Metabolism of xenobiotics

INTRODUCTION

 Human beings are continuously

exposed to several foreign compounds such as
drugs, pollutants, food additives, cosmetics,
pesticides, etc., certain unwanted compounds are
produced in the large intestine by the bacteria
which enter the circulation. They are called as
Xenobiotics.
Xenos means stranger- it is a Greek word.
 INTRODUCTION

 Whether they are ingested accidently or in some

other way, they may be absorbed from the GI
track, and enters into the organs and tissues of
the body. Hence these compounds should be
removed from the body quickly.
 DEFINITION

 All the biochemical reactions involved in the

conversion of foreign, toxic, and water insoluble
molecules to non-toxic, water soluble and
excretable forms are called as Detoxification or
Biotransformation reactions.
 The purpose of which is to increase their water
solubility (polarity) and thus excretion from the
body.
 In some cases, these reactions may instead
increase the toxicity of a foreign compound,
then these reactions are called as Entoxification
reactions.
 Medical Importance

 Detoxification is a process by which we reduce

the level of toxins in our body.
 It protects the body and its organs from
deleterious effects of toxins.
 Reducing the levels of toxins in our body,
reduces the strain on the immune system.
 It removes most of the drugs consumed from the
body.
 Medical Importance

 Occasionally, detoxification may generate

toxic substance, from a relatively non-toxic
substance
 It can cause changes to our cells and DNA.
 Toxins can be a major causative factor for
the incidence of cancer.
 TYPES OF XENOBIOTICS

Xenobiotics can be
Endogenous Exogenous

Drugs, food
Bilirubin, bile acids, additives,pollutants,
steroids, eicosanoids, insecticides, chemical
and certain fatty acids. carcinogens. etc.
 METABOLISM OF XENOBIOTICS
 It occurs in to two phases.
 which may occur together or separately
Phase I Phase II
Oxidation
is followed conjugation
Oxidation,
by reactions
Reduction,
and Hydrolysis Compounds involved in conjugations

Glucuronic acid,
Amino acids
(glycine)
Glutathione,
Sulphate, acetate,
and methyl group.
 Purpose and consequences

 Purpose consequence

• Changes in solubility
characteristics
• converts lipophilic to • Detoxification
hydrophilic. • Metabolic activation.
• Facilitates excretion.
 Overview of biotransformation reactions

 Phase I reactions can also convert xenobiotics

from inactive to biologically active
compounds.(Metabolic Activation).
 In these cases the original xenobiotics can be
called as prodrugs or procarcinogens.
 Overview of biotransformation reactions

 Phase II-conjugation reactions can convert the

active products of Phase I reactions to active or
inactive species. Which are then excreted in
urine or bile.
 In few cases, the conjugation may actually
increase the biological activity of xenobiotic
(Metabolic action)
 Overview of detoxification reactions
Some drugs
directly enter
phase II
metabolism.

Oxidation,
DRUG Phase I Phase II Conjucated
Reduction, and
(or) Hydrolysis. Product

Following phase I the

drug may be activated, Conjugated drug is
unchanged, or most usually inactive.
often , inactivated.
Role of liver
 BIOTRANSFORMATION
Potentially toxic xenobiotic Relatively harmless

Detoxification Metabolic Activation

Inactive metabolite Reactive intermediate

 Comparing Phase I and Phase II
Particulars Phase I Phase II
Types of reactions. Oxidation. Conjugations.
Reduction.
Hydrolysis.
Increase in
hydrophillicity. Small . Large.

General Exposes functional Polar compounds

mechanisms. groups. added to
functional group.
Consequences. May result in Facilitates
metabolic excretion.
activation.
 Factors affecting detoxification of drugs

 Prior administration of drugs or co-

administration of other drugs.
 Diet.
 Hormonal status.
 Genetics.
 Diseases (decrease in cardiac and pulmonary
disease)
 Age and developmental status.
 Functional status of liver and kidney.
 EFFECT OF XENOBIOTICS

 Cyto P450 GSH S Transferase or

Reactive
Xenobiotic
 B Epoxide hydrolase Non toxic
Metabolite Metabolite

Covalent binding to Macromolecule

Cell injury Hapten Mutation

Antibody Production Cancer

Cell injury
 Phase I reactions

 Phase I reactions are :

