Introduction

to
Drug
Metabolism

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Contents
 Introduction
 Phase1 reactions
 Phase2 reactions
 Factor affecting biotransformation of
drugs

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 METABOLISM
OR
BIOTRANSFORMATION
 1.Biotransformation means chemical alteration of the drug in
the body that convert nonpolar(lipid soluble) to polar(lipid
insoluble) compounds.

 2.Consequences of biotransformation
 (i)Active drug-Inactive metabolite(pharmacological
inactivation)
 (ex.chloramphenicol)
 (ii)Active drug-Active metabolite(toxicological activation)
 (ex.Phenacetin)
 (iii)Inactive drug-Active toxic metabolite(pharmacological
activation)
 (ex.levodopa)
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SITES OF DRUG METABOLISM
1.The major site of drug metabolism is
Liver.

2.Other sites are GI mucosa, lungs, skin

and kidneys

3.Every tissue has some ability to

metabolize drugs.

4.Some drug, after oral administration

(clonazepam) are extensively
metabolized in GI or liver,
 Before reaching systemic circulation
called first pass metabolism, it reduces
their bioavailability.

 5.In the liver cells metabolic enzymes

are located in the smooth microsomes of
endoplasmic reticulum.

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 DRUG METABOLISING
ENZYMES
 1.Microsomal-The smooth endoplasmic reticulum of liver
and other tissues contains a large variety of enzymes, together
called microsomal enzymes.
 They catalyse glucuronide conjugation, most oxidative
reactions and some reductive and hydrolytic reactions.
 The glucuronyl transferase are important microsomal enzymes.

 2. Non-microsomal-Enzymes occuring in sites other than

endoplasmic reticulum are called non-microsomal enzymes.
 These are usually present in the cytoplasm, mitochondria and occur
mainly in the liver, GI tract, plasma.

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 PHASES OF
METABOLISM
1.Phase1(Non-synthetic reactions)-
 (i) Functionalisation reactions.
 (ii)A change in drug molecules generally
result in the introduction of functional
group in to molecules or the exposure of the
new functional groups of molecule.
 (iii) Small polar functional groups like
OH,NH2,COOH etc are either added or
unmasked(if already present)

 2.Phase2(synthetic reactions)-
 (i)Conjugation reactions.
 (ii)Involve covalent attachment of small
polar endogenous molecules like glucuronic
acid,sulphate,amino acids to the phase1
metabolite .

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 PHASE 1 REACTION
 1.Oxidative
 2.Reduction
 3.Hydrolytic
 4.Alkylation
 5.Dealkylation
 6.Isomerization
 7.Decarboxylation
 8.Dimerization

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OXIDATIVE REACTIONS
 1.Oxidative reactions are the most important metabolic
reactions.

 2.Oxidative reactions are important for drugs because they

increase hydrophilicity of drugs by introducing polar
functional groups such as OH.

 3.Oxidation of drugs is non-specifically catalysed by a

number of enzymes primarly located in the
microsomes.some oxidative reaction is catalysed by non-
microsomes.

 4.The most important component of MFO is cytohrome P-

450 because it binds the substrate and acctivates the
oxygen.
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OXIDATIVE PHASE1 REACTION

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Cytochrome p450 oxidation-reduction cycle

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 REDUCTIVE REACTION
 1.Reduction are also capable of generating polar functional
groups such as hydroxy and amino which can undergo further
biotransformation or conjugation. A number of reductive
reactions are exact opposite of oxidation.

 2. cytochrome P450 system is involved in some reaction but

other reaction catalyzed by reductases enzymes present in
different sites within the body.

 3. Reductive reactions,which usually involve addition of

hydrogen to the drug molecule,occur less frequently than the
oxidative reaction.

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REDUCTIVE AND HYDROLYTIC
PHASE1 REACTION

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 HYDROLYTIC REACTION
1. It is the reaction between compound and water.
2. The addition of water across a bond also gives more polar
metabolites.
3. Different enzymes catalyse the hydrolysis of drugs.
(i) esterase enzymes
(ii) amidase enzymes.
 4. A number of functional groups are hydrolysed like
ethers,ether,amides etc.

