1 | Biochemistry Notes by Dr.

M A Bari Siddiqui Drmentors

XENOBIOTICS
XENOBIOTICS: A xenobiotic (Gk xenos “stranger”) is a compound that is foreign to the body.
The principal classes of xenobiotics of medical relevance are
 drugs,
 chemical carcinogens
 polychlorinated biphenyls (PCBs) and
 certain insecticides
BIOTRANSFORMATION: It is conversion of lipophilic substances to water soluble ones catalysed by enzymes in
the liver and other tissues.
DETOXIFICATION: The biochemical changes proceeding in the body, which convert foreign molecules into less
toxic and more soluble substances that can easily be excreted or metabolized. It is the metabolism of
xenobiotics taking place inside the body
It takes place in two phases
 In phase 1, the major reaction involved is hydroxylation, catalyzed mainly by members of a class of
enzymes referred to as monooxygenases or cytochromes P450. Reduction and hydrolysis are the other
two reactions in phase 1. Phase 1 metabolism renders compounds more reactive, introducing groups
that can be conjugated with glucuronic acid, sulfate, acetate, glutathione, or amino acids in phase 2
metabolism
 In phase 2 reactions, these derivatives are conjugated with molecules such as glucuronic acid, sulfate, or
glutathione. This renders them even more water soluble, and they are eventually excreted in the urine
or bile.
 2 | Biochemistry Notes by Dr. M A Bari Siddiqui Drmentors

Phase I:
1. Hydroxylation is the chief reaction involved in phase1. The responsible enzymes are called
monooxygenases or cytochrome P450s.
Cytochrome P450:
 Is a heme enzyme with molecular mass of about 55 kDa.
 When exposed to carbon monoxide exhibit an absorption peak at 450 nm.
 In mammals, cytochromes P450 are present in highest amount in liver cells and enterocytes but are probably
present in all tissues. In liver and most other tissues, they are present mainly in the membranes of the
smooth endoplasmic reticulum or in mitochondria of Adrenal Glands.
 Often exhibit broad substrate specificity, thus acting on many compounds; consequently, different P450s may
catalyze formation of the same product
 Extremely versatile catalysts, perhaps catalyzing about 60 types of reactions. However, basically they catalyze
reactions involving introduction of one atom of oxygen into the substrate and one into water.
 Their hydroxylated products are more water-soluble than their generally lipophilic substrates, facilitating
excretion
 Many are inducible, resulting in one cause of drug interactions.
 Many are inhibited by various drugs or their metabolic products, providing another cause of drug interactions
 Some exhibit genetic polymorphisms, which can result in atypical drug metabolism.
 Their activities may be altered in diseased tissues (eg, cirrhosis), affecting drug metabolism
 Isoforms of Cytochrome P450 Make Up a Superfamily of Heme-Containing Enzymes. They are classified based
on amino acid sequence homology of the enzymes.

Functions of cyt p450: Approximately 50% of the common drugs that humans ingest are metabolized by isoforms of
cytochrome P450. They also act on steroid hormones, carcinogens, and pollutants. The major cytochromes P450 in drug
metabolism are members of the CYP1, CYP2, and CYP3 families. In addition to their role in metabolism of xenobiotics,
cytochromes P450 are important in the metabolism of a number of physiological compounds—for example, the synthesis
of steroid hormones and the conversion of vitamin D to its active metabolite, calcitriol.

Mech of axn: The overall reaction

catalyzed by a cytochrome P450 is:
RH + O2 + NADPH + H+ R-OH +
H2O + NADP
The role of NADPH is to reduce
cytochrome P450; the reduced
cytochrome then reduces oxygen to
water and the hydroxyl group that is
introduced into the substrate. The
reaction mechanism is complex . Using
18O2 it has been shown that one atom
of oxygen forms the hydroxyl group of R–
OH and the other forms water. This dual
fate of the oxygen accounts for the
former naming of monooxygenases as
“mixed-function oxidases.”