 Oxidation.
 Reduction.
 Hydrolysis reactions.
 They are also known as hydroxylation reactions.
Since they introduce or expose a functional
group that serves as the active center for
sequential conjugation in phase II reactions.
 Enzymes of phase I reactions

 Mainly catalyzed by members of the enzymes known

as monooxygenases, mixed function oxidases,
(or)cytochrome P450s.
 Other important enzymes are
 Aldehyde and alcohol dehydrogenase
 Deaminases.
 Esterases.
 Amidases.
 Epoxide hydrolases.
 Oxidation
 Most of the foreign substances are detoxified by
oxidation.

 These includes alcohols, aldehydes, amines, aromatic

hydrocarbons and sulphur compounds.

 Aliphatic compounds are more easily oxidized than

aromatic ones.
 Oxidation

 Indole and Skatole are produced from

tryptophan, by the action of microbes.
 They are responsible for the dis agreeable odour
of the feces.
 They undergo oxidation.
 Indole → Indoxyl.
 Skatole → Skatoxyl.
 OXIDATION OF ALCOHOLS

 Both aromatic and aliphatic alcohols

undergo oxidation to form their
corresponding acids.
 Ex:
Methanol → formaldehyde → formic acid.

Ethanol → Acetaldehyde → acetic acid

Benzyl alcohol →Benz aldehyde → benzoic

acid.
 Methanol toxicity

 It has relatively low toxicity.

 It is metabolized in the liver.

 In the first step of degradation, methanol is

transformed to formaldehyde via enzyme ADH

 Transformation of formaldehyde to formic acid

via aldehyde dehydrogenase is faster.
 Methanol toxicity

 The metabolism of formic acid is very slow.

 Thus it often accumulates in the body ,which results

in metabolic acidosis.

 The major damage occurs to the optic nerve.

 Ethanol is given as an antidote, since it is the

substrate of ADH, methanol is spared.
 Methanol Toxicity

ADH FDH
Methanol Formaldehyde Formic Acid CO2 + H2o

* ADH – Alcohol Dehydrogenase

* FDH – Formaldehyde Dehydrogenase Metabolic Acidosis and
Tissue injury
 Oxidation of aldehydes
 Aldehydes are oxidised to corresponding acids.
 Acid thus formed is further conjugated in phase II
 Ex: Benz aldehyde → benzoic acid.
Benzoic acid is conjugated with glycine to form
Hippuric acid.
This reaction is exclusively takes place in the
liver.
Hippuric acid excretion test is used to
determine the detoxification functions of liver.
Normal level of excretion - 0.5 – 1.0 g/day.
Oxidation of aromatic hydrocarbons

 Aromatic hydrocarbons are oxidized to phenolic

compounds.
 Which can further conjugated with glucuronic
acid (or )sulphuric acid in phase II reactions, so
as to be excreted through urine.
Benzene Mono, Di, Tri hydroxy
phenol
 Oxidation of amines

 Many primary aliphatic amines undergo

oxidation to form the corresponding acids and
nitrogen is converted to urea.

 EX: Benzyl amine → Benzoic acid + urea.

 While aromatic amines are oxidised to phenols.

Aniline → P- Amino phenol.
Oxidation of anilids and sulphur containing
compounds
 Anilids are oxidized to corresponding phenols.

 Acetanilide is oxidized in the body to form P-

acetyl amino phenol.

 Acetanilide is a constituent of analgesic drug.

 Sulfur compounds:
 Organic sulfur is oxidized to sulfuric acid.
 Oxidation (entoxification)

 Oxidation of certain compounds may also result

in the production of more toxic
compounds(entoxification).
 Hence their formation is prevented.
 EX: Methanol-------------->Formic Acid.
Halagenated Alcohol------> Halogenated
Acid.
 Ethylene glycol -------------> oxalic Acid.
 Cytochrome P450.

