300

Biotransformation
XENOBIOTICS AND DETOXIFICATION
Questions:
1. Detoxification of xenobiotics april 2001
2. Detoxification by conjugationapril 2001
3. Cytochrome P 450april 2004
4. How are Xenobiotics detoxified by conjugation? Pon Nov 2011
5. Detoxification reactions. Pon May 2014
6. How do the following Xenobiotics undergo biotransformation? Pon May
2004
a. Methanol
b. Picric acid

1. Detoxification refers to the process whereby the noxious substances in human body
are rendered innocuous and water soluble for elimination. The term detoxification is
replaced by biotransformation or metabolism of xenobiotics.
2. Biotransformation is transformation of one chemical to other within the body by a
chemical reaction within the body.
3. Xenobiotics are compounds, which may be accidentally ingested as drugs, or those
produced in vivo by bacterial metabolism. Eg:
a. Accidental ingestants: preservatives, food additive, and adulterants.
b. Drugs for therapeutic purposes: eg. acetaminophen taken in toxic dose gets
converted to a more toxic metabolite.
c. In vivo:
i. Bilrubin, and steroids from metabolism
ii. In the large intestine by the bacteria which enter the circulation. These
include indole from tryptophan, cadaverine from lysine, tyraminef rom
tyrosine,p henolf rom phenylalanin
4. Site of detoxification
a. The detoxification reactions are carried out mainly in the liver which is equipped
with the enzyme machinery.
b. Kidney and other organs may sometimes be involved.
c. The products formed by detoxification are mostly excreted by the kidneys, less
frequently excreted via feces or expired air.
5. Mechanism of detoxification: It is divided into two phases. One or both phases may
be involved in detoxification.
a. Phase 1: the reactions are oxidation, reduction and hydrolysis.
b. Phase 2: The reactions are conjugation with groups like glucuronic acid, amino
acid, lutathione, sul fate, acetate, and methyl groups.
6. Oxidation: Alcohols, aldehydes, amines, etc undergo oxidation.
c. Methanol > Formic acid
d. Ethanol > Acetic acid
e. Benzaldehyde > Benzoic acid
f. Aliphatic amine > Aliphatic acid
g. Aniline > p-Amino phenol.
h. Sulphur > Sulfuric acid
i. Drugs: Mebromamate > Hydroxymepbrobamate
2. Reduction: examples: Picric acid is reduced picramic acid. Chloral is reduced to
Trichloro ethanol.

301

3. Hydrolysis: Hydrolysis of esters, glycoside, amide are examples. Aspirin, atropine,

procaine are examples of detoxification by hydrolysis.
4. Conjugation:
a. Conjugation is the process in which a foreign compound combines with a
substance
produced in the body.
b. At least 8 different conjugating agents have been identified in the body:
Glcuronic acid, glycine, Cysteine, glutamine, methyl groups, sulfate, acetic acid,
and thiosulfates
c. Conjugation with glucuronic acid is the most common.The active form of
glucuronic acid is UDP-glucuronic acid produced in the uronic acid pathway. The
microsomal enzymes UDP glucuronyl transferase participate in glucurnide
formation. Eg. Bilirubin (fat soluble) bilirubin diglucuronide (water soluble).
d. Many aromatic carboxylic acids (e.g. benzoic acid, phenylacetic acid) are
conjugated with glycine. Hippuric acid is formed when glycine is conjugated with
benzyl CoA.
e. Alkyl or aryl halides, alkenes, nitro compounds and epoxides get conjugated with
cysteine of glutathione.
f. Phenylacetic acid is conjugated with glutamine to form phenylacetyl glutamine.
g. The methyl group (-CH3) of S-adenosylmethionine is frequently used to
methylate certain xenobiotics. This is catalysed by the enzyme
methyltransferase.
Methyltransterase
S-Adenosylmethionin + X-OH------------------------ S-Adenosylhomocysteine + XO-CH3
h. Sulfate: Several aliphatic and aromatic compounds undergo sulfation. Phenol
phenyl sulfate
i. Acetyl CoA: sulfanilamide are converted to acetyl sulfanilamide
j. Thiosulfate : The highly toxic cyanides are conjugated with thiosulfate to form
less toxic thiocyanate.
5. Detoxification by drugs:
a. The toxic effects of certain metals such as arsenic, mercury and cadminum could
be overcome by administering BAL (British antilewisite). This compound was
developed during the World War ll and was cused as a detoxifying agent for
certain war poisons.
b. The mechanism of action of BAL is not clearly known. lt is believed that BAL
readily combines with metals and gets easily excreted into urine.
Cytochrome P450
1. Role of P-450 Enzyme systems in biotransformation: (S.N)
2. What is the function of cytochrome P450 in the body? MGR Feb 2014
3. Cytochrome P 450.
1. Cytochrome P450s (CYPs) are actually a superfamily of related, heme-containing
monooxygenase enzymes that participate in a broad variety of reactions. The name
P450 reflects the absorbance at 450 nm by the protein. They are heme containing
enzymes, localized in the endoplasmic reticulum of liver.
2. They are monooxygenases. NADPH (and not NADH) is the co-enzyme for all the P450
enzymes. The pverall reaction is:
3. R-H + O2 + NADPH + H+ R-OH + H2O + NADP+
4. Almost all common drugs are metabolized by the P450 system. There are about 150
isoforms of the P450 enzymes.They are inducible enzymes. Phenobarbital causes
increased activity of P450.
5. The P450 enzymes are seen in many tissues, including adrenal glands, where they are

302

present both in mitochondria and in microsomes.

6. The mitochondrial P450 enzymes utilize NADPH linked flavoprotein, adrenodoxin
reductase, and a non-heme iron-sulfur protein, adrenodoxin. They are mainly involved in
steroid biosynthesis.
7. The liver uses this same system in bile acid synthesis and in the hydroxylation of
cholecalciferol to 25-hydroxycholecalciferol (vitamin D3 , see p. 386), and the kidney
uses it to hydroxylate vitamin D3 to its biologically active 1,25-di - hydroxylated form.
8. Microsomal cytochrome P450 monooxygenase system found in the liver is responsible
for the detoxification of foreign compounds (xenobiotics).
Detoxificationby P-450:
1. Most of the oxidation reactions of detoxification are catalysed by monooxygenase or
cytochrome P-450
2. P-450 is a cytochrome involved in biotransformation reactions. They are heme
containing proteins present in endoplasmic reticulum of liver. They absorb light at 450
nm when exposed to carbon monoxide and hence the name.
3. It is an inducible enzyme. Phenobarbital causes increased activity of P-450. They exist
in isoforms, some with low catalytic activity which explains individual variations in drug
response.
4. Most of the reactions of cytochrome P-450 involve the addition of a hydroxyl group to
aliphatic or aromatic compound.
5. Multiple forms of cytochrome P450 (6 species) have been isolated. They are all
hemoproteins, containing heme as the prosthetic group.
6. Cytochrome P 450 species are found in the highest concentration in the microsomes of
liver. In the adrenal gland, they occur in mitochondria.
7. The mechanism of action of cytochrome P 450 is complex and is dependent on NADPH.
8. The phospholipid-phosphatidyl choline is a constituent of cytochrome P 450 system
which is necessary for the action of this enzyme.
9. A distinct species namely cytochrome P 448 is specific for the metabolism of polycyclic
aromatic hydrocarbons, hence it is also known as aromatic hydrocarbon hydroxylase.