabetic patients are at an increased risk for
vitamin A deficiency.45 This deficiency may manifest as night blindness, immune
deterioration, birth defects, or decreased red blood cell production.46 Purported
therapeutic uses for vitamin A include diseases of the skin, acute promyelotic
leukemia, and viral infections.
Retinoids (a class of chemical compounds that are related chemically to
vitamin A) have been used as pharmacologic agents to treat disorders of the skin.
Psoriasis, acne, and rosacea have been treated with natural or synthetic retinoids.
Moreover, retinoids are effective in treating symptoms associated with congenital
keratinization disorder syndromes. Therapeutic effects stem from its antineoplas-
tic activity.47 Patients suffering from these illnesses may be supplementing with
vitamin A and their dosages should be explored.
Vitamin A may increase anticoagulant effects of warfarin.48 This interaction
could increase the risk of bleeding complications in surgical patients, which can
be avoided by informing the patient of this effect preoperatively
Excess vitamin A intake during pregnancy, as well as deficiency, may lead
to birth defects. For this reason, pregnant woman who are not vitamin A deficient
should not consume more than 2600 IU/day of supplemental retinol.49 Patients
using isotretinoin and pregnant women taking valproic acid, likewise, are at
increased risk for vitamin A toxicity.46,50 Finally, alcohol consumption decreases
the liver toxicity threshold for vitamin A, thereby narrowing its therapeutic win-
dow in alcoholics.51
7.3.2 VITAMIN B12

Vitamin B12, the largest and most complex of all vitamins, is unique in that it
contains cobalt, a metal ion. B12 deficiency may affect up to 10 to 15% of people
over the age of 60.52 B12 deficiency manifests as pernicious anemia. This syndrome
includes a megaloblastic anemia as well as neurologic symptoms. The neurologic
manifestations result from degeneration of the lateral and posterior spinal columns
and include symmetrical paresthesis with loss of proprioception and vibratory

sensation, especially involving the lower extremities.46
The most documented use of vitamin B12 is in the treatment of pernicious
anemia. Many of the neurological, cutaneous, and thrombotic clinical manifes-
tations have been successfully treated with oral or intramuscular cyanocobal-
amin.53 A commonly used anesthetic, nitrous oxide, inhibits both vitamin B12-
dependent enzymes and may produce clinical features of deficiency, such as
megaloblastic anemia and neuropathy. Some experts believe that vitamin B12
deficiency should be ruled out before the use of nitrous oxide, since many elderly
surgical patients have this deficiency.52,54
The drugs colchicines, metformin, phenformin, and zidovudine (AZT) may
decrease the levels of vitamin B12 in a patient.55–58 Histamine2 (H2) receptor
blockers and proton pump inhibitors may decrease absorption of vitamin B12 from
food, but not absorption from dietary supplements.59–61
7.3.3 VITAMIN C
Ascorbic acid, also known as vitamin C, is an essential water-soluble vitamin.
The symptoms of scurvy, which include bleeding and easy bruising, can be
prevented with as little as 10 mg of vitamin C due to its association with collagen,
but it can also be used to prevent a host of other disease processes.62
Numerous people moderately supplement their diet with vitamin C in order
to prevent infection from viruses responsible for the common cold, yet research
reviews over the past 20 years conclude that there is no significant impact on the
incidence of infection.63 However, there are a few studies that show that certain
groups of people who are susceptible to low dietary intake of vitamin C, such as
marathon runners, may be less susceptible when supplementation is used. Fur-
thermore, vitamin C may decrease the duration or severity of colds via an anti-
histamine effect when taken in large doses.64
There is some evidence that patients taking vitamin C supplements may have
a reduced anticoagulant effect from warfarin or heparin. Increased doses of these
anticoagulants might be advised to achieve therapeutic levels.65,66 It is recom-
mended that patients on anticoagulation therapy should limit vitamin C intake to
1 g/day. As always, the precise dosage regimen must be monitored by the appro-
priate lab studies. Since high doses may also interfere with certain laboratory
tests, such as serum bilirubin, creatinine, and stool guaiac assay, it is crucial to
inquire about any over-the-counter supplementation with the vitamin.6 There is
evidence that vitamin C may increase the inotropic effect of dobutamine in
patients with abnormal left ventricular function. Infusion of vitamin C into indi-
viduals with normal heart function was shown to increase contractility of the left
ventricle.67 High doses of vitamin C may increase acetaminophen levels, while
aspirin and oral contraceptives may lower serum levels of vitamin C.68–70
7.3.4 VITAMIN D
Vitamin D deficiency does occur in the elderly and shows increased incidence in
people that live in Northern latitudes.71,72 The main function of this vitamin is in
calcium homeostasis. Individuals with osteoporosis frequently have a deficiency
in vitamin D.73 With increasing age, vitamin D and calcium metabolism increase
the risk of deficiency. Studies show a clear benefit of vitamin D and calcium
supplementation in older postmenopausal women. Supplementation results in
increased bone density, decreased bone turnover, and decreased nonvertebral
fractures, as well as decreases in fall risk and body sway.74
Hypervitaminosis D can occur with high doses of the vitamin. Symptoms
include nausea, vomiting, loss of appetite, polydipsia, polyuria, itching, muscular
weakness, joint pain, and, in some cases, can lead to coma and death.46 In order
to prevent the syndrome, the Food and Nutrition Board has set an upper limit of
supplementation at 2000 IU/day for adults.75
The cardiac patient taking calcium channel blockers may present for surgery
while concurrently taking supplemental vitamin D and calcium. The combination
of vitamin D and calcium may interfere with calcium channel blockers by antag-
onizing its effect. Hypercalcemia exacerbates arrhythmias in patients taking dig-
italis. A state of hypercalcemia may be induced by the concomitant use of thiazide
diuretics with vitamin D, which may lead to these complications. Conversely,
anticonvulsants, cholesterol-lowering medications, and the fat substitute olestra
may decrease the absorption of vitamin D.76
7.3.5 VITAMIN E
Antioxidant properties define the primary function of vitamin E. Dietary defi-
ciency is quite prevalent even in the developed world; therefore, supplementation
is reasonable.77 The anesthesiologist must be keenly aware of vitamin E supple-
mentation, as it may increase the effects of anticoagulant and antiplatelet drugs.
Concomitant use of vitamin E with these drugs may increase the risk of hemor-
rhage.78 Further, preliminary evidence suggests that type 2 diabetics may have
an increased risk of hypoglycemia since vitamin E may enhance insulin sensitivity
and, therefore, adjustment of oral hypoglycemics would be advisable.79,80
Cholestyramine, colestipol, isoniazid, mineral oil, orlistat, sucralfate, and the fat
substitute olestra may possibly decrease the absorption of vitamin E, leading to
decreased levels in the serum.6
7.3.6 FOLATE
Folic acid and folate have been used interchangeably, although the most stable
form that is used by the human body is folic acid. This water-soluble, B-complex
vitamin occurs naturally in foods and in metabolically active forms.81 Since 1998,
the fortification of cereal with folate has significantly decreased the prevalence
of folate deficiency.82
Excess folate intake has not been associated with any significant adverse
effects. Patients taking large amounts of nonsteroidal antiinflammatory drugs
(NSAIDs), such as aspirin or ibuprofen, experience interference in folate metab-
olism, although regular use shows no significant changes. Patients suffering from
seizures who use phenytoin for therapy may report decrease in seizure threshold
when taking folate supplements.83 The body’s ability to absorb or utilize folate
may be decreased if taking nitrous oxide, antacids, bile acid sequestrants, H2
blockers, certain anticonvulsants, and high-dose triamterene. Supplementation of
folic acid may also correct for megaloblastic anemia due to B12 deficiency, but
the neurological damage will not be prevented. In these cases, one must be careful
to pinpoint the true cause of the anemia to prevent neurological complications.43
7.4 HERBALS
7.4.1 SAW PALMETTO
Saw palmetto is used mainly for treatment of benign prostatic hyperplasia with
free fatty acids and sterols being the main components.84 Despite an uncertain
mechanism, the literature does demonstrate antagonism at the androgen receptor
for dihydrotestosterone and 5α-reductase enzyme.84 Though prostate size and
prostate-specific antigen level are not decreased by saw palmetto, biopsies have
demonstrated decreases in transitional zone epithelia in prostates of men treated
with this agent compared to a placebo.84 When compared with finasteride, a 5α-
reductase inhibitor, saw palmetto use resulted in fewer side effects and increased
urine flow.84 However, a study of patients with prostatitis/chronic pelvic pain
syndrome that evaluated the safety and efficacy of saw palmetto compared to
finasteride reported that at the end of the investigation, more patients opted to
continue finasteride treatment rather than the saw palmetto treatment. The
researchers found that in patients with the studied condition, saw palmetto had
no appreciable long-term improvement and, with the exception of voiding,
patients on finasteride experienced significant improvement in all other analyzed
parameters.85
Adverse reactions to saw palmetto are rare, but there are reports of mild
gastrointestinal symptoms and headaches.84 Results of a recent investigation
indicated that recommended doses of saw palmetto are not likely to alter the
pharmacokinetics of co-administered medications dependent on the cytochrome
P-450 isoenzymes CYP2D6 or CYP3A4, such as dextromethorphan and alpra-
zolam.86 Further, there are few herbal–drug interactions in the literature regarding
saw palmetto, but, as always, care and responsibility should be exercised when
taking this agent.84
7.4.2 ST. JOHN’S WORT

St. John’s wort is used to treat anxiety, mild-to-moderate depression, and sleep-
related disorders.84,87 Other uses have included treatment of cancer, fibrositis,
headache, obsessive-compulsive disorder, and sciatica.88 Active compounds in the

agent include the naphthodihydrodianthrones, hypericin and pseudohypericin, the
flavonoids, quercitrin, rutin, and hyperin, and the xanthones.84,89
It is thought that extracts of St. John’s wort, such as WS 5570, are widely
used to treat mild-to-moderate depression.90,91 Such extracts are standardized
based on their hypericin content and have demonstrated an effectiveness superior
to placebo and potentially as great as selective serotonin reuptake inhibitors and
low-dose tricyclic antidepressants.88
The exact mechanism of action of St. John’s wort remains controversial. This
herbal substance demonstrates irreversible inhibition of monoamine oxidase in
vitro, but such inhibition has yet to be observed in vivo.92 In the feline lung
vasculature, St. John’s wort exhibited a vasodepressor effect that was mediated
or modulated by both a gamma-aminobutyric acid (GABA) receptor and an L-
type calcium channel sensitive mechanism.93 Studies performed in vitro have
demonstrated GABA receptor inhibition by hypericum perforatum. This finding
may indicate that a GABA inhibitory mechanism is responsible for the antide-
pressant effect.94,95 However, other theorized pathways include inhibition of sero-
tonin, dopamine, and norepinephrine reuptake in the central nervous system, thus
making its mechanism of action somewhat similar to traditionally used antide-
pressant medications.84
Regarding side effects, St. John’s wort is typically well tolerated.84 Associated
side effects may include photosensitivity, restlessness, dry mouth, dizziness,
fatigue, constipation, and nausea84,87 (Table 7.1). Other noteworthy side effects
of St. John’s wort include its induction of the cytochrome P-450 system (CYP
34A), thus affecting serum levels of cyclosporine in patients after organ trans-
plantation, and the potential threat of serotonergic syndrome in patients concur-
rently taking prescription antidepressants.84 The serotonergic syndrome is char-
acterized by hypertonicity, myoclonus, autonomic dysfunction, hallucinosis,
tremors, hyperthermia, and potentially death.96,97 Specifically, use of St. John’s
wort is not recommended with photosensitization drugs such as tetracyclines,
antidepressants such as monoamine oxidase inhibitors and selective serotonin
reuptake inhibitors, and β-sympathomimetics such as ephedra and pseudoephe-
drine hydrochloride. Finally, there is little to no data regarding the potential
anesthetic–St. John’s wort interactions.
7.4.3 ECHINACEA
Echinacea is part of the daisy family found throughout North America. There are
nine species of echinacea in total and the medicinal preparations are derived from
three of these: Echinacea purpurea (purple coneflower), Echinacea pallida (pale
purple coneflower), and Echinacea angustifolia (narrow-leaved coneflower).96,98,99
Echinacea is recommended as a prophylactic and treatment substance for upper
respiratory infections. However, data are insufficient at present to support the
former.84 It has alkylamide and polysaccharide substances that possess significant
TABLE 7.1
Herbal Agents, Potential Side Effects, and Anesthesia Considerations
Herbal
Agents Potential Side Effects Anesthesia Considerations
Echinacea Unpleasant taste, tachyphylaxis, Can potentiate barbiturate toxicity

affects cytochrome P-450 enzyme,
hetatotoxicity
Ephedra Hypertension, tachycardia, Can interact with anesthetics, i.e.,
(Ma Huang) cardiomyopathy, stroke, cardiac halothane, and cause cardiac
arrythmias dysrhythmias
Feverfew Aphthous ulcers, gastrointestinal Can increase risk of intraoperative
irritability, headache hemodynamic instability
Garlic Halitosis, increases in bleeding time, Can increase risk of intraoperative
hypotension, affects cytochrome hemodynamic instability
P-450 enzyme
Ginger Increases in bleeding time Can increase risk of intraoperative
hemodynamic instability
Gingko biloba Platelet dysfunction Can increase perioperative bleeding
tendencies and decrease
effectiveness of intravenous
barbiturates
Ginseng Hypertension, increases in bleeding Can increase risk of intraoperative
time, hypoglycemia, insomnia, hemodynamic instability
vomiting, epistaxis
Kava kava Dermopathy, affects cytochrome Can potentiate the effect of
P-450 enzyme, hepatotoxicity barbiturates/benzodiazepines
resulting in excessive sedation
St. John’s wort Dry mouth, dizziness, affects Pseudoephedrine, MAOIs, SSRIs
cytochrome P-450 enzyme, should be avoided
constipation, nausea, serotonergic
syndrome
Note: MAOIs = monoamine oxidase inhibitors, SSRIs = selective serotonin reuptake inhibitors
Source: Modified from Kaye AD, Clarke RC, Sabar R, et al. Herbal medicines: current trends in
anesthesiology practice — a hospital survey. J Clin Anest 12: 468–471, 2000.
in vitro and in vivo immunostimulation properties due to enhanced phagocytosis

and nonspecific T-cell stimulation.100
The consumption of echinacea at the onset of symptoms has been clinically
shown to decrease both the severity and duration of the cold and flu. Employing
quantitative polymerase chain reaction (PCR) to identify in vivo alterations in the
expression of immunomodulatory genes in response to echinacea has been per-
formed.101 Investigations conducted on in vivo gene expression within peripheral
leukocytes were evaluated in six healthy nonsmoking subjects. Blood samples
were obtained at baseline and on subsequent days following consumption of a
commercially blended echinacea product. The overall gene expression pattern

between 48 hours and 12 days after taking echinacea was consistent with an
antiinflammatory response. The expression of interleukin-1beta, intracellular
adhesion molecule, tumor necrosis factor-alpha, and interleukin-8 was modestly
depressed up through day 5, and returned to baseline by day 12. Further, the
expression of interferon-alpha consistently increased through day 12, thus indi-
cating an antiviral response. Therefore, initial data yielded a gene expression
response pattern consistent with the ability of echinacea to decrease both the
intensity and duration of cold and flu symptoms.101
Aside from the effects of echinacea on innate immunity, few studies are
available that have examined the ability for enhancement of humoral immunity.
Although a study using female Swiss mice as the model found support for the
use of Echinacea purpurea, as suggested by anecdotal reports, and demonstrated
potential enhancement of humoral immune responses, in addition to innate
immune responses.102 However, it is important to note that the use of Echinacea
purpurea, as dosed in one study, was not effective in treating upper respiratory
tract infections and related symptoms in pediatric patients, aged 2 to 11. Further,
the consumption of Echinacea purpurea was associated with an increased risk
of rash.103
Regarding side effects, echinacea is often well tolerated with the most com-
mon side effect being its unpleasant taste.84,104 Extended use of echinacea for
more than 2 months may lead to tachyphylaxis.105 Anaphylaxis has also been
reported with a single dose of this herbal agent.96 Further, echinacea use has been
associated with hepatatoxicity if taken with hepatotoxic agents, including ana-
bolic steroids, amiodarone, ketoconazole, and methotrexate.106 Further, flavinoids
from Echinacea purpurea can affect the hepatic cytochrome P-450 and sulfotrans-
ferase systems.107,108 For example, one investigation found that echinacea
decreased the oral clearance of substrates of the cytochrome P-450 1A2 system,
but not the oral clearance of substrates of the 2C9 and 2D6 isoenzymes in vivo.
The herbal also selectively modulates the activity of the cytochrome P-450 P3A
isoenzyme at both hepatic and intestinal sites. The researchers, therefore, urged
caution when echinacea is combined with medications dependent upon the cyto-
chrome P-450 3A or 1A2 systems for elimination.109 Drug levels may become
elevated with concomitant use of echinacea. Some drugs that are metabolized by
the cytochrome P-450 3A enzyme include lovastatin, clarithromycin, cyclospor-
ine, diltiazem, estrogens, indinavir, triazolam, and numerous others. Taking mida-
zolam and echinacea together seems to increase levels of the sedative.109 Finally,
echinacea use should exceed 4 weeks and it should not be used in patients with
systemic or autoimmune disorders, patients who are pregnant, or patients who
are immunocompromised.84
The immunostimulatory effects of echinacea may antagonize the immuno-
suppressive actions of corticosteroids and cyclosporine.110 Echinacea may also
lead to inhibition of the hepatic microsomal enzyme system and, as such, its use
with drugs, such as phenobarbital, phenytoin, and rifampin, which are metabo-
lized by these enzymes, should be avoided as toxicity may result.
7.4.4 FEVERFEW
Feverfew is used to treat headache, fever, menstrual abnormalities, and prevent
migraines.111 The name is derived from the Latin word febrifugia, which means
“fever reducer.”112 Although feverfew is commonly used for migraine headaches,
the literature is inconclusive regarding its efficacy.113,114 In a study reviewing
evidence from double-blind, randomized controlled trials of the clinical efficacy
of feverfew vs. placebo for migraine prophylaxis, investigators found insufficient
evidence to suggest a benefit of feverfew over placebo for the prevention of
migraines.115 As with most herbal compounds, analyses of feverfew-based prod-
ucts have yielded significant variations in the parthenolide contents, which are
believed to be the active ingredients.116
Regarding the effects of the antiinflammatory lactone parthenolide, a German
study indicated that parthenolide may support T-cell survival by downregulating
the CD95 system. The CD95 system is a critical component of the apoptotic, or
programmed cell death pathway of activated T-cells. Further, the authors reported
that pathenolide can have therapeutic potential as an antiapoptotic substance
blocking the activation-induced cell death of T-cells.117
Feverfew also has demonstrated inhibition of serotonin release from aggre-
gating platelets. This mechanism may be related to the inhibition of arachidonic
acid release via a phosholipase pathway.118–120 It has also been found that feverfew
has decreased approximately 86 to 88% of prostaglandin production without
exhibiting inhibition of the cyclooxygenase enzyme.121
Adverse reactions to feverfew include apthous ulcers, abdominal pain, nausea,
and vomiting. A rebound headache may occur with abrupt cessation of this
herbal.111–112 Better tolerance to feverfew has been suggested when compared to
conventional migraine medications because in studies feverfew use resulted in
no alteration in heart rate, blood pressure, body weight, or blood chemistry like
conventional migraine medications.111 A condition known as “post-feverfew syn-
drome” can occur in long-term users, which manifests as fatigue, anxiety, head-
aches, insomnia, arthralgias, and muscle and joint stiffness.111,122
Feverfew may inhibit platelet action; therefore, it is reasonable to avoid the
concomittant use of this herb in patients taking medications, such as heparin,
warfarin, NSAIDs, aspirin, and vitamin E.123,124 Furthermore, herbs, like feverfew,
can interact with iron preparations, thereby reducing the bioavailability of that
substance.106
7.4.5 EPHEDRA
Since the U.S. government’s ban on ephedra-based products, there has been an
obvious decline in its prevalent use in this country. However, patients may still
present for perioperative assessment with a history of use of ephedra. Ma huang,
an ephedra-based alkaloid, is similar in structure to amphetamines and is tradi-
tionally indicated for the treatment of various respiratory disorders, such as the
flu, the common cold, allergies, and bronchitis. Additionally, it is commonly used
as an appetite suppressant.84 Ma huang, or ephedra, acts as a sympathomimetic

agent and exhibits potent positive ionotropic and chronotropic responses. In
addition to its antitussive actions, ephedra may also possess bacteriostatic prop-
erties.112 As a cardiovascular and respiratory sympathomimetic, it utilizes an α-
or β-adrenergic sensitive pathway.125 Recent laboratory data using the cat lung
vascular bed indicate that ephedra-mediated pulmonary hypertension is dependent
upon α(1)-adrenoreceptor-sensitive mechanisms.126
The appetite suppressant and metabolic enhancer effects of ma huang made
it a potent ingredient of various over-the-counter weight loss compounds. How-
ever, even prior to the U.S.’s federal ban on ma huang, many herbal manufacturers
were already promoting their ephedra-free supplements due to the numerous
reported adverse effects of ephedra.
Dangerous side effects of ma huang administration include hypertension,
tachycardia, cardiomyopathy, cardiac dysrrhythmias, myocardial infarction,
stroke, seizures, psychosis, and death.84 Many of these complications have been
attributed to a lack of standardization in its formulation.127,128 Before the U.S. ban
of ma huang, approximately 16,000 cases of adverse events, including 164 deaths,
had been reported to the U.S. Food and Drug Administration from 1994.129
Further, the Bureau of Food and Drug Safety of the Texas Department of Health
reported eight ephedra associated fatalities during a 21-month period between
1993 and 1995; seven of the fatalities were due to myocardial infarction or
stroke.89 Patients at highest risk of side effects include those who are pregnant,
have hypertension, coronary vascular disease, seizures, glaucoma, anxiety, or
mania.84
The use of ma huang is highly relevant to the perioperative period. The
possibility of hypertension causing myocardial ischemia or stroke needs to be
considered. Further, ephedra can potentially interact with general anesthetic
agents, such as halothane, isoflurane, desflurane, or cardiac glycosides, like dig-
italis, to cause cardiac dysrrhythmias. Patients taking ephedra for prolonged
periods of time can also deplete their peripheral catecholamine stores. Therefore,
under general anesthesia, these patients can potentially experience profound intra-
operative hypotension, which can be controlled with a direct vasoconstrictor (e.g.,
phenylephrine) instead of ephedrine. Finally, use of ephedra with phenelzine or
other monoamine oxidase inhibitors may result in insomnia, headache, and trem-
ulousness, and concurrent use with the obstetric drug oxytocin has resulted in
hypertension.130
7.4.6 GINGER
Ginger has been used for the treatment of nausea, vomiting, motion sickness, and
vertigo.87 A study of the effects of ginger on subjects with vertigo found that no
subjects experienced nausea after caloric stimulation of the vestibular system, in
contrast to those treated with a placebo.131 It is postulated that ginger may
be superior to dimenhydrinate in decreasing motion sickness.132 For vomiting
episodes, this herbal has also been effective in decreasing symptoms associated
with hyperemesis gravidarum.133
The effect of ginger on the clotting pathway has also been investigated. Ginger
has exhibited potent inhibition of thromboxane synthetase and this results in an
increased bleeding time.134 The ability of ginger constituents and related sub-
stances to inhibit arachidonic acid-induced platelet activation in human whole
blood has also been investigated. The data from that study revealed ginger com-
pounds and derivatives are more potent antiplatelet agents than aspirin under
conditions employed in the study. [8]-Paradol, a constituent of ginger, was iden-
tified the most potent antiplatelet aggregation agent and cyclooxygenase 1 inhib-
itor.135 In another study, administration of ginger has also resulted in decreases
in blood pressure, serum cholesterol, and serum triglycerides in diabetic rats.136
Thus, further investigation into these effects in this disease is warranted.
Adverse effects of ginger include bleeding dysfunction and its use is con-
traindicated in patients with coagulation abnormalities or those on anticoagulant
medications such as NSAIDs, aspirin, warfarin, and heparin.87 Ginger may
increase bleeding risk, enhance barbiturate effects, and, as a result of an inotropic
effect, interfere with cardiac medications. Large quantities of ginger may also
cause cardiac arrhythmias and central nervous system depression.137
7.4.7 GARLIC
Garlic’s use is prevalent and is available in powdered, dried, and fresh forms.84
Allicin, the main active ingredient in garlic, contains sulfur, and crushing the
clove activates the enzyme allinase, thus facilitating the conversion of alliin to
allicin.96
Recommended uses for garlic have focused on treating hypercholesterolemia,
hypertension, and cardiovascular disease and studies have targeted its hypo-
cholesterolemic and vasodilatory activity.84,138–142 Investigations have found that
garlic may lead to inhibition of the HMG-CoA reductase and 14α-demethylase
enzyme systems, thereby exerting a lipid-reducing effect.84 Garlic may also be
used for its antiplatelet, antioxidant, and fibrinolytic actions.140,143,144 There is
minimal data present to support the use of garlic for hypertension, as its depressor
effects on systolic and diastolic blood pressure appear to range from minimal to
modest.84,96
Chronic oral use of garlic has been reported to augment the endogenous
antioxidants of the heart.146 A recent study hypothesized that garlic-induced
cardiac antioxidants may provide protection against acute adriamycin-induced
cardiotoxicity. Using the rat model, researchers discovered an increase in oxida-
tive stress as evidenced by a significant increase in myocardial thiobarbituric acid
reactive substances (TBARS) and a decrease in myocardial superoxide dismutase
(SOD), catalase, and glutathione peroxidase activity in the adriamycin group.
However, in the garlic treated rats, the increase in myocardial TBARS and a
decrease in endogenous antioxidants by adriamycin was significantly attenuated.
Therefore, one may conclude garlic administration may help prevent this form
of drug-induced cardiotoxicity.145 The effects of allicin in the feline and rat lung
vasculature have also been studied. Allicin has shown significant vasodepressor
activity in the pulmonary vascular bed of the rat and cat.146 Although allicin has
been found to lower blood pressure, insulin, and triglycerides levels in fructose-
fed rats, it has also been considered important to investigate its effect on the
weight of animals.
Recent data indicate garlic may be an effective treatment against methicillin-
resistant Staphylococcus aureus (MRSA) infection. In a study using mice, inves-
tigators demonstrated that the garlic extracts, diallyl sulphide and diallyl disul-
phide, showed protective qualities against MRSA infection. Such conclusions,
coupled with further investigation, may result in the use of such extracts in MRSA
infection treatment.147
Side effects of garlic are minimal, with odor and gastrointestinal discomfort
being the most commonly reported.84 Induction of the cytochrome P-450 system
may occur as evidenced by reduction of serum levels of one medication.84 Anes-
thesiologists must be aware that garlic may augment the effects of warfarin,
heparin, aspirin, and may result in an abnormal bleeding time. This effect can
result in increased risk of peri-operative hemorrhage.148 Thus, monitoring of
cardiovascular function in this period is crucial.
7.4.8 GINGKO BILOBA

