Describe the role of

xenobiotics in diseases
Insecticide
 COSMETICS

Food additives
Food additives
PRESERVATIVES
ENVIRONMENT POLLUTANTS
Cigarette smoke
And most importantly DRUGS
Substance that has no useful function in the living organism is
called xenobiotics.

Xenobiotics Xenobiotics

a. Natural a. Exogenous
b. Man-made b. Endogenous
Xenobiotics

a. Natural :Toxins from animals and plants

b. Man-made : There are more than 200,000 man made

chemicals. Such as
Food additives
Food preservative
Insecticides
Pesticides
EXOGENOUS XENOBIOTICS

Food additives
Food preservatives
Insecticides
Pesticides
Environmental pollutants
Chemicals (CO, SO2)
Heavy metals
Radiations (UV and cosmic rays)
Drugs
ENDOGENOUS XENOBIOTICS

Are the compound produced in the body

Bilirubin

Steroids

Eicosanoids

Certain fatty acids

  Cancer
 Autoimmunity
 Cell Injury
Substances that need to be eliminated/detoxified are called
Xenobiotics

Drugs and other xenobiotics when gain access into human

biological system they undergo chemical alteration a process
called Biotransformation
Biotransformation and detoxification of xenobiotics

Biotransformation is a process by which a substance is changed

from one chemical form to another by chemical reactions within the
body.

Metabolites of some drugs have therapeutic effect rather than

absorbed drug.

Drugs and other xenobiotics are inactivated by series of enzymatic

reactions and excreted from the body.

The process is known as DETOXIFICATION.

 (PROCARCINOGENS)

Bioactivation

(CARCINOGENS)
 PHASE I REACTIONS

In phase 1 reactions xenobiotics are converted into more water

soluble (polar)

Thus, facilitating their excretion.

Hydroxylation : Catalyzed by monooxygenases / Cyt P450 is

the chief reaction of Phase 1
 Cytochrome P450

It occur in all living organisms from bacteria to mammals.

All the cells except

Cells

contain CytP450
Highest amount occur in LIVER
They are heme containing enzymes
Cytochrome P450 occur in the smooth endoplasmic reticulum of
(liver) cells and most other tissues
Cytochrome P450 is mostly associated with phospholipids
(lecithin)
Cytochrome P450 is so called because it absorbs light at 450 nm
when complexed with carbon monoxide.

It is the chief enzyme system responsible for the metabolic

clearance of drugs consumed by humans.

150 isoforms of cytochrome P450 altogether in different organisms

There are approx. 57 isoforms in humans

 Most of them have molecular weight of about 55kDa

 It is abbreviated as CYP

Most of them are inducible

 CYP 1A1
1 Family
A Subfamily
1 Individual Enzyme
Thus CYP 1A1
 CYP 2E1
Consumption of ethanol causes the induction of CYP 2E1

It is known to metabolize compounds found in tobacco smoke

Most of which are procarcinogens

Converting then into carcinogens

Risk of Carcinogenicity will increase by the consumption of

ethanol
 CYP 1A1

Is involved in the metabolism

Converting then into potential carcinogens

Polymorphism of cytochrome P450 explains the variations in drug
response among various individuals.

Variants with low catalytic activity will have slower metabolism of

the substrate therefore prolonged drug action and accumulation
Polymorphism is of CYP 2A6

Which is involved in the metabolism of tobacco to cotinine

 Three alleles of CYP 2A6

one wild type two null or inactive alleles

Individuals with null alleles will have impaired metabolism of nicotine.

Protected from becoming tobacco dependent smokers.

Individuals with null alleles will smoke less.

 Because their blood and brain level of nicotine will remain

elevated for longer than those with wild type allele.
CYP2D6: It metabolizes approx. 50% of
drugs consumed by humans.
 Mechanism of action Cytochrome P450

Cyt P450 is the most versatile biocatalyst.

It is hydroxylation through which bulk of xenobiotics are

metabolized.

Substrates are hydrophobic and are rendered hydrophilic by

hydroxylation reactions
X – H + O2 + NADPH + H+

Cyt P450 / monooxygenases

X – OH + H2O + NADP+
They are called monooxygenases because they catalyze reaction
where only one of the two oxygen atom of O2 is incorporated into

the substrate.

The other oxygen is reduced to H2O

 It requires two substrates to act as reductant for two oxygen

atoms of O2

The main substrate (X – H) accept one oxygen atom.

Co substrate (NADPH + H+) furnishes hydrogen atom to reduce

other oxygen to H2O.
The role of NADPH is to reduce cytochrome P450 .

