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• A xenobiotic is a foreign chemical substance

Metabolism / Biotransforamtion / found within an organism that is not normally
Bioaccumulation of Toxicants naturally produced by or expected to be present
(Xenobiotics) within that organism.
• It can also cover substances which are present in
much higher concentrations than are usual.

Metabolism of drugs and xenobiotics

※Endogenous: Pigments , hormone
※nonendogenous : Such as drugs , food additives,
pollutants, toxin, etc.
 Most of these compounds are subject to
metabolism (biotransformation) in human
body.
Functional significance:
● inactivation and facilitated elimination
of drugs and xenobiotics
● activation of prodrugs
● formation of active metabolites with
similar or novel activity
● detoxification of toxic xenobiotics
● toxification of non-toxic xenobiotics

Enzyme specificity in drug metabolism Product of Metabolism

● key problem: a limited number of
enzymes must cope with an unlimited
Xenobiotics
number of substrates Must be converted to a
water-soluble substance
● many drug-metabolizing enzymes have (hydrophilic)
fairly broad specificities Prodrugs/Metabolites
Are mostly lipophilic or
● enzyme specificities overlap—many lipid-soluble
compounds
drugs give rise to multiple metabolites Metabolism Is also called DETOXIFICATION or DETOXICATION

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Metabolism Important Points to Remember

• Elimination of drugs and other foreign compounds
• Prodrugs – active drug
• Xenobiotics – foreign compound
Most drugs entering the
body are lipophilic
two categories:
Catabolism breaks down organic matter
Anabolism uses energy to build up or construct Drug molecules easily diffuse
components of cells such as proteins and through the lipophilic membranes
nucleic acids.
of the GIT

Important Points to Remember Definition of the biotransformation

Some of the Prodrugs or  Conversion of lipophilic xenobiotics to water-

Xenobiotics are NOT soluble chemicals by a process catalyzed by
completely excreted in enzymes in the liver and other tissues.
the urine due to the  In most cases, biotransformation lessens the
toxicity of xenobiotics, but many must undergo the
reabsorption in the renal process to exert their toxic effects.
tubules

Purpose of biotransformation Sites of biotransformation

• Liver
1. facilitates excretion: Converts lipophilic to hydrophilic
• Primary site! Rich in enzymes
compounds
• Acts on endogenous and exogenous compounds
2. Detoxification/inactivation: converts chemicals to less toxic
• Extrahepatic metabolism sites
forms
• Intestinal wall
3. Metabolic activation: converts chemicals to more toxic
• Sulfate conjugation
active forms • Esterase and lipases - important in prodrug
metabolism
• Lungs, kidney, placenta, brain, skin, adrenal glands

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Lipophilic toxicant

Biotransformation
SITES DRUG BIOTRANSFORMATION
of
LIVER The most important organ in drug
Hydrophilic metabolite
metabolism

 Contains almost all drug

metabolizing enzyme

INTESTINAL MUCOSA
 Contains CYP3A4 isoenzyme
and P-glycoprotein

Metabolite excreted

Product of Metabolism
Introduction of Functional Polar Groups to Xenobiotics
The product of metabolism must become hydrophilic or
converted to a water-soluble substance for elimination
Functional Group

Xenobiotic

ELIMINATION

Forms inactive and non-toxic Unwanted biological effect

substance

Introduction of Functional Polar Groups to Xenobiotics

Introduction of Functional Polar Groups to Xenobiotics

Reduction of Ketones & Aldehydes to Alcohol Oxidation of Alcohol to Acid

of the of the

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Introduction of Functional Polar Groups to Xenobiotics Introduction of Functional Polar Groups to Xenobiotics

Hydrolysis of Ester & Amide to Acid Reduction of Nitro compounds to form NH2 moiety
of the of the