There are many active components present in gingko, including the flavinoid
glycosides and terpenoids. The flavinoids have demonstrated antioxidant activity
while the terpenoids have shown antagonism to platelet action.84 Gingko has been
used to treat intermittent claudication, vertigo, and enhance memory.89 Subjects
using this herbal have reported decreased pain in the affected lower extremities
and increased symptom-free distance in ambulation. In addition to inhibiting the
platelet-activating factor, gingko may also mediate nitric oxide release and
decrease inflammation.84,149–154
To evaluate the efficacy of gingko on dementia, a double-blind and placebo-
controlled randomized trial using the extract EGB761 was performed. It was
found that EGB761 had the potential to stabilize and modestly improve cognitive
performance and social functioning.84,155 In addition, the improvement in cogni-
tion was comparable to the effect of donazepil on dementia.84 This effect on
cognition function and memory may be related to activation of cholinergic neu-
rotransmitters. It is important to note, however, that data are inconclusive regard-
ing the ability of this herbal to improve memory in subjects without dementia.84
Although the pathogenesis of acute pancreatitis is not well understood, there
are numerous data that suggest a role for oxygen free radicals in the progression
and complications of pancreatitis. The effects of EGB761 have shown a positive
effect on acute pancreatitis and this effect may be linked to a free radical scavenger
effect by gingko.156
Gingko is generally well tolerated in healthy adults for about 6 months.84
However, aside from the mild gastrointestinal distress, the potential of gingko on
antiplatelet activating factor has resulted in gingko biloba-induced spontaneous

hyphema (bleeding from the iris, the anterior chamber of the eye), spontaneous
bilateral subdural hematomas, and subarachnoid hemorrhage.84,87,157–160 Therefore,
the use of anticoagulants and gingko should be strictly monitored and possibly
avoided.84
Regarding the effects of gingko on pharmacokinetics, an open-labeled and
randomized crossover trial was conducted on healthy human volunteers to deter-
mine if ginkgo alters the pharmacokinetics of digoxin. The investigators found
the concurrent use of orally administered gingko and digoxin did not seem to
have a significant effect on the pharmacokinetics of digoxin in healthy volun-
teers.161 Therefore, one may conclude that concurrent use of gingko biloba with
aspirin, NSAIDs, warfarin, and heparin is not recommended as gingko may
increase the potential for bleeding in these patients. It is also advisable to avoid
use of gingko with anticonvulsant drugs, such as carbamazapine, phenytoin, and
phenobarbital, as the herbal may decrease the effectiveness of these medica-
tions.106 Concurrent use of gingko and tricyclic antidepressants also is not advised
because of the potential to lower the seizure threshold in these patients.106
7.4.9 KAVA KAVA

Kava kava, an extract of the Piper methysticum plant, is employed for its proposed
anxiolytic, antiepileptic, antidepressant, antipsychotic, and sedative proper-
ties.162–164 Some of the active ingredients of kava kava include the lactones or
pyrones, kawain, methysticin, dihydrokawain, dihydromethysticin.165,166 Kava
extracts available commercially are usually found to contain approximately 30
to 70% kava lactones.165
The extract WS 1490 has been investigated to determine its effectiveness in
the treatment of anxiety.166 WS 1490 has been shown effective in anxiety disorders
as a treatment alternative to benzodiazepines and tricyclic antidepressants, and
reported not to have the problems associated with those two classes of drugs.166
However, therapeutic effect may take up to 4 weeks and data have indicated
treatment for 1 to 8 weeks to obtain significant improvement.165,167
Although the exact mechanism of kava kava’s effects on the central nervous
system is largely unknown, the pyrones have demonstrated competitive inhibition
of the monoamine oxidase B enzyme.165 Inhibition of this enzyme may result in
the psychotropic effects related to kava kava use, as this enzyme is responsible
for the breakdown of amines that play a role in psychoses.168
Regarding adverse effects, patients that experience hepatic adverse reactions
are known as “poor metabolizers.” Typically, these patients have a deficiency in
the cytochrome P-450 2D6 isozyme.165 Therefore, it is recommended that patients
who use kava kava receive routine liver function tests to monitor for the develop-
ment of hepatotoxicity.165 Furthermore, there have been 24 documented cases of
hepatotoxicity following the use of kava kava and, in some cases, death or liver
transplant occurred after 1 to 3 months of use.165 In countries, such as Germany
and Australia, kava kava use longer than 3 months is not recommended.167 Other
side effects of kava kava use include visual changes, a pellagra-like syndrome with
characteristic ichthyosiform dermopathy, and hallucinations.165,169,170
Regarding drug interactions, kava kava may react adversely with the benzo-
diazepine alprazolam, other central nervous system depressants, statins, alcohol,
and levodopa, consequently resulting in excessive sedation, among other side
effects; therefore, the supplement should be avoided in those patients with endog-
enous depression.165,171–173 Finally, kava kava may also affect platelets in an
antithrombotic fashion by inhibiting cyclooxygenase and, thus, attenuating throm-
boxane production.165 Pain relief mechanisms utilized by the herbal may be
similar to local anesthetic responses and could be dependent on a nonopiate
sensitive pathway.174,175
7.4.10 GINSENG
There are three main groups of ginseng that are classified based on their geo-
graphic origin.84 These are Asian ginseng, American ginseng, and Siberian gin-
seng, with the pharmacologically active ingredient in ginseng being ginsenos-
ides.84,89,112 Asian and American ginsengs have been used to increase resistance
to environmental stress, promote diuresis, stimulate the immune system, and aid
digestion.176,177 Further, while Asian ginseng has shown promise in improving
cognition when combined with the herbal agent gingko, American ginseng has
been studied for its potential to stimulate human tumor necrosis factor alpha
(TNF-alpha) production in cultured human white blood cells.177,178 American
ginseng may also possess hypoglycemic activity.179,180 Such effects have been
observed in both normal and diabetic subjects and may be attributed to ginseng’s
components, specifically ginsenoside Rb2 and panaxans I, J, K, and L.181–185
Typically ginseng is well tolerated, but side effects, such as bleeding abnor-
malities secondary to antiplatelet effects, headache, vomiting, Stevens-Johnson
syndrome, epistaxis, and hypertension have been reported.186–192 Drug interactions
between Asian ginseng and calcium channel blockers, warfarin, phenelzine, and
digoxin have also been noted.84 It may be advisable for patients on anticoagulant
medications, such as warfarin, heparin, aspirin, and NSAIDs, to avoid ginseng.
Further, because of ginseng’s association with hypertension and the deleterious
outcomes linked to chronic hypertension, the anesthesiologist should be aware
of whom and for how long patients may have been taking this herbal product.
Since many anesthetic agents can cause generalized vasodilation, hemodynamic
lability may be seen.
Regarding ginseng’s interaction with antidepressants, such as monoamine
oxidase inhibitors, concurrent use of ginseng with phenelzine sulphate should be
avoided as manic episodes have been reported with routine use of both.193,194
Finally, as a result of ginseng’s potential to cause decreased blood glucose levels,
it should be used cautiously in diabetic patients on insulin or other oral hypogly-
cemic agents, and blood glucose levels should be monitored.
7.5 CONCLUSION
The growing use of supplements, such as minerals, vitamins, and herbals, in the
world warrants a more comprehensive understanding of these agents by the
medical community. It is important for the anesthesiologist to recognize certain
facts regarding these supplements. For example, there are about 1300 g of calcium
in a 70 kg adult and the mineral magnesium activates approximately 300 enzyme
systems in the human body; most of these systems are involved in energy metab-
olism.195 Aside from these, the anesthesiologist must regulate the patient’s phys-
iological functions and appreciate the effect of these supplements on such func-
tions during various operative procedures. As demonstrated in this chapter, the
use of these compounds may prove beneficial for some patients, but result in
alterations in normal physiologic functions in others, thus potentially resulting
in deleterious consequences. These agents, in addition to all other medications
taken by the patient, should be screened by medical practitioners, in particular
anesthesiologists, as some of these compounds may interact with chosen anes-
thetics during the stages of anesthesia. Furthermore, education of patients regard-
ing the serious potential drug-supplement interactions should be an integral com-
ponent of the preoperative assessment. Specifically, the American Society of
Anesthesiologists (ASA) recommends that all herbal medications be discontinued
2 to 3 weeks prior to elective surgery.
Due to current lax regulations in some countries, some of these agents are
poorly categorized and standardized, thus resulting in a high risk of adverse effects
when used by an uninformed or misinformed public. Within the past few decades,
hundreds of deaths have been linked to the use of these agents, specifically the
herbals. Data also suggest that less than 1% of adverse effects associated with
herbals are reported. In general, whether the patient is taking minerals, vitamins,
and/or herbals, one thing is for certain, an open line of communication between
medical practitioner and patient should exist regarding all of these agents. This
communication is essential to ensure quality patient treatment, a stable and secure
rapport, and a properly informed and educated general public.
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8 Nutrient- and Drug-

Responsive Polymorphic
Genes as Nutrigenomic
Tools for Prevention of
Cardiovascular Disease
and Cancer
Kevin Wood and Marica Bakovic
CONTENTS
8.1 Introduction..............................................................................................238
8.2 Genetic Polymorphisms and CVD..........................................................239
8.2.1 Apolipoprotein E and CVD ........................................................240
8.2.2 APOE Genotype and Saturated Fat Intake .................................240
8.2.3 APOE Genotype and Sucrose Intake ..........................................241
8.2.4 Methylenetetrahydrofolate Reductase and CVD ........................241
8.2.5 MTHFR 677 C→T Polymorphism, Folate Intake, and CVD ....242
8.2.6 Hepatic Lipase and CVD ............................................................243
8.2.7 HL-514 C→T Polymorphism and CVD.....................................244
8.2.8 Statins, Niacin, and Polymorphisms Related to the
Lowering of Cholesterol..............................................................244
8.3 Genetic Polymorphisms and Cancer .......................................................246
8.3.1 Vitamin D and the Vitamin D Receptor .....................................247
8.3.2 VDR Polymorphisms, Vitamin D Intake, and
Colorectal Cancer ........................................................................248
8.3.3 Methylenetetrahydrofolate Reductase, Folate Status,
and Cancer ...................................................................................249
8.3.4 MTHFR 677 C→T Polymorphism, Folate Intake, and
Colorectal Cancer ........................................................................250
8.3.5 MTHFR 677 C→T and 1298 A→C Polymorphisms and
Acute Leukemia...........................................................................250
237
8.3.6 MTR 2756 A→G Polymorphism and Cancer ............................251

8.4 Conclusions..............................................................................................251
References .........................................................................................................252
8.1 INTRODUCTION
Nutrigenomics is an integrative science combining biotechnology, molecular
medicine, and pharmacogenomics, which is revolutionizing how nutrition and
diet are viewed.1 A working definition of this new branch of genomics, by Kaput
and Rodriguez, is that “it seeks to provide a genetic understanding for how dietary
chemicals can affect the balance between health and disease by altering the
expression of an individual’s genetic makeup.”2 In an attempt to clarify the term,
they have proposed the following five tenets:
1. Common dietary compounds act on the human genome, directly or

indirectly, to alter gene expression or structure.
2. In some individuals under certain circumstances, diet can be a serious
risk factor for a number of diseases.
3. Some diet-regulated genes (and their normal, common variants) are
susceptibility genes and likely to play a role in the onset, incidence,
progression, and/or severity of chronic diseases.
4. The degree to which diet influences the balance between healthy and
disease states depends on an individual’s genetic makeup.
5. Dietary interventions based on knowledge of nutritional requirement,
nutrition status, and genotype (i.e., “individualized nutrition”) can be
used to prevent, mitigate, or cure chronic disease.
The possible applications of nutrigenomics could be numerous. Genomic

information could be used to understand the basis of individual differences in
response to dietary patterns.1 Nutrigenomic data could be gathered to provide a
sound basis for the development of safe and effective diet therapies for individuals
or subgroups in specific populations. Therefore, the main goal of nutrigenomics
is to understand each individual’s responses to micro- and macronutrients via
genetic screening in order to create safe and effective dietary treatments for them.
Nutrigenomics can establish more flexible, long-term dietary strategies for early
prevention of chronic diseases, based on genetic predispositions and the knowl-
edge of specific dietary requirements of individuals and particular (ethnic) groups,
relying less on the entire population.3
In particular, single nucleotide polymorphisms (SNPs) will be very important
in identifying individual variations in responses to nutrients. SNPs are changeable
sites within a genome, occurring once every 1000 to 2000 nucleotides,4 where
the rarer nucleotide is present at a frequency of greater than 1% in a population.5
SNPs are different than sporadic mutations, which are germline polymorphisms
occurring with a frequency of less than 1%.5 These genetic variations alter protein
Nutrient- and Drug-Responsive Polymorphic Genes 239
structure and function when the nucleotide base substitution occurs in a gene’s
coding region.6 Alternatively, a polymorphism may have no effect on the protein
product because the base substitution occurs in a DNA region that is not involved
in gene transcription or translation; however, it could be a part of regulatory
promoters.
Molecular techniques for detecting SNPs (polymerase chain reaction, restric-
tion enzyme detection) have been used extensively in pharmaceutical, toxicolog-
ical, and clinical research disciplines, but not so much in nutritional sciences.4
The recent development of extensive genomic databases and high throughput
genetic screening made it possible for the study of the interactions of interindi-
vidual variation and nutrition. The greatest potential for benefit from those studies
will be for health maintenance by preventing or slowing the early stages of diet-
related diseases. Nutrigenomics could also provide the means to develop better
molecular biomarkers of early changes between health maintenance and disease
progression.
The most common diseases, such as cardiovascular disease (CVD) and cancer,
are known to be polygenic, meaning that the incidence of the disease is attribut-
able to polymorphisms of multiple interacting genes.6,7 These multifactorial dis-
eases involve the interaction of many genes as well as gene–environment and
gene–diet interactions. Complex interactions of multiple internal and external
factors determine the probability of developing these multifactorial diseases.
Since the interactions can make it increasingly difficult to identify and charac-
terize what combinations of genetic variants are relevant to CVD and cancer, the
study of gene–diet interactions can help to establish the strongest relationships
between gene variations and phenotypes in healthy individuals in order to relate
their specific genotype to biomarkers of disease risk.5 Defining these pathophys-
iologic mechanisms in each individual can aid in identifying individuals at risk
for disease and to apply diet and lifestyle changes to modify disease risk factors.6
This chapter focuses on certain polymorphisms where the intake of a specific
nutrient related to that polymorphism affects the pathogenesis of CVD and/or
cancer.
8.2 GENETIC POLYMORPHISMS AND CVD

Our health is maintained through the integration of numerous complex homeo-
static physiological processes operating at the cellular, tissue, and organ level.8
Genes encode the proper amount of functional proteins and enzymes necessary
for these processes and, hence, a disease such as CVD may be thought of as a
result of failure of adequate homeostasis within a physiological system. This
failure may result from failure at the genetic level or from environmental exposure
or from an imbalance of the two. CVD most often does not result from a single
mutation in one gene, but since it is polygenic, it is more likely that CVD risk
is a result of many interacting mutations in several different genes (except for
single autosomal dominant inheritance, such as familial hypercholesterolemia).
A single polymorphism in a single gene likely plays a small role in CVD, but it
is the combination of several polymorphic genes that determines susceptibility

to CVD. Even though pharmaceutical agents can successfully normalize plasma
lipid concentrations, dietary modification is currently the cornerstone of primary
CVD prevention.9 The success of this approach is known to vary and depend on
individual responses to recommended dietary changes, which is determined by
gene–nutrient interactions. Hence, exposure to the same level of a dietary factor
can decrease the risk in one person, but not in another, depending on specific
gene variants. There are many genes involved in CVD pathogenesis that have
been documented; however, they are either not affected by diet or are still not
well established as risk factors.
8.2.1 APOLIPOPROTEIN E AND CVD

Apolipoprotein E (APOE), a constituent of triglyceride-rich chylomicrons, very
low-density lipoprotein (VLDL) particles, and their remnants,10 is a protein asso-
ciated with the metabolism of triacylglycerol-rich lipoproteins and high-density
lipoprotein (HDL).11 It also serves as a ligand for low-density lipoprotein (LDL)
receptors and LDL-receptor related proteins and, therefore, mediates chylomicron
and VLDL clearance from plasma.11,12 The APOE gene, located on human chro-
mosome 19, is 3.7kb in length and contains four exons. Genetic variation at the
APOE gene locus is an important determinant of serum LDL-cholesterol con-
centrations.13
Three common alleles of the APOE gene have been described: E2, E3, and
E4, which produce three isoforms of the protein E2, E3, and E4. These isoforms
differ in amino acid sequence at positions 112 and 158.14 E3 has a cysteine at
112 and an arginine at 158. E2 has a cysteine at both positions and E4 has an
arginine at both positions. Population studies have shown that plasma total cho-
lesterol and LDL cholesterol (LDL-C) concentrations are lowest in subjects with
the E2 allele, intermediate in those with the E3 allele, and highest in those with
the E4 allele.11 APOE allele frequencies vary, with the major allele in all popu-
lations being E3 (gene frequency of 0.49 to 0.91), followed by E4 (gene frequency
0.06 to 0.37) and E2 (gene frequency 0 to 0.15).12 The E4 variant has been
associated with increased cholesterol absorption, increased LDL production from
VLDL, and decreased bile acid synthesis and LDL clearance from plasma. The
E4 allele is also associated with a greater risk of coronary artery disease in both
men and women.
8.2.2 APOE GENOTYPE AND SATURATED FAT INTAKE

In Western societies, where diets are high in saturated fat and cholesterol, studies
have shown that the E4 allele is associated with higher LDL-C and triglyceride
and lower HDL cholesterol (HDL-C) concentrations.12 In 2001, Campos et al.
studied how the genetic variants of APOE would interact with habitual saturated
fat intake to affect the plasma lipid profile. Higher saturated fat intake was
associated with higher VLDL cholesterol, lower HDL-C, and smaller LDL
particles in E2 carriers, and the opposite or no effect was observed in the E4 and
E3 carriers. This suggests that carriers of the E2 variant allele may actually be
particularly susceptible to coronary disease when exposed to high saturated fat
diets. It was found that the E2 allele has a more detrimental influence on the
lipoprotein profile in individuals eating a high saturated fat diet compared to low
saturated fat, and the mechanism for this response may be due to the effect of
the E2 allele on receptor-mediated VLDL clearance. Compared to E3, E2 binds
poorly to lipoprotein receptors leading to delayed clearance and increased accu-
mulation of chylomicron and VLDL remnants.11
8.2.3 APOE GENOTYPE AND SUCROSE INTAKE

Erkkila et al.11 conducted a cross-sectional study to investigate the interaction
between APOE genotype and dietary intake of fat and carbohydrate (sucrose),
on serum cholesterol and triacylglycerol concentrations in patients with coro-
nary artery disease. The allele frequencies for E2, E3, and E4 were 0.039, 0.757
and 0.204, respectively. Patients had to complete 4-day food records and all
amounts and qualities of foods in the records were checked by a clinical
nutritionist for completion. Results showed that LDL-C and total cholesterol
were lower in patients with the E2 allele than in patients with the E3 and E4
allele, while serum triacylglycerol levels tended to be higher in E2 carriers. In
addition, a high intake of sucrose was associated with high serum triacylglycerol
only in patients with the E2 allele, while high triacylglycerol content was more
observed in E4 carriers with a low sucrose intake. The investigators concluded
that coronary artery disease patients with the E2 allele will likely have a greater
triacylglycerol response to high dietary sucrose intake than will patients with
the E3 or E4 allele.
8.2.4 METHYLENETETRAHYDROFOLATE REDUCTASE AND CVD

The enzyme methylenetetrahydrofolate reductase (MTHFR), a 70 kDA flavopro-
tein is a member of the MTHFR family.15 MTHFR catalyzes the conversion of
5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate (Figure 8.1), which
is a methyl donor group in the methylation of homocysteine to methionine.4–6,16,17
The human MTHFR gene is located on chromosome 1p36.3 and has a coding
region length of 1.980 bp.16 A common polymorphism of the MTHFR gene has
been associated with CVD risk and is well documented.18 The polymorphism,
which consists of a C→T change at nucleotide 677, results in an amino acid
substitution of alanine in place of valine.19 This amino acid substitution produces
an enzyme with decreased activity (approximately 35% of the value in individuals
without the mutation) and decreased thermal stability. The TT genotype is pre-
dictive of increased plasma homocysteine and an increased risk for CVD.20
Elevated plasma homocysteine has also been identified as a risk factor for car-
diovascular-related diseases of the coronary, cerebral and peripheral arteries, and
venous thrombosis.21 Although there have been some studies to show the contrary,
Deoxyuridylate
Diet Intake Folate Dihydrofolate
dUMP
Serine
Dimethyl Methionine Tetrahydrofolate Glycine

Glycine DNA
SAM RNA
Betaine Methyl protein MS
5,10-Methylene
Transferase Tetrahydrofolate
SAH B12
SCH3
MTHFR
Homocysteine 5-Methyltetra-
Betaine
hydrofolate Thymidylate
Cystathionine (circulating folate) dTMP
Choline B Synthase
B6
Cysteine DNA Synthesis
FIGURE 8.1 Schematic representation of folate/homocysteine/methionine metabolism in

DNA methylation and DNA synthesis. (Adapted from Steenge et al. (2003) Betaine
supplementation lowers plasma homocysteine in healthy men and women. J Nutr 133:
1291–1295; and Ma et al. (1999) A polymorphism of the methionine synthase gene:
association with plasma folate, vitamin B12, homocysteine, and colorectal cancer risk.
Canc Epidemiol Biomark Prev 8: 825-829.) Abbreviations: MS = methionine synthase,
SAM = S-adenosyl methionine, SAH = S-adenosyl homocysteine, MTHFR = methylene-
tetrahydrofolate reductase, dUMP = deoxyuridine 5′-monophosphate, dTMP = deoxythy-
midine 5′-monophosphate.
total homocysteine does seem to be a predictor of CVD, particularly in high-risk

populations, but is only weakly associated with risk in otherwise healthy sub-
jects.16 Homocysteine concentrations can be modulated by genetic and physio-
logical factors, diet, folate supplementation, riboflavin, choline, and vitamins B6
and B12.17
8.2.5 MTHFR 677 C→T POLYMORPHISM, FOLATE INTAKE,

AND CVD
Observational studies have shown that when folate status is less than adequate,
individuals with the MTHFR 677 TT genotype have lower blood folate and higher
plasma homocysteine concentrations than those with the MTHFR 677 CC gen-
otype.22–26 One of the most recent studies in this area, by Shelnutt et al.,27 recruited
only 677 CC and TT, nonpregnant healthy women, aged 20 to 30 years old and
placed them on folate deficient and folate rich diets. The folate depletion diet
provided approximately 115 µg dietary folate equivalents (DFE) per day for 7
weeks, and the folate repletion diet consisted of a combination of the depletion
diet, plus 400 µg DFE/d for 7 weeks. Serum folate concentrations did not differ
between the two genotypes at baseline. Results showed that TT individuals were
less capable of maintaining normal blood folate concentration during the deple-
tion phase than CC individuals.27 Mean plasma homocysteine concentration was
~3 µmol/l higher in TT subjects with low folate status postdepletion than in CC

subjects. Seven weeks of folate repletion was also insufficient to decrease the
mean plasma homocysteine concentration in women with the TT genotype.
Hence, this study, in which nutrient intake was controlled, supports the conclusion
that folate deficiency alone is sufficient to negatively affect plasma homocysteine
concentration in TT individuals.27
Studies by Ashfield-Watt et al.28 and de Bree et al.24 have both shown similar
results for men and women. Ashfield-Watt et al. placed subjects on three different
diets, each for 4 months. The “exclusion diet” was a usual diet, but folate-fortified
foods were replaced with unfortified foods, and a placebo tablet taken daily. The
“folate-rich diet” was a usual diet plus additional folate-fortified foods and nat-
urally folate-rich foods to reach a total of at least 400 µg/d. The ”supplement
diet” was the exclusion diet plus a 400 µg folate supplement daily instead of the
placebo. The folate supplement did have a significantly greater effect on plasma
folate than the folate-rich diet did, but the supplement had minimal additional
homocysteine-lowering capacity compared with diet alone.28 The folate-rich and
supplement interventions significantly increased plasma folate in TT individuals
above the baseline concentrations observed in those with the CC genotype. The
TT genotypes showed a greater homocysteine-lowering response to active folate
interventions than subjects with the other genotypes.
The study by de Bree et al. also supports this result. They showed that at any
folate intake, TT subjects have lower plasma folate than CT and CC subjects, yet
at high plasma folate concentrations, the homocysteine concentrations in TT
subjects are as low as those in CT and CC subjects. Overall, findings show TT
homozygotes have a greater requirement for folate to achieve homocysteine
concentrations comparable with the prevalent concentrations seen in CT and CC
individuals.28 These studies provide very useful data on folic acid fortification
for public health reasons and of supplementation, which is relevant for CVD
prevention.
8.2.6 HEPATIC LIPASE AND CVD