 Most of the oxidation reactions of detoxification are

catalysed by monooxygenase(or )cytochrome P450.
This enzyme is also known as mixed function
oxidases, which is associated with the microsomes of
the liver.
 Most of the reactions of cytochrome P450 involve the
addition of a hydroxl group to aliphatic or aromatic
compounds which may be represented as
 RH+ O2+NADPH+H+----------- >OH+H2O+NADP
 RH represents a wide variety of xenobiotics which
may include drugs, carcinogens, pesticides
pollutants etc.,
 Properties of human cytochrome P450.

 They are all hemoproteins.

 Exhibit broad substrate specificity.(act on many
compounds).
 Versatile catalysts(since it catalyse 60 types of
reactions).
 Larger amount is found in microsomes of liver.
But found in other tissues such as intestine,
lung, brain etc.
 Properties of human cytochrome P450.
 It is located in smooth ER.
 They are NADPH dependent enzymes.
 In some cases, their products are mutagenic (or)
carcinogenic.
 They are inducible enzymes. Its synthesis is
increased by administration of drugs such as
phenobarbitol.
 A phospholipid– phosphotidyl choline is a
constituent of cytochrome P450 system and is
necessary for the action of this system.
 Reduction

 Reduction is less common and less important

than oxidation in human beings.

 Picric acid -----------> Picramic acid

 Reduction of aldehydes:

 EX: chloral hydrate----------> Trichloroethanol.

 Chloral hydrate is a sedative.
 Reduction

 It is excreted after conjugation with D-

glucuronic acid as corresponding glucuronide.

 Reduction of Nitro compounds:

 P- nitrobenzene--------> P- aminobenzene.
 P- nitrophenol----------> P- aminophenol.
 Hydroxylation
 Detoxification of a number of drugs and steroids
occur by hydroxylation.

 These reactions are catalyzed by cytochrome

P450. dependent monooxygenases.

 Pentobarbital cytP450 Hydroxy phenobarbital.

 phenobarbital an anticonvulsant drug.

 Hydroxylation

 Meprobamate undergo hydroxylation

 Meprobamate → P-OH meprobamate.
 Meprobamate is a tranquilizer.
Trichloro acetic acid
Chloral
Tri chloro ethanol

Chloral used as a Hypnotic.

 Hydrolysis

 Certain therapeutic compounds undergo hydrolysis.

 Hydrolysis of the bond such as ester,glycoside, and
amide is important in the metabolism of xenobiotics.
 EX: Aspirin H20 Acetic acid + salicylic acid.
 (Acetyl salicylic acid ).(hydrolysis in the ester bond)
 Atropine Hydrolised Tropic acid +Tropine.
 Atropine is a psychoactive drug.
 Acetanilide H2O Aniline + Acetic acid.
 Procaine ----- > P-aminobenzoic acid + diethyl
amino ethanol.
 Phase II Reactions of xenobiotics.
Conjugation
 Conjugation is a process by which the foreign
molecules and their metabolites are coupled with a
conjugating agent and are converted to soluble, non-
toxic derivatives which are easily excreted in urine.
 Conjugation reactions can occur either
independently (or) can follow phase I
(hydroxylation) reactions.
 Conjugation takes place in liver. But it can also
occur in kidney.
 After conjugation the products are generally
rendered non-toxic but in certain condition they are
left unchanged (or) become more toxic.
 Conjugation

 Types of phase II reactions:

 Glucuronidation.
 sulfation.
 Acetylation.
 Methylation.
 Conjugation with amino acids.
 Conjugation withG-SH.
 Glucuronidation.

 It is the most common Conjugation reaction.

 UDP-glucuronic acid, is the glucuronyl donor,
which is formed in the uronic acid pathway of
glucose metabolism.
 Glucuronyl transferase, present in both the ER
and cytosol are the catalysts.
 The glucuronide may be attached to oxygen,
Nitrogen, (or) sulphur groups of the substrates.
 Glucuronidation.