General Metabolic Pathways

• Most metabolic products are less pharmacologically active
 Approximately 30 different enzymes catalyze Important exceptions:
reactions involved in xenobiotic metabolism; • Where the metabolite is more active
however, this note will only cover a selected (Prodrugs, e.g. Erythromycin-ethylsuccinate (less irritation of GI) -->
group of them. Erythromycin)
• Where the metabolite is toxic (acetaminophen)
• It is convenient to consider the metabolism • Where the metabolite is carcinogenic
of xenobiotics in two phases
• phase Ⅰand phase Ⅱ

toxic metabolites Where the metabolite is carcinogenic

BIOACTIVATIONS

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Phases of Drug Metabolism

Close relationship between the
biotransformation of drugs and normal
biochemical processes occurring in the body:
• Metabolism of drugs involves many pathways associated with the
synthesis of endogenous substrates such as steroid hormones,
cholesterol and bile acids
• Many of the enzymes involved in drug metabolism are principally
designed for the metabolism of endogenous compounds
• These enzymes metabolize drugs only because the drugs resemble
the natural compound
• Phase I Reactions
• Convert parent compound into a more polar (=hydrophilic) metabolite by adding or
unmasking functional groups (-OH, -SH, -NH2, -COOH, etc.)
• Often these metabolites are inactive
• May be sufficiently polar to be excreted readily
• Phase II Reactions
• Conjugation with endogenous substrate to further increase aqueous solubility
• Conjugation with glucoronide, sulfate, acetate, amino acid
• Phase I usually precede phase II reactions

Drug Metabolism - Phase I

Liver is principal site of drug metabolism: • Phase I Reactions
• Other sites include the gut, lungs, skin and kidneys • Oxidation
• For orally administered compounds, there is the • Reduction
• Hydrolytic cleavage
“First Pass Effect” • Alkylation Purpose
• Intestinal metabolism , Liver metabolism, Enterohepatic recycling (Methylation)
Introduction of polar functional groups in
and Gut microorganisms - glucuronidases • Dealkylation
a molecules
• Ring cyclization ♣ Increase a molecule’s polarity
• N-carboxylation ♣ Does provide a site for phase II
• Dimerization metabolism
• Transamidation
• Isomerization
• Decarboxylation

Drug Metabolism - Oxidation

Phase I in Action

Imipramine Two types of oxidation reactions:

• Oxygen is incorporated into the drug molecule (e.g. hydroxylation)
• Oxidation causes the loss of part of the drug molecule
(e.g. oxidative deimination, dealkylation)

4-hydroxy
N
imipramine CH2
(cardiotoxic)
CH2 desmethyl
imipramine
N (antidepressant)
CH3 CH3

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Drug Metabolism - Oxidation

Microsomal Mixed Function Oxidases (MFOs)
• MFO consists of two enzymes:
• “Microsomes”
form in vitro after cell homogenization and fractionation of ER – Flavoprotein, NADPH-cytochrome c reductase
• One mole of this enzyme contains one mole each of flavin mononucleotide (FMN) and
• Rough microsomes are primarily associated with protein synthesis flavin adenine dinucleotide (FAD)
• Smooth microsomes contain a class of oxidative enzymes called • Enzyme is also called NADPH-cytochrome P450 reductase

• “Mixed Function Oxidases” or “Monooxygenases” – Cytochrome P450

• These enzymes require a reducing agent (NADPH) and molecular oxygen • named based on its light absorption at 450 nm when complexed with carbon monoxide
• is a hemoprotein containing an iron atom which can alternate between the ferrous (Fe ++)
(one oxygen atom appearing in the product and the other in the and ferric (Fe+++) states
form of water) • Electron acceptor
• Serves as terminal oxidase
• its relative abundance compared to NADPH-cytochrome P450 reductase makes it the
rate-limiting step in the oxidation reactions

Drug Metabolism - Oxidation Cytochrome P450 dependent

Mixed Function Oxidases
• Drug oxidation requires:
–Cytochrome P450
DRUG METABOLITE
–Cytochrome P450 reductase =DRUG+O
–NADPH O2
Liver
–Molecular oxygen microsomes
NADPH NADP+

H+ WATER

• The cycle involves four steps:

1. Oxidized (Fe3+) cytochrome P-450 combines with a drug substrate to form a binary complex.
 Phase Ⅰ: Oxidation
2. NADPH donates an electron to the cytochrome P-450 reductase, which in turn reduces the
oxidized cytochrome P-450-drug complex. 1. Hydroxylation
3. A second electron is introduced from NADPH via the same cytochrome P-450 reductase, which
serves to reduce molecular oxygen and form an "activated oxygen"-cytochrome P-450-
substrate complex.
RH + O2 + NADPH + H+  R-OH + H2O + NADP+
4. This complex in turn transfers "activated" oxygen to the drug substrate to form the oxidized
product. The potent oxidizing properties of this activated oxygen permit oxidation of a large Addition of an oxygen atom or bond
number of substrates.
Require NADH or NADPH and O2 as cofactors

RH: drugs, cacinogens, pesticides, petroleum products,

pollutants, steroids, eicosanoids, fatty acids, retinoids, etc.
 Enzyme:
Cytochrome P450s-dependent monooxygenase

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Hydroxylation：O2 Drug Metabolism - Oxidation

Aromatic hydroxylation:
* It has been shown by the use of O2 that one atom of
oxygen enters R-OH and one atom enters H2O.