Hepatic lipase (HL), a lipolytic enzyme synthesized primarily in hepatocytes,
located on human chromosome 15q21, plays a critical role in lipid metabolism,
particularly in HDL metabolism.29–32 HL attaches to the sinusoidal endothelial
surface after being secreted from hepatocytes and hydrolyzes triglycerides and
phospholipids in plasma lipoproteins.30,31,33 HL has been implicated in regulating
HDL cholesterol levels, although the precise mechanism by which HL modulates
lipoprotein metabolism has yet to be characterized.34 It is thought that increased
HL activity is linked with generating small LDLs and contributes to its association
with low HDL because HL can also increase the hepatic uptake of HDL lipids.35
There have been several polymorphisms identified in the HL gene, including
some causing a rare HL deficiency condition.36,37 Recent studies have also shown
that polymorphisms in the HL gene promoter region are related to variations in
plasma HDL concentrations and that there are associations between HL gene
promoter polymorphisms and HL activity.38–41 A polymorphism in the HL gene,

which causes a C→T substitution at nucleotide 514, is favorable.38,39 The T allele
is associated with decreased HL activity, therefore, increasing HDL concentra-
tions.38,39 Hence, there is a good deal of research supporting the fact that rare
alleles of common promoter polymorphisms, such as the nucleotide 514 C→T
substitution, are associated with a significant decrease in plasma HL activity and
increased HDL levels.33,39,42
8.2.7 HL-514 C→T POLYMORPHISM AND CVD

Using a multiethnic population in Singapore and subjecting them to a highly
Westernized lifestyle, Tai et al.9 investigated the possible gene–nutrient interac-
tions between the −514 C→T polymorphism and dietary fat on HDL metabolism.
When there was no stratification by fat intake considered, the T allele was
associated with higher plasma HDL concentrations, higher triglyceride (TG)
concentrations and higher HDL/TG ratios. The TT subjects showed higher TG
levels only when fat intake was greater than 30% of total energy. An earlier study
by Ordovas et al.43 supports the results of Tai et al. using subjects from the
Framingham Study. They were also able to show that the T allele was associated
with significantly greater HDL concentrations only in those subjects consuming
less than 30% of energy from fat. When the total intake of fat was greater than
30% of energy, the mean HDL concentrations were lowest among those individ-
uals with the TT genotype, and no differences were observed between CT and
CC individuals.43 Overall, both studies indicate that the TT genotype is associated
with a more atherogenic lipid profile when subjects consume diets with a fat
content of at least 30% of total energy.9,43
8.2.8 STATINS, NIACIN, AND POLYMORPHISMS RELATED TO THE

LOWERING OF CHOLESTEROL
For individuals with lipoprotein abnormalities, lowering LDL-C continues to be
the primary target of cholesterol-lowering therapy. Lifestyle modification is the
first means of lowering LDL-C and drug therapy is reserved for those where
lifestyle modification is ineffective. 44 The most commonly used hypo-
cholesterolemic drugs are hydroxyl methylglutaryl coenzyme-A (HMG CoA)
reductase inhibitors, known as statins. The use of statins has become popular
since the 1990s due to their high efficacy and low secondary effects.45 Their
principal action is to lower plasma lipids via a considerable decrease in plasma
LDL-C as well as a slight decrease in triglycerides and a modest increase in HDL-
C.46–48 However, hypolipidemic agents show considerable individual variation in
their effects. The differences may be attributable to the interaction of environ-
mental and genetic factors.44,45 Among the genetic factors that may be responsible
for this individual variation, the APOE locus has been the most widely stud-
ied.44,49,50 There is a significant difference in response to statins associated with
the APOE locus. Subjects carrying the E2 allele have a significantly higher
response to LDL-C lowering and those carrying the E4 allele have a significantly
lower response.51 Plasma total cholesterol, LCL-C, and triglyceride responses
were significantly greater for E2 subjects as compared to E3 and E4 subjects
when taking atorvastatin, one of the most effective statins available.
Other polymorphisms involved in the individual response to statins are the
PvuII and AvaII polymorphisms of the LDL-receptor gene. These polymorphisms
have been shown to influence the levels of LDL-C in both normo- and hyperc-
holesterolemic subjects and may influence the response to treatment with statins.52
There has been minimal work done on these polymorphisms with conflicting
results. Studies have reported that subjects with the P2 allele of the PvuII poly-
morphism have a better response than those homozygous for the P1 allele, but
this has yet to be reproduced.45,53 Lahoz has shown a significant trend in the AvaII
polymorphism showing that those homozygous for the (−) allele responded less
than heterozygotes, who in turn responded less than homozygotes for the (+)
allele. Clinical importance for these polymorphisms is currently unknown and
requires further studies to determine an effect.
The −514C/T hepatic lipase and apolipoprotein A-1 promoter polymor-
phisms can also influence the effectiveness of statin therapy. Individuals who
are homozygous for the C allele had scarce modification of HDL-C with
pravastatin treatment while in carriers of the T allele, the increase in HDL-C
was almost 7%.54 However, it has been reported that those with the CC genotype
may benefit more from statins since they appear to have greater improvement
in coronary lesions than CT and TT genotypes. Carriers of the A allele for the
apoA-1 polymorphism showed HDL-C concentrations significantly higher than
G allele carriers with pravastatin treatment. The mechanisms by which the G/A
polymorphism influences HDL-C levels and its response to pravastatin are not
known. This is the case for many of the above-mentioned polymorphisms
interacting with statins. The value of determining genetic polymorphisms in
order to predict the effects of statin treatment is still unclear, but will improve
with further research.
Niacin (nicotinic acid) has been used as a lipid lowering agent to treat
dyslipidemia since 1954.55–57 Niacin is one of the most effective agents available
for raising HDL-C and lowering triglycerides and, hence, adding niacin to statin
therapy may be desirable. There has been reluctance to take immediate-release
niacin due to possible side effects of flushing and skin rash, but extended-release
niacin is both better tolerated and safer.58 Many studies suggest that combination
niacin–statin therapy leads to improved lipoprotein profiles and clinical outcomes
over monotherapy using either niacin or a statin.59,60 The addition of 2000 mg/d
of extended-release niacin to patients on statin therapy can decrease total choles-
terol an additional 21%, LDL-C by 31%, triglycerides by 27%, and increase
HDL-C by an additional 27%. Therefore, drug–nutrient interactions need to be
considered for each individual and these drug therapies could be more effective
if they are tailored better to individual needs.
8.3 GENETIC POLYMORPHISMS AND CANCER

There has been a wealth of evidence from epidemiological, clinical, and labora-
tory studies to suggest that nutrition and dietary factors can influence the risk for
the development of cancer, prognosis after diagnosis, as well as the quality of
life during treatment.61 It is estimated that many of the cancers in the U.S. are
related to nutritional and dietary factors.61,62 The molecular pathologies of cancer
are known to involve many steps involving the culmination of genetic changes
resulting from the interaction between genetics and the environment.61,63,64 The
great challenge has been to show the actual causative relationships and underlying
mechanisms that link dietary constituents to cancer risk.61 Although it has been
difficult to define relationships between nutritional factors and disease due to the
limitations of methodologies for assessing exposure to these factors, complexity
of dietary patterns, as well as the trouble in untangling the influence of dietary
constituents, it has been recognized that not all persons exposed to the same risk
factors will develop the associated disease.61,63,65,66 This is why it is now clear
that differential genetic susceptibility may explain the variation in response seen
among those with apparently similar diets or exposure to nutritional factors.
There are many genetic factors that can increase the risk for cancer, but the
most well-known are the highly penetrant, dominant mutations that epidemiolog-
ical studies show to have very high relative risks. However, in contrast with the
highly penetrant dominant mutations are the genetic mutations, which are not
sufficient to cause disease, but may affect cancer susceptibility or the response
to environmental exposure. These mutations can actually have a much greater
effect on a population, despite posing lower individual risk.64 The most studied
of these mutations are gene variants that influence metabolic activation, detoxi-
fication, or elimination of carcinogens. Some of these gene variants/polymor-
phisms can be relatively common in the general population (up to 53%) and are
known to play a role in the ability of an individual to withstand exposure to
carcinogens or to inhibit the initiation, promotion, or proliferation in carcinogen-
esis.67,68 With much speed, nutrition and cancer research is transitioning from an
observational approach to more of a molecular approach as the knowledge of
genomics and new technologies grows.69 Not only will nutritional genomics
determine which nutrient-related genetic and epigenetic changes cause pheno-
typic changes that influence cancer risk, but it will also establish which interac-
tions are the most important under certain circumstances.68 Although this area is
receiving more and more attention, it is still not entirely clear as to how genetic
polymorphisms are related to the impact of diet on the cancer process, but it is
plausible that polymorphic differences have been a contributing factor in the
inconsistencies surrounding diet and health.70 This section focuses on certain
genes and nutrients that are known to play a role in mitigating cancer based on
specific variations in diet–gene interactions and gene functions.
8.3.1 VITAMIN D AND THE VITAMIN D RECEPTOR

Vitamin D is an essential nutrient that is mainly produced in the skin as a
photoproduct of 7-dehyrocholesterol, or obtained through the diet via such foods
as dairy products (milk, cheese, butter) and seafood (fish, oysters).71,72 Other
sources of vitamin D include fortified milk products, breakfast cereals, and
vitamin D-containing multivitamins and supplements. Vitamin D is a prohormone
necessary for the regulation of serum calcium and phosphorous. One of the active
hormonal forms, 1,25-hydroxycholecalciferol [1,25(OH)2D] produced in the kid-
ney, enhances calcium absorption from the small intestine and is tightly regulated
to ensure calcium homeostasis. Vitamin D does undergo several hydroxylations
to become biologically active.73 The first hydroxylation occurs in the liver, at
carbon 25, to yield 25-hydroxyvitamin D [25(OH)2D]. Since this reaction is not
tightly regulated, 25(OH)D levels reflect exposure to sunlight and dietary intake
and, hence, plasma 25(OH)D is the best indicator of nutritional vitamin D
status.72–74
1,25(OH)2D is a specific ligand for the nuclear vitamin D receptor (VDR),
which is expressed in different tissues, especially in the intestine, bone, pancreas,
breast, prostate, pituitary, gonads, mononuclear cells, activated T lymphocytes,
and skin.73 Once 1,25(OH)2D binds with VDR, the resulting complex translocates
to the nucleus and interacts with respective vitamin D-responsive elements
(VDRE) within regulatory (promoter) regions; hence, initiating or repressing
transcription of various target genes. When VDR is bound to the 1,25(OH)2D
form, it transactivates genes that inhibit proliferation,75 promote differentia-
tion,76,77 or induce apoptosis.78,79 Vitamin D and its various forms have been
investigated as dietary anticarcinogens, and observational and cohort studies have
supported an inverse relationship between vitamin D and prostate, colorectal, and
breast cancers.65,72
Biological activity of circulating 1,25(OH)2D is modulated through variants
in the VDR gene, which was found to be polymorphic at multiple sites.80 These
polymorphisms include FokI (FF, Ff, ff), BsMI (BB, Bb, bb), TaqI (TT, Tt, tt),
and a 3’-untranslated region poly-A site (SS, Ss, ss) of the VDR gene.68,71,73,81
Except for FokI, all polymorphisms are in strong linkage disequilibrium with
each other and may potentially influence the expression or function of the VDR
protein. The FokI polymorphism influences the development of colorectal ade-
nomas.81 The BsMI B polymorphism and short poly-A polymorphisms have been
associated with increased breast cancer risk.68,80 BsMI has also been associated
with decreased risk of prostate cancer and benign prostatic hyperplasia.68,81 The
TaqI polymorphism may reduce the risk of lymph node metastasis, inhibit tumor
progression, and increase survival among ER (+), tamoxifen-treated women. TaqI
has also shown reductions in prostate cancer risk.71,83
8.3.2 VDR POLYMORPHISMS, VITAMIN D INTAKE, AND

COLORECTAL CANCER
Vitamin D had first been implicated as protective against cancer by ecological

studies linking colorectal cancer mortality to latitude, which correlates with
sunlight exposure.81,84 The ecological studies were then followed by analytic
epidemiologic studies of vitamin D and cancer, most of which found weak inverse
relationships between vitamin D status and colorectal cancer risk.81,85,86 It has
been suggested that vitamin D reduces epithelial cell proliferation and promotes
differentiation in various cell cultures.87,88 Other studies have also shown that
vitamin D induces apoptosis in colorectal tumor cell lines and premalignant
adenoma cells.89 Clinical evidence has shown that a high level of VDR expression
is associated with a favorable prognosis of colorectal cancer.90
Ingles et al.81 examined whether the 5′ start codon FokI polymorphism and
the 3′ UTR BsMI VDR polymorphism are associated with risk of colorectal
adenoma in a multiethnic population of men and women aged 50 to 74 with no
history of invasive cancer or inflammatory bowel disease. Cases had a first-time
diagnosis of one or more colorectal adenomas, confirmed by histology, and
controls had no past adenoma and were matched to cases by gender, age, and
date of sigmoidoscopy. The frequency of the FokI F allele was highest in African
Americans (74%) and the frequency of the BsMI B allele was highest in Cauca-
sians (42%). Only large, not small, adenomas were associated with the FokI
genotype, suggesting that this genotype may influence the progression of ade-
nomas to a more malignant phenotype. No association between the BsMI geno-
type and the risk of large adenoma was seen.81
A study by Peters et al.85 investigated the association of a VDR polymorphism
with colorectal adenomas and the interactions with vitamin D and calcium in
subjects between the ages of 18 and 74. Detailed information about sun exposure,
use of vitamin and mineral supplements, medical history, physical activity, and
alcohol and tobacco consumption was collected from each subject, along with
blood samples for measuring serum 25(OH)D levels and for FokI genotyping.
The results showed no evidence for an overall effect of the FokI polymorphism
and colorectal adenoma nor interaction with either serum vitamin D or calcium
intake; however, an inverse association of serum vitamin D with colorectal ade-
noma was found. A similarly designed study by Slattery et al.91 also found no
evidence for an inverse association with the FokI polymorphism and colon cancer,
but did find inverse associations for the SS poly-A, BsMI and TaqI polymor-
phisms, and colon cancer. Wong et al.84 have reported that the FokI variant actually
increases the risk of colorectal cancer in a Singapore Chinese population. Com-
pared to those individuals carrying the FF genotype, those with the Ff genotype
had a 51% increase in risk and those with the ff genotype had an 84% increase
in risk.
There are studies, though, that show a decrease in colon cancer risk based
on vitamin D-VDR interaction. Slattery et al.92 found the greatest reduced risk
for colon cancer was observed for individuals with a high intake of calcium,
vitamin D, and low-fat dairy products, and with the SS poly-A or BB BsMI
genotypes. Also, calcium, vitamin D, and low-fat dairy products were inversely
associated with rectal cancer in women, but not in men. These results are sup-
ported by Kim et al.93 who showed that those with the BB BsMI genotype and
the lowest calcium and vitamin D intake were at a reduced risk of colorectal
adenomas relative to those with the bb BsMI genotype and the highest levels of
intake, although this interaction wasn’t statistically significant.92 It has been
proposed that the antineoplastic activity of the BB and SS alleles is due to the
increased 1,25(OH)2D levels associated with those alleles.92 It may also be pos-
sible that different VDR genotypes interact differently with VDREs, which could
lead to altered ability to transcriptionally regulate target genes.
8.3.3 METHYLENETETRAHYDROFOLATE REDUCTASE, FOLATE

STATUS, AND CANCER
As mentioned earlier, MTHFR irreversibly converts 5,10- methylenetetrahydro-
folate to 5-methyl tetrahydrofolate, which is the major form of circulating folate
in plasma71 (see Figure 8.1). Besides being the major circulating form of intra-
cellular folate, 5,10-methylenetetrahydrofolate is also the cofactor for methylating
deoxyuridylate to produce thymidylate, the rate-limiting nucleotide in DNA syn-
thesis (see Figure 8.1). Decreasing the availability of MTHFR increases the
availability of methylenetetrahydrofolate and, therefore, reduces the chances of
insufficient thymidylate and misincorporation of uracil into DNA.71 The reduced
incorporation of uracil into DNA leads to less chromosome breaking, and possibly
a lower cancer risk.94,95
There are many common genetic polymorphisms that appear to modulate
cancer risk through their influence on folate metabolism. Two polymorphisms in
the MTHFR gene, one consisting of a C to T substitution at nucleotide 677
(alanine to valine), and another consisting of an A to G substitution at nucleotide
1298 (glutamate to alanine) both result in a reduction in MTHFR activity.
Methionine synthase (MS) catalyzes the transfer of a methyl group from 5-methyl
tetrahydrofolate to homocysteine, producing methionine and tetrahydrofolate (see
Figure 8.1), which is necessary for maintaining an adequate level of intracellular
methionine, the precursor of the methyl donor S-adenosyl methionine (SAM).71,96
SAM is an important methyl group donor that is involved in over 100 methylation
reactions that includes DNA methylation. A polymorphism of the methionine
synthase gene (MTR), consisting of an A to G substitution at nucleotide 2756,
results in a conversion of aspartate to glycine and reduces MS activity. A lack of
sufficient SAM can lead to DNA hypomethylation and modify cancer risk.96 These
polymorphisms can have different effects on cancer pathogenesis depending on
individual folate status.
8.3.4 MTHFR 677 C→T POLYMORPHISM, FOLATE INTAKE,

AND COLORECTAL CANCER
Because colorectal carcinomas are derived from rapidly proliferating tissues, they
have a great requirement for DNA synthesis and are affected by the metabolic
fate of folic acid. Epidemiological studies have shown that colorectal cancer was
reduced (~50%) in individuals with the MTHFR 677 TT genotype when folate
intake was adequate, compared to those with the MTHFR 677 CC genotype.71
Slattery and Potter96 analyzed a mixed-ethnicity population of individuals aged
30 to 79 who were diagnosed with first primary colon cancer matched with control
subjects. Dietary food frequency questionnaires were distributed. After genotyp-
ing for the MTHFR 677 C→T polymorphism, frequencies of the CC, CT, and
TT genotypes among all cases were 45.9%, 44.7%, and 9.5%, respectively, and
were very similar for the controls. High intakes of folate were associated with a
30 to 40% risk reduction in colon cancer among those with the TT genotype
relative to those with low folate intake who were CC genotypes.97 Shannon et
al.98 provided further support by showing that the MTHFR TT genotype conferred
an increased risk of colorectal cancer in subjects older but not younger than 70
years of age. Hence, the variant TT form of the MTHFR gene appears to be
slightly inversely associated with colon cancer, while the heterozygote CT and
wild-type CC genotypes appear to have similar associations with colon cancer.97
8.3.5 MTHFR 677 C→T AND 1298 A→C POLYMORPHISMS

AND ACUTE LEUKEMIA
The etiology of most types of leukemia is still unknown, but it is unlikely to be

caused by a single genetic defect, and is most likely a result of adverse gene-
environment interactions, with susceptibility being related to polymorphisms in
multiple genes.95 Skibola et al. have hypothesized that there may be a correlation
between functional polymorphisms in the MTHFR gene and leukemogenesis due
to the association between folate status and susceptibility to genetic damage in
dividing cells. Since leukemias, like colorectal carcinomas, are also derived from
rapidly proliferating tissues, they have a great requirement for DNA synthesis
and are likely to be affected similarly by the metabolic fate of folic acid. These
investigators designed their study to evaluate what role, either individually or
combined, the 677 C→T and 1298 A→C MTHFR polymorphisms play in leu-
kemia susceptibility. They hypothesized that the carriers of the variant alleles
may have a protective advantage against leukemia. Case individuals consisted of
those diagnosed with acute leukemia and were matched with controls. After
genotyping, the MTHFR 677 CC, CT, and TT genotype frequencies for cases
and controls were 57.9%, 32.5%, and 9.6%, and 53.2%, 35.0%, and 11.8%,
respectively. The MTHFR 1298 AA, AC, and CC genotype frequencies for cases
and controls were 49.8%, 41.9%, and 8.3%, and 45.0%, 44.3%, and 10.7%,
respectively. Results demonstrated that individuals with at least one MTHFR
mutation at 677 or 1298 were less likely to develop acute lymphocytic leukemia.
When examining joint effects between the two polymorphisms, it was found that
the double heterozygotes (677 CT/1298 AC) were approximately five times less
likely than 677 CC/1298 AA individuals to develop acute lymphocytic leukemia.95
8.3.6 MTR 2756 A→G POLYMORPHISM AND CANCER

The MTR 2756 GG genotype has been reported to be associated with a reduced
risk of adenomas in colorectal cancer, up to a 41% lower risk than in MTR
2756A.71,96,99 The most recent study, by Ulvik et al., genotyped 2179 subjects with
colorectal cancer matched with controls for age and gender. In this Norwegian
population, the MTR variant form GG was associated with a significant risk
reduction of more than 25%, and the same was seen for the variant MTFHR 677
TT genotype. Specifically for the MTR AG genotype, the risk was considered
intermediate. A similar study by Ma et al.96 found that the MTR GG genotype
was associated with a 40% decreased risk of colorectal cancer, but was not
associated with plasma levels of folate or total homocysteine. A possible expla-
nation for this lack of association is that homocysteine may be remethylated to
methionine through an alternative pathway by betaine methyltransferase96 (see
Figure 8.1). This pathway adds yet another nutrient, choline, to the equation,
which could have a role in modulating the processes involved with folate defi-
ciency, DNA methylation, and cancer. This study, however, took into account
alcohol intake since ethanol can interfere with folate and methyl group metab-
olism as well as MS activity. Colorectal cancer risk conferred by high alcohol
intake may overcome the protective effect of these polymorphisms. Out of 10
cases and 21 controls that had the MTR GG genotype, the risk was 10-fold higher
in those men who drank one or more drinks per day than those who drank less.96
8.4 CONCLUSIONS
In this new millennium, research in nutrition and disease prevention must now
prioritize in understanding the molecular basis by which nutrients affect the
disease process.68,100,101 A well-coordinated, multidisciplinary effort must be made
amongst all types of scientists — nutritional scientists, molecular biologists,
geneticists, statisticians, and clinical cancer researchers — in order to move this
research forward. There are many more genes related to the pathogenesis of
cancer and CVD that have not been included in this paper because there hasn’t
yet been information for those genes showing relevance for dietary interventions.
For each gene there may be one or more polymorphisms associated with it, hence,
creating even more possible interactions between genetic polymorphisms and
diet. This creates an ongoing challenge in trying to solve multifactorial, polygenic
diseases. The individualization of treatments has been one of the strongest ther-
apeutic tools, yet also one of the biggest challenges of the pregenomic era.
However, now, with the availability of comprehensive genetic and molecular
profiles and with improved molecular techniques, it will be possible to respond
to diseases with individual strategies based on genetic profiles.16 Nutrigenomics
has a powerful message — food and nutrients have the potential to affect the
balance between health and disease by affecting gene expression and function.102
Exploring the role that functional gene polymorphisms play in determining
risk and in determining the levels of intermediate phenotypes is important to our
understanding of the key metabolic pathways and physiology in both the disease
and disease-free states.7 More research in this area is still required, as there are
many more genetic polymorphisms and nutrient interactions that are related to
cancer and CVD. As was seen for the added benefit of including niacin with
statins for treating hypercholesterolemia, there is great potential to improve treat-
ment by adding a simple nutrient to drug therapy. These benefits will vary
according to each individual’s genotype. Yet, also, there is the potential for adverse
effects with adding a nutrient to a drug therapy, which can also vary depending
on one’s genotype; hence, caution is required as well. It seems that only the
surface has been scratched in the pursuit of solving the genetic–nutrient–drug
mechanisms of CVD and cancer, but a great deal of research will undoubtedly
follow. The key now is for researchers to always think of nutrient, drug, and gene
interactions when designing experiments because the three are no longer solely
independent of each other, but inevitably linked.
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Available online: http://www.dietitians.ca
9 Nutrigenomics and
Pharmacogenomics of
Human Cancer: On the
Road from Nutrigenetics
and Pharmacogenetics
Alexandre Loktionov
CONTENTS
9.1 Introduction..............................................................................................262
9.2 Diet, Genetic Background, and Cancer Risk..........................................263
9.2.1 Xenobiotic Metabolism-Controlling Pathways ...........................264
9.2.2 Folate Metabolism and DNA Methylation .................................271
9.2.3 Alcohol Metabolism ....................................................................272
9.2.4 DNA Repair.................................................................................273
9.2.5 Energy Balance Regulation .........................................................274
9.2.6 Other Physiological Pathways, Genes, and
Research Perspectives..................................................................274
9.3 Advances in Cancer Pharmacogenomics and
Links to Nutrigenomics...........................................................................276
9.3.1 Drug-Metabolizing Phase I and Phase II Enzymes....................276
9.3.2 Enzymes of Purine/Pyrimidine Metabolism ...............................290
9.3.3 Enzymes and Regulatory Factors of Folate Metabolism............291
9.3.4 DNA Repair Effects on Cancer Treatment .................................291
9.3.5 ATP-Binding Cassette Transporters ............................................292
9.3.6 Other Drug Targets and Recent Advances..................................293
9.4 Conclusion ...............................................................................................294
References .........................................................................................................295
261
9.1 INTRODUCTION
Recent rapid progress in the determination of the human genome sequence has
strongly stimulated the development of genomic approaches to the investigation
of a broad spectrum of medical problems.1,2 Functional genomics is now expand-
ing into several bioscientific domains, which previously had little relation to
genetic research. Emerging amalgamation of functional genomics with nutritional
sciences has recently been called nutrigenomics; however this new discipline is
just entering its embryonal state, being mainly discussed in numerous reviews
devoted to future opportunities rather than existing reality.3–6 This dream-like
state of the subject has brought about a very vague concept of suggested bound-
aries of nutrigenomics, which often tend to incorporate transcriptomics, proteom-
ics, and, eventually, metabolomics. However, the integrative ideas, more appro-
priately attributable to the systems biology, still have little impact on our
knowledge regarding the relationship between human genome, nutritional factors,
and genesis of major human chronic diseases, such as cardiovascular disease and
cancer.
Cancer remained one of the most enigmatic conditions still challenging sci-
entists and physicians at the beginning of the new millennium. Although it is
generally accepted that genetic component plays a leading role in the pathogenesis
of neoplastic disorders, only a minor fraction of all human cancers (i.e., hereditary
cancer syndromes accounting for 5 to 10% of all cancer cases) inevitably results
from specific deteriorating mutations transmitted through germ line.7,8 In contrast,
the development of sporadic tumors is regarded as a complex pathogenetic process
involving interactions of multiple contributing factors, both internal (intrinsic host
factors) and external.9 In this context, diet obviously constitutes an omnipresent
combination of external influences potentially important for all malignancies at
different stages of neoplastic growth. Furthermore, dietary factors can interact
with anticancer drugs used at later stages of the disease, thus affecting chemo-
therapy outcome.
Although it would be extremely tempting to have a chance to look at cancer
pathogenesis and medicamentous treatment through the prism of genome-gov-
erned network of interactions between nutrition, chemotherapeutic agents, and
biological systems in all their complexity, the existing knowledge is obviously
insufficient to allow any fact-based analysis of this type. In the context of this
chapter, it appears to be justified to use a limited meaning of the term “nutrige-
nomics,” analogous to “pharmacogenomics,” which is often defined as a field
aiming to investigate the genetic basis for interindividual differences in drug
response using genome-wide approaches.10 It is, however, clear that the effects
of drugs, which are administered as pure compounds in precise doses, are much
easier to assess than combined and interfering effects of numerous food constit-
uents.4 Nevertheless, this chapter is an attempt to analyze the present state of
nutrigenomic and pharmacogenomic research in relation to human cancer.
Nutrigenomics and Pharmacogenomics of Human Cancer 263
9.2 DIET, GENETIC BACKGROUND, AND CANCER RISK

