Xenobiotics: A xenobiotic (Gk xenos “stranger”) is a compound that is foreign to the body.

Classes of xenobiotics:
A. Drugs
B. chemical carcinogens
C. naturallyoccurring compounds in plant foods
D. insecticides and other pesticides.

Site of metabolism of xenobiotics: the liver

Steps of metabolism of xenobiotics:
Phase 1: It involves hydroxylation (mainly), deamination, dehalogenation, desulfuration, epoxidation,
peroxygenation, and reduction catalyzed by monooxygenases or cytochromes P450 and hydrolysis (eg, catalyzed by esterases) and
certain other nonP450-catalyzed reactions.

Phase 2: Products produced in phase 1 goes through conjugation with glucuronic acid, sulfate, acetate,
glutathione, or amino acids to produce polar compounds, soluble in water, to be

excreted from the body with bile or urine.

xenobiotics Phase1 Phase2

Prodrug and carcinogen from inactive of biologically More inactive
active compounds.
Prodrug and carcinogen From active to less active or More inactive
inactive forms
xenobiotics Converts into active products Less active or inactive PHASE 1
xenobiotics Converts into inactive More active
products
Cytochrome P450:
It is a heme enzyme.

Genes: 57 genes in human are related to Cyt P450 synthesis.

Mystery of naming: it was originally discovered when it was noted that preparations of microsomes (fragments of the
endoplasmic reticulum) that had been chemically reduced and then exposed to carbon monoxide exhibited an absorption peak at
450 nm.

Functions of the Cyt P450:

A. metabolism of xenobiotics.
B. the synthesis of steroid hormones.
C. the conversion of vitamin D to its active metabolite, calcitriol

The overall reaction catalyzed by a cytochrome P450 is: RH + O2 + NADPH + H+ → R-OH + H2

O + NADP

Abundant specific to tissue: liver contains a lot of Cyt P450 (20% of proteins of hepatocytes) in the membranes of
the smooth endoplasmic reticulum, which constitute part of the microsomal fraction when tissue is subjected to subcellular
fractionation. Moreover, it is also present in other body but in small amount.

Most important Cyt P450 members:

Family Function Members
CYP1 Drug and steroid (especially 3 subfamilies
estrogen) metabolism
CYP2 Drug and steroid metabolism 13 subfamily
CYP3 Drug and steroid (including 1 subfamily
testosterone) metabolism

Special for CYP 450:

A. In the adrenal gland, it is found both in SER and in mitrochondria.
B. Mitrochondrial and Microsomal CYP differs by using an NADPH-linked flavoprotein, adrenodoxin reductase, and a
nonheme iron-sulfur protein, adrenodoxin by mitrochondria.

Steps of hydroxylation:
NADPH

NADPH-cytochrome P450 reductase

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Induction of P450:

REACTION
Glucuronidation

Sulfation

Conjugation With
Glutathione

Acetylation
Methylation
cytochrome P450

Reductive activation of molecular oxygen

Hydroxylation of substrates

Example-1: Phenobarbital>>induces> >CYP2C0>>>CYP2C9>>>Increase the metabolism of warfarin

Dose of Warfarin must be increased.

Example-2: Ethanol>>induces> >CYP2E1>>> Metabolization of widely used solvents and compounds found in tobacco smoke.
Result- increases the risk of carcinogenicity.

Example-3: Grapefruit>>>furanocoumarins>>induces>>CYP 450>>affects>>statins, omeprazole, antihistamines and

benzodiazepine antidepressants

Polymorphism of P450 to abolish smoking: CYP2A6 has three alleles- one wild type and other two is null-
which is responsible for metabolism of nicotine.

So, person’s having null alleles will face an elevated level of nicotine in their brain and blood which will inhibit them from smoking
much.

Consequently, it has been speculated that inhibiting CYP2A6 may provide a novel way to help smoking cessation.

NAME OF THE ENZYME

glucuronosyltransferas
e

Glutathione S-
transferases

acetyltransferases
methyltransferases
PHASE 2
GROUP DONOR

UDP-glucuronic acid

adenosine 3′-
phosphate-5′-
phosphosulfate
(PAPS)

acetyl-CoA
S-adenosylmethionine
COMPOUDS

2-acetylaminofluorene
(a carcinogen),
aniline, benzoic acid,
meprobamate (a
tranquilizer), phenol,
and many steroids
Some alcohols,
arylamines, and
phenols

substrates,bilirubin,
steroid hormones and
some carcinogens
and their
metabolites(ligandin)
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Flow charts: Further fate of substrates conjugated with the Glutathione.

Gospel of glutathione:
d
In addition to its role in phase 2 metabolism, glutathione has a number of other roles in metabolism:

o1. It provides the reductant for the reduction of potentially toxic hydrogen peroxide to water in the reaction catalyzed by glutathione
peroxidase.

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2. It is an important intracellular reductant and antioxidant, helping to maintain essential -SH groups of enzymes in their reduced
state. Its involvement in the hemolytic anemia caused by deficiency of glucose-6-phosphate dehydrogenase is discussed in Chapters
20 and 53.

n3. A metabolic cycle involving GSH as a carrier has been implicated in the transport of some amino acids across membranes in the
kidney. The first reaction of the cycle is shown below.

Amino acid + GSH → γ-Glutamyl amino acid + Cysteinylglycine

This reaction transfers amino acids across the plasma membrane, the amino acid being subsequently hydrolyzed from its complex
with glutamate and the GSH being resynthesized from cysteinylglycine.

The enzyme catalyzing the above reaction is f-glutamyltransferase (GGT).

It is present in the plasma membrane of renal tubular cells and bile ductule cells, and in the endoplasmic reticulum of hepatocytes.

The enzyme has some diagnostic value because it is released into the blood from hepatic cells in various hepatobiliary diseases.

Factors responsible for affecting enzymes of the xenobiotics:

A. Species
B. Age
C. Gender
D. Gene
E. Disease (cirrhosis of liver)
F. Metabolites of some xenobiotics can inhibit or stimulate the activities of xenobiotic-metabolizing enzymes.

Responses to xenobiotics:
A. Pharmacologic response:
Covalent binding of xenobiotic metabolites to macromolecules including DNA
DNA repair mechanisms of the cell are activated
Transfer of multiple ADP-ribose units onto DNA binding proteins
Depletion of NAD as it is the source of ADP-ribose
Severely impaired ATP formation
Cell death
B. Immunological response:
Xenobiotic may bind to a protein
Acting as a hapten alters its antigenicity
 Stimulate antibody production only by binding to a protein
Antibodies react not only with the modified protein but also with the unmodified protein
Imitating autoimmune disease

C. Carcinogenic response:
a. Some chemicals (eg, benzo[α]pyrene) require activation by cytochrome P450 in the endoplasmic reticulum to become
carcinogenic (they are thus called indirect carcinogens).
b. The enzyme epoxide hydrolase is of interest because it can exert a protective effect against some carcinogens. The
products of the action of cytochrome P450 on some procarcinogen substrates are epoxides. Epoxides are highly reactive
and mutagenic or carcinogenic. Epoxide hydrolase is present in the membranes of the endoplasmic reticulum like
cytochrome P450. It acts on these compounds, converting them into much less reactive dihydrodiols.