METABOLISM OF XENOBIOTICS

Raymond Oliver A. Cruz, MD raymondolivercruz.wordpress.com

WHAT IS A XENOBIOTIC?
A xenobiotic is a compound that is foreign to the body It may include drugs, chemical carcinogens, and various other compounds The xenobiotic may be excreted unchanged, or it may me metabolized by the bodys enzymes to other compounds

Metabolism of Xenobiotics
Occurs in mostly in liver (enzymatic processes)
Convertion into more hydrophilic substances - excretion via urine May convert procarcinogenics into cytotoxic, mutagenic compounds Different persons may have differences in metabolism (genetic diff., physiol. factors)

Metabolism of one xenobiotic may influence metabolism of another

Xenobiotics Drugs Other foreign non-essential compounds Metabolism in non-hepatic tissue Intestinal mucosa Kidney Lung Bacteria in GI-tract

First-pass metabolism: Xenobiotic metabolized before reaching general circulation

2 PHASES OF XENOBIOTIC METABOLISM

1. Hydroxylation / Biotransformation 2. Conjugation

Pathways of metabolism Phase 1: Biotransformation Attachment of new functional groups, transformation of existing func. grps, oxidation, reduction, hydroxylation, hydrolysis etc. Phase 2: Conjugation. Masking of an existing functional group by acetylation, glycosylation, attachment of an amino acid, or other mechanism

More hydrophilic drug

Renal excretion
CH3

NH

NH

Demethylation
HO O OH

Conjugation glucuronic acid

HO O OH

Phase 1

Phase 2

HO

HO

OH O OH CO2H

Morphine

HYDROXYLATION
Catalyzed by cytochrome P450, a monooxygenase Hydroxylation may sometimes terminate the action of a drug Other phase 1 reactions may include deamination, dehalogenation, desulfuration, epoxidation, peroxygenation, and reduction Reactions involving hydrolysis also occur in phase 1

CYTOCHROME P 450
The reaction catalyzed by a monooxygenase is as follows: RH + O2 +NADPH + H+ R-OH + H2O + NADP It is sometimes referred to as a Mixed Function Oxidase Approximately 50% of the drugs humans ingest are metabolized by isoforms of cytochrome P450

Participation of the CYP Enzymes in Metabolism of Some Clinically Important Drugs

CYP Enzyme Examples of substrates
1A1
1A2 2A6

Caffeine, Testosterone, R-Warfarin

Acetaminophen, Caffeine, Phenacetin, R-Warfarin 17-Estradiol, Testosterone

2B6
2C-family

Cyclophosphamide, Erythromycin, Testosterone

Acetaminophen, Tolbutamide (2C9); Hexobarbital, SWarfarin (2C9,19); Phenytoin, Testosterone, R- Warfarin, Zidovudine (2C8,9,19);

2E1
2D6 3A4

Acetaminophen, Caffeine, Chlorzoxazone, Halothane

Acetaminophen, Codeine, Debrisoquine Acetaminophen, Caffeine, Carbamazepine, Codeine, Cortisol, Erythromycin, Cyclophosphamide, S- and RWarfarin, Phenytoin, Testosterone, Halothane, Zidovudine

Adapted from: S. Rendic Drug Metab Rev 34: 83-448, 2002

CONJUGATION
Compounds produced in phase 1 are converted by specific enzymes to various polar metabolites by conjugation with glucuronic acid, sulfate, acetate, glutathione, or certain amino acids, or by methylation

GLUCURONIDATION
UDP-glucuronic acid is the glucuronyl donor, and glucuronosyltransferases are the catalysts This is the most frequent conjugation reaction

SULFATION
Sulfate donor is adenosine 3-phosphate5-phosphosulfate (PAPS), called active sulfate.