It is generally accepted that diet can affect cancer risk in humans. The relationship
had been well demonstrated by epidemiological studies; however, precise bio-
logical mechanisms involved in these interactions remain obscure since simulta-
neous determination of various biological effects exerted by thousands of food
components ingested daily is a formidable problem.11–13 At this level of complex-
ity, application of the traditional reductionist approach attempting to separately
analyze effects of individual diet-derived bioactive substances could easily lead
to flawed conclusions. Persistent lack of clear understanding of the cancer patho-
genesis further aggravates the situation.
Figure 9.1 roughly illustrates the present state of general scientific knowledge
regarding the relationship between diet and neoplastic growth (the scheme also
includes the main therapeutic strategies presently applied in the management of
cancer). It is transparent that the existing views on the diet–cancer relationship
are mainly focused on the very beginning of the disease, i.e., cancer initiation,
presuming that dietary composition shifts towards patterns corresponding to either
“protection/prevention” or “high-risk/predisposition” can respectively reduce or
increase diet-related risk of tumor development. This concentration of attention
on the borderline separating physiological (normal) conditions from neoplasia
has an obvious positive side, strongly stimulating research in the direction of
dietary cancer prevention. At the same time, relatively little is known about effects
of diet in cancer patients with the exception of some described interactions
between dietary components and chemotherapeutic drugs, which are going to be
considered from a nutrigenomic/pharmacogenomic point of view later in this
chapter.
Identification of dietary components affecting cancer risk in humans has
always been a difficult task. It was widely accepted for decades that excessive
consumption of fat and red meat and insufficient intake of fruits, vegetables,
and dietary fiber increase cancer risk.11 However, discrepancies between positive
results of earlier case-control studies (e.g., those indicating risk-increasing
effect of dietary fat and protective action of fruits, vegetables, and dietary fiber)
and failure to confirm these findings in recent large-scale prospective trials11–15
create new uncertainties in the field. Among “traditional” dietary risk factors
only relationship between consumption of red meat and increased risk of distal
colon cancer has been clearly confirmed by several large-scale prospective
studies.16–18 Unexpected results of the prospective studies have already gener-
ated intense discussions with regard to the use of different methodologies of
data collection in human nutrition studies.19,20 In the meanwhile, new aspects
of the problem emerge. Links between overeating/obesity and risk of cancer of
several sites (colon, breast, esophagus, endometrium, kidney) have been repeat-
edly reported.11,12,21–23 The latter area is especially important in view of the
obesity “epidemic” rapidly spreading in the developed countries of Western
Europe and North America.24
Dietary influences
Carcinogenic Protective
Effects on the malignant growth are largely unknown Life support
Protective Carcinogenic (interactions with chemotherapeutic drugs possible) (anti-cahexia
measures)
Cancer Early cancer Cancer Advanced Terminal

initiation (cancer promotion) progression cancer cancer
HEALTH N E O P L A S T I C G R O W T H
Period of efficient surgical treatment
Main therapy Palliative

therapy
Adjuvant
Chemotherapy
therapy
and
Radiotherapy
FIGURE 9.1 Schematic representation of neoplastic growth dynamics and influences of

diet and anticancer treatments at its different stages.
The absence of complete clarity with regard to cancer-affecting dietary com-

ponents makes application of systems biology approaches to nutritional science
a challenging task for the future. The present knowledge in the field widely
defined as nutrigenomics (see above) is limited by often patchy information on
certain human genetic variants influencing (patho)physiological responses to the
action of bioactive food components. Analysis of these (essentially nutrigenetic)
observations is presented below.
9.2.1 XENOBIOTIC METABOLISM-CONTROLLING PATHWAYS

The problem of associations between common gene polymorphisms, dietary
influences, and cancer risk has attracted researchers since the early 1980s, when
discovery of polymerase chain reaction made real analysis of human genetic
variants possible. The prevailing idea of the field could be briefly summarized as
the search for common gene variants responsible for deficient or altered metab-
olism of food-derived carcinogenic substances. Enzymatic pathways controlling
metabolism of xenobiotics (including carcinogens) had been the main targets of
these studies.25 Special attention was paid to the genes encoding enzymes partic-
ipating in Phase I and Phase II reactions. Reactions of Phase I, which often lead
to procarcinogen activation, are mostly catalyzed by the cytochrome P450 (CYP)
superfamily of enzymes providing catalytic hydroxylation of diverse compounds
linked only by their lipophilic nature.26,27 Numerous single nucleotide polymor-
phisms (SNPs) in the CYP genes (CYP1A1, CYP1A2, CYP1B1, CYP2A6
CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4, and CYP3A5) have been
studied in relation to cancer risk; however, no firm evidence in favor of this link
has been produced.28 It should be noted that the initial concept of the P450 family,
regarding it mainly as a hepatic xenobiotic detoxification system, is now being
expanded upon the discovery of multiple additional functions of these
enzymes.26,27 It also has become clear that some CYPs are closely involved in
the metabolism of anticancer chemotherapeutic agents.28 These interactions are
going to be addressed later in this chapter.
Available information on interactions of dietary factors with polymorphic
variants of the CYP genes is presented in the beginning of Table 9.1. Although
the number of studies approaching gene–nutrient interactions as a factor in human
cancer development is limited, it is obvious that polymorphic genes of the P450
family have been investigated mostly in the context of association between cooked
red meat consumption and colorectal neoplasia. Several reports indicate possible
risk-modulating effects of CYP gene variants (see Table 9.1), which might be in
line with the idea of enhanced activation of meat processing-generated carcino-
gens, such as polycyclic aromatic hydrocarbons (PAH), heterocyclic amines
(HCA), and N-nitroso compounds by some variant CYP enzymes.70–72
However, reported effects and interactions are usually weak and often difficult
to reproduce. Moreover, the complexity of human diets and abundant presence
of multiple bioactive dietary components makes investigation of isolated
gene–nutrient interactions notoriously difficult. Furthermore, effects of different
gene polymorphisms often overlap as several examples given in Table 9.1 show.
For instance it is very difficult to separate influence of CYP family (especially
CYP1A1 and CYP1A2) and N-acetyltransferase (NAT1 and NAT2) genes on
colorectal cancer risk associated with cooked meat consumption. CYP family
genes encode numerous xenobiotic-metabolizing enzymes, but CYP3A4, being
the most abundant P450 enzyme in the human liver and gut73–75 and a major factor
in drug metabolism, deserves particular attention. Although over 70 SNPs are
recently described in the corresponding gene,75 knowledge of their physiological
significance is still very scarce. Correlations between CYP3A4 gene polymor-
phisms and cancer risk only start to emerge,75 and there is little doubt that further
investigation of this gene can produce abundant information on carcinogenesis,
responses to cancer treatment, and modulation of these processes by active com-
ponents of diet (see also Chapter 2, Hypolipidemic Therapy: Drugs, Diet, and
Interactive Effects).
Whereas polymorphic xenobiotic-metabolizing enzymes of Phase I are
mostly responsible for a generation of active carcinogenic substances, Phase II
metabolism is directed on detoxification of carcinogenic and mutagenic sub-
stances through conjugation reactions producing soluble and easily excretable
products. Inherited deficiency of some Phase II enzymes, such as “Null” variants
of glutathione S-transferases M1 (GSTM1) and T1 (GSTT1) caused by long
deletions in the corresponding genes, has repeatedly been reported to be associ-
ated with increased risk of cancers of different sites.76 GST family enzymes are
strongly involved in the metabolism of PAH,77 thus changes in their activity
are likely to affect the probability of cancer initiation. Table 9.1 shows that
266
TABLE 9.1
Interactions between Common “Metabolic” Gene Variants and Dietary Factors in Cancer Risk Modulation
Gene Tumor Site/Type Dietary Factor(s) Interactive Effects Refs.
Genes Encoding Phase I Metabolism Enzymes

CYP1A1 Ovarian cancer Caffeine Elevated ovarian cancer risk related to increased caffeine intake was observed (29)
in individuals with the CYP1A1*2C allele.
Colorectal cancer White meat No clear pattern of CYP1A1 polymorphism influence on colorectal cancer (30)
risk related to white meat consumption revealed.
CYP1A2 Colorectal cancer Cooked red meat Well-done red meat consumption was associated with an increased risk of (31)
colorectal cancer among ever-smokers with the CYP1A2 rapid phenotype
(and NAT2 rapid).
CYP1A2 rapid phenotype is associated with certain genetic variants, e.g., (32)
CYP1A2*F.
Ovarian cancer Caffeine The associations of caffeine and coffee intake and ovarian cancer risk were (33)
stronger in women with CYP1A2 (high-rapid) genotype.
CYP2A6 Colorectal cancer Cooked red meat CYP2A6 high activity was associated with an increased risk of colorectal (34, 35)
cancer, particularly among consumers of well-done meat (see also effect
of GSTA1 genotype).
Information on the role CYP2A6 gene variants in the enzyme activity (36)
modulation is scarce; however, the effect appears to be likely.
CYP2E1 Colorectal cancer Red meat and Individuals with a 5’ 96-bp promoter region insertion variant of the CYP2E1 (37)
processed meat gene had an increased rectal cancer risk associated with high red meat or
processed meat consumption.
Incomplete intestinal Salted food (salted Association between incomplete intestinal metaplasia and salted food intake (38)
metaplasia (stomach meat, dehydrated was observed among subjects bearing homozygous c1/c1 CYP2E1 variant
cancer precursor) salted vegetables, (defined by RsaI polymorphism in 5′ region). (See also effects of GSTM1
salted raw fish) and GSTT1 genotypes.)
Upper aerodigestive Alcohol The highest risk of oral cavity/pharyngeal cancer was observed among the (39)
tract cancer heaviest drinkers with either the CYP2E1 C (defined by DraI
polymorphism in intron 6) or the CYP2E1 c2 (defined by RsaI
polymorphism in 5′ region) allele.
Myeloperoxidase Breast cancer Antioxidant-rich fruits The A-allele (G463A promoter region SNP) was associated with reduced (40)
(MPO) and vegetables breast cancer risk in women (especially premenopausal) who consumed
higher amounts of fruits and vegetables.
Genes Encoding Enzymes with Dual (Predominantly Phase II, but Some Phase I Metabolism) Function
N-acetyltransferase 1 Breast cancer Cooked red meat Elevated breast cancer risk related to increased well-done red meat (41, 42)
(NAT1) consumption was observed in women with putative rapid acetylation-
associated NAT1*11 or NAT1*10 alleles.
Colorectal adenomas Cooked red meat Stronger association of well-done red meat consumption with colorectal (43)
adenoma development was observed in individuals with rapid acetylation-
related NAT1 genotypes.
N-acetyltransferase 2 Colorectal cancer Cooked red meat Well-done red meat consumption was associated with an increased risk of (31)
(NAT2) colorectal cancer among ever-smokers with NAT2 rapid phenotype (and
Nutrigenomics and Pharmacogenomics of Human Cancer
CYP1A2 rapid phenotype).

Colorectal adenoma Cooked red meat Frequent meat consumption combined with the NAT2 slow acetylation (44)
slightly increased colorectal adenoma risk.
Breast cancer Cooked red meat Rapid/intermediate NAT2 genotype combined with consumption of well- (45)
done meat was associated with a nearly eight-fold increase of breast cancer
risk.
No NAT2 genotype or meat consumption effect on breast cancer risk found. (46)
267
268

Hepatocellular cancer Red meat A trend of increased hepatocellular carcinoma risk with increasing red meat (47)
intake was observed in rapid acetylators (defined here as subjects
homozygous for the wild type NAT2*4 allele).
Sulfotransferase 1A1 Breast cancer Red meat The risk of breast cancer was elevated with increasing doneness of red meat (48)
(SULT1A1) intake among women with the Arg/Arg or Arg/His genotype, but not with
the His/His homozygocity (lower enzyme activity-associated) at codon 213.
Prostate cancer An increased risk of prostate cancer was associated with the presence of the (49)
*1/*1 (His/His at 213) SULT1A1 genotype; however, there was no clear
correlation among genotype, meat consumption, and cancer risk.

Microsomal epoxide Colorectal adenomas Cooked red meat Elevated adenoma risk associated with meat consumption and high predicted (50)
hydrolase (EPHX1) EPHX1 activity (Tyr at codon 113 and Arg at codon 139).
Increased adenoma risk was found to be associated with high meat intake (51)
and His 113 genotype.
Lung cancer Intake of protein, Among smokers with high animal fat and protein intake and low (52)
carbohydrates, carbohydrate and fiber intake (“unhealthy” dietary pattern) individuals with
animal fat, and His/His 113 homozygocity were at significantly lower lung cancer risk.
dietary fiber
Genes Encoding Phase II Metabolism Enzymes

Glutathione Colorectal cancer Well-done meat intake GSTA1*B allele, which causes lower expression of the enzyme, was (34)
S-transferase A1 associated with an increased risk of colorectal cancer, especially among
(GSTA1) well-done meat consumers.
Glutathione Lung cancer Intake of Increased intake of isothiocyanates was associated with cancer-protective (53–55)
S-transferase M1 isothiocyanate-rich effect in GSTM1-Null individuals.
(GSTM1) cruciferous
vegetables (esp.
broccoli)
Increased intake of isothiocyanates was associated with cancer-protective (56)
effect in GSTM1-positive individuals.
Colorectal cancer Increased intake of cruciferous vegetables was associated with cancer risk (57)
reduction in GTSM-Null individuals.
Increased intake of isothiocyanate-rich cruciferous vegetables was associated (58)
with cancer risk reduction in individuals with the GSTM1-Null and GSTT1-
Null combination.
Colorectal adenomas Protective effect of broccoli was observed in subjects with GSTM1-Null (59)
genotype.
Prostate cancer Men with GSTM1-positive genotype had the greatest reduction of cancer (60)
risk.
Head and neck cancer Consumption of raw cruciferous vegetables was associated with cancer (61)
incidence reduction only in GSTM1-Null individuals.
Breast cancer Consumption of cruciferous vegetables, particularly broccoli, was (62)
marginally protective, but risk was unaffected by GSTM1 genotype.
Glutathione Lung cancer Red meat Among smokers with high animal fat and protein intake and low (52)
S-transferase P1 carbohydrate and fiber intake (“unhealthy” dietary pattern) individuals with
(GSTP1) Ile/Ile 105 homozygocity were at significantly higher lung cancer risk.
Glutathione Lung cancer Intake of Low intake of isothiocyanates was associated with marginally increased (53)
S-transferase T1 isothiocyanate-rich cancer risk in GSTT1-Null individuals.
(GSTT1) cruciferous
vegetables (esp.
broccoli)
269
270

Increased intake of isothiocyanates was associated with cancer-protective (54, 55)

effect in individuals with simultaneous homozygous deletion of both
GSTM1 and GSTT1.
Colorectal cancer Increased intake of isothiocyanate-rich cruciferous vegetables was associated (58, 63, 64)
with cancer risk reduction in individuals with the GSTM1-Null and GSTT1-
Null combination.
Manganese superoxide Breast cancer Antioxidant-rich fruits Women homozygous for the Ala(-9) allele (Val-9Ala polymorphism in the (65, 66)
dismutase (MnSOD) and vegetables and signal sequence of the MnSOD protein), especially those with lower
dietary supplements antioxidant intake, were at higher risk of developing breast cancer.
No correlation of breast cancer risk with either MnSOD genotype or (67)
antioxidant intake.
Prostate cancer Beta-carotene Homozygocity for the Ala(-9) in combination with high antioxidant levels (68)
treatment, was associated with lower relative risk for both total and fatal prostate
antioxidants cancer.
Catalase (CAT) Breast cancer Fruit and vegetable The high-activity (wild type, CC) catalase homozygocity in combination (69)
consumption with higher vegetable and, especially, fruit consumption was associated
with breast cancer risk reduction.
interactions of food components with genetic variants of these enzymes were

mostly approached from the point of view of nutritional cancer prevention. Efforts
of investigators were concentrated predominantly on cancer-protective effects of
isothiocyanate-rich cruciferous vegetables, which are believed to influence both
Phase I and Phase II xenobiotic metabolism; in particular, through induction of
GSTM1 and GSTT1.78,79 Most of the cited studies revealed stronger protective
effect of isothiocyanates in GST-deficient individuals (GSTM1-Null and GSTT1-
Null genotypes); therefore, it can be suggested that this effect can be explained
by compensatory activation of other GST family enzymes (e.g., GSTA1) provid-
ing an “overlapping” detoxifying effect.80
UDP-glucuronosyltransferases (UGTs) constitute another group of Phase II
enzymes responsive to dietary effects.81,82 Highly polymorphic UGTs exert con-
jugation of a variety of xenobiotics, including carcinogenic HCA and PAH, and
complex dependence of enzyme activity of nutritional factors and polymorphisms
in the genes encoding UGT1A181,83–85 and UGT1A786 has already been reported.
Genes encoding enzymes that participate in the generation of reactive oxygen
species (e.g., myeloperoxidase) and protection from its effects (superoxide dis-
mutases, catalase, glutathione peroxidase) are considered here, together with
those controlling Phase I and Phase II reactions, since oxidative stress is a known
factor in carcinogenesis.87 First studies linking variants of these genes with dietary
factors indicate the importance of fruit- and vegetable-derived antioxidants in
modulating cancer risk in a genotype-dependent manner (see Table 9.1).
It should be repeated that the modest size of Table 9.1 reflects the fact that,
despite abundance of epidemiological studies approaching relationship between
“metabolic” (i.e., Phase I and II metabolism-related) gene polymorphisms and
cancer risk, relatively few authors have addressed the possible contribution of
dietary factors to this relationship.
9.2.2 FOLATE METABOLISM AND DNA METHYLATION

Genes that encode enzymes of folate metabolism are especially interesting in
view of their contribution to DNA methylation, which directly influences gene
expression and, therefore, the processes of normal and neoplastic cell growth and
differentiation.88 At the same time, folate and related nutrients (group B vitamins,
methionine, choline), being dietary constituents, provide a direct link to nutrition.
Abundance of useful information regarding this metabolic pathway is not sur-
prising. Considerable attention has been paid to the role of two common SNPs
(C677T and A1298C) in the sequence of the methylenetetrahydrofolate reductase
(MTHFR) gene in cancer susceptibility. It has been reported that interaction of
folate status determined by dietary intake with the C677T polymorphism influ-
ences human DNA methylation,89 whereas mechanisms behind possible effects
of the A1298C SNP remain to be explored further.90 This may depend generally
on the genetic linkage between the two alleles,91 since the MTHFR variant
encoded by the C1298 (Ala429) appears to be functionally indistinguishable from
the wild-type enzyme.92
There are numerous publications on colorectal cancer risks in relation to the

T/T677 MTHFR homozygocity considered in detail in several recent reviews.93–96
Briefly, the T/T677 genotype effect on colorectal cancer risk strongly depends
on adequate folate status. The T/T homozygocity is associated with reduced risk
of colorectal cancer in individuals with high folate intake and low alcohol con-
sumption, whereas combination of low folate and high alcohol consumption
increases colorectal cancer risk in this genotype group, even in comparison with
individuals bearing the C/T or C/C677 allele combinations.93–96 Essentially a
similar pattern has been demonstrated in several breast cancer studies97–99; how-
ever, other investigators failed to reveal an effect of folate consumption, while
observing reduction of breast cancer risk in postmenopausal women with T677
homozygocity and receiving hormone replacement therapy.100 There are also
reports indicating that the combination of T/T677 homozygocity and low folate
intake may be regarded as a risk factor for cervical101 and bladder102 cancer.
It is likely that polymorphic variants of other genes encoding enzymes
involved in folate metabolism (methionine synthase, methionine synthase reduc-
tase, cystathionine β-synthase, thymidylate synthase) can affect interaction
between consumption of folate and associated nutrients and cancer risk,95 but
available information is not sufficient to allow arriving at firm conclusions. Nev-
ertheless, a common polymorphism in methionine synthase gene (Asp919Gly)
was shown to affect colorectal adenoma risk through interaction with methionine
and alcohol intake.103 Likewise, there are reports on the diet-dependent influence
of thymidylate synthase gene promoter polymorphisms on colorectal adenoma104
and lung cancer105 risks, as well as of methionine synthase reductase A66G SNP
on lung cancer risk.106
9.2.3 ALCOHOL METABOLISM

Alcohol consumption is a common dietary habit repeatedly shown to be associ-
ated with tumors of several sites.107 As it has already been mentioned, alcohol
consumption is known to interact with folate intake and genotype of folate-
metabolizing enzymes, presumably through alterations in one-carbon metab-
olism.108 It is also known that polymorphic variants of alcohol dehydrogenases
(ADHs) and aldehyde dehydrogenases (ALDHs), playing central roles in the
metabolism of ethanol and its main metabolite, acetaldehyde, can influence risk
levels for alcohol drinking-related cancers.109 It is, however, difficult to distinguish
between effects of different alleles of genes encoding these enzymes on carcino-
genesis per se and their influence on drinking habits.110 Indeed, ADH2*1 allele
encoding less active variant of the enzyme is associated with increased risk for
both alcohol dependence and upper gastrointestinal tract cancer.111–113
Similarly, the inactive ALDH2 allele (ALDH2*2), the presence of which
greatly decreases tolerance to alcohol (homozygous individuals are normally
unable to drink alcohol), is associated with excessive risk of esophageal and head
and neck cancer in drinking heterozygote carriers.110–114 It should be noted that
these variants of the ADH2 and ALDH2 enzymes are especially common in
China, Japan, and other populations of East Asia. Studies of other genes of this
group have shown that the presence of the low activity allele of the ADH3
(ADH3*2), in combination with alcohol drinking, appeared to result in elevated
risk for the development of upper digestive tract tumors.114–116 However, other
investigators reported no effect.117 Also, a higher risk of developing breast
cancer118 and colorectal adenomas119 was observed in individuals homozygous
for the ADH3*1, which is associated with increased activity of the enzyme. It
was also found that the presence of ADH1C “rapid” allele ADH1C*1 in alcoholics
was associated with increased risk of upper aerodigestive tract cancer.120
9.2.4 DNA REPAIR

It is generally accepted that insufficient or inadequate DNA repair is an important
factor in cancer initiation. Interactions between dietary factors and DNA repair
mechanisms are normally regarded from the point of view of effects of mutagenic
derivatives of some food constituents at the level of DNA (e.g., DNA adduct
formation) and their elimination by DNA repair systems. These mechanisms are
often analyzed together with xenobiotic metabolism pathways discussed above.
However, some authors directly addressed concerted influence of nutrients and
polymorphic genes-encoding DNA repair enzymes on cancer risk, demonstrating,
in particular, effects of variants of OGG1 (8-oxoguanine glycosylase I)121 and
XRCC3 (x-ray repair cross complementation group 3)122 genes on colorectal
cancer risk related to meat consumption.
Several groups tried to approach possible relationship of DNA repair enzyme
genotypes and protective effects of antioxidant-rich fruits and vegetables. The
presence of the Trp194 allele of XRCC1 gene was shown to enhance the breast
cancer-protective action of high intake of fruits, fruit juices, vegetables, and
antioxidant supplements,123 whereas the presence of the XRCC1 Gln399 allele
was associated with increased prostate cancer risk among men with low dietary
intake of vitamin E or lycopene.124 In a recent study, an inverse association
between consumption of fruits and vegetables and breast cancer risk was observed
among women homozygous for the Leu84 allele or bearing at least one Val143
allele of MGMT (O-6-methylguanine DNA methyltransferase) gene.125
Moreover, it has been shown that daily green tea consumption reduces lung
cancer risk in individuals with the Cys326 allele of OGG1(8-oxoguanine-DNA
glycosylase) gene126 The latter group of studies highlights a possibility of inter-
action between DNA repair systems and dietary factors exerting protective influ-
ences against cancer. It is well established that calorie restriction inhibits exper-
imental carcinogenesis and may be beneficial in humans.127,128 Effects of calorie
restriction include stimulation of nuclear DNA repair129,130; however, this phe-
nomenon observed in experimental studies requires further investigation in
humans. It is also likely that some dietary constituents can modulate DNA repair
as it has already been demonstrated for selenomethionine in vitro.131,132 The idea
of active dietary interventions directed on DNA repair enhancement133–135 appears
to be very attractive, but its realization still belongs to the future.
9.2.5 ENERGY BALANCE REGULATION
There is no doubt that obesity and related disorders, especially metabolic syn-
drome,9,136 may increase cancer risk. While caloric restriction is believed to be
cancer-protective,127 overeating is regarded as a major avoidable cause of cancer,
and there is sufficient evidence linking obesity with cancer of the colon, breast,
endometrium, kidney, and esophagus.23,137 Although regulation of energy balance
now attracts considerable attention of the scientific community,9,138 the knowledge
of its involvement in cancer pathogenesis remains at best fragmentary.
Recent studies of genes encoding factors regulating food/energy intake have
revealed the existence of multiple polymorphic variants.9 Publications suggesting
impact of some of these variants in cancer risk determination are just starting to
emerge. Among polymorphic genes encoding factors regulating food/energy
intake leptin gene variants have been demonstrated to be associated with suscep-
tibility to prostate cancer139 and non-Hodgkin lymphomas;139,140 in the latter case,
a leptin receptor gene SNP has been shown to be interactive.140,141 Polymorphisms
in the ghrelin and neuropeptide Y genes also affected non-Hodgkin lymphoma
risk.142 Variation of the gene encoding melanocortin-1 receptor (an important
factor in central regulation of energy intake) is proven to be a factor in skin cancer
and melanoma risk,143 but these links may reflect the key role of the gene in skin
pigmentation regulation. Moreover, there is a growing body of evidence linking
polymorphisms in the insulin and, especially, insulin-like growth factor pathway
genes with colorectal,144,145 breast,146 lung,147 prostate,148,149 and oral cavity150
cancers.
Modulation of cancer risk has also been reported for genes encoding proteins
involved in energy expenditure control; however this effect may not be directly
related to nutritional factors. Briefly, there were reports linking polymorphisms
in the 2- and 3-adrenergic receptor genes with susceptibility to breast,151 colorec-
tal,152 and endometrial153 cancer. Multifunctional factors of the arachidonic acid
pathway, in particular, peroxisome proliferator-activated receptor gamma (PPAR)
can be mentioned here as well, since PPAR polymorphisms have been shown to
affect risk of colorectal tumours,154–156 bladder,157 renal,158 and endometrial159
cancer. Interestingly, there appeared to be colorectal cancer risk-affecting inter-
actions between polymorphic PPAR gene variants and diet155,156; however, these
findings require confirmation.
Limits of this chapter do not allow extensive analysis of this complex
field, ramifications of which expand far beyond the area of nutrigenetics and
nutrigenomics.
9.2.6 OTHER PHYSIOLOGICAL PATHWAYS, GENES, AND