* This dual fate of the oxygen accounts for the former

naming of monooxygenases as “mixed-function Aliphatic hydroxylation:

oxidases”.
RH + O2 + NADPH + H+  R-OH + H2O + NADP+

Drug Metabolism - Oxidation Drug Metabolism - Oxidation

Epoxidation: O-demethylation: S-demethylation:

Dealkylation: N-oxidation: N-hydroxylation:

Cytochrome P450s-dependent monooxygenase

CYP or Cytochrome P-450 1. Cytochrome P450: Isozymes
★ Isozymes - multiple forms of an enzyme
★ Heme proteins
★ Supergene family
★ Iron containing porphyrin - binds O2
What is a Heme Protein?
- More than 8,000 P450 genes
- More than 368 gene families, 814 subfamilies
- Human: 18 families, 43 subfamilies, 57 sequenced
Cytochrome P-450, Hemoglobin, & Myoglobin ALL Heme Proteins! genes
★ Nomenclature
★ The name cytochrome P450 is derived from the spectral
CYP1A2
properties of this hemoprotein  in its reduced (Fe2+) form, it
family individual member
binds CO to give a complex that absorbs light maximally at 450 subfamily
of that subfamily
nm

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Cytochrome P450 Substrates

CH

• This enzyme is very important. approximately 50 % of H2

C
Allyl
the drugs that humans ingest are metabolized by
isoforms of cytochrome P450.
Vinyl

• These enzymes also act on various carcinogens and Aromatic Benzyl

pollutants.




Aliphatic Alicyclic

Drug Metabolism - Oxidation Other (non-microsomal)

Phase I reactions

Oxidation reactions NOT catalyzed by Cytochrome P450:
Flavin containing monoxygenase system
• Present mainly in liver but some is expressed in gut and lung
• Located in smooth endoplasmic reticulum
• Oxidizes compounds containing sulfur and nitrogen
• Uses NADH and NADPH as cofactors
Hydrolysis in plasma by esterases (suxamethonium by
cholinesterase)
Alcohol and aldehyde dehydrogenase in liver cytosolic
(ethanol)
Monoamine oxidase in mitochondria (tyramine,
noradrenaline, dopamine, amines)
Xanthine oxidase (6-mercaptopurine, uric acid production)
Enzymes for particular substrates (tyrosine hydroxylase,
dopa-decarboxylase etc.)

Monoamine oxidase, MAO ADH and ALDH

RCH2NH2+O2+H2O2 RCHO+NH3+H2O
ADH alcohol dehydrogenase

★ MAO catalyze the oxidative deamination of monoamines. ALDH aldehyde dehydrogenase

★ Oxygen is used to remove an amine group from a molecule, resulting in • Alcohol Dehydrogenase belongs to the oxidoreductase
the corresponding aldehyde and ammonia. family of enzymes.

★ MAO are found bound to the outer membrane of mitochondria in most • high concentrations within the liver and kidney.
cell types in the body. They belong to protein family of flavin containing
amine oxidoreductases.

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ADH
Function ALDH

NAD+ NADH NAD+ NADH

• The primary and most common role of ADH in humans R CH2OH R CHO
is to detoxify incoming ethanol by converting it into
R CO2H
aldehyde.
During this reaction, hydrogen is removed from the
• The resulting aldehyde, a more toxic molecule than alcohol and transferred to a molecule called nicotinamide
ethanol, is quickly converted into acetate by aldehyde adenine dinucleotide (NAD), converting it to reduced NAD
dehydrogenase (ALDH) and other molecules easily (NADH).
utilized by the cell.
NADH participates in numerous other metabolic
ALDH
reactions, passing on the hydrogen to other compounds
ADH
NADH NADH
or electron transfer chain.
NAD+ NAD+