The reduced cytochrome P450 reduces oxygen to H2O

And other oxygen is introduced into the main substrate

Due to this dual fate of oxygen

It is also called mixed function oxidases/ mixed function

oxygenases

To indicate that they oxidizes two different substrates

simultaneously
In addition to their role in metabolism of xenobiotics, Cyt P450 are
important in the metabolism of number of physiological compounds e.g.

•Synthesis of steroid hormones

•Conversion of Vit. D to its active metabolite, calcitriol

 PHASE I REACTIONS
(FUNCTIONALIZATION)

Hydroxylation : Catalyzed by monooxygenases / Cyt P450 is the chief

reaction of Phase 1

Other reactions include

Oxidation, reduction, hydrolysis, deamination, desulfuration,

epoxidation etc.

Phase I metabolized compounds may be directly excreted

Can undergo further metabolism by Phase II reactions

Hydroxylation or Oxidation
Xenobioics are mostly metabolized by hydroxylation or oxidation in
phase I

Hdroxylation

Cyt P450
Toluene benzyl alcohol
+O2

NADP+
NADPH +H+
 Oxidation

Benzyl alcohol Benzoic acid +gly

Hippuric acid
 Alcohol dehydrogenase
Ethyl alcohol CH3CHO

NADH +H+
NAD +

Aldehyde dehydrogenase
CH3 CH3COOH
CHO

NAD+ NADH +H+

Methanol Formic acid

Ethylene glycol Oxalic acd

 REDUCTION

Para-nitrophenol para –aminophenol

Nitrobezene Aminobenzene
( Aniline)

CCl3 CHO CCl3CH2OH

Chloral Trichloroethanol
 REDUCTION

Para-nitrophenol para –aminophenol

Nitrobezene Aminobenzene
( Aniline)

CCl3 CHO CCl3CH2OH

Chloral Trichloroethanol
 HYDROLYSIS
Hydrolysis is a chemical reaction in which the toxic molecules in the
body are broken down into smaller molecules

The hydroxyl and hydrogen ions are transferred to different fragments

of substrate

X – Y + H2 O X – OH + Y – H

Hydroxyl Hydrogen ion

Esters, Amides, Hydrazides, Glycosidic , Peptide, Carbamates etc are

biotransformed by hydrolysis with various enzymes such as (Esterase,

Amidase, Peptidase etc. )

Esterase
+ H2O

+ CH3COOH
 PHASE II
(Conjugation)

A xenobiotic metabolized into polar compounds by phase I

reactions may not be sufficiently polar to be eliminated from the
body.

Phase I intermediates usually undergo biotransformation by

Phase II reactions

Phase II reactions mostly metabolizes phase I byproducts (–OH,

COOH, -NH2).

Into more hydrophilic and nontoxic compounds so that it can be

eliminated from the body.
 PHASE II reactions
(CONJUGATION)

Conjugation reactions

Molecules normally present in the body are added to the reactive

site (such as –OH, -COOH, -NH2) of phase I metabolite

Certain xenobiotics (phenol ) are directly conjugated without

undergoing phase I reaction.
 Conjugation
1.Glucuronic Acid
Glucuronic acid conjugation catalyzed by UDP-glucuronyl transferase

Is the most common phase II reaction

UDP-glucuronic acid is glucuronyl donor.

Glucuronic acid can conjugate

Hydroxyl (both phenolic , alcoholic), carbonyl, sulphydryl

and amino compounds.

Aniline, morphine (analgesic), many steroids, acetylaminofluorine

(a carcinogen) are all excreted as glucuronide derivative
Bilirubin, a hydrophobic molecule is

Conjugated and excreted as glucuronide derivative

Which is more water soluble

Bilirubin + UDP – Glucuronate

UDP – glucuronyl
transferase

Bilirubin diglucuronate + UDP

(conjugated bilirubin)
2. Glutathione

Glulathione (γglutamyl-cysteinyl-glycine)

Conjugates number of xenobiotics and many drugs

Enzyme catalyzing the conjugation is Glutathione-S-tranasferase

Which is found in high concentration in liver

Glutathione is abbreviated as GSH

Because it is the sulfhydryl group that take part in the reaction

 glutathione S-tranasferase
X + GSH X---S-G
Glutathione conjugate

GGT
X---S-G + Acceptor cysteinyglycine-X
+
gammaglutamyl acceptor
dipeptidase
cysteinyl glycine-X Cysteine-X + Glycine

N-acetyl Transferase Mercapturic acid

Cysteine-X + acetyl CoA
+ CoA
Cysteine conjugates are N-acetylated to yield mercapturic
acid
 3. Glycine