RESEARCH PERSPECTIVES
Other regulatory and metabolic systems are certainly involved in cancer patho-
genesis, but it is very difficult to analyze them in relation to gene–nutrient
interactions due to the lack of available information. For instance, dietary

approaches to cancer angiogenesis modulation definitely deserve intense devel-
opment. It is known that plant-derived polyphenols can inhibit tumour angio-
genesis in experimental conditions,159,160 but adequate biomarkers (including
relevant genes) of their effects for human studies remain to be identified despite
discussions on developing strategies of dietary cancer prevention based on the
use of products rich in these compounds.160,161 Often mechanisms of dietary
interventions are difficult to interpret, such as results indicating that calorie
restriction inhibits tumor-associated endothelial growth in experimental mam-
mary carcinogenesis, whereas specificity of its influence on tumor angiogenesis
remains unclear.162 It is also evident that effects of dietary factors almost always
have multiple biological endpoints. Indeed, the same polyphenolic flavonoids
exert their anticancer influence not only through angiogenesis inhibition, but
also by increasing apoptosis rate, inhibiting cell proliferation and tumor inva-
sion through multiple molecular mechanisms, including modulation of signal
transduction pathways.159–161,163
This chapter does not specifically address the relationship of dietary influ-
ences with regulatory systems mediated by hormones and vitamins, which can
strongly affect risk of some cancers, but the importance of genetic diversity in
these systems should not be forgotten. For example, polymorphic variants of the
vitamin D receptor gene have repeatedly been shown to be associated with altered
susceptibility to several types of malignancies164–167; however, correct interpreta-
tion of these findings is difficult given apparent multiplicity of both external
factors having effects on this system (intakes of calcium, vitamin D, dairy prod-
ucts, sun exposure) and its involvement in a wide variety of biological pro-
cesses.166 Likewise, mechanisms of action of potentially cancer-protective
carotenoids168 or ω-3 polyunsaturated fatty acids169 are complex and not entirely
understood. It is impossible to disagree with the conclusion that in the past many
studies in the domain of diet and cancer research were hampered by inadequate
design, use of invalid biomarkers, and problems with exposure monitoring.13
Development of nutrigenomics of cancer on this shaky basis is hardly possible;
thus, fundamental revision of research approaches is ongoing.
Complexity of dietary influences involving simultaneous diverse effects of
multiple bioactive agents often makes their investigation a hard puzzle to solve.
Therefore, a somewhat similar but much better defined field of cancer pharma-
cogenomics will be addressed in the next part of this chapter. Thorough investi-
gation of interactions between well-characterized drugs and their biological tar-
gets, with a close view on the significance of genetic background in these
interactions, has already resulted in much better understanding of both efficiency
and complications of cancer chemotherapy. This experience can be used in the
development of nutrigenomic approaches in cancer research. In addition, nutri-
tional influences often interact with chemotherapeutic schemes; thus, analysis of
recent developments in the field of pharmacogenetics/pharmacogenomics appears
to be entirely justified.
9.3 ADVANCES IN CANCER PHARMACOGENOMICS

AND LINKS TO NUTRIGENOMICS
Pharmacogenomics is a discipline that has recently arisen from pharmacogenetics
after the groundbreaking event of human genome sequence determination.
Although the main goal of revealing the genetic mechanisms behind interindi-
vidual variability of responses to pharmaceutical agents10 remains the same,
possibility of applying genome-wide research approaches can further accelerate
the development of the field. However, efficient introduction of innovative
research technologies requires time; therefore, pharmacogenetic approach
remains dominant in the field. This section is devoted to characterization of the
current state of pharmacogenomics of cancer with regard to possible links with
nutrigenomics.
Cancer pharmacogenomics is understandably focused mostly on cytotoxic
chemotherapy and, to some extent, radiotherapy of advanced tumors (including
relatively early adjuvant therapy — see Figure 9.1), which commonly results in
severe toxicity and side effects. For this reason, the main goal of pharmacoge-
nomic studies in this field is the search for individual genetically determined
patterns of sensitivity and responses to cytotoxic agents. This knowledge will
allow the selection of the most efficient (in terms of their specific aggressiveness
against malignancy) and, at the same time, the least harmful (in terms of general
toxicity) schemes of treatment on the individual basis. Several authors reviewed
main achievements in the field in general,170–172 so it is logical to concentrate on
the last advances.
Table 9.2 provides a more detailed outlook of recent studies in the area of
cancer pharmacogenomics. It is remarkable that even superficial comparison of
Table 9.1 and Table 9.2 shows considerable similarity of genes and metabolic
pathways involved in control of nutrient and drug effects. Further discussion is
subdivided into a few topics reflecting main biological mechanisms affected by
genetic variability.
9.3.1 DRUG-METABOLIZING PHASE I AND PHASE II ENZYMES

Drug metabolism represents a particular case of xenobiotic metabolism, thus it
is natural that Phase I and II enzymatic systems mentioned above are going to
be discussed here. The reactions of Phase I mediated by cytochrome P450 family
enzymes are very important for initial biotransformation of anticancer agents,
often resulting in the activation of inactive prodrugs with production of highly
cytotoxic (and, usually, generally toxic) active compounds. This mechanism has
been described for such widely used medicines as cyclophosphamide, doxorubi-
cine, thiotepa, and tegafur.266 At the same time, P450s act as deactivators of other
antitumor drugs, such as taxanes (paclitaxel, docetaxel), Vinca alkaloids (vinblas-
tine, vincristine), camphotecins (irinotecan, topotecan), antiestrogens (tamox-
ifen), and some other agents.266
It is now established that the CYP3A subfamily of enzymes is especially

important for Phase I drug metabolism, being the most abundant P450 in the
human liver73,267 and intestine.74,268 Numerous anticancer drugs are known to be
substrates of the widely expressed CYP3A4266; however, it is likely that the
contribution of the CYP3A5 may be more important than previously suggested.269
Although considerable interindividual variability in the expression of CYP3A
subfamily enzymes is a well-known fact, and its implications for drug metabolism
are difficult to underestimate, mechanisms of this variability are complex and not
entirely clear. Recent studies have shown the presence of multiple polymorphisms
in genes encoding CYP3A473,270–272 and CYP3A5,73,273,274 the latter being increas-
ingly regarded as a major polymorphic contributor to drug metabolism.269,273 The
role of the CYP3A family gene polymorphisms in relation to anticancer therapy
remains poorly investigated; however, first studies in this direction are starting
to emerge (see Table 9.2).
The CYP3A family is especially interesting in the context of this chapter
because these enzymes are known to be targets of interaction with certain food
components; in particular, grapefruit juice, oral intake of which selectively inhib-
its CYP3A4 activity in the small intestine275,276 and liver277 resulting in supression
of the oxidative metabolism of relevant substrates. It is believed that furanocou-
marins and isoflavones present in grapefruit juice act as CYP3A4 inhibitors (see
Chapter 2, Hypolipidemic Therapy: Drugs, Diet and Interactive Effects). It is also
intriguing that a popular herbal product, St. John’s wort (Hypericum perforatum),
has been shown to induce CYP3A4 expression.278 Given the role of the CYP3A4
in transformation of a number of anticancer agents,266 these interactions poten-
tially may be important, especially in altering acute toxic reactions on the admin-
istration of cytotoxic drugs. It should be noted that information regarding possible
influence of natural food components and common herbs on both efficiency and
side effects of cancer chemothetapy is almost nonexistent. Only one pilot study
described reduced bioavailability of the anticancer agent, etoposide, following
grapefruit juice ingestion.279
Several other polymorphic Phase I enzymes are important for anticancer
therapy. Pharmacogenomic approaches look promising for CYP1A2 and fluta-
mide, CYP2A6 and tegafur, CYP2B6 and cyclophosphamide, CYP2C8 and pacli-
taxel, and CYP2D6 and tamoxifen,266,280 but further studies are required for the
identification of clinically relevant gene variants. Likewise, multiple polymor-
phisms have been identified in genes encoding carboxylesterases (CESs) 1 and
2281; however, their physiological significance remains to be explored. Polymor-
phic CESs participate in drug metabolism acting as Phase I enzymes participating
in the activation of such anticancer agents as irinotecan (CPT-11), paclitaxel, and
capecitabine282; therefore, information on the presence of CES variants may be
used for the development of individualized chemotherapy schemes.
The importance of polymorphisms in genes encoding Phase II enzymes for
cancer chemotherapy is well exemplified by the effects of variants of hepatic
UDP-glucuronosyltransferase 1A1 (UGT1A1) in treatment of advanced solid
(mostly colorectal) tumors with topoisomerase I inhibitor irinotecan. UGT1A1
278
TABLE 9.2
Polymorphic Genes Encoding Factors Affecting Outcome of Cancer Chemotherapy and Radiotherapy
Therapeutic Agents Reported Clinical Effects or Functional Importance
Gene Malignancy Site and Type (Chemo- or Radio-) (References given in parentheses)
Polymorphic Genes Encoding Drug-Metabolizing Phase I and Phase II Enzymes

CYP1A1 Childhood acute lymphoblastic Multiagent chemotherapy The presence of the C6235 allele (T6235C SNP in the
leukemia (ALL) 3’ flanking region) was associated with increased risk
of relapse (173).
CYP2B6 Haematological malignancies Cyclophosphamide (CP) The presence of the T516 allele (T516C SNP) was
associated with increased rate of formation of CP-
active metabolite,4-hydroxycyclophosphamide (174).

CYP3A4 Breast cancer Anthracycline-based adjuvant Patients who carried the CYP3A4*1B in combination
chemotherapy followed by high-dose with CYP3A5*3 and functional variants of GSTM1 and
multiagent chemotherapy GSTT1 had significantly lower probability of survival
(175).
Childhood acute lymphoblastic Multiagent chemotherapy Patients with the CYP3A4*1B genotype had decreased
leukemia (ALL) risk of peripheral nephropathy caused by chemotherapy
(176).
CYP3A5 Breast cancer Anthracycline-based adjuvant Patients who carried the CYP3A5*3 in combination with
chemotherapy followed by high-dose CYP3A4*1B and functional variants of GSTM1 and
multiagent chemotherapy GSTT1 had significantly lower probability of survival
(175).
Childhood acute lymphoblastic Multiagent chemotherapy Patients with the CYP3A5*3 genotype had decreased
leukemia (ALL) risk of peripheral nephropathy caused by chemotherapy
(176).
UDP-glucuronosyl-transferase Cancer of several sites, mostly Irinotecan The presence of the UGT1A1*28 (7 TA repeats instead
1A1 (UGT1A1) advanced colorectal tumors of 6 in the promoter region) allele was associated with
severe toxicity (177-180) and impaired glucuronidation
of the active metabolite of irinotecan, SN-38 (178, 181,
182).
The presence of the UGT1A1*27(Pro229Gln) alleles
was associated with severe toxicity in a few cases (177).
The presence of the A-3156 allele (G-3156A promoter
SNP) was associated with the severe neutropenia (183).
The presence of the G-3279 allele (T-3156G promoter
region SNP) was associated with severe toxicity; this
SNP was also shown to be linked to the UGT1A1*28
variant (184).
UDP-glucuronosyl-transferase Colorectal cancer Irinotecan Genotypes UGT1A7*2/*2 and UGT1A7*3/*3 encoding

1A7 (UGT1A7) low activity variants of the enzyme were associated with
better antitumor response and lower gastrointestinal
toxicity; however, study size was small (185).
UDP-glucuronosyl-transferase Colorectal cancer Irinotecan UGT1A9 genotype 118 (dT)(9/9) was associated with
1A9 (UGT1A9) better antitumor response and lower gastrointestinal
toxicity; however, study size was small (185).
UDP-glucuronosyl-transferase Breast cancer Tamoxifen Tamoxifen-treated patients with high enzyme activity-
2B15 (UGT2B15) associated UGT2B15 genotypes had increased risk of

recurrence and poor survival (186)
Glutathione-S-transferase M1 Breast cancer CP, 5-FU (5-fluorouracil), doxorubicin Combination of GSTM1-Null and GSTT1-Null
(GSTM1) genotypes was associated with better survival (187).
Anthracycline-based adjuvant Patients who carried the GSTM1-Null and GSTT1-Null
chemotherapy plus high-dose in combination with low activity variants of CYP3A4
multiagent chemotherapy and CYP3A5 had significantly lower probability of
treatment failure (175).
279
280

Ovarian cancer Cisplatinum, alkylating agents Combination of GSTM1-Null and GSTT1-Null was
associated with better responses to chemotherapy (188).
Paclitaxel, cisplatinum The GSTM1-Null genotype was associated with
significantly longer survival time (189).
Several chemotherapy schemes Patients carrying the GSTM1-Null were less likely to
have disease progression (190).
Malignant glioma Nitrosoureas The GSTM1-Null genotype was associated with better
survival, but higher probability of adverse effects (191).

Childhood acute lymphoblastic Multiagent chemotherapy GSTM1-Null genotype was associated with a decreased
leukemia (ALL) risk of relapse following cytotoxic chemotherapy (192,
193).
No evidence of GSTM1 genotype (alone or combined
with GSTT1 genotype) effect was found (194).
Multiagent chemotherapy Simultaneous presence of GSTM1-Null and GSTT1-
Null genotypes was associated with early relapses
(195).
Adult acute myeloid leukemia Multiagent chemotherapy The presence of either GSTM1-Null or its combination
(AML) with GSTT1-Null was associated with poor survival
following chemotherapy (196).
Glutathione-S-transferase T1 Breast cancer CP, 5-FU, doxorubicin Combination of GSTT1-Null and GSTM1-Null
(GSTT1) genotypes was associated with better survival (196).
Anthracycline-based adjuvant Patients who carried the GSTT1-Null and GSTM1-Null
chemotherapy plus high-dose in combination with low activity variants of CYP3A4
multiagent chemotherapy and CYP3A5 had significantly lower probability of
treatment failure (175).
Ovarian cancer Cisplatinum, alkylating agents Combination of GSTM1-Null and GSTT1-Null was
associated with better responses to chemotherapy (188).
Childhood acute lymphoblastic Multiagent chemotherapy Combination of GSTT1-Null and GSTM1-Null
leukemia (ALL) genotypes was associated with a decreased risk of
relapse following cytotoxic chemotherapy (189).
Simultaneous presence of GSTT1-Null and GSTM1-
Null genotypes was associated with early relapses
(195).
Childhood acute myeloid leukemia Multiagent chemotherapy Children with GSTT1-Null genotype had greater toxicity
(AML) and reduced survival after chemotherapy (197).

Adult acute myeloid leukemia Multiagent chemotherapy GSTT1-Null genotype was associated with an increased
(AML) toxicity of combined chemotherapy and poor survival
(198).
The presence of either GSTT1-Null or its combination
with GSTM1-Null was associated with poor survival
Glutathione-S-transferase P1 Breast cancer Several chemotherapy schemes Women homozygous for the Val105 allele (Ile105Val
(GSTP1) polymorphism) had a 60% reduction in mortality risk

(199).
Colorectal cancer 5-FU/oxaliplatin chemotherapy The presence of the Val105/Val105 homozygocity was
associated with increased survival (200, 201).
Non-small lung carcinoma Platinum-based chemotherapy The presence of the Val105 allele was associated with
significantly improved survival (202).
Ovarian cancer Several chemotherapy schemes Patients carrying the Val105 allele were less likely to
have disease progression (190).
281
282

Stomach cancer 5-FU/cisliplatinum chemotherapy The presence of the Val105 allele was associated with
significantly improved survival (203).
Malignant glioma Nitrosoureas Homozygous simultaneous presence of Ile105 and
Ala114 (Ala114Val) in combination with GSTM1-Null
was associated with better survival, but higher
probability of adverse effects (191).
Childhood acute lymphoblastic Multiagent chemotherapy The presence of the Val105 homozygocity was associated
leukemia (ALL) with a decreased risk of relapse following cytotoxic

chemotherapy (192, 204).
Therapy-related acute myeloid Several chemotherapy schemes The presence of the Val105 allele was associated with
leukemia (t-AML) an increased risk of developing t-AML after cytotoxic
chemotherapy (205).
Glutathione-S-transferase A1 Breast cancer Cyclophosphamide-containing The presence of the T-69 homozygocity (C-69T SNP in
(GSTA1) combined chemotherapy the promoter region) was associated with better survival
NAD(P)H: quinone Childhood acute lymphoblastic Multiagent chemotherapy The presence of the NQO1*2 allele (609T allele of
oxidoreductase (NQO1) leukemia (ALL) C609T SNP) was associated with increased risk of
relapse (173).
Superficial bladder cancer Mitomycin C chemotherapy The presence of the NQO1*2 allele was associated with
reduced response to mitomycin C (207).
Sulfotransferase 1A1 Breast cancer Tamoxifen The presence of the SULT1A1*2/*2 homozygocity
(SULT1A1) (His/His of Arg213His polymorphism) was associated
with poor survival following tamoxifen therapy (208).
Sulfotransferase 1E1 Breast cancer Several chemotherapy schemes The presence of the C-1653 allele (T-1653C SNP in the
(SULT1E1) promoter region) was associated with significantly
higher risk of recurrence (209).
Polymorphic Genes Encoding Enzymes of Purine/Pyrimidine Metabolism

Thiopurine Acute lymphoblastic leukemia 6-Mercaptopurine, The presence of alleles causing TPMT deficiency
S-methyltransferase (TPMT) (ALL) and acute myeloid 6-thioguanine, azathiopurine (TPMT*2, TPMT*3A, TPMT*3C, TPMT*7) was
leukemia (AML) in children. associated with intolerance of conventional therapeutic
doses (170, 210–212). Genotype-dependent dose
adjustment provided better chemotherapy tolerance
(210, 212, 213).
Dihydro-pyrimidine Cancer of several sites 5-FU-based chemotherapy The presence of the DPYD*2A allele causing a reduced
dehydrogenase (DPD or DPD activity was associated with toxic effects of 5-
DPYD) fluorouracil (5-FU) chemotherapy (214–216).

Cytidine deaminase (CDA) Cancer of several sites Gemcitabine, cisplatin Severe chemotherapy toxicity was observed in a patient
homozygous for the Thr70 allele (Ala70Thr
polymorphism); however, only a few patients were
analyzed (217).
Deoxycytidine kinase Acute myeloid leukemia (AML) 1-β-arabinofuranosyl-cytosine (AraC) The presence of the haplotype combining C-360
(dCK) homozygocity (C-360G SNP in the 5’ flanking region)
with C-201 homozygocity (C-201T in the 5’ flanking
region) resulted in a poor response to chemotherapy (218).
Polymorphic Genes Encoding Enzymes and Regulatory Factors of Folate Metabolism

Methylenetetrahydrofolate Colorectal cancer Fluoropyrimidine-based chemotherapy The T677 homozygocity (C677T SNP) was associated
reductase (MTHFR) with better chemotherapy response rates (219–221).
Ovarian cancer Methotrexate The T677 homozygocity was associated with increased
toxicity of methotrexate therapy (222).
Acute leukemia Methotrexate The T677 homozygocity was associated with increased
toxicity of methotrexate chemotherapy (223).
283
284

Childhood acute lymphoblastic Methotrexate The presence of the T677T + A1298 (A1298G SNP)
leukemia (ALL) haplotype (especially in combination with TS high-
activity genotype) was associated with reduced event-
free survival (224).
The presence of the T677 allele was associated with
higher risk of relapse (225).
Thymidylate synthase (TS) Colorectal cancer 5-FU-based chemotherapy schemes, The presence of TS genotypes associated with high activity
capecitabine of the enzyme (those with 3R or three 28-bp repeats in

the 5′ promoter/enhancer region, G variant of the G12G
SNP in the repeat sequence, and a 6bp insertion in the 3′-
untranslated region) was associated with poorer prognosis
and shorter survival (201, 203, 226–229).
5-FU-based chemotherapy Simultaneous presence of 2R (low activity-related) and
6bp ins in the 3′-UTR (high activity-related) was
associated with severe side effects (230).
The presence of high activity-related TS genotypes was
associated with longer disease-free survival (231).
Stomach cancer 5-FU-based chemotherapy The presence of low activity-related TS genotypes was
associated with longer disease-free and overall survival
(232)
Non-small-cell lung cancer 5-FU-based chemotherapy The presence of TS genotypes associated with low
activity of the enzyme was associated with better
survival (233).
Childhood acute lymphoblastic Methotrexate The presence of TS genotypes associated with high
leukemia (ALL) activity of the enzyme in combination with either
MTHFR T677A1298 haplotype or MTHFD1 A1958
allele was associated with reduced event-free survival
(224).
Methylenetetra-hydrofolate Childhood acute lymphoblastic Methotrexate The presence of the A1958 allele (G1958A SNP) in
dehydrogenase (MTHFD1) leukemia (ALL) combination with TS high-activity genotype was
associated with reduced event-free survival (224).
Methionine synthase (MTR) Esophageal cancer 5-FU-based radiochemotherapy The presence of the G2756 allele (A2756G SNP) was
associated with significantly longer survival (234).
Reduced folate carrier (RFC1) Childhood acute lymphoblastic Methotrexate The A80 allele (G80A SNP) was associated with reduced
leukemia (ALL) event-free survival (235).
No effect of RFC1 gene variants revealed (236).
Polymorphic Genes Encoding DNA Repair Enzymes

Excision repair cross Non-small-cell lung cancer Cisplatin combination chemotherapy The presence of the AAC codon 118 allele (Codon 118
complementation group 1 AAC→AAT silent SNP) was associated with better
(ERCC1) survival (237).
Docetaxel-cisplatin The presence of the AAC codon 118 allele was associated
with better survival (238).
Platinim-based chemotherapy The presence of the C8092 allele (C8092A SNP shown
to be linked to the codon 118 SNP) was associated with

better survival (239).
Colorectal cancer Platinum-based chemotherapy The presence of the C8092 allele was associated with a
significantly increased risk of grade 3 or 4
gastrointestinal toxicity. Codon 118 SNP did not affect
toxicity (240).
The presence of the AAC codon 118 allele was associated
with better survival (241).
285
286

Oxaliplatin/5-FU The presence of the AAT codon 118 allele was associated
with higher chemotherapy response rate (242).
Xeroderma pigmentosum Non-small-cell lung cancer Platinum-based chemotherapy The presence of the Asn312 allele (Asp312Asn XPD
group D (XPD or ERCC2) polymorphism) was associated with reduced survival
(243).
Colorectal cancer Oxaliplatin/5FU The presence of the Lys751 homozygocity (Lys751Gln
XPD polymorphism) was associated with better
survival (244).
Acute myeloid leukemia Multiagent chemotherapy The presence of the Lys751 homozygocity was
associated with better survival (245).
Excision repair cross Superficial bladder cancer Bacillus Calmette–Guerin treatment The presence of the Val1097 allele (Met1097Val ERCC6
complementation group 6 polymorphism) was associated with reduced
(ERCC6) recurrence-free survival (246).
Xeroderma pigmentosum Superficial bladder cancer Bacillus Calmette–Guerin treatment The presence of the A-4 allele (G-4A SNP in the 5′
group A (XPA) untranslated region) was associated with reduced
recurrence-free survival (246).
X-ray repair cross Colorectal cancer Oxaliplatin/5-FU The presence of the Gln399 allele (Arg399Gln XRCC1
complementation group 1 polymorphism) was associated with resistance to
(XRCC1) chemotherapy and reduced survival (247).
Breast cancer Radiotherapy The presence of the Trp(194) allele (Arg194Trp XRCC1
polymorphism) was associated with adverse responses
to radiotherapy (248).
The presence of the Gln399 allele was associated with
protection against adverse effects of radiotherapy (249).
Non-small-cell lung cancer Radiotherapy Combined analysis of polymorphisms at codons 194,
280, and 399 has shown correlations between XRCC1
haplotypes and survival (250).
Therapy-related acute myeloid Radiotherapy The presence of the Gln399 allele was associated with a
leukemia (t-AML) decreased risk of developing t-AML (251).
Apurinic/apyrimidinic Breast cancer Radiotherapy The presence of the Glu148 allele (Asp148Glu APE1
endonuclease 1 polymorphism) was associated with protection against
(APE1/APEX1) the development of acute adverse effects of
radiotherapy (249).
DNA mismatch repair gene Therapy-related acute myeloid Cyclophosphamide procarbazine The presence of the (-6)C allele (T-6C SNP at the 3′
hMSH2 leukemia (t-AML) splice acceptor site of exon 13) was associated with an
increased risk of developing t-AML (252).
Polymorphic Genes Encoding ATP-Binding Cassette Transporters (ABC)

Multi-drug resistance (MDR-1 Acute myeloid leukemia (AML) Multiagent chemotherapy The presence of the C3435 homozygocity (C3435T SNP)
or ABCB1) was associated with an increased probability of relapse
and reduced overall survival (253).
Breast cancer Anthracyclines or anthracyclines The presence of the T3435 homozygocity was associated
combined with taxanes with an increased probability of favorable clinical
response to preoperative chemotherapy (254).