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PHASE 2 REACTIONS
1. Conjugation with glucuronic acid
2. Conjugation with sulphate moieties
3. Conjugation with alpha amino acid
4. Acetylation reaction
5. Methylation reactions

•The endogenous substances for conjugation reactions are

derived mainly from carbohydrates or amino acid and
usually possess large molecular size.

•The molecular weight of the conjugate is important for

determining its route of excretion.

•High molecular weight conjugates are excreted

predominantly in bile while low molecular weight
conjugates are excreted in the urine.
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CONJUGATION WITH GLUCURONIC
ACID
 Conjugation with glucuronic acid is the most common and
important phase2 reaction in vertebrates.

 The biochemical donor of glucuronic acid is uridine

diphosphate –D-glucuronic acid and the reaction is carried
out by enzyme uridine diphosphate-glucuronyl transferase.

 Glucuronyl transferase is present in microsomes of most

tissues but liver is the most active site of glucuronide
synthesis.

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Glucuronide formation occurs in 2 steps-
1.Synthesis of an activated coenzyme uridine-5’-diphospho-
alpha-D-glucuronic acid(UDPGA) from UDP-
glucose(UDPG). The coenzyme UDPGA acts as the donor
of glucuronic acid. UDPG is synthesized by interaction of
alpha-D-glucose -1-phosphate with uridine
triphosphate(UTP).
2. Transfer of the glucuronyl moiety from UDPGA to the
substrate RXH in presence of enzyme UDP-glucuronyl
transferase to form the conjugate.

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The steps involved in glucuronide synthesis are given
below-

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An example of glucuronidation of benzoic
acid-

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CONJUGATION WITH SULPHATION

 1.Sulphation is similar to glucuronidation but

it is catalysed by non- microsomal enzymes
and occur less commonly as the moiety that
transfer sulphate to the substrate.

 2.The endogenous donor of the sulphate group

is 3’phosphoadenosine-5’-phosphosulphate
and enzyme catalysing the reaction is
sulphotransferase.
3.Functional groups capable of forming sulphate
conjugates include phenols,alcohols.

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Sulphation also occur in 2 steps-
1.Synthesis of an activated coenzyme 3’-phosphoadenosine-
5’-phosphosulphate(PAPS) which act as a donor of
sulphate to the substrate.2 steps occur-
(i) An initial interaction between the sulphate and the
adenosine triphosphate(ATP) to yield adenosine-5-
’phosphosulphate(APS).
(ii) Activation of APS to PAPS.
2. Transfer of sulphate group from PAPS to the substrate
RXH in presence of enzymes sulphotransferase and
liberation of 3’-phosphoadenosine-5’-phosphate(PAP).

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The steps are summarized in the equation-

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CONJUGATION WITH ACETYLATION

 1.Conjugation with acetylation is an important metabolic

pathway for drugs containing the amino group.

 2.The cofactor for these reactions is acetyl coenzymesA

and the enzymes are non- microsomal N-acetyl
transferase,located in the soluble fraction of cells of
various tissues.

 3.Acetylation is not a true detoxification process.

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CONT………
 Two categories of inducers have been
defined-
 1.Phenobarbital type inducers: include several drugs and
pesticides which increase the rate of metabolism of a large
number of drugs.
 2.polycyclic hydrocarbon type inducers: such as 3-methyl
cholanthrene and cigarette smoke which stimulate the
metabolic rate of few drugs.
 Some drugs such as
carbamazepine,meprobamate,rifampicin etc..stimulate
their own metabolism the phenomenon called self
induction
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 BIOLOGICAL FACTORS
 Species difference-
 Difference in both phase 1 and phase2 reactions. In phase1
reactions, both qualitative and quantitative variation in the
enzyme and their activity have been observed.
 In phase2, the variations are mainly qualitative and
characterized either by the presence of conjugating
enzymes.
 Strain difference-A study of inter-subject
variability in drug response is called pharmacogenetics.
 Sex difference-regulation by sex harmones and
generally observed following puberty.

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CONT…….
 Age -In neonates : microsomal enzyme system not fully
developed… ex. caffiene
 In infants: same as neonates.
 In children: metabolise certain drugs much more rapidly than
adults.
 Diet-the enzyme content and activity is altered by a number
of dietary components-
 (i).protein-carbohydrate ratio.
 (ii).starvation
 (iii)grapefruit
 (iv)fat free diet
 (v)malnutrition in women
 (vi)alcohol ingestion

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