Cholic acid + Glycine Glyco-cholic acid

Deoxycholic acid +Glycine Glycodeoxycholic acid

Chenodeoxy + Glycine Glyco-chenodeoxycholic acid

cholic acid

Lithocholic acid +Glycine Glycolithocholic acid

 3. Glycine
Potentially harmful substances are converted to harmless
derivatives and are excreted in urine

e.g. benzoic acid (a food preservative)

Benzoic Acid + Glycine

Hippuric acid

(Benzoyl glycine)
4. Glutamine

Phenyl acetic acid + Glutamine

Phenyl acetyl glutamine

5. Sulfation
Alcohols, steroids, arylamines and phenols

Sulfotransferase
R – OH + PAPS RO – SO3 + PAP
3'-phospho adenosine 5'
phospho sulfate (PAPs)

Sulfotransferase
Phenol + PAPS Phenyl sulfate + PAP

Sulfotransferase
Indole + PAPS Indoxyl sulfate + PAP
6. Acetylation

Acetly
Isoniazid + Acetylated isoniazed
transferase

Acetly
Sulfanilamide + acetyl CoA Acetylated sulfanilamide
transferase
7. Methylaton
Amino, hydroxyl or thiol

X – OH + SAM

methyl transferases

S-adenosyl homocystine + XO – CH3

In phase III

Xenobiotic conjugates are further metabolized and excreted

e.g. processing of glutathione conjugates to acetylcystine

conjugates
S.NO. PHASE I PHASE II
1. Hydroxylation, oxidation, Conjugations
reduction, hydrolysis etc.

2. Degradative Synthetic
3. Introduction or unmasking of Generally conjugates phase I
functional groups(-OH, _NH2, - metabolites with glucuronic
SH, -O, -COOH) acid, sulphate, methyl, acetyl,
amin acids

4. Functionalization Conjugation

5. Increase in hydrophilicity is Increase in hydrophilicity is

small large.

6. Metabolites formed may be Metabolites usually larger,

smaller, polar/non polar, polar, water soluble and
active/inactive inactive
Phase I and phase II reactions of xenobiotics occur primarily in
liver

Once biotransformed in liver it enter the blood stream. It is

removed from the body by
Factors that affect the level of xenobiotics in the body include

 Rate of biotransformation

1.Species variation: Human CYP450 1A2 has 10-fold

higher catalytic activity than rat CYP450 1A2

2. Inter individual variations: Among human individuals, the

activity of CYP450 1A2 may vary.
 3. Enzyme Induction/Inhibition: Induction /Inhibition will
affect rate of clearance

4. Age and sex: The expression of CYP450 enzymes is age and
sex dependent.

5. Hormone and cytokines: CYP450 enzyme expression can

also be regulated by hormones and cytokines-mainly
through nuclear receptors.

 Polarity of the parent xenobiotics

 Rate of excretion by the liver or kidneys

REALTIONSHIP BETWEEN XENOBIOTICS EXCRETION
AND ASSOCIATED TOXICITY

Most xenobiotics are biotransformed into products that are more

polar and thus more readily excreted than the parent molecules.

Therefore, the rate of metabolism is a critical determinant of toxic

potential of xenobiotics.

Compounds that are readily metabolized are usually readily

excreted and thus are proportionally less prone to accumulate in
tissues.
Some tissues can accumulate certain xenobiotics and release
them
slowly.

For example, lead stored in bone does not produce toxicity

but can be mobilized from bone and enter into soft tissues and
cause toxicity.

Pb can also enter into RBC and cause hemolysis--may produce
anemia
Scheme showing how metabolism of Xenobiotics
can result in cell injury/autoimmune diseases or
Cancer
Failure to detoxify CYP450 metabolized intermediates into non-toxic
metabolites by conjugation.

Gives them chance to covalently react with cellular macromolecules

e.g. proteins, nucleic acids, lipids.

Causing cell injury, cell death, mutation, cancer and/or autoimmune

diseases.
The generalization that phase 1 and 2 are detoxifying and
chemopreventive might not be true under all circumstances.
Dubin-Johnson Syndrome

Defective transport of conjugated xenobiotics may cause its

accumulation in the body.

An example is of Dubin-Johnson Syndrome. Which is caused due to

inability of liver cells to excrete conjugated bilirubin.
Toxic Effects of some Xenobiotics on Humans

Food additives (Monosodium Glutamate & artificial sweeteners)

Immediate effect: Headache, altered concentration

Long term effects: Risk of infertility, cancer and CVD

Chemicals (Bisphenol A)

Hormonal Imbalance

Malabsorption syndrome

Heavy metals in water (For example Arsenic)

Arsenic produces variety of lesions on hands and foot

It may also cause cancer