Cancer of several sites Irinotecan The presence of the C1236 allele (C1236T SNP) was
associated with significantly increased exposure to
irinotecan and its active metabolite SN-38 (178).
Childhood acute lymphoblastic Multiagent chemotherapy The presence of the T3435 allele was associated with
leukemia (ALL) reduced probability of CNS relapse (204).
287
288

The presence of the C3435 homozygocity was associated

with reduced event-free survival and overall survival
(255).
Nasopharyngeal carcinoma Irinotecan Irinotecan clearance was lower in patients homozygous
for the C3435 allele (256).
Breast cancer resistance protein Cancer of several sites Irinotecan Extensive accumulation of SN-38 was observed in a
(BCRP or ABCG2) patient homozygous for the A421 allele (C421A SNP)
(257).
Nasopharyngeal carcinoma Irinotecan Higher irinotecan to SN-38 conversion of was related to
the absence of CTCA deletion in the 5′ flanking region
(256).
Polymorphic Genes Encoding Other Drug Targets (Receptors, Regulatory Factors, etc.)
Epidermal growth factor Colorectal cancer (rectum) Adjuvant and neoadjuvant Patients with Arg497 homozygocity (Arg497Lys SNP)
receptor (EGFR or HER-1) chemoradiation or lower number (<20) of CA repeats (Inthron 1 CAn
repeat polymorphism) had a higher recurrence risk
(258).
Colorectal cancer 5-FU/oxaliplatin Patients with less than 20 CA repeats were more likely
to show disease progression (259).
HER-2 receptor (HER-2, c-erb- Ovarian cancer Cisplatin, paclitaxel The presence of Val655 homozygocity (Ile655Val
B2 or neu) polymorphism) was associated with reduced overall
survival (260).
Immunoglobulin G Fc receptor Follicular lymphoma Rituximab The presence of His131 homozygocity (Arg131His
II (FcRIIa, CD32) polymorphism) was associated with better response rate
and freedom from progression (261).
Immunoglobulin G Fc receptor Follicular lymphoma Rituximab The presence of Val158 homozygocity (Phe158Val
III (FcRIIIa, CD16) polymorphism) was associated with better response rate
and freedom from progression (261).
Id vaccination The presence of Val158 homozygocity was associated
with longer progression-free survival (262).
Interleukin-1β (IL-1B) Stomach cancer Palliative chemotherapy The presence of T-511 (C-511T SNP) and C-31 (T-31C
SNP) was associated with reduced progression-free and
overall survival, but only in patients with wild-type
genotype of interleukin-1 receptor antagonist (IL-RN)
(263).
Interleukin-6 (IL-6) Breast cancer Anthracycline-based adjuvant The presence of the C-174 allele (G-174C SNP) was
chemotherapy and multiagent high- associated with better disease-free and overall survival
dose chemotherapy (264).
Interleukin 8 receptor CXCR1 Colorectal cancer 5-FU/oxaliplatin Patients with CXCR1 2607 heterozygocity (G2607C
SNP) were more likely to show disease progression
(254).
Interferon-γ (IFN-γ) Melanoma Cisplatin, vinblastine, and dacarbazine The presence of T834 allele (A874T SNP) was
combined with interleukin-2 and associated with better response, progression-free, and
interferon-α and in some cases overall survival (265).

tamoxifen
289
exerts glucuronidation of SN-38, the active metabolite of irinotecan, producing

an inactive and nontoxic glucuronic acid conjugate SN-38G (178). It has repeat-
edly been shown that impaired activity of the enzyme caused by the presence of
variant alleles, in particular the UGT1A1*28 causing reduced gene expression
and enzyme activity, can result in severe toxicity (see Table 9.2). Although little
is known about interactions between nutritional factors and UGTs, there are
reports indicating that isothiocyanate-rich cruciferous vegetables can affect
enzyme activity in individuals with the UGT1A1*28 variant,84 and that curcumin
transiently inhibits UGT activities in cell culture.82
Enzymes of the glutathione-S-transferase family have also been shown to be
involved in Phase II drug metabolism, detoxifying drugs and their toxic deriva-
tives by glutathione conjugation.76 The presence of homozygous-long deletions
in GSTM1 and GSTT1 genes (“Null” variants) is often associated with higher
chemotherapy efficiency for solid tumors (breast, ovary); however, controversial
results were obtained for leukemias (see Table 9.2). The presence of the Val105
allele of the GSTP1 gene was also associated with better outcomes of chemo-
therapy and improved survival in most studies. It is important to remember that
activities of the GST family enzymes can be modulated by dietary intake of
cruciferous vegetables; however, this type of diet–gene interaction has not been
investigated in relation to anticancer chemotherapy.
9.3.2 ENZYMES OF PURINE/PYRIMIDINE METABOLISM

Thiopurine-S-methyltransferase (TPMT) is one of the key enzymes in the
biotransformation of aromatic and heterocyclic sulfhydril compounds, including
anileukemia agents 6-mercaptopurine, 6-thioguanine, and azathiopurine. The
TPMT is the catalyst for the S-methylation of these drugs leading to the formation
of inactive metabolites.170 It is now evident that several variants of the highly
polymorphic TPMT gene encode the enzyme molecules with decreased stability
and lower specific activity (especially TPMT*3A, TPMT*3C, TPMT*5).283 Indi-
viduals homozygous for these variants as well as compound heterozygotes are
TPMT-deficient and application of conventional doses of thiopurine chemother-
apy can cause severe hematopoietic toxicity in such patients.170,210–212 It has
already been shown by several studies that prospective genetic screening can
identify patients requiring lower thiopurine doses, thus increasing therapy toler-
ance.210,212,213 Although influence of genetic variation on TPMT expression is well
established, a possibility of contribution of other factors, including those of
dietary origin, remains to be investigated.
Numerous studies have been devoted to the characterization of genetic factors
affecting 5-fluorouracil (5-FU)-based chemotherapy. Dihydropyrimidine dehy-
drogenase (DPD) is the initial rate-limiting enzyme in the degradation of 5-FU.
The enzyme activity is highly variable and its partial loss is observed in at least
3% of individuals.170 DPD is encoded by a highly polymorphic DPYD gene and
the DPYD*2 allele is associated with a reduced enzyme activity and high risk of
5-FU toxicity.214–216 Nevertheless, other factors (in particular, aberrant methylation
of the DPYD gene promoter region284 and transcription regulation by a specific

activator protein285) appear to be involved in variations in DPD activity; thus, the
predictive value of DPYD genotype determination remains unclear. Nothing is
known about effects of dietary factors on DPD activity.
9.3.3 ENZYMES AND REGULATORY FACTORS OF FOLATE

METABOLISM
The folate system has already been briefly considered in this chapter as a good
example of diet–gene interactions modulating cancer risk. Several cytotoxic
agents employed in cancer chemotherapy schemes, including fluoropyrimidines
(5-FU, capecitabine) and methotrexate, inhibit folate metabolism acting at its
different stages.171 For this reason polymorphic genes encoding enzymes of this
metabolic pathway attracted considerable attention of investigators. Analysis of
the effects of the methylenetetrahydrofolate reductase (MTHFR) gene variants
revealed different response patterns in different malignancies. Several studies in
colorectal cancer patients show that the presence of the T667 (Val222) allele was
associated with better responses to fluoropyrimidine therapy,219–221 whereas the
same allele appeared to be linked with excessive toxicity and poor response during
methotrexate therapy of ovarian cancer222 and leukemias.223–225
Thymidylate synthase (TS) is another key polymorphic enzyme of folate
metabolism. TS is one of the main targets of fluoropyrimidines, which inhibit its
activity.286 It is known that genetic variation (variable number of 28-bp repeats)
in the promoter/enhancer region of the TS gene results in two- to fourfold changes
in gene expression defining “rapid” and “slow” alleles (see Table 9.2). In most
clinical studies, the presence of the rapid genotype was associated with poor
prognosis in fluoropyrimidine-treated patients (see Table 9.2).
Several reports describe possible effects of other polymorphic genes encoding
factors participating in folate metabolism regulation (methylenetetrahydrofolate
dehydrogenase, methionine synthase, reduced folate carrier), but all of this infor-
mation requires confirmation by further studies.
Antifolate chemotherapeutic agents are widely used in cancer treatment, but
the role of dietary folate in these conditions is not completely clear. It could be
assumed that high folate intake may decrease the effectiveness of this type of
chemotherapy, but this assumption has not been corroborated in studies of breast
cancer patients.287 In contrast, folate supplementation has recently been suggested
as a way to increase chemotherapy efficiency288; however, the problem still
requires further research.
9.3.4 DNA REPAIR EFFECTS ON CANCER TREATMENT

Most cytostatic therapies are based on inducing irreversible lethal damage in
tumor cell DNA; therefore, DNA repair systems, which serve as a protective
mechanism at initial stages of carcinogenesis, may contribute to intrinsic
drug resistance during anticancer chemotherapy. This phenomenon appears to be
especially graphic in relation to platinum-based cytotoxic therapy, the treatment

of choice for lung and ovarian cancer, and some colorectal tumors.289 Platinum
derivatives exert their cytotoxic effect through formation of platinum DNA
adducts, which then become legitimate targets of the system of nucleotide exci-
sion repair.289,290 Successful removal of platinum adducts can lead to reduced
efficiency of the therapy and is believed to be the main mechanism responsible
for the development of resistance to platinum therapy.
The excision repair, cross-complementation group 1 (ERCC1) gene plays the
leading role in the nucleotide excision repair system, and the presence of a silent
SNP (AACAAT, both coding for asparagine) in codon 118 of this gene has been
found to cause reduced ERCC1 expression.291,292 Several recent studies clearly
show that this variant is also associated with higher response rate to platinum-
based chemotherapy and better survival (see Table 9.2). These observations can
be regarded as the most convincing evidence linking polymorphisms in DNA
repair genes with the effectiveness of cytotoxic anticancer chemotherapy.
Although numerous polymorphic variants are described for other genes encoding
factors of DNA repair systems (XPD, ERCC2, XPA, XRCC1, APE1, hMSH2),
and some of these variants have been associated with gene expression changes,
their clinical importance needs to be confirmed.
Little is known about possible dietary effects on DNA repair with regard to
anticancer chemotherapy. Recent discovery of DNA repair modulation by seleno-
methionine,131,132 abundantly present in Brazil nuts,293 suggests that interactions
are not excluded, but investigation of this area, again, belongs to the future.
9.3.5 ATP-BINDING CASSETTE TRANSPORTERS

Membrane-bound transporter proteins have recently emerged as a major defensive
system that exerts protection from toxic influences through control of xenobiotic
uptake and excretion.170,294 The ATP-binding cassette transporter (ABC) gene
superfamily appears to be the most important from the point of view of cancer
chemotherapy since expression of its members has been implicated in tumor cell
resistance to cytostatic agents, altered drug disposition, and toxic effects of
drugs.294 P-glycoprotein encoded by the MDR-1 (ABCB1) gene is the most
extensively studied among ABC transporters. P-glycoprotein is closely involved
in the regulation of drug bioavailability through interplay with CYP3A4.295 Mul-
tiple polymorphisms have been determined in the sequence of the MDR-1
gene,296,297 and their functional significance is now being investigated. Attempts
to assess the role of some of these SNPs in response to cancer chemotherapy
have been undertaken, but results of several studies published so far look con-
flicting (see Table 9.2). Although information on numerous polymorphisms in
other ABC transporter genes (ABCC1, ABCC2, ABCC3, ABCC4, ABCC5,
ABCC6, ABCG2) is now available,294 their potential validity as biomarkers pre-
dicting response to drugs remains to be clarified.
ABC transporters are very intriguing as a known target for diet–drug inter-
actions. Grapefruit juice components have already been mentioned as potent
inhibitors of CYP3A4; however, they have also been shown to suppress P-

glycoprotein transporter activity.298 Another recent report describes inhibition of
P-glycoprotein-mediated transport by curcuminoids.299 On the other hand, induc-
tion of P-glycoprotein by St. John’s wort has been demonstrated as well.300 These
facts again highlight the necessity of thorough investigation of diet–drug inter-
actions in the context of anticancer treatment.
9.3.6 OTHER DRUG TARGETS AND RECENT ADVANCES

Table 9.2 indicates several reports implicating other polymorphic drug targets,
including receptors and cytokines on the effects of anticancer chemotherapy.
Some of these targets appear to be especially interesting due to their significance
for innovative anticancer treatment methods based on the use of monoclonal
antibodies301,302 or small molecule inhibitors of the epidermal growth factor recep-
tor (EGFR) tyrosine kinase activity.302,303 Recently introduced monoclonal anti-
bodies include anti-HER2/neu trastuzumab (Herceptin®), anti-VEGF bevaci-
zumab (Avastin®), anti-EGFR cetuximab (Erbitux®), and anti-CD20 rituximab
(Rituxan®).301,302 Tyrosine kinase inhibitors comprise imatinib (Gleevec™), gefi-
tinib (Iressa™), and erlotinib (Tarceva®).302,303 The importance of pharmacoge-
nomic approaches to these new therapeutic strategies is already highlighted by
the association between polymorphisms of immunoglobulin GFc receptors and
clinical response to rituximab (see Table 9.2). It is very likely that variants of
highly polymorphic targets of other new agents304 will be found to be important
for therapeutic efficiency as well. These modern directions of antitumor therapy
have emerged very recently, so it is not surprising that information regarding
possible impact of dietary factors is not yet available.
Pharmacogenomics of cancer is often related to later stages of the disease,
when malignant process becomes irreversible and can be only temporarily con-
tained by chemotherapy. At these stages, oncological patients are at a very high
risk of developing cachexia, which is characterized by weight loss with depletion
of host resources of adipose tissue and skeletal muscle.305 About half of all cancer
patients suffer from this syndrome.305 It is known that the development of cachexia
is related to profound deregulation of the central control of energy homeostasis
followed by cytokine production, release of lipid-mobilizing and proteolysis-
inducing factors, and eventually severe general metabolic alterations.305–308 Given
the presence of numerous polymorphic genes encoding regulatory proteins con-
trolling these pathways,9 it is impossible to exclude that the genetic background
may determine predisposition or resistance to cachexia. Correlations of the pres-
ence of polymorphisms in the genes encoding interleukin-1β,263,309 interleukin-1
receptor antagonist,263 and interferon-γ310 with survival of patients with advanced
gastric263 and pancreatic309,310 tumors, which are typically aggravated by fatal
cachexia, have already been reported suggesting that this is another nutrition-
related problem requiring urgent investigation.
9.4 CONCLUSION
The presented overview of pharmacogenomic studies highlights a few impressive
examples of successful utilization of information regarding genetic profiles of
oncological patients in the development of individual-oriented (“tailored”) che-
motherapeutic strategies. These examples include interactions between UGT1A1
polymorphisms and treatment with irinotecan as well as TPMT variants and
thiopurine chemotherapy. Furthermore, promising results have already emerged
from studies of the effects of thymydilate synthase alleles on fluoropyrimidine-
based chemotherapy, influence of ERCC1 genotype on platinum resistance, and
effects of GST-family gene genotypes in different schemes of cytostatic treatment.
This chapter was focused predominantly on the role of inherited structural DNA
variations (polymorphisms); however, it should be understood that other mecha-
nisms, especially changes in gene expression in the malignant tissue, are not less
important in terms of the development of individually tailored schemes of cancer
therapy. There is no doubt that clinical significance of pharmacogenomic infor-
mation is going to increase rapidly with introduction of modern genome-wide
analytical approaches.
In contrast, nutrigenomics of cancer is only starting to come into existence.
Although serious attempts to reveal links between carcinogenesis and nutrition
have been undertaken for decades, making firm conclusions remains difficult.
Cooked red meat consumption appears to be the only dietary influence, which
can be regarded as a proven risk factor for several types of cancer. A number of
nutrients have been shown to have cancer-preventive potential, but further studies
are needed to develop reliable strategies for their wide application. Introduction
of nutrigenomic approaches into this area has brought little clarification so far;
thus, full realization of potential strengths of this approach belongs to the future.
Parallel analysis of genetic factors involved in the metabolism of nutritional
agents and anticancer drugs reveals a striking similarity, which is not surprising,
given the plant-derived nature of many anticancer agents.266 This similarity results
in multiple opportunities for drug–nutrient interactions that is further modulated
by variants of gene structure and expression (see schematic representation in
Figure 9.2). Some situations of nutrient–drug interactions are recognized and
partially investigated, which is exemplified by studies on modulation of drug
transport and metabolism (MDR-1 and P-glycoprotein system and CYP3A
enzymes, respectively) by bioactive substances present in grapefruit juice or St.
John’s wort. However, significance of these known interactions for anticancer
therapy remains obscure. Only further studies in this direction will help to elu-
cidate their importance.
Although it is obvious that successful development of nutrigenomics is com-
plicated by many unsolved problems, the breathtaking speed of the technical
progress introduces numerous new powerful research tools in the field. This
innovation, along with the revision of some inefficient old investigative
approaches in nutritional science, brings expectations that serious breakthroughs
await us in the near future.
Biosystem (organism)
Genetic background
Targets
E
DIET
E
E
T1 T2
Nutrient–drug Nutrient–drug Nutrient–drug
Phase I Phase II
interactions interactions interactions
T1 T2
DRUG E
E
E
Targets
Genetic background
FIGURE 9.2 Diet–drug interactions and their genetic modulation. Abbreviations: T1 —

systems of initial transport of xenobiotics including drugs, also providing some reverse
transport of unmetabolized agents: “Phase 0”; Phase I — Phase I metabolism of xenobi-
otics and drugs; Phase II — Phase II metabolism of xenobiotics and drugs; T2 — systems
of eventual transport (secretion/excretion) of xenobiotic end metabolites (conjugates):
“Phase III”; E — effects of drugs; Targets — biomolecular targets of drugs and food-
derived bioactive substances and their metabolites; symbols representing genetic back-
ground and targets are duplicated to simplify the scheme. It should be understood that in
reality drugs and food-derived substances interact with the same targets regulated by the
same genes.
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308. Rubin H. Cancer cachexia: its correlations and causes. Proc Natl Acad Sci USA.
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DK5836_Index.fm Page 319 Monday, May 22, 2006 2:32 PM
Index
A Alzheimer’s disease (AD), 130, See also

Neurodegenerative disorders
Acarbose, 6, 10–12 amyloid peptides, 146–148
ACAT inhibitors, 37 antioxidants and, 133
Acetaminophen, 68, 79, 214 apoptotic mechanisms, 131
Acetylsalicylic acid, See Aspirin dietary silicon and, 133–134
Activated protein-1 (AP-1), 104 epidemiology, 146
Activity-dependent neutrotrophic factor lifestyle and dietary habits and, 134
(ADNF), 145 omega-3 fatty acids and, 149
Acute lymphoblastic leukemia (ALL), See oxidative stress and, 130, 146–149
Leukemia susceptibility gene, 146
Adenosine triphosphate (ATP) wine consumption and risk, 133, 157
cassette transporter, See ATP-binding Amino acids, See also specific amino acids
cassette transporter antidepressant interactions, 182–186, 199
flavonoid binding, 142 chemotherapy and, 106, 107
Adhyperforin, 194 nitrosamine formation, 66
Adjuvant chemotherapy, See Cancer Amiodarone, 18, 46, 219
chemotherapy, adjuvant agents Amitriptyline, 181, 183
Adriamycin, 222 Amlodipine, 46
Amyloid peptides, 146–148
Aflatoxin, 65, 67, 70
Amyotrophic lateral sclerosis (ALS), 130, 154,
Age-related macular degeneration, 143
See also Neurodegenerative
Age-related neurodegenerative disorders, See
disorders
Neurodegenerative disorders;
apoptotic mechanisms, 131
specific disorders
epidemiology and genetics, 154
Aging, neurodegenerative disorder risk factor,
oxidative stress and, 130
130–131
Anabolic steroids, 219
Alcohol dehydrogenases, 272–273
Anesthesiology and supplements, 48, 209
Alcohol (ethanol) interactions, See also Red
herbals, 216–225
wine; Wine consumption
minerals, 210–212
cancer nutrigenomics, 272
recommendations, 226
dietary carcinogens and, 68–69 vitamins, 213–215
head and neck cancer, 67 Anorexia-cachexia syndrome, 99–100, See also
kava kava, 225 Cachexia
liver cancer, 65 Antacids, 212, 216
metformin and, 9 Anthocyanins, 41, 135, 136
sulfonylureas and facial flushing, 8 Anthracyclines, 101, 278, 279, 287, 289
vitamin A hepatotoxicity, 213 Antibiotics
Aldehyde dehydrogenases, 272–273 garlic, 223
Aldehydes, 132 mineral interactions, 210–212
Allicin, 42, 43, 222–223 Anticoagulants, See specific drugs
Allyl sulfides, 79 Anticonvulsants, 215, 216, 224
Alpha-linoleic acid (LNA), 108 Antidepressants, See Depression
Alprazolam, 225 pharmacotherapy; specific types
319
320 Nutrient Drug Interactions
Antiemetics, 221 garlic and, 223

Antioxidants, 137, See also specific antioxidants gingko and, 224
anti-atherosclerosis benefits, 43 ginseng and, 225
biomarkers, 74 NFκB inhibition and, 145
cancer preventive/protective effects, 73–75, vitamin C interactions, 214
99 Atherogenic dietary factors, 31–32, 39, 41, See
chemotherapy outcomes and, 101–102 also Cardiovascular disease;
nutrigenomics, 270, 271, See also Dietary fats and cholesterol
Cancer nutrigenomics Atorvastatin, 17, 33, 45, 46, 245
dietary flavonoids, 135–140, See also ATP-binding cassette transporter, 37, 102,
Flavonoids 287–288, 292–293
dementia risk and, 133 Atypical antidepressants, 181, 195
polyphenol metabolism and activity, Avasimibe, 37
140–142, See also Polyphenols Azathiopurine, 283, 290
putative health benefits, 142–143 AZT, 214
garlic and, 222
hypolipidemic/cardioprotective effects, 39,
41 B
multi-nutrient cocktails, 106, 156
neurodegenerative diseases and, 130, 133 Bacillus Calmette–Guerin treatment, 286
Alzheimer’s disease, 147–149 Baicalein, 76
inhibition of NFκB activation, 144 Barbecued meats, 82
Parkinson’s disease, 153 Barnidipine, 46
research and therapeutic implications, Basic fibroblast growth factor (BFGF), 106
156–157 Bcl-2, 104, 133
Anxiety treatment, 224 Bcl-XL, 104
Apigenin, 76, 77–78, 103 B-complex vitamins, neurodegenerative
Apolipoprotein E, 146, 240–241, 244–245 disorders and, 149
Apoptosis Benzo(a)pyrene (BaP), 65, 72, 78
neurodegenerative diseases and, 130, 131 Bergamottin, 45
NFκB activation, 145 Beta blockers, 210, See also Calcium channel
oxidative stress and, 130, 131, 132 blockers
Parkinson’s disease and, 152–153 grapefruit juice and, 46
vitamin D and cancer, 248 resin interactions, 18
Apples, 135, 136–137 Beta-carotene, See β-Carotene
Apricots, 138 Bevacizumab, 293
Apurinic/apyrimidinic endonuclease 1, 287 Bezafibrate, 35
1-β-Arabinofuranosyl-cytosine (AraC), 283 Biguanides, See Metformin
Arabinosylcytosine, 112 Bile acid sequestrants, 18, 35–36, 216
AraC, 113 Bilobalide, 195
Arachidonic acid (AA), 100, 105, 108, 109, 111, Bioavailability, 2, See also specific drugs,
220 chemicals, and nutrients
Arginine, 106, 107 cancer chemotherapeutic agents, 103
Aromatase, 79 grapefruit juice and, 3, 5, 17, See also
Aryl hydrocarbon receptor (AhR), 66, 72, 82 Grapefruit juice
carcinogen complex and activation, 73 Bipolar disorder, 191
phytochemical activation, 75–77 Bisphosphonates, 210
phytochemical antagonism, 77–78 Black tea, 78, 135
Ascorbic acid, See Vitamin C Blackberries, 136
Aspartame, 84 Blackcurrants, 136
Aspirin (acetylsalicylic acid) Bladder cancer, 64, 106, 272, 282, 286
“alcohol flush” symptom treatment, 8 Blood pressure, 196
feverfew interactions, 220 Blueberries, 138
folate and, 216 Brain, antioxidant status, 131–132
Index 321
Brain-derived neurotrophic factor (BDNF), 149 gene polymorphisms and dietary factors,
Brazil nuts, 292 265–271, See also Cancer
Breast cancer, 64, 96 nutrigenomics
antioxidants and, 101 green tea phytochemicals, 82
COX-2 overexpression and, 105 epidemiology, 96–97
diet and lifestyle risk factors, 97 oxidant stress and, 75
fatty acid supplementation effects on patient weight loss, 97
chemotherapy, 112 phytochemical induction of Phase II
flaxseed phytoestrogens and, 113 enzymes, 80–81
genetic polymorphisms and dietary factors, procarcinogenic phytochemicals, 77–78
267–270 public perception of risk, 83–85
mushroom polysaccharides and, 108 Cancer chemotherapy, 95–96, 106–107,
polymorphic genes and chemotherapy 263–264
outcomes, 278–282, 286, 287, 289 antioxidants and, 101–102
vitamin D and, 247 chemosensitizers, 102–103
Breast cancer resistance protein (BCRP), 288 drug resistance development, 102
Broccoli, 76, 80, 135, 138 genetic polymorphisms, drug interactions
Brussels sprouts, 76 and outcomes, 276–290, See also
Bupropion, 181 Cancer pharmacogenomics and
Buthionine sulphoximine, 131 pharmacogenetics
Butyrate, 105 interindividual response variation, 100
in vitro and in vivo bioavailability, 103
nutrigenomics, See Cancer nutrigenomics
C nutritional status and treatment outcomes,
97, 99–102
Cabbage, 76 beneficial dietary deficiencies, 114
Cachexia, 99–100, 107, 293 pharmacogenomics, 276–294, See also
Caffeic acid, 141 Cancer pharmacogenomics and
Caffeine, 79, 266 pharmacogenetics
Calbindins, 133 Cancer chemotherapy, adjuvant agents, 98–99
Calcium/calmodulin-dependent protein kinase antifolate chemotherapeutic agents, 291
II (CaMKII), 182, 191 flaxseed phytoestrogens, 113–114
Calcium channel blockers, 210, 215, See also gastrointestinal toxicity treatment, 100–101
Beta blockers hematopoietic toxicity treatment, 100
cancer chemotherapy chemosensitizer, 102 immunomodulators, 107–108
ginseng and, 225 monoclonal antibodies, 293
grapefruit juice and, 45, 46 NSAIDs, 105, 110
Calcium supplements, 210, 215 nutritional cocktails, 106
Caloric restriction, cancer preventive/protective pharmacogenomics, See Cancer
effects, 273, 274 pharmacogenomics and
Camptothecin, 113 pharmacogenetics
Cancer, 262, See also specific types polyunsaturated fatty acids, 108–113
angiogenesis modulation, 275 targets
body composition changes, 99 COX inhibition, 105–106, 110
cachexia, 99–100, 107, 293 drug transporting glycoprotein
calorie restriction and, 273, 274 interactions, 102–104
changes in nutritional status, 97, 99–101 transcription factors, 104–105
COX-2 overexpression and, 105–106 Cancer nutrigenomics, 246–252, 271–272
diet and lifestyle risk factors, 64, 96–97 alcohol/aldehyde dehydrogenase
dietary chemopreventative/chemoprotective polymorphisms, 272–273
factors, 73–75, 95–96, 263, See alcohol metabolism, 272–273
also Cancer chemotherapy, angiogenesis modulation, 275
adjuvant agents; specific nutrients diet, genetics, and risk, 263–264
or phytochemicals DNA repair enzymes, 273
energy balance regulation, 274 Cardiovascular disease, 31–32

