3 Metabolism

Metabolic Changes of Drugs and Related Organic Compounds
Kristine Mae F. Gante, RPh

Pharmacy Department School of Health Sciences Saint Paul University Philippines
Topic Outline
General
Pathways of Drug Metabolism Sites of Drug Metabolism Roles of Cytochrome P-450 Monooxygenase in Oxidative Biotransformation Phase 1 Phase 2
Introductory Concepts
Biochemically
speaking: Metabolism means Catabolism (breaking down of substances) + Anabolism (building up or synthesis of substances)
But
when we speak about drug metabolism, it is only catabolism. That is drug metabolism is the break down of drug molecules
So
what is the term used to describe building the drug molecules? We use the word synthesis, then Drugs are synthesized in laboratory and thus is not an endogenous event
What Roles are Played by Drug Metabolism?

One
of four pharmacokinetic parameters, i.e., absorption, distribution, metabolism and excretion (ADME) of Drugs:
Metabolism and excretion together are elimination.
Elimination

Excretion
physically removes drugs from the body:

The major excretory organ is the kidney. The kidney is very good at excreting polar and ionized drugs without any major metabolism. The kidney is unable to excrete drugs with high LWPC.

In
general, by metabolism drugs become more polar, ionizable and thus more water soluble to enhance elimination.

It
also affect deactivation and thus detoxication or detoxification. Many drugs are metabolically activated (Prodrugs) Sometimes drugs become more toxic and carcinogenic
General Pathways of Drug Metabolism

2 Categories Phase I Functionalization Phase II Conjugation

2 Categories Phase I Functionalization
Oxidation Reduction Hydrolysis

2 Categories Phase I Functionalization
The purpose of these reactions is to introduce a functional polar group(s) [e.g. OH, COOH, NH2, SH] into the xenobiotic molecule to produce a more water-soluble compounds.

2 Categories Phase I metabolism can be achieved by:
Direct introduction of the functional group (e.g. aromatic and aliphatic hydroxylation)

Phase
I metabolism can be achieved
by:
Modifying or unmasking existing functionalities e.g.: reduction of ketones and aldehydes to alcohols, oxidation of alcohols to carboxylic acids, hydrolysis of esters and amides

Phase
I metabolism
Phase 1 reactions may not produce sufficiently hydrophilic or inactive metabolites, but they generally tend to provide a functional group or handle the molecule that can undergo subsequent