3. Bio-transformation
• DEFINITION : Chemical alteration of the drug in the body.
Non-polar compounds to polar (lipid-insoluble) compounds.
• SITES :
liver kidney intestine lungs
4. • ACTIONS
1. Inactivation
EG : Ibuprofen, lidocaine, propranolol
2. Active metabolite from active drug
Active drug Active metabolite
Morphine Morphine-6-glucoronide
Codeine Morphine
Amitryptiline Nortriptiline
Spironolactone Canrenone
Imipramine Desimipramine
5. 3. Active metabolite from inactive drug
Prodrug Active form
Levodopa Dopamine
Enalapril Enalaprilat
Prednisone Prednisolone
Sulfasalazine 5-aminosalicylic acid
Acyclovir Acyclovir triphosphate
6. Bio-transformation Phases
Phase 1 Phase 2
• Non-Synthetic/functionalization
reactions
• Metabolite may be active or inactive
1. Oxidation
2. Reduction
3. Hydrolysis
4. Cyclization
5. De-cyclization
(On Race Honda CD)
• Synthetic / conjugation reaction
• Mostly inactive
1. Glucuronide conjugation
2. Acetylation
3. Methylation
4. Sulphate conjugation
5. Glycine conjugation
6. Glutathione conjugation
7. Ribonucleotide/nucleotide synthesis
(Get A Motor Scooter, Go Get Racing)
7.
8. ENZYME INDUCTION
Microsomal Non-microsomal
Smooth endoplasmic reticulum( in liver,
kidney, intestine, lungs)
Cytoplasm and m itochondria (in liver)
Eg : Monooxygenases, CYP450, UGT,
epoxide hydrolases etc.
Esterases, amidases, some flavoprotein,
most conjugases
Catalyzes oxidation, reduction,
glucoronide conjugation etc.
All conjugation except glucoronidation
Inducible Non-inducible
9. Microsomal Enzymes
• Microsomes : ER is isolated by homogenization and fractionation of cells
reform into
Vesicles known as microsomes
10. • They are inducible by drugs and environmental factors and some dietary
constituents.
• For e,g
1. A component of grapefruit juice inhibits drug metabolism (leading to potentially
disastrous consequences, including cardiac dysrhythmias)
2. Brussel sprouts and cigarette smoke induce P450 enzymes.
3. Components of the herbal medicine St John’s wort induces CYP450 isoenzymes as
well as P-glycoprotein (P-gp)
12. Cytochrome P450
• Cytochrome P450 enzymes are heme proteins, comprising a large family (‘superfamily’)
each referred to as CYP followed by a defining set of numbers and a letter.
• P450 because reduced heme protein binds with CO to form a complex that absorbs light
maximally at 450nm.
• Not all 57 human CYPs are involved in drug metabolism.
• CYP enzymes in families 1–3 mediate 70%–80% of all drug metabolism.
• Twelve CYPs accounted for 93.0% of drug metabolism.
14. • CYP Isoforms found in liver are
CYP1A2,
CYP2A6, CYP2B6, CYP2C9, CYP2C18, CYP2C19, CYP2D6, CYP2E1
CYP3A4, CYP3A5
CYP4A11
CYP7
• The most active CYPs for drug metabolism are of CYP2c, CYP2d, and CYP3a
subfamilies.
18. Drug interaction due to enzyme induction
Auto-induction : Because the inducing agent is often itself a substrate for the induced
enzymes, the process can result in slowly developing tolerance
Eg: carbamazepine, rifampicin, nevirapine dose needed to be doubled after 2 weeks.
Enzyme induction can increase toxicity of an active metabolite of paracetamol by its
metabolite N-acetyl-P-benzoquinone imine (NAPQI).
The risk of serious hepatic injury following paracetamol overdose is increased in
patients in whom CYP has been induced, for example, by chronic alcohol Consumption.
Antibiotic rifampicin, given for 3 days, reduces the effectiveness of warfarin as an
anticoagulant.
20. Therapeutic Uses Of Enzyme Induction
• Non-hemolytic anemia : Its due to deficient glucuronidation of bilirubin,
phenobarbitone hastens its clearance.