folate metabolism and DNA methylation, antioxidant vitamin supplements and, 39,
249–251, 271–272 41, 43
oxidant/antioxidant enzyme diet-heart hypothesis, 31–32
polymorphisms, 270, 271 hypolipidemic/cardioprotective natural
vitamin D and vitamin D receptor, 247–248, dietary components, 39–44
275 hypolipidemic drugs, 31–38, See also
xenobiotic-controlling pathways, 264–271 Hypolipidemic and
dual Phase I/Phase II enzyme hypocholesterolemic drugs
polymorphisms, 267–268 metabolic syndrome, 32
Phase I (CYP enzyme) polymorphisms, orlistat and, 14
264–266 Cardiovascular disease, genetic polymorphisms
Phase II (glutathione S-transferase) and nutrigenomics, 239–245
polymorphisms, 266, 268–269, apolipoprotein E, 240–241
271 saturated fat intake, 240
Cancer pharmacogenomics and sucrose intake, 241
pharmacogenetics, 276–290 hepatic lipase, 243–244
ATP-binding cassette transporter, 287–288, methylenetetrahydrofolate reductase,
292–293 241–242
CYP enzyme gene polymorphisms, niacin, 245
277–278 statins, 244–245
DNA repair effects, 285–287, 291–292 Carmustine, 114
drug-metabolizing enzymes, 276–290 β-Carotene
folate metabolism enzymes, 283–285, 291 age-related cognitive impairment and, 148
glutathione S-transferase gene cancer risk and, 96, 99
polymorphisms, 279–282, 290 chemopreventative effects in smokers,
purine/pyrimidine metabolism enzymes, 74–75
283, 290–291 chemotherapy outcomes and, 101
receptor and regulatory factor gene neurodegenerative diseases and, 134
polymorphisms, 288–289, 293 orlistat and, 15
sulfotransferase gene polymorphisms, Carotenoids, 131
282–283 antioxidant activity, 135–136
UDPGT polymorphisms, 277, 279, 290 hypolipidemic/cardioprotective effects, 41
Cantharidin, 76 neurodegenerative diseases and, 134
CAPE, 156 Carvedilol, 46
Capecitabine, 291 Caspase inhibitors, 155–156
Capsaicin, 79, 104 Catalase, 155, 270
Captopril, 212 Catechins, 135, 138
Carbamazapine, 224 cardiovascular protective effects, 142
Carbidopa, 212 chemopreventative effects, 75
Carbohydrate malabsorption, α-glucosidase neurodegenerative diseases and, 143
inhibitors and, 11–12 Celecoxib, 113
Carboxyfullerenes, 155 Celiprolol, 46
Carcinogen dose–response curves, 83 Cerivastatin, 18, 33
Carcinogens, cooked or processed meat, 63–70, Cervical cancer, 64, 106, 113, 272
See also Heterocyclic amines; Cetuximab, 293
Meat-associated carcinogens; Chemotherapy, See Cancer chemotherapy
Nitrosamines; Polycyclic aromatic Chinese medicine, 106, 107
hydrocarbons Chlorophyll, 78
Carcinogens, food spoilage, See Aflatoxin Chlorpropamide, 6–8
Carcinogens, hormesis (beneficial low-level Chocolate, 135
effects), 83–84 Cholesterol, dietary, See Dietary fats and
Carcinogens, tobacco smoke, 66, 69, 75 cholesterol
Index 323
Cholesterol absorption inhibitors, 18–19, 37, ALS and, 156

See also Hypolipidemic and Alzheimer’s disease model, 147
hypocholesterolemic drugs cancer preventive/protective effects, 75, 98,
Cholesterol ester transfer protein (CETP) 103, 105, 106
inhibitors, 37 P-glycoprotein transporter inhibition and,
Cholestyramine, 18, 35, 215 293
Chromium, 210–211 transcription factors and, 104
Chrysin, 76 xenobiotic metabolism and, 76, 77–78, 79
Ciprofibrate, 35 Cyanidin, 41, 136
Cisplatin, 114, 280, 281, 282, 283, 285, 288, 289 Cyanocobalamin, 214
Citalopram, 181, 183, 192 Cyclic AMP-dependent protein kinase (PKA),
Clarithromycin, 219 182
Clomipramine, 181, 183 Cyclic GMP, 112
Cobalt, 213 Cyclooxygenase enzyme inhibitors, 105–106,
Colchicines, 214 141, See also COX-1; COX-2
Colesevelam, 35–36 cancer chemoprevention/chemotherapy, 110
Colestipol, 18, 35, 215 Cyclophosphamide, 112, 113, 276, 277, 278,
Colorectal cancer, 64 282, 287
cooked meat carcinogens and, 265, 266 Cyclosporin A, 102
COX-2 overexpression and, 105 Cyclosporine, 217, 219
fatty acid supplementation effects on CYP1A
chemotherapy, 113 cooked meat carcinogen metabolism, 69–70,
genetic polymorphisms and dietary factors, 82
266–270 phytochemical induction, 75–77
genetic polymorphisms and risk, 250–251 phytochemical inhibition, 79, 80
mushroom polyphenols and, 107 CYP1A1
polymorphic genes and chemotherapy BaP metabolism, 72
outcomes, 279, 281, 283–286, 288, polymorphisms and cancer risk, 266
289 polymorphisms and chemotherapy
vitamin D and, 247–249 outcomes, 276–278
Complex carbohydrates, 11 CYP1A2
Constitutive androstane receptor (CAR), 72 aflatoxin metabolism, 70
Cooking effects, fruits and vegetables, 137 BaP metabolism, 72
Cooking effects, meat, See Meat-associated echinacea and, 219
carcinogens polymorphisms and cancer risk, 266
Coronary heart disease, See also Cardiovascular polymorphisms and chemotherapy
disease outcomes, 277
hypolipidemic therapy, 33, 38, See also CYP2A6, 266, 277
Hypolipidemic and CYP2B, 72
hypocholesterolemic drugs CYP2B1, 80
red wine protective effects, 142 CYP2B6, 277, 278
Corticosteroids, 219 CYP2C8, 277
COX-1, 105 CYP2D6, 224, 277
ginger and, 222 CYP2E1, 68–69, 80
COX-2 phytochemical inhibition, 80
inhibition and cancer polymorphisms and cancer risk, 266
chemoprevention/chemotherapy, CYP3A
111 cancer pharmacogenomics and, 277
neurodegenerative diseases and, 133 herb interactions, 5
overexpression and cancer progression, SXR receptor, 72
105–106 CYP3A4, 6
phytochemical inhibitors, 106 aflatoxin metabolism, 70
Cranberries, 105 antidepressant metabolism, 194
Curcumin, 135 chemotherapeutic pharmacogenomics, 277
fibrate metabolism, 17 monoamine hypothesis, 180, 182, 198

gingko biloba and, 195 nutrient and phytochemical interactions,
grapefruit juice and, 17, 45, 277 182–198
hypolipidemic drug interactions, 44 amino acids, 182–186, 199
juice–drug interactions, 47 folate, 186–190, 199–200
phytochemical inhibition, 80 gingko biloba, 195, 201
polymorphisms and cancer risk, 265 ginseng, 195, 201, 225
polymorphisms and chemotherapy homocysteine, 186–190
outcomes, 277, 278 omega-3 fatty acids, 197–198, 202
St. John’s wort and, 72, 104 one-carbon metabolism, 186–190
statins and, 33 S-adenosylmethionine, 190–191
CYP3A5, 277 St. John’s wort, 193–194, 201, 217
Cytidine deaminase (CDA), 283 summary table, 199–202
Cytochrome P450 system, 2, 16–19, 71, See tyramine, 196, 202
also specific CYP subfamily types vitamin B6, 195–196, 201
cancer pharmacogenomics and, 276–277 vitamin B12, 186–188
dietary carcinogen exposure and, 70–73, zinc, 192–193, 200, 201
265–266 protein phosphorylation, 182
garlic and, 223 relapse prevention, 184, 187
gingko biloba and, 195 serotonin syndrome, 180, 185, 194, 217
grapefruit juice and, 17, 18, 45, 277 Desflurane, 221
meglitinide metabolism, 12 Desipramine, 181, 184, 187
phytochemical exposure and, 75–77 DHA, See Docosahexaenoic acid
phytochemical inhibition, 79–80 Diabetes
polymorphisms and cancer risk, 264–266 chromium and, 211
polymorphisms and chemotherapy combined lifestyle/drug intervention, 9
outcomes, 276–278 dietary intervention, 2, 3
procarcinogenic phytochemicals, 77–78 fiber, 4–5
receptor binding, 72, See also specific healthy diet, 3–4
receptors supplements and herbal products, 5
secondary carcinogen metabolism, 66–70 glucose-lowering agents, 6–13, See also
St. John’s wort and, 5 Glucose-lowering agents
statins and, 17, 33 lipid-lowering agents, 16–19, See also
thiazolidinedione metabolism, 9 Hypolipidemic and
hypocholesterolemic drugs
vitamin A deficiency, 213
D Diallyl disulfide, 81
Dietary fats and cholesterol
Dacarbazine, 289 apolipoprotein E polymorphisms and,
Daidzein, 76, 79 240–241
DATATOP, 153 diabetic energy intake and, 4
Dementia, See also Alzheimer’s disease fish oil supplements, 111, 113, 149
age correlation, 146 healthy diet guidelines, 3
gingko and, 223 hepatic lipase polymorphisms and, 244
lifestyle and dietary habits and, 134 hypolipidemic drug interactions, 38, 44–48
Deoxycytidine kinase (dCK), 283 lipid homeostasis and, 38–44
Deprenyl, 152 omega fatty acids, See Omega-3 (ω-3) fatty
Depression, 179–182 acids
lipid status and, 197 polyunsaturated fatty acid benefits, 39, See
methylation metabolism, 186–190 also Polyunsaturated fatty acids
Depression pharmacotherapy, 179–182, 198, Dietary fiber, See Fiber
See also specific antidepressants Dietary guidelines, 3–4
antidepressant types, 181 Dietary protein, cancer risk and, 67
bipolar contraindications, 191 Diet–heart hypothesis, 31–32
Index 325
Digitalis, 210, 215, 221 chemotherapy outcomes and, 107

Digoxin, 18, 224, 225 depression and, 197, 202
Dihydropyrimidine dehydrogenase (DPD), 283, metabolite anti-inflammatory properties,
290–291 109
Dihydrotestosterone, 216 wound healing, 100
Diindolylmethane (DIM), 75 Electroconvulsive shock therapy, 192
Diltiazem, 46, 219 Ellagic acid, 98, 104–105
DIM, 78 Elm bark, 101
Dimenhydrinate, 221 Endometrial cancer, 106
7,12-Dimethylbenz(a)-anthracene, 96 Energy intake
Dimethylnitrosamine, 66 acarbose treatment for diabetes and, 11
Diosmin, 76, 79 apolipoprotein E polymorphisms and, 241
Dioxins, 72, 73, 83 caloric restriction and cancer risk, 273, 274
Disopyramide, 46 low calorie diet and neurodegenerative
DMN-induced carcinogenesis, 78, 79 disorder risk, 130
DNA methylation, 249–251, 271–272 sibutramine and, 15–16
DNA oxidation, 132, 147 Ephedra, 217, 220–221
DNA repair Epicatechin, 135, 136, 143
cancer pharmacogenomics and, 285–287, Epidermal growth factor (EGF), 104, 288, 293
291–292 Epigallocatechin-3-gallate (EGCG), 75, 78,
polymorphic genes and cancer risk, 273 106, 135, 143, 156
Dobutamine, 214 Epileptic seizures, 145
Docetaxel, 276, 285 Epinephrine, depression pharmacotherapy and,
Docosahexaenoic acid (DHA), 108 182
Alzheimer’s disease model, 149 Epothiline, 113
cancer chemoprevention/chemotherapy, 99, Eriodictyol, 138
111–112, 113 Erlotinib, 293
depression and, 197, 202 Escitalopram, 181
wound healing, 100 Esophageal cancer, 64, 66, 285
Donazepil, 223 Estrogens, 219
Dopamine Ethanol interactions, See Alcohol (ethanol)
depression pharmacotherapy and, 182 interactions
NFκB and apoptosis induction, 145 Ethoxyquin, 79
Parkinson’s disease and, 149–153 Excision repair cross complementation group 1
polyphenol-associated inhibition of (ERCC1), 285, 292
oxidation of, 141 Excision repair cross complementation group 6
St. John’s wort and, 194 (ERCC6), 286
Doxorubicin, 101, 103, 112, 114, 276, 280 Ezetimibe, 18–19
Drug-metabolizing reactions, See Xenobiotic
metabolism
Drug resistance, 102 F
polymorphic genes and chemotherapy
outcomes, 287 Familial ALS, 154
transporters, 102 Farnesoid X receptor (FXR), 38, See also
Duloxetine, 181 specific receptors
Fat excretion, 14
Fat intake, See Dietary fats and cholesterol
E Fatty acid binding proteins, 109–110
Fecal fat excretion, 14
Echinacea, 217–219 Felodipine, 5, 45, 46
EGB761, 143, 223 Fenofibrate, 17–18, 35, 47
Eicosapentaenoic acid (EPA), 40, 108 Feverfew, 220
cancer chemoprevention/chemotherapy, Fiber, 4–5
99–100, 111–112, 113 cancer risk and, 263
hypolipidemic/cardioprotective effects, 42, depression pharmacotherapy and, 186–190,

43 199–200
Fibrates, 17–18, 34–35 genetic polymorphisms and cancer risk,
grapefruit juice and, 47 249–251, 271–272
statins and, 47 genetic polymorphisms and cardiovascular
Fibroblast growth factor-2 (FGF2), 104 health, 242
Fibrosarcoma, 112 metformin and, 8
Finasteride, 216 NSAID interactions, 216
Fish resin interactions, 18
fermented, nitrosamines in, 66 Follicular lymphoma, 289
omega fatty acids and, 111, 113, 149, 197 Food spoilage carcinogens, See Aflatoxin
Fish oil supplements, 111, 113, 149 Free radicals, 131, See also Oxidative stress;
Flavanols, 40, 136, 138, 142 Reactive oxygen species
Flavanones, 41, 135, 138, 140 French paradox, 142–143
Flavones, 135, 138, 140 Fruit juice–drug interactions, 47, See also
Flavonoids, 135–142, See also specific Grapefruit juice
substances Fruits and vegetables, See also Flavonoids;
age-related diseases and, 130 specific foods
anticancer mechanisms, 275 age-related diseases and, 130
antioxidants, 131, See also Antioxidants cancer prevention/protection effects, 73–75,
cancer chemotherapy chemosensitizers, 95–96, 273, See also specific
102–103 nutrients or phytochemicals
cooking/processing effects, 137 cooking/processing effects, 137
COX-2 inhibitors, 106 deficiency and cancer risk, 263
CYP1A induction, 76 healthy diet guidelines, 3–4
cytoprotective effects, 142 induction of Phase II enzymes, 80–81
echinacea, 219 natural antioxidants, 135–143, See also
Antioxidants; specific types
enzymes inhibited by, 141
neurodegenerative disease risk and, 134
gingko biloba, 223
procarcinogenic phytochemicals, 77–78
glutathione synthesis upregulation, 74
public risk perceptions, 85
juice–drug interactions, 47, See also
Furanocoumarins, 45
Grapefruit juice
metabolism of, 135–136
neurodegenerative diseases and, 134
putative health benefits, 142–143
G
red wine protective effects, 142 Galangin, 76
structures, 136–137 Gallic acid equivalent, 155
subtypes, 135 Gallocatechin, 135
xenobiotic metabolism and, 79 Gamma-aminobutyric acid (GABA), 195, 217
Flavonols, 40, 135, 138, 140 Gamma-glutamylcysteine synthase, 74, 80
Flaxseed, 98, 104, 113–114 Garlic, 222–223
Fluoropyrimidine, 283 antioxidant action, 222
Fluoroquinolones, 213 hypolipidemic/cardioprotective effects, 42,
5-Fluorouracil, 101, 108, 113, 114, 279–291 43
Fluoxetine, 181, 183, 184–185, 187 xenobiotic metabolism and, 80–81
Flutamide, 277 Gastrointestinal toxicity, 100–101
Fluvastatin, 17, 33, 45, 46 Gastrointestinal tract cancers, See also Stomach
Fluvoxamine, 183 cancer
Folate and folic acid, 215–216 mushroom polyphenols and, 107–108
antifolate chemotherapeutic agents, 291 phytochemicals and risk, 81
cancer pharmacogenomics and, 283–285, polymorphic genes and chemotherapy
291 outcomes, 282, 284
deficiency and cancer risk, 272 Gefitinib, 293
Index 327
Gemcitabine, 100, 113, 283 β-Glucuronidase, 140

Gemfibrozil, 17–18, 33, 35, 47 Glutamine, cancer chemotherapy and, 101, 107
Genetic polymorphisms, 238–239, 265 Glutathione (GSH), 74, 131
alcohol/aldehyde dehydrogenases, 272–273 acetaminophen metabolism and, 68
cancer and, 246–251, 263–293, See also conjugation, 66, 68, See also Phase II
Cancer nutrigenomics; Cancer xenobiotic conjugation
pharmacogenomics and Parkinson’s disease and, 151–152
pharmacogenetics selenium and, 212
cardiovascular disease and, 239–245, See Glutathione peroxidase, 74
also Cardiovascular disease, Glutathione S-transferase (GST), 44, 69–70, 73
genetic polymorphisms and gene polymorphisms and cancer risk, 265,
nutrigenomics 268–269, 271
DNA repair enzymes, 273 genetic polymorphisms and chemotherapy
energy balance regulation, 274 outcomes, 279–282, 290
oxidant/antioxidant enzyme phytochemical induction, 80
polymorphisms, 270, 271 Goldenseal, 80
receptors and regulatory factors, 288–289, Grape seed extract, 98, 104
293 Grapefruit, flavonoids in, 138
xenobiotic-controlling pathways, 264–265 Grapefruit juice
dual Phase I/Phase II enzyme CYP enzyme inhibition, 17, 18, 277
polymorphisms, 267–268 hypolipidemic drug interactions, 45–47
Phase I (CYP enzyme) polymorphisms, metabolic disorders and, 5–6
264–266, 276–278 P-glycoprotein transporter inhibition and,
Phase II (glutathione S-transferase) 293
polymorphisms, 266, 268–269, statin bioavailability and, 17, 45, 46
271 Grapes, 135
Genetics, neurodegenerative disorders, 146, Green tea, 135
150, 154 cancer prevention/protection effects, 75, 98,
Genistein, 76, 79, 98, 103 103–104, 106, 273
Ghrelin, 274 flavonoids, 135
Ginger, 221–222 immunomodulatory activity, 107
Gingko biloba, 143, 195, 201, 223–224 polyphenols, 143, See also Polyphenols
Ginseng, 225 processed meat carcinogens and, 82–83
antidepressant interactions, 195, 201, 225 transcription factors and, 104
complementary cancer treatment, 103 xenobiotic metabolism and, 76, 78
Ginsenosides, 103 GTP-binding proteins, 107
Glibenclamide, 6, 7 Guar gum, 5, 8
Gliclazide, 6, 7–8 Guggulipid, 44
Glimepiride, 6, 7
Glioblastoma multiforme, 105
Glioma, 280, 282 H
Glipizide, 6, 7
Gliquidone, 7 Halothane, 221
Glitazones, 6, 8 Hematopoietic toxicity, 100
Glucose-lowering agents, 6–13 Heparin, 214, 220, 223, 224, 225
α-glucosidase inhibitors, 6, 10–12 Hepatic lipase, 243–244
magnesium interactions, 211 Hepatitis B, 65
meglitinide derivatives, 6, 12–13 HER-2/neu, 104, 105, 112, 288, 293
metformin, 6, 8–9 Hesperetin, 138, 140, 142
sulfonylureas, 6–8 5-HETE, 109–111
thiazolidinediones, 6, 9–10 12-HETE, 109–110
Glucose tolerance, 210–211 15-HETE, 100
α-Glucosidase inhibitors, 6, 10–12 Heterocyclic amines (HCAs), 63–70, 265
Glucosinolates, 75–76 animal-based research, 68
cooked/processed meat, 65–66, 82 sterols and stanols, 19

experimental models and future research, transcription regulators, 38
81–83 vitamin D and, 215
green tea phytochemicals and, 82–83
metabolism of, 69–70
phytochemical inhibition of metabolism, 79 I
phytochemicals and formation during meat
processing, 82 Ibuprofen, 216
tobacco smoke, 66 Id vaccination, 289
xenobiotic metabolism, 64–68 Imatinib, 293
High-density lipoprotein (HDL) cholesterol Imipramine, 181, 190, 192
apolipoprotein E polymorphisms and, 240 Immunoglobulin GFc receptor, 289, 293
CETB inhibitors and, 37 Immunomodulatory agents, 107
hepatic lipase polymorphisms and, 243–244 echinacea and, 217–219
niacin and, 36 Indinavir, 219
Histamine2 (H2) receptor blockers, 214 Indole-3-carbinol, 73, 76, 77, 78, 85, 114
HMG CoA reductase inhibitors, See Statins Insulin-like growth factor (IGF-1), 104, 274
Homocysteine, 186–190, 199–200, 241–243 Insulin resistance, 211
Hops, 79 Insulin secretion stimulation, 12–13
Hormesis, 83–85 Interferon-γ (IFN-γ), polymorphic genes and
Hot dogs, 66, 68, 82 chemotherapy outcomes, 289, 293
Hydrogen peroxide, 74, 131–131, 152 Interleukin-1 (IL-1), 111
Hydroxyl radical, 131 NFκB activation, 144
7-Hydroxymatairesinol, 113 polymorphic genes and chemotherapy
β-Hydroxy-β-methylbutyrate (HMB), 100 outcomes, 289, 293
Hydroxytirosol, 156 Interleukin-6 (IL-6), 289
4-Hydroxy-2-trans-nonenal (4-HNE), 132 Interleukin-8 (IL-8), 108, 289
5-Hydroxytryptamine (5-HT), See Serotonin Intestinal P-glycoprotein, See P-glycoprotein
5-Hydroxytryptophan (5-HTP), 184–185 Irinotecan, 113, 276, 279, 287, 288, 290
Hyperemesis, 222 Iron
Hyperforin, 47, 72, 104, 194, See also St. John’s deficiency, 212
wort drug interactions, 212
Hyperglycemia pharmacotherapy, See Glucose- feverfew interactions, 220
lowering agents oxidative stress and, 151
Hypericin, 47 Isoflavones, 41, 43, 45, 135, 138
Hypertensive crisis, 196 Isoflurane, 221
Hypolipidemic and hypocholesterolemic drugs, Isoniazid, 215
16–19, See also specific drugs Isoprostanes, 109
ATP-binding cassette transporter, 37 Isothiocyanates, 76, 79, 290
bile salt binders (resins), 18, 35–36 Isoxanthohumol, 76
cholesterol absorption inhibitors, 18–20, 37
dietary component interactions, 38, 44–48
emerging and future directions, 37–38 J
ezetimibe, 18–19
fat-soluble vitamin absorption and, 14–15 JNK activation, 142
fibrates, 17–18, 34–35 JTT-705, 37
garlic and, 222 Juice–drug interactions, 47, See also Grapefruit
genetic polymorphisms and cardiovascular juice
health, 245
grapefruit juice and, 6, 45–47
HMG CoA reductase inhibitors, See Statins K
niacin, 36
orlistat, 13–14 Kaempferol, 76, 78, 138
statins, See Statins Kaki, 136
Index 329
Kale, 138 hypolipidemic therapy, See Hypolipidemic

