Xenobiotic metabolism
Cytochrome P 450 enzyme
Lecture outlines
• General overview about CYP 450
• Biological role of CYP 450
• Nomenclature and classifications
• Cytochrome P450 catalysis
• CYP inducers and inhibitors
• Effects of xenobiotic metabolism
Cytochrome P450 (CYP) overview
• Cytochrome P 450 enzymes are a superfamily of heme enzymes
(many isoforms)
• Can catalyze different reactions types, mainly hydroxylation.
• It act as a first line of defense against toxins, it is a powerful
detoxification system
• CYP has a wide substrate specifity-advantage.
• Can be induced or inhibited
• Occurs in most of tissue with highest amount in liver followed by
small intestine but probably present in all tissue (except of muscles and
Biological role of Cytochrome P450 (CYP)
• CYP enzymes is the first defense mechanism in detoxification
phase 1 xenobiotic metabolism
• CYP enzymes present in the adrenal gland are responsible for
various hydroxylases which is the play key roles in production of
steroid hormones, vitamins A and D, lipid-like eicosanoid
molecules involved in signaling.
• Metabolism of fatty acids and eicosanoids e.g. CYP51, essential
in eukaryotic sterol biosynthesis.
Major role of cytochrome P450
• Major role in drug metabolism and detoxification
• type CYP3A4 estimated to act on ~ 50% of known drugs
e.g. the antibiotic erythromycin
• Human has 57 cytochromes P450s, which belong to 18 families and 41
subfamilies.
• One of the p 450 which exhibit polymorphism is CYP2D6,it catalyze
the metabolism of a large number of clinically important drugs like
antidepressant
Nomenclature and Classifications
The Cytochrome P 450 name derived from:
• Membrane bound within a cell(cyto) + heme pigment (chrome and P)+
• Absorbs light at 450 nm when exposed to CO in spectrophotometer.
Nomenclature and Classifications
• CYP enzymes classified according to their amino acid
sequence:
1.Families proteins with 40% amino acid sequence, it consists of serial
numbers
e.g. CYP2, CYP3
2. Subfamilies members of family have 55% amino acid sequence, it
consists of letter from A to D
e.g. CYP2D, CYP3A
3. Individual genes
e.g. CYP2D6, CYP3A4
Nomenclature and Classifications
Nomenclature and classifications
CYP1A2
Individual member
Family Subfamily of that subfamily
Nomenclature and classifications
• CYP(gene family)(subfamily)(individual gene)
• CYP1A2: metabolizes caffeine
• CYP3A4: most abundant CYP with broad substrate-specificity
• CYP2E1: metabolizes acetaminophen and ethanol
CYPs catalyze several types of oxidation reactions including:
• Hydroxylation of an aliphatic or aromatic carbon
• Epoxidation of a double bond
• Heteroatom (S-, N-, and I-) oxygenation and N-hydroxylation
• Oxidation/reduction
• Cleavage of esters
• Dehydrogenation
• Dealkylation
Cytochrome P450 catalysis
• Overall reaction
NADPH + H+ + O2 + R-H NADP+ + H2O + R-OH
• O2 is activated and cleaved; one to the product, the other to water.
• Electron transport systems in endoplasmic reticulum (microsomal; 50 of 57
isoforms) and mitochondria (7 of 57 isoforms)
• NADPH is a two-electron donor, but the heme iron can accept only one electron
at a time (Fe3+
• Fe2+).
• Electron transfer to cytochrome P450 is by NADPH cytochrome P450 reductase
relays the electron from NADPH to cytochrome P450 one at a time.
• Role of cytochrome b5 is not understood and varies among the different P450s
• Hydroxylation is the chief reaction in phase 1
• The enzymes responsible for hydroxylation reactions are
monooxygenase enzyme or cytochrome P450.
• Example:
CYP enzymes induction and inhibition
• Role in metabolism of drugs means sensitivity to level of enzymatic
activity of cytochrome P450s
• Xenobiotics/drugs induce expression of the cytochrome P450 that
metabolizes that compound
• Particular xenobiotics/drugs can also inhibit certain cytochrome P450s
• Unintended effects on one drug can occur due to another drug
inducing/ inhibiting P450 levels
CYP enzyme induction
• CYP enzyme inducers increase the rate of hepatic metabolism, usually
through increased transcription of mRNA, and decrease serum
concentrations of other drugs metabolized by the same hepatic
isoenzyme.
• E.g.
• Rifampin and rifabutin are classic examples of enzyme inducers that
decrease plasma concentrations of coadministered CYP substrates.
CYP enzyme inducers
Warfarin
Inactive
• Coadministration of warfarin and phenobarbital reduces the effect of
warfarin
• The dose of warfarin must be increased to achieve therapeutic effect
• Ethanol induction of CYP2E1 procarcinogen carcinogen
CYP enzyme inhibition
• Drug interactions due to drug metabolism inhibition are common
because, CYP450-mediated metabolism is the major route of
elimination for a large number of drugs.
• Mechanisms of CYP450 inhibition can be categorized as:
Reversible or Irreversible
1. Reversible inhibition is a result of rapid association and dissociation
between the substrate drugs and the enzyme and can be categorized
as competitive or non-competitive.
Examples:
sertraline, montelukast, sulfaphenazole, nootkatone, quinidine and,
ketoconazole
2. Irreversible inhibition it generally derives from the activation of a
substrate drug by a CYP450 isoform into a reactive metabolite, which
binds to the enzyme heme prosthetic site (part of the active site),
resulting in irreversible long-lasting loss of enzyme activity
e.g: Nerve gases
You have to think
What is the drug- drug interactions?
Summary
• P450s are a large class of heme proteins with absorbance at 450 nm,
divided into families and subfamilies
• P450s play many biological roles
• P450s are involved in numerous drug interactions, and their gene
expression is affected by xenobiotics/drugs
• P450s catalyze a wide range of chemical reactions on a large set of
substrates. The reaction involves O2 and electron transfer from a
second enzyme called NADPH-cytochrome P450 reductase
• P450 reactions of endogenous substrates are diverse and involved in
different metabolic processes e.g. sterol biosynthesis
• P450 reactions of exogenous substrates have 3 types of outcome