R CH2OH R CHO R CO2H

Absorption  In people who consume alcohol at moderate levels and/or only

occasionally, most of the alcohol is broken down by ADH and ALDH.
20%
• Soluble in water
after higher alcohol consumption, The MEOS plays a role in
alcohol metabolism.
• Small size - penetrates everywhere,
easily crosses all bio membranes MEOS
CH3CH2OH + NADPH + O2 + H+ CH3CHO
+ NADP+ + 2H2O
• Rapidly absorbed from GI
80% ALDH
CH3CHO CH3COOH

MEOS: Microsomal Ethanol-Oxidizing System , is also called Cytochrome

P450-dependent Microsomal Ethanol Oxidizing System. converts alcohol to
acetaldehyde

MEOS Because the MEOS metabolizes not only alcohol but also
CH3CH2OH + NADPH + O2 + H+ CH3CHO + NADP+ + 2H2O
other compounds (certain medications), enhanced MEOS
activity resulting from high alcohol consumption also can
This reaction also relies on oxygen and NADPH, and
results in the formation of NADP and water. alter the metabolism of those medications.

◆consume oxygens of liver and NADPH

◆ As byproducts of these reactions, oxygen radicals or
This may contribute to harmful interactions between
reactive oxygen species (ROS) are generated. These ROS can alcohol and those medications or otherwise influence the
contribute to liver damage through a variety of mechanisms. activity of those medications.

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Alcoholism leads to fat accumulation in the liver,

hyperlipidemia, and ultimately cirrhosis.

ALCOHOL  Phase Ⅰ: Reduction

Mechanism of Fatty Liver Nitro and Azo Reduction
• The likelihood of hypoglycemia is also increased in alcoholics when
they fast, as they often have low hepatic stores of glycogen because • NADPH dependent microsomal and nitro-reductase
of poor nutrition. enzymes.
• Bacterial reductases play a role in enterohepatic
• The shift in the NADH/NAD+ ratio also inhibits β-oxidation of fatty
acids and promotes triglyceride synthesis; this increases hepatic recirculation of nitro or azo containing drugs.
synthesis of VLDL, and the remaining excess triglyceride is deposited
in the liver. + O
Ar N Ar N O Ar NHOH Ar NH2
O

Ar N N Ar' Ar N N Ar' H2N Ar + H2N Ar'

H H

sudan red incident sudan red (Azo dye)

chili patse

nitroreductase

Azo dyes that are toxic only after reduction and cleavage of the
azo linkage to give aromatic amines, mostly via intestinal
anaerobic bacteria. The aromatic amines are metabolically
oxidized to reactive electrophilic species that covalently bind chloromycetin (chloramphenicol)
DNA .

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 Phase Ⅰ: Reduction
Hydrolysis Drug Metabolism - Reduction
Substrates: esters , amide , glycoside, etc. Azo-reduction:

 Catalyzed by widely distributed hydrolytic enzymes

 Hydrolysis of esters  major metabolic pathway for Nitro-reduction:
ester drugs
☻Non-specific esterases (liver, kidney, and intestine)
☻Plasma pseudocholinesterases also participate

Dehalogenation:
CO2H CO2H
HO CH3
Acetylsalicylic O CH3 esterase OH
Acid, ASA O O
salicylic acid
ASA

Drug Metabolism - Reduction

Hydrolysis reactions
• Almost any drug can undergo modifications by drug-metabolizing enzyme
systems
Ester hydrolysis:

• Drugs can be subjected to several Phase I pathways

• These reactions create functional groups that place the drugs in a correct
chemical state to be acted upon by Phase II conjugative mechanisms
Amide hydrolysis: • Main function of phase I reactions is to prepare chemicals for phase II
metabolism and subsequent excretion
• Phase II is the true “detoxification” step in the metabolism process.