GLUTATHIONE
Glutathione (-glutamyl-cysteinylglycine) is a tripeptide consisting of glutamic acid, cysteine, and glycine Electrophilic xenobiotics (such as certain carcinogens) are conjugated to the nucleophilic GSH: R+GSHRSG The enzymes catalyzing these reactions are called glutathione S-transferases and are present in high amounts in liver cytosol

ACETYLATION
X+Acetyl-CoAAcetyl-X + CoA Catalyzed by acetyltransferases present in the cytosol of various tissues, particularly the liver The drug isoniazid, used in the treatment of tuberculosis, is subject to acetylation

ACETYLATION
Polymorphic types of acetyltransferases exist, resulting in individuals who are classified as slow or fast acetylators Slow acetylators are more subject to certain toxic effects of isoniazid because the drug persists longer in these individuals

METHYLATION

Conjugation Reactions Glucuronidation

CO2H O OH HO O OH O P O P O CH2 OH O OH ON O NH O

+ ROH or R 3N UGT

CO2H OO R OH OH OH

O-glucuronide
CO2H R + N R O R OH OH OH

UDP- -D-glucuronic acid

N+-glucuronide

Liver has several soluble UDP-Gluc-transferases

HO 3 N O N CH3
6

N N

CH3

HO

Morphine

Am itriptyline

Cotinine

Glucuronic acid conjugation to phenols, 3-amines, aromatic amines

Conjugation Reactions Sulfation

R OH
NH2 N N

+
N N H O H OH O OH O P O S O O

O R O S OH O

(PAPS, 3-phosphoadenosine5-phosphosulfate)

H HO

H OH

Examples: ethanol, p-hydroxyacetanilide, 3-hydroxycoumarin

H2N N O

H2N O HO S O N

NH2

NH

Minoxidil

Minoxidil-sulfate

Sulfation may produce active metabolite

Conjugation Reactions Acetylation

O Ar NH2 CoA S O Ar N H R N H CH3 CH3 O R S R O R NH2 R OH R SH

O CH3 O CH3

Acetyl transferase

Examples: Procainamide, isoniazid, sulfanilimide, histamine

NAT enzyme is found in many tissues, including liver

PURPOSE OF XENOBIOTIC METABOLISM

To increase the water solubility of the xenobiotic and facilitate excretion from the body If xenobiotic is not metabolized, it would remain in the nonpolar state and would be stored indefinitely in the adipose tissues

A METABOLIZED XENOBIOTIC MAY HAVE THE FOLLOWING FATES:

It may be converted from an inactive to a biologically active compound (prodrug or procarcinogen) Additional phase 1 reactions may convert the active compounds to less active or inactive forms prior to conjugation.

A METABOLIZED XENOBIOTIC MAY HAVE THE FOLLOWING FATES:

In other cases, it is the conjugation reactions that convert the active products of phase 1 reactions to less active or inactive species, which are subsequently excreted in the urine or bile In a very a few cases, conjugation may increase the biologic activity of a xenobiotic

PROPERTIES OF HUMAN CYTOCHROME P450

All are hemoproteins Exhibit broad substrate specificity Found mostly in the liver but is also present in other tissues A mitochondrial or endoplasmic reticulum enzyme Activity may be induced or depressed by certain drugs, causing drug interactions

Ex. Warfarin, used to prevent blood clotting, is metabolized by a cytochrome which is induced by Phenobarbital. Thus when both drugs are given, warfarin dose must be increased to achieve therapeutic doses.

PROPERTIES OF HUMAN CYTOCHROME P 450

Exhibits genetic polymorphism, so different individuals metabolize drugs differently Activity is affected also by tissue or organ disease (e.g. Cirrhosis) NADPH, not NADH, is involved in the reaction mechanism The preferred lipid in its structure is phosphatidylcholine

CYTOCHROME NOMENCLATURE
CYP cytochrome Arabic number family (40% or more sequence identity) Capital letter subfamily (>55% sequence identity) Individual P450s are given arabic numerals Ex. CYP1A1 means it is a member of family 1 subfamily A and is the first individual member of the subfamily

RESPONSE TO XENOBIOTICS
May be affected by age, sex, and genetic factors (Pharmacogenomics) Reaction to drugs: Pharmacogenetics May result in cell injury, allergies, or carcinogenesis