• Cushings syndrome: Phenytoin may reduce the manifestation by enhancing
degradation of adrenal steroids.
• Chronic poisoning
• Liver diseases.
23. Drug Interaction Due To Enzyme Inhibition
• Several inhibitors of drug metabolism influence the metabolism of different
stereoisomers selectively.
• Examples of drugs that inhibit the metabolism of the active (S) and less active (R)
isomers of warfarin
Inhibition of metabolism drugs
Stereoselective for ( S ) isomer Phenylbutazone
Metronidazole
Sulfinpyrazone
Trimethoprim–
sulfamethoxazole
Disulfiram
Stereoselective for ( R ) isomer Cimetidine
Omeprazole
Non-stereoselective amiodarone
25. UDP Glucoronosyl Transferases(UGT)
• Catalyse phase II reaction - Glucuronidation
• Glucoronides excreted via – Intestine via bile (majority)
Kidney (minority)
• Glucoronides are cleaved by beta-glucuronidase found in bacteria of lower GIT –
enterohepatic circulation of drugs eg. OCPs
• UGT 1 – drugs metabolism
UGT 2 – endogenous substrate metabolism
26. Flavin Monooxygenases
• Phase I reaction
• 6 families are present
• FMO 3 is most abundant in human liver
• They are not induced or inhibited by any drug
Not involved in drug-drug interaction
27. Epoxide Hydrolases
• Carry out hydrolysis of epoxide, many of which are produced by CYP
• It metabolises very few drugs eg. Carbamazepine
28. Glutathione S Transferases (GST)
• Found in both ER and cytosol
• Microsomal form – metabolism of endogenous leukotrienes and prostaglandins
• Cytosolic form – conjugation, reduction, isomerization reaction of drug metabolism.
29. Factors Affecting Drug Metabolism
Genetic Factors Non-genetic Factors
Genetic polymorphism Commensal Gut Microflora
Diet And Envt Factors
Age And Sex
Concurrent Exposure To Inhibitors or Inducers
Diseases
30. Genetic Polymorphism
• Definition : Occurrence of a variant allele of a gene at a population frequency more
than 1%, resulting in altered expression of functional activity of the gene product, or
both.
• Based on metabolic ratio, individuals are divided into 1- Poor metabolisers
2- Extensive metabolisers
3- Ultra rapid metabolisers
• Metabolic Ratio = Percentage of dose excreted as unchanged drug
Percent of dose excreted as metabolite in urine
31. Conclusion
• Understanding drug metabolism and interaction within the body allows principles of
biotransformation to be applied in better designing and therapeutic uses of drug.
• Increased understanding of biotransformation based on pharmacogenomics will also
render pharmacologic treatment of disease more individualised, efficacious and safe.
Phase 1 reactions often introduce a reactive group, such as hydroxyl, into the molecule, a process known as ‘functionalisation’. This group then serves as the point of attack for the conjugating system to attach a substituent such as glucuronide explaining why phase 1 reactions so often precede phase 2 reactions
Both microsomal and non-microsomal enzymes are deficient in new born ESP PREMATURE, hence making them more susceptible to many drugs.
Each of the pink or blue rectangles represents one single molecule of cytochrome P450 (P450) undergoing a catalytic cycle. Iron in P450 is in either the ferric (pink rectangles) or ferrous (blue rectangles) state. P450 containing ferric iron (fe3+) combines with a molecule of drug (‘DH’), and receives an electron from NADPH– P450 reductase, which reduces the iron to fe2+. This combines with molecular oxygen, a proton and a second electron (either from NADPH–P450 reductase or from cytochrome b5) to form an fe2+ooh–dh complex. This combines with another proton to yield water and a ferric oxene (feo)3+–dh complex. (Feo)3+ extracts a hydrogen atom from DH, with the formation of a pair of short-lived free radicals (see text), liberation from the complex of oxidised drug (‘DOH’), and regeneration of P450 enzyme.
(When starting treatment with the antiepileptic drug carbamazepine. Treatment starts at a low dose to avoid toxicity (because liver enzymes are not induced initially) and is gradually increased over a period of a few weeks, during which it induces its own metabolism)