Drug Metabolism - Phase II

• Conjugation reactions
xenobiotic
• Glucuronidation by UDP-Glucuronosyltransferase:
Phase I
(on -OH, -COOH, -NH2, -SH groups)
Phase II • Sulfation by Sulfotransferase:
(on -NH2, -SO2NH2, -OH groups)
Protection • Acetylation by acetyltransferase:
excrection Elimination
(on -NH2, -SO2NH2, -OH groups)
Reactive • Amino acid conjugation
metabolite nontoxic (on -COOH groups)
metabolite • Glutathione conjugation by Glutathione-S-
transferase:
Cell injury Antibody product mutation (to epoxides or organic halides)
• Fatty acid conjugation
(on -OH groups)
cancer
Cell injury • Condensation reactions

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Drug Metabolism - Glucuronidation Drug Metabolism - Glucuronidation

• Glucuronidation ( = conjugation to a-d-glucuronic acid) • Glucuronidation – requires creation of high energy intermediate:
UDP-Glucuronic Acid:
• Quantitatively the most important phase II pathway for drugs and
endogenous compounds
• Products are often excreted in the bile.
• Enterohepatic recycling may occur due to gut glucuronidases
• Requires enzyme UDP-glucuronosyltransferase (UGT):
• Genetic family of enzymes
• Metabolizes a broad range of structurally diverse endogenous and
exogenous compounds
• Structurally related family with approximately 16 isoforms in man

Drug Metabolism - Glucuronidation

• Glucuronidation Pathway and Enterohepatic Recirculation Drug Metabolism - Glucuronidation
• N-glucuronidation:
• Occurs with amines (mainly aromatic )
• Occurs with amides and sulfonamides

Drug Metabolism - Glucuronidation

• O-glucuronidation:
Drug Metabolism - Acylation
• Occurs by ester linkages with carboxylic acids Acetylation:
• Occurs by ether linkages with phenols and alcohols • Common reaction for aromatic amines and sulfonamides
• Requires co-factor acetyl-CoA
• Responsible enzyme is N-acetyltransferase
• Takes place mainly in the liver
• Important in sulfonamide metabolism because acetyl-sulfonamides are less
soluble than the parent compound and may cause renal toxicity due to
precipitation in the kidney

Fatty Acid Conjugation:

• Stearic and palmitic acids are conjugated to drug by esterification reaction
• Occurs in liver microsomal fraction
(Cannabinols are metabolized in this fashion => long half-life)

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Drug Metabolism - Other conjugations

Amino Acid Conjugation:
• ATP-dependent acid:CoA ligase forms active CoA-amino acid conjugates which then
react with drugs by N-Acetylation:
– Usual amino acids involved are:
• Glycine. Glutamine, Ornithine, Arginine

Glutathione Conjugation:
• Tripeptide Gly-Cys-Glu; conjugated by glutathione-S-transferase (GST)
• Glutathione is a protective factor for removal of potentially toxic compounds
• Conjugated compounds can subsequently be attacked by
g-glutamyltranspeptidase and a peptidase to yield the cysteine conjugate =>
product can be further acetylated to N-acetylcysteine conjugate

2. Sulfate Conjugation
Sulfate Conjugation
• Some alcohols, arylamines, and phenols are sulfated. O
O
• Catalyzed by sulfotransferases
• liver, kidney and intestine ＋PAPS
+PAP
• Sulfate donor: adenosine 3’-phosphate-5’-phosphosulfate
(PAPS); this compound is called “active sulfate.” HO HO 3SO

• Leads to inactive water-soluble metabolites estrone estrone sulfate

• Glucuronate conjugation often more competitive process
O
CH3 CH3
HX Drug HO -O S O
CO2H O CO2H
sulfotransferase O H2N H2N
HO HO
O O -O S X Drug
alpha-Methyldopa
-O S O P O O
PAPS Adenine
Aldomet ® - Merck
O OH O Antihypertensive

H2O3PO OH

3. Conjugation with glutathione Glutathione (GSH) Conjugation

• DETOXIFICATION of electrophiles!
• Electrophilic chemicals cause:
• Tissue necrosis
• Carcinogenicity
glutamic acid, cysteine, glycine • Mutagenicity
• Teratogenicity
GST
R + GSH R-S-G • The thiol (SH group) ties up potent electrophiles
where R= an electrophilic xenobiotics
R: epoxides and halogenides
GST: Glutathione S-Transferases (Liver and kidney)

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Glutathione S-transferase Glutathione (GSH) Conjugation

Epoxide or an
Arene Oxide
oxidation
(+)benzo[a]pyrene-
7,8-dihydrodiol- HSG SG
9-10-epoxide O
Benzene OH
H+
HO