2. reduction: detoxification in few 3. hydrolysis: glycosidic, amide and ester bonds undergo hydrolysis
cases is seen in for detoxification.

Picric acid  picramic acid Eg:

Aspirin  salicylic acid + acetic acid
Nitrobenzene  amino benzene Atropine  tropic acid + tropine
Actanilide  aniline + acetic acid
 3 | Biochemistry Notes by Dr. M A Bari Siddiqui Drmentors

Phase II:
1. Glucuronidation: Xenobiotics are glucuronidated using UDP-glucuronic acid, catalyzed by a
variety of glucuronosyltransferases, present in both the endoplasmic reticulum and cytosol.
Molecules such as 2-acetylaminofluorene (a carcinogen), aniline, benzoic acid, meprobamate (a
tranquilizer), phenol, and many steroids are excreted as glucuronides. The glucuronide may be
attached to oxygen, nitrogen, or sulfur groups of the substrates. Glucuronidation is probably the
most frequent conjugation reaction.
2. Sulfation: Some alcohols, arylamines, and phenols are sulfated. The sulfate donor in these and
other biologic and sulfation reactions (eg, sulfation of steroids, glycosaminoglycans, glycolipids,
and glycoproteins) is adenosine 3′-phosphate-5′-phosphosulfate (PAPS) so-called “active
sulfate.”
3. Conjugation With Glutathione: The tripeptide glutathione (γ-glutamylcysteinylglycine) is
important in the phase II metabolism of electrophilic compounds, forming glutathione S-
conjugates that are excreted in urine and bile. The reaction catalyzed by glutathione S-
transferases is:
R + GSH →R−S−G
Where R is an electrophilic compound. There are four classes of cytosolic glutathione S-
transferase and two classes of microsomal membrane-bound enzyme, as well as a structurally
distinct kappa class that is found in mitochondria and peroxisomes. Glutathione S-transferases
are homo- or heterodimers of at least seven different types of subunit, and different subunits are
induced by different xenobiotics. Because glutathione S-transferases also bind a number of
ligands that are not substrates, including bilirubin, steroid hormones and some carcinogens and
their metabolites, they are sometimes known as ligandin. Glutathione S-transferase binds
bilirubin at a site distinct from the catalytic site, transporting it from the bloodstream to the liver,
then to the endoplasmic reticulum for conjugation with glucuronic acid, and excretion in the bile.
Binding of carcinogens sequesters them, so preventing their actions on DNA. The liver has a very
high activity of glutathione S-transferase; in vitro the entire pool of glutathione can be depleted
within minutes on exposure to xenobiotic substrates.The activity of glutathione S-transferase is
upregulated in many tumors, leading to resistance to chemotherapy. Glutathione conjugates may
be transported out of the liver, where they are substrates for extracellular γ-
glutamyltranspeptidase and dipeptidases. The resultant cysteine S-conjugates are taken up by
other tissues (especially the kidney) and N-acetylated to yield mercapturic acids (N-acetyl
cysteine S-conjugates) which are excreted in the urine. Some hepatic glutathione S-conjugates
enter the bile canniculi, where they are broken down to cysteine S-conjugates that are then taken
up into the liver for N-acetylation, and re-excreted in the bile.
4. Acetylation—Acetylation is represented by
X + Acetyl-CoA → Acetyl-X + CoA
Where X represents a xenobiotic. As for other acetylation reactions, acetyl-CoA is the acetyl
donor. These reactions are catalyzed by acetyltransferases present in the cytosol of various
tissues, particularly liver. The drug isoniazid, used in the treatment of tuberculosis, is subject to
acetylation. There is polymorphism of acetyltransferases, resulting in individuals who are
classified as slow or fast acetylators. Slow acetylators are more subject to the toxic effects of
isoniazid because the drug persists longer in these people.
5. Methylation—A few xenobiotics are subject to methylation by methyltransferases, employing S-
adenosylmethionine as the methyl donor.