 Compounds conjugated with glucuronic acid are :

 Bilirubin.
 Aromatic acid---Benzoic acid.
 Phenols, secondary, tertiary aliphatic alcohols.
 Antibiotics like chloramphenicol.
 Hormones--- Thyroid hormone, derivatives of
corticosteroids and sex hormones metabolites.
 2-acetaminofluorene (a carcinogen).
 Aniline.
 Meprobamate (a tranquilizer).
Conjugation with bilirubin
 Conjugation with bilirubin

 Most of the bilirubin excreted in the bile is in

the form of bilirubin diglucuronide.
 Bilirubin-UGT activity can be induced by
certain drugs.
 Which includes phenobarbital when these drugs
are administered, it increase the formation of
glucuronide.
 Sulfation

 Compounds which are conjugated with sulphate are

phenols, cresols, indole, steriods,oestrogen and
androgen etc.,
 The active form of sulfate is 3’phosphoadenosine 5-
phosphosulfate (PAPS) involve in conjugation
reactions and the enzyme sulfotransferase catalyze the
process.
 PAPS donate the sulphate group.
 Sulfation

 Phenol+PAPS sulphotransferase phenyl

sulphate + PAP

 Skatoxyl PAPS
Ethereal sulphate+PAP
 Indoxyl
 Acetylation
 Compounds which are conjugated with acetic acid
are sulphanilamide, PABA, Isoniazid.
 Acetyl coA is the active form of acetic acid.
 These reactions are catalyzed by Acetyl transferase.
 Which are present in the cytosol of various tissues
particularly liver.
 Sulfanilamide +Acetyl coA Acetyl transferase
+acetyl sulfanilamide + coASH.
 PABA Acetyl coA Acetylated PABA.
 Methylation

 Methylation is limited in the body.

 Compounds conjugated with methyl group are

Nicotinamide, p-methyl amino azobenzene, P-
dimethyl amino benzene, estrogen,
Catecholamine, etc.,

 S-adenosyl methionine -----methyl donor.(active

form)
 Methylation

 Reactions ------ >transmethylation

 Enzyme involved----->Methyl transferase.

 Nicotinamide CH3 N-Methyl
Nicotinamide
 p-Methyl amino CH3 P- Dimethyl
azobenzene amino azobenzene
(hepatic Carcinogen)
 Conjugation with amino acids

 Compounds which are conjugated with glycine

 are benzoic acid, phenyl acetic acid, cholic acid, and
deoxy cholic acid.
Ex: Benzyl coA + Glycine----- > Hippuric acid.
Phenyl acetic acid + Glycine---- > Phenyl aceturic
acid.
Cholic acid + Glycine---- > Glycocholic acid.

Deoxycholic acid + Glycine---Glycodeoxycholic

acid.
 Conjugation with glutathione

 Glutathione -- > a tripeptide consist of Glutamic

acid, cysteine, and glycine( active conjugating
agent).
 Organic compounds such as alkyl or aryl
halides, alkenes, nitro compounds and epoxides
get conjugated with cysteine of glutathione.
 The glutamate and glycine of glutathione are
removed and an acetyl group is added to the
cysteine residue.
 Results in formation of mercapturic acid.
 Detoxification by drugs

 Some of the drugs are administered to detoxify

foreign substances .
 The toxic effects of certain heavy metals such as
arsenic, mercury, and cadmium could be
overcome by administering BAL(British Anti
Lewisite). 2,3 Mercapto propanol.
 This drug was developed during II world War
and was used as a detoxifying agent for certain
war poisons.
 Detoxification by drugs

 BAL readily combines with metals and gets

easily excreted into urine.
 Metals binds with – SH group of enzymes, there
by inactivating them.
 BAL acts as an antidote by virtue of pulling
away the SH groups from the heavy metals.
 Such a liberation helps in the detoxifying
mecahnism.
 SUMMARY

 Xenobiotics are the chemical compounds

foreign to the body, such as drugs, food
additives, and environmental pollutants.
 Xenobiotics are metabolised in two phases.
 The major reactions of phase I is hydroxylation
catalyzed by a variety of monooxygenases, also
known as cytochrome P450.
 In phase II the hydroxylated species are
conjugated with a variety of hydrophilic
compounds such as glucoronic acid, sulfate
 SUMMARY

Glutathione,etc.
 The combined operation of these two phases
renders lipophilic compounds into water soluble
compounds that can be eliminated from the
body.
 Xenobiotic can produce a variety of biologic
effects, including pharmacologic responses,
toxicity, immunologic reactions and cancer.