Kava, 80, 224–225 and hypocholesterolemic drugs
Keratinocyte growth factor, 101 oxidation and atherosclerosis, 39, See also
Ketoconazole, 219 Lipid oxidation
Krestin, 98, 108 statins and, 33
LOX inhibitors, 110
Ltb4, 111
L Lung cancer, 64, 96
COX-2 and, 106
α-Lactalbumin, 184–185, 199 dietary chemopreventative agents, 74–75, 96
Lazaroids, 147 genetic polymorphisms and dietary factors,
L-DOPA (levodopa), 152, 212, 225 268–269
Leeks, 138 mushroom polysaccharides and, 108
Legume protein, 42 nutrition and treatment outcomes, 99–100
Lemons, 138 phytochemicals and risk, 81
Lentinan, 107 polymorphic genes and chemotherapy
Leptin gene polymorphism, 274 outcomes, 281, 284, 285, 286, 287
Leucovorin, 113 Lutein, 41, 78, 98
Leukemia Luteolin, 76, 78
antioxidant vitamins and, 101–102 Lycopene, 41, 79, 134
gene polymorphisms and cancer risk, Lysine, chemotherapy adjuvant, 106
250–251
gene polymorphisms and chemotherapy
outcomes, 280–287 M
omega fatty acid supplementation effects,
Ma huang, 220–221
112
Macrocytic anemia, 188
Levothyroxine, 210
Macular degeneration, 143
Lewy body, 151
Magnesium, 211
Licorice root, 101
Maitake mushroom, 114
Linoleic acid, 108
Major depressive disorder, See Depression
Lipase inhibitor, 13–14
Malignant glioma, 280, 282
Lipid homeostasis, dietary factors in, 38–44, See Marshmallow root, 101
also Dietary fats and cholesterol
Matrix metalloproteinases (MMPs), 104
hypolipidemic and cardiprotective nutrients, Meat-associated carcinogens, 63–70, 82, 263,
38–44 See also Heterocyclic amines;
Lipid-lowering agents, See Hypolipidemic and Nitrosamines; Polycyclic aromatic
hypocholesterolemic drugs hydrocarbons
Lipid mobilizing factor (LIF), 99 animal-based research, 68
Lipid oxidation, 132, 147 chemopreventative dietary components, 75
Lipids, dietary, See Dietary fats and cholesterol experimental models and future research,
Lipoxygenase, 141 81–83
Lithium, 187 gene polymorphisms and cancer risk,
Liver cancer, 64, 65, 70, 268 265–268
Liver X receptors (LXRs), 38 green tea phytochemicals and, 82–83
Losartan, 46 xenobiotic metabolism and, 70–73, 265–266
Lovastatin, 17, 33, 45, 46, 219 Megaloblastic anemia, 9
Low density lipoprotein (LDL) cholesterol Meglitinide derivatives, 6, 12–13
apolipoprotein E polymorphisms and, Melanoma, polymorphic genes and
240–241, 244–245 chemotherapy outcomes, 289
bile salt binders (resins) and, 18 6-Mercaptopurine, 283, 290
cholesterol absorption inhibitors and, Metabolic syndrome, 1–2, 32
18–19, 37 dietary interventions, 2, 3
diet–heart hypothesis, 31 lipid-lowering agents, 16–19
Metformin, 5, 6, 8–9, 214 Muscle relaxants, 211

Methionine restriction, 114 Mushroom polysaccharides, 98, 107–108, 114
Methionine synthase gene (MTR), 249, 272, Myeloperoxidase, 267, 271
285 Myricetin, 76, 78
Methotrexate, 219, 283, 284–285, 291
Methyldopa, 212
Methylenetetrahydrofolate reductase N
(MTHFR), 186, 241–242,
249–251, 271–272, 283, 285, 291 N-acetylcysteine supplementation, 69, 155
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine N-acetyltransferase, 81, 265, 267
(MPTP), 150–151 NAD(P)H:quinone oxidoreductase (NQO1),
Mevalonate, 106–107 282
Mevastatin, 107 Naringenin, 47, 76, 78, 79, 103, 138, 142
Microsomal epoxide hydrolase, 268 Naringin, 79
Microsomal triglyceride transport protein Nasopharyngeal cancer, 288
(MTP) inhibitors, 37 Nateglinide, 6, 12–13
Microtubule-associated protein 2 (MAP2), 182 National Cholesterol Education Program, 3
Midazolam, 219 Natural killer (NK) cell stimulation, 108
Miglitol, 6, 10 Nausea treatment, 221
Migraine treatment, 220 Necrosis, 131
Mineral oil, 215 Nerve growth factor (NGF), 145
Minerals, See also specific minerals Neurodegenerative disorders, 129–131, See also
dietary fiber and, 4 Alzheimer’s disease; Amyotrophic
supplements, 210–212 lateral sclerosis; Parkinson’s
Mint, 138 disease
Mirtazapine, 181 apoptosis, 130–131
Mistletoe lectins, 107 dietary silicon and, 133–134
Mitochondrial dysfunction, Parkinson’s disease epidemiological studies, 133–134
and, 151–153 oxidative stress and, 130, 143–145
Mitogen-activated protein kinase (MAP kinase), red wine protective effects, 143
141 research and therapeutic implications,
Mitomycin C, 112, 282 156–157
Mivacuronium, 211 Neurofibrillary tangles (NFTs), 146
Moclobemide, 183 Neuropeptide Y, 274
Monoamine hypothesis of major depression, NFκB, See Nuclear transcription factor κB
180, 182, 198 Niacin, 36, 245
Monoamine oxidase B (MAO-B) inhibition, Nicardipine, 46
152, 153, 224 Nicotonic acetylcholine receptors (nAChRs),
Monoamine oxidase inhibitors (MAOIs), 193
180–181, See also Depression Nifedipine, 46
pharmacotherapy Nimoldipine, 46
ephedra and, 221 Nisoldipine, 46
ginseng and, 195, 225 Nitrendipine, 46
serotonin syndrome, 180, 185 Nitric oxide synthase (NOS), 112
St. John’s wort and, 217 Nitrosamines (NAs), 63–70, 265
tryptophan and, 183 animal-based research, 68
tyramine and, 196 cooked/processed meat, 65, 66, 82
vitamin B6 and, 195–196 experimental models and future research,
Monoclonal antibodies, 293 81–83
Monounsaturated fatty acids (MUFAs), 39, 40 green tea phytochemicals and, 82–83
Mood disorders, See Depression metabolism of, 68–69
Mucositis, 100–101 phytochemical inhibition of metabolism, 79
Multidrug resistance proteins, 102–103, 287, phytochemicals and formation during meat
292 processing, 82
Index 331
tobacco smoke, 66, 75 glucose-lowering agents, See Glucose-

xenobiotic metabolism, 64–68 lowering agents
Nitrosoureas, 280, 282 lipid-lowering agents, 16–19, See also
Nitrotyrosine, 132 Hypolipidemic and
Nitrous oxide, 214, 216 hypocholesterolemic drugs
N-methyl-D-aspartic acid (NMDA), 145, 182, orlistat, 13–15
192–193 sibutramine, 15–16
N-nitrosamides, 66 6-OHDA, 143
NNK, 69, 75, 79 Olestra, 215
Non-Hodgkin lymphoma, 274 Omega-3 (ω-3) fatty acids
Nonsteroidal anti-inflammatory drugs cancer treatment potential, 99, 108–112
(NSAIDs), See also specific drugs depression and, 197–198, 202
ascorbic acid interactions, 214 eicosanoid product balance, 109
food sources, 39
chemopreventive effects, 105, 110
feverfew interactions, 220
40
folate and, 216
neurodegenerative diseases and, 149
gingko and, 224
wound healing, 100
ginseng and, 225
Omega-6 (ω-6) fatty acids
neurodegenerative diseases and, 133 beneficial effects, 40
Norepinephrine, 182–186 eicosanoid product balance, 109
depression pharmacotherapy and, 180, 181, improving ω-3/ω-6 ratio, 39
182 Onions, 135, 138
St. John’s wort and, 194 cooking/processing effects, 137
Nortriptyline, 181, 183 induction of Phase II enzymes, 81
Nuclear transcription factor κB (NFκB), 104, Oral contraceptives, 214
106, 112 Orange juice, 138
neurodegenerative diseases and, 130 Orlistat, 13–15, 215
ALS, 154 Oronasopharyngeal cancers, 64
Alzheimer’s disease, 147–148 Osteoporosis, 215
oxidative stress and activation, 132–133, Osteosarcoma, 106
143–145 Ovarian cancer, 64, 266, 280–281, 283, 288
Parkinson’s disease, 152 Oxaliplatin, 113, 281, 286, 289
Nutrigenomics, 262, See also Genetic Oxidative stress, 131–132, See also
polymorphisms Antioxidants; Reactive oxygen
applications, 238 species
cancer, See also Cancer nutrigenomics apoptosis induction, 130, 131, 132
genetic polymorphisms, 246–251 cancer and, 75
pharmacogenomics and, 276–294, See chemotherapy mechanism and, 101
also Cancer pharmacogenomics functional benefits of low-level stress, 74
and pharmacogenetics glutathione synthesis pathway, 74
cardiovascular disease, 239–245, See also iron and, 151
Cardiovascular disease, genetic neurodegenerative diseases and, 130
polymorphisms and nutrigenomics ALS, 154–156
working definition, 238 Alzheimer’s disease, 146–149
NFκB activation, 132–133, 143–145
Parkinson’s disease, 151–153
O research and therapeutic implications,
156–157
Obesity and overweight, 1–2, 9–10, 13–14 pancreatitis and, 223
cancer risk and, 263 polyphenol cytoprotective effects, 142
dietary interventions, 2, 3 8-Oxoguanine glycosylase I, 273
pharmacological intervention Oxytocin, 221
P Phenformin, 214
Phenobarbital, 72, 219, 224
Paclitaxel, 276, 277, 280, 288 Phenytoin, 216, 219, 224
Pancreatic cancer, 106 Phloretin, 76
Pancreatitis, 223 Phosphoinositide 3-kinase (PI 3-kinase), 141
Paradol, 222 Phospholipase A2, 141
Parkinson’s disease (PD), 130, 149, See also pH-sensitive drug absorption, 7
Neurodegenerative disorders Phytoestrogens, 113–114, 138, See also
antioxidants and, 134, 153, 157 Isoflavones
apoptotic mechanisms, 131, 152–153 Pioglitazone, 6, 9–10
cytoskeletal pathology, 151 Pitavastatin, 33, 45, 46
environmental factors and, 150–151 Platelet-derived growth factor (PDGF), 106
epidemiology and genetics, 150 Platinum-based chemotherapy, 281, 285, 286,
lifestyle and dietary habits and, 134 292, See also specific agents
mitochondrial dysfunction, 151 Policosanol, 43
neuroimaging and autopsy data, 153 Polycyclic aromatic hydrocarbons (PAHs),
oxidative stress and, 130, 151–153 63–70, 265
Paroxetine, 181 animal-based research, 68
Parsley, 138 cooked/processed meat, 65, 82
Parthenolide, 220 experimental models and future research,
PBN, 147 81–83
PCBs, 72 green tea phytochemicals and, 82
Pecans, 105 metabolism of, 69–70, 265
PEITC, 79 phytochemicals and formation during meat
Penicillamine, 155, 212, 213 processing, 82
Pernicious anemia, 213–214 tobacco smoke, 66, 75
Peroxisome proliferator-activated receptor, 9, xenobiotic metabolism, 64–68
34–35, 38, 72, 274 Polymorphic genes, See Genetic
Peroxynitrite, 131, 141 polymorphisms
P-glycoprotein, 45 Polyphenols, 131, 135, See also Flavonoids
chemopreventive agents and, 96 absorption and metabolism, 140–141
herb interactions, 5 apples, 136–137
phytochemical interactions and cancer biological actions, 141–142
chemotherapy, 102–104, 292–293 “French paradox,” 142–143
Pharmacogenomics, 262, 276, See also Cancer green tea, 143, See also Catechins
pharmacogenomics and red wine protective effects, 142
pharmacogenetics structures, 136
Pharyngitis treatment, 101 Polyunsaturated fatty acids (PUFAs), See also
Phase I xenobiotic metabolic reactions, 44–45 Omega-3 (ω-3) fatty acids;
cancer nutrigenomics and, 264–268 Omega-6 (ω-6) fatty acids
cancer pharmacogenomics and, 276 adjuvant chemotherapy, 108–113
meat carcinogen bioactivation, 64 beneficial effects, 39
phytochemical exposure and, 75–77 mood disorders and, 197
Phase II xenobiotic conjugation neurodegenerative diseases and, 149
cancer nutrigenomics and, 266–271 Potatoes, 137
cancer pharmacogenomics and, 276–290 Pramipexole, 153
hypolipidemic drug interactions, 44–45 Pranidipine, 46
phytochemical exposure and, 75–77 Pravastatin, 5, 17, 33, 45, 46, 47, See also Statins
phytochemical induction, 80–81 Pregnancy
secondary carcinogen metabolism, 66–70 iron levels and, 212
xenobiotic exposure and regulation of, magnesium-anesthetic interactions, 211
72–73 vitamin A and, 213
Phenelzine, 181, 195–196, 221, 225 8-Prenylnaringenin, 76
Phenethyl isothiocyanate, 76 Proanthocyanidins, 136, 138
Index 333
Procyanidins, 98, 104, 194 Red wine, 135, 138, 156, See also Wine
Proline, 106 consumption
Propafenine, 46 ALS and, 155
Propofol, 210 cardiovascular protective effects, 142
Prostaglandin D2, 36 flavonoids in, 135, 138
Prostaglandin E2, 105, 106, 111 neurodegenerative diseases and, 143
Prostate cancer, 64 stilbenes, 136
COX-2 and, 106 Red yeast rice, 44
DNA repair enzymes and, 273 Reduced folate carrier 1 (RFC1), 285
fatty acid metabolism, 110 Reductase inhibitors, 216
fatty acid supplementation effects on Repaglinide, 6, 12, 18
chemotherapy, 113 Resins (bile salt binders), 18
genetic polymorphisms and dietary factors, Resveratrol
268, 269, 270 ALS and, 156
leptin gene and, 274 chemotherapy adjuvant, 98, 106
mushroom glucan and, 114 protein kinase inhibition, 142
patient use of alternative medicine, 97 red wine protective effects, 142
vitamin D and, 247 stilbenes, 136
Protein, dietary, cancer risk and, 67 transcription factors and, 104
Protein kinase B, 141 xenobiotic metabolism and, 76, 77
Protein kinase C (PKC), 141, 182 Retinoid X receptor (RXR), 72
Protein kinases, 141, 182 Retinoids, 213
Protein oxidation, 132 Rifampin, 219
Proteolysis inducing factor (PIF), 99–100 Riluzole, 155
Pseudoephedrine hydrochloride, 217 Rituximab, 289, 293
Psoralens, 45 Rosiglitazone, 6, 9–10
Public risk perceptions, 83–85 Rosuvastatin, 17, 33, 45, 46
Purine/pyrimidine metabolism, 290–291 Rotenone, 151
Rutin, 79
Q
S
Quercetin, 76, 78, 79, 103, 135, 136, 138, 141,
142, 156 S-adenosylmethionine (SAM), 186, 190–191,
Quinidine, 46 200, 249
Quinolone, 210, 211 Sage, 78
Quinone reductase, 80 Sausages, 66, 82
Saw palmetto, 216
Scutellaria baiacalensis, 106
R Secoisolariciresinol diglycoside (SDG), 113
Selective serotonin reuptake inhibitors (SSRIs),
Radiotherapy 181, 184–185, See also Depression
hormesis (beneficial low-level effects), 83 pharmacotherapy; specific drugs
polymorphic genes and chemotherapy St. John’s wort and, 194, 217
outcomes, 286–287 tryptophan and, 183
Ras mutation activation, 107 Selegiline, 148, 155
Rasayanas, 114 Selenium, 212
Raspberries, 105 Selenomethionine, 292
Reactive oxygen species (ROS), 131, See also Serotonin (5-HT)
Oxidative stress depression pharmacotherapy and, 183, 199
antioxidant defenses, 74, See also major depression etiology, 180
Antioxidants St. John’s wort and, 194
phase II xenobiotic conjugation and, 73 zinc and brain uptake, 193
Serotonin and norepinephrine reuptake Sucrose intake, cardiovascular health and, 241
inhibitors (SNRIs), 15–16, 181, Suicide victims, 193
183, 184, See also Depression Sulfonylureas, 6–8, 211
pharmacotherapy; specific drugs Sulforaphane, 76, 79–80, 98
Serotonin syndrome, 180, 185, 194, 217 Sulfotransferase, 81, 268, 282–283
Sertraline, 181 Superoxide, 131
Shiitake mushrooms, 107 Superoxide dismutase, 132, 270
Sibutramine, 15–16 mutation and ALS, 154–156
Silicon, dietary, neurodegenerative disorders Synapsin, 191
and, 133–134 Synaptotagmin, 182
Simvastatin, 5, 17, 33, 45, 46, 47, See also α-Synuclein, 151
Statins
Single nucleotide polymorphisms, See Genetic
polymorphisms T
Skin cancer, 274
Skin disorders, 213 Tamoxifen, 108, 276, 277, 279, 282
SN-38G, 290 Tannic acid, 79
Soy, 41, 43, 76, 81, 98, 138, See also Daidzein; Tannins, 136
Genistein TCDD, 75, 78
Spinach, 78 Tea, See Black tea; Green tea
Squamous cell carcinoma, 106 Tegafur, 276, 277
St. John’s wort, 216–217 Tetracyclines, 210, 211, 213
antidepressant interactions, 193–194, 217 Tetrahydrobiopterin (BH4), 189
cancer chemotherapy and, 104 Theaflavins, 79, 135
drug metabolism interactions, 5, 72 Thearubigins, 135
hypolipidemic drug interactions, 47 Thiazide diuretics, 18, 210, 215
P-glycoprotein activity and, 293 Thiazolidinediones (TZDs), 6, 9–10, 35
xenobiotic metabolism and, 80, 104, 277 Thiobarbituric acid reactive substances
Stanols, 19, 42, 43 (TBARS), 147, 222
Statins, 18, 33–34, See also specific drugs Thiocyanates, 76
antitumor activity, 106–107 6-Thioguanine, 283, 290
cytochrome P450-mediated metabolism, 17 Thiopurine S-methyltransferase (TPMT), 283,
ezetimibe and, 37 290
fibrate interaction, 47 Thiotepa, 276
food effects on bioavailability, 17 Throat Coat, 101
genetic polymorphisms and cardiovascular Thromboxane A2 (TXA2), 106
health, 244–245 Thromboxane synthetase, 222
grapefruit juice and, 17, 45, 46 Thymidylate synthase, 283, 291
herb interactions, 5 Thyroid hormone, 212
kava interactions, 225 Thyroxine, 18
niacin and, 245 Tobacco smoke, carcinogens in, 66, 69, 75
Phase II glucuronidation, 44 Tobacco smokers, chemopreventative dietary
St. John’s wort and, 47 components, 74–75
Steroid and xenobiotic receptor (SXR), 72 α-Tocopherol, See Vitamin E
Sterol regulatory element-binding proteins Tolbutamide, 6–7
(SREBs), 38 Tomatoes, 137, 138
Sterols, 19, 42, 43 Topotecan, 276
Stilbenes, 136 Torcetrapib, 37
Stomach cancer, 64, 75, See also Trabectidin, 114
Gastrointestinal tract cancers Trans-fats, 3
polymorphic genes and chemotherapy Transcription regulators, hypolipidemic therapy
outcomes, 284, 289 applications, 38
Strawberries, 105, 135 Tranylcypromine, 181
Sucralfate, 215 Trastuzumab, 293
Index 335
Trazodone, 181, 195 Vitamin B complex, neurodegenerative

Triamterene, 216 disorders and, 149
Triazolam, 219 Vitamin B6, 195–196, 201
Tricyclic antidepressants, 181, See also Vitamin B12, 213–214
Depression pharmacotherapy depression pharmacotherapy and, 186–188,
depression pharmacotherapy and, 184 199–200
gingko and, 224 metformin and, 8–9
SAM and, 190–191 Vitamin C (ascorbic acid), 131, 214
St. John’s wort and, 194 antioxidant activity, 135–136
tryptophan and, 183 brain tissue, 131–132
Trientine, 155 cancer and, 99, 101, 106
Tryptophan, 182–186, 199 deficiency, 214
Tryptophan hydroxylase, 189, 190 dementia risk and, 133
Tryptophan pyrrolase, 184, 186
41, 43
Tubocurarine, 211
neurodegenerative diseases and, 133–134,
Tumor necrosis factor (TNF-β), 111
149
ginseng and, 225
NSAID interactions, 214
NFκB activation, 144–145
Vitamin D, 215, 247
TUNEL, 153
cancer and, 247–248
Turmeric, 78, 135, See also Curcumin deficiency, 215
Tyramine, 196, 202 receptor polymorphisms and cancer risk,
Tyrosine, 182–184, 199 247–249, 275
Tyrosine hydroxylase, 189, 190 Vitamin E (α-tocopherol), 131, 215
Tyrosine kinases, 141, 293 antioxidant activity, 135–136
brain, 132
cancer and, 99
U chemotherapy outcomes and, 101
fatty acid supplementation effects, 112
Ubiquitin, 151
feverfew interactions, 220
UDP-glucuronyltransferase (UDPGT), 44, 70,
hypolipidemic/cardioprotective effects, 41
73, 80, 271, 277, 279, 290
neurodegenerative diseases and, 133–134
Unipolar depression, 179–182
Alzheimer’s disease, 147–149
antidepression therapy, See Depression dl-α-tocopherol, 148
pharmacotherapy
γ- vs. α-tocopherol forms, 133
lipid status and, 197
Parkinson’s disease, 153, 157
methylation metabolism, 186–190 orlistat and, 14–15
monoamine hypothesis, 180, 182, 198 superoxide dismutase defect model and, 155
Urokinase-type plasminogen activator (uPA), supplement cardiovascular protective
104 effects, 39, 43
Voglibose, 10
V
W
Vascular endothelial growth factor (VEGF), 104
Venlafaxine, 181 Walnuts, 105
Verapamil, 46, 210 Warfarin, 18, 213, 214, 223, 224, 225
Very low density lipoproteins (VLDL) Watercress, 80
apolipoprotein E polymorphisms and, 240 Weight-reducing drugs, 15–16
statins and, 33 Wine consumption, See also Red wine
Vinblastine, 276, 289 neurodegenerative diseases and, 134, 157
Vincristine, 114, 276 neurological disorders and, 143
Vitamin A, 15, 135, 213 WS 1490, 224
X secondary carcinogen metabolism, 66–70

St. John’s wort and, 80, 104, 277
Xanthine oxidase, 141 Xeroderma pigmentosum groups, 286
Xenobiotic metabolism, See also Cytochrome X-ray repair cross complementation group 1
P450 system (XRCC1), 286
cancer nutrigenomics and, 264–271
X-ray repair cross complementation group 3
cancer pharmacogenomics and, 276–290
(XRCC3), 273
chemoprotective phytochemicals and, 64
dietary carcinogens and, 64–68, 70–73, 265
dual Phase I/Phase II enzyme
polymorphisms, 267–268
Z
Phase I polymorphisms, 264–266 Zidovudine (AZT), 214
Phase II (glutathione S-transferase)
Zinc, 192–193, 201
polymorphisms, 266, 268–269,
deficiency, 212
271
phytochemical induction, 75–77, 80–81 supplementation, 212–213
phytochemical inhibition, 79–80

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DK5836_title 4/10/06 9:28 AM Page 1

Boca Raton London New York

CRC is an imprint of the Taylor & Francis Group,

Editorial Advisory Board

FRANK GREENWAY, M.D.

KHURSHEED N. JEEJEEBHOY, M.D.

MULCHAND S. PATEL, PH.D.

KATHLEEN M. RASMUSSEN, PH.D.

1. Genomics and Proteomics in Nutrition, edited by

2. Perinatal Nutrition: Optimizing Infant Health and Development,

3. Soy in Health and Disease Prevention, edited by Michihiro Sugano

4. Nutrition and Cancer Prevention, edited by Atif B. Awad

5. Cancer Prevention and Management through Exercise

6. Nutritional Strategies for the Diabetic/Prediabetic Patient,

7. Nutrient–Drug Interactions, edited by Kelly Anne Meckling

Introduction to Clinical Nutrition: Second Edition, Revised

Pediatric Gastroenterology and Nutrition in Clinical Practice,

Nutrients and Cell Signaling, edited by Janos Zempleni

Mitochondria in Health and Disease, edited by Carolyn D. Berdanier

Thiamine, edited by Frank Jordan and Mulchand Patel

Phytochemicals in Health and Disease, edited by Yongping Bao

Handbook of Obesity: Etiology and Pathophysiology, Second Edition,

Handbook of Obesity: Clinical Applications, Second Edition,

No claim to original U.S. Government works

International Standard Book Number-10: 1-57444-915-X (Hardcover)

Library of Congress Cataloging-in-Publication Data

Nutrient drug interactions / edited by Kelly Anne Meckling.

Visit the Taylor & Francis Web site at

DK5836_Discl.indd 1 5/25/06 7:58:32 AM

Kelly Anne Meckling, Ph.D.

Alan D. Kaye Kevin Wood

1 Diabetes, Obesity, and

Diabetes, Obesity, and Metabolic Syndrome 3

FIGURE 1.1 Potential food–drug or nutrient–drug interactions of clinical interest in

1.2 DIET INTERVENTION

1.2.1 HEALTHY DIET

1.2.2 SPECIAL FOODS AND NUTRIENTS

People are encouraged to choose a variety of fiber-containing foods, such as

Diabetes, Obesity, and Metabolic Syndrome 5

1.2.2.2 Dietary Supplements and Herbal Products

1.2.2.3 Grapefruit Juice

The incidental discovery in a pilot study of a single volunteer demonstrating that

The potential interaction of grapefruit juice has been particularly investigated

1.3 PHARMACOLOGICAL INTERVENTION

Diabetes, Obesity, and Metabolic Syndrome 7

Metformin is a biguanide compound that exerts complex metabolic effects.64

Diabetes, Obesity, and Metabolic Syndrome 9

1.3.1.3 Thiazolidinediones (TZDs)

reported,39 to our knowledge, clinically relevant food–drug interactions have not

1.3.1.4 α-Glucosidase Inhibitors

α-glucosidase inhibitors are pharmacological agents that competitively and

Diabetes, Obesity, and Metabolic Syndrome 11

Second, if given in excessively high dosage, α-glucosidase inhibitors could result

double-blind, 24-week Finnish study, no significant differences in nutrient intakes

1.3.1.5 Meglitinide Derivatives