HO GST
+ glutathione HO
OH

DNA reactive; Inactive

lung and skin
tumors

DETOXIFICATION aflatoxin b-1 epoxide

4. Acetylation
X + Acetyl-CoA - - - - >Acetyl-X + CoS
sulfanilamide
where X represents a xenobiotics.
(for: aromatic amines)
• Important for drugs with primary amino groups
• Enzyme: acetyltransferases • Generally, metabolites are nontoxic and inactive
• present in the cytosol of various • Acetylation does NOT increase water solubility
tissues, particularly in liver. • Detoxification or termination of drug activity

isoniazid

Methylation Metabolism via Methylation

• A few xenobiotics are subject to
methylation by methyltransferase,
emplyoing S-adenosylmethione(SAM) as the methyl
donor.
SAM
• Key for biosynthesis of many compounds
• Important in the inactivation of physiologically
active biogenic amines  neurotransmitters
• norepinephrine, dopamine, serotonin, histamine
• Minor pathway in the metabolism of drugs
• Methylation does NOT increase water solubility
• Most methylated products are inactive

catechol

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PHASE 1 & PHASE 2 REACTION

Drug Metabolism - Phase I & II
Oxidation & Conjugation Reaction
Phase I and II - Summary: Phase 1 & Phase 2 reactions complement one another in
detoxifying & facilitating the elimination of drugs &
• Products are generally more water soluble Xenobiotics
• These reactions products are ready for (renal) excretion
• There are many complementary, sequential and competing CH3 CH2OH
pathways
• Phase I and Phase II metabolism are a coupled interactive
system interfacing with endogenous metabolic pathways

Allylic Hydroxylation of Marijuana (Δ1 tetra Hydrocannabinol)

PHASE 1 & PHASE 2 REACTION PHASE 1 & PHASE 2 REACTION

Oxidation & Conjugation Reaction Oxidation & Conjugation Reaction

H
H-C-H (CH3) CH2OH
COOH
H (CH2OH)
H-C-OH
oxidation
CH3
CH3

CH3
CH3

11- Hydroxy Δ9 tetra Δ9 TH Cannabinoic Acid

Hydrocannabinol
More polar

Allylic Hydroxylation of Marijuana (Δ1 tetra Hydrocannabinol)

PHASE 1 & PHASE 2 REACTION

PHASE 1 & PHASE 2 REACTION
Oxidation & Conjugation Reaction
Oxidation & Conjugation Reaction

COOR
 Water-soluble
OR compound

CH3
 eliminated
completely in the
CH3 urine
Δ9 TH 11-Cannabinoic Acid

Subsequent conjugatinon with Glucoronic acid

Subsequent conjugatinon with Glucoronic acid
Glucuronide Conjugate at either the COOH group or the Phenolic OH Group Glucuronide Conjugate at either the COOH group or the Phenolic OH Group

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Comparing Phase I & Phase II Factors Affecting Metabolism

Enzyme Phase I Phase II

Types of reactions Hydrolysis Conjugations Age (reduced in aged & children)
Oxidation older people less efficient at metabolism
Reduction
Increase in Small Large Sex (women more sensitive to ethanol, Linked to hormonal differences)
hydrophilicity
Species (phenylbutazone 3h rabbit, 6h horse, 8h monkey, 18h mouse, 36h
General mechanism Exposes functional Polar compound added
group to functional group man)
Race (fast and slow isoniazid acetylators, fast = 95% Eskimo, 50% Brits,
Consquences May result in Facilitates excretion
metabolic activation 13% Finns 13% Egyptians)
Clinical or physiological conditions
• Disease states
• Liver, cardiac, kidney disease

AGE
• Fetus & Neonate : CP
SPECIES
UDPGT
• Lower drug metabolizing capacity
compare to adult
Conjugated-CP • Rate of Hexobarbital metabolism
• Sensitive to drug,
Species Sleeping Time (min)
• Long duration of action
Excretion
Mice 12
• Glucoronosyltransferase (UDPGT) Rat 90
Dog 315

GREY BABY SYNDROME

Coumarin : 7-Hydroxylation found in only human,
(toxicity of chloramphenicol
rabbit, cat but not in rat, mice
in newborn baby)
Volmiting
Therefore animal used for toxicity test must has the
irregular & rapid respiration
drug metabolism process similar to human
Cyanosis

Birth Adult

GENETICS
• Activity of xenobiotic metabolizing enzymes
can be vary between individual

• Population can be divided to RAPID

METABOLIZER and SLOW METABOLIZERS

Variation in toxicant metabolism

Variation of toxicant level in the body

Variation of toxicant response/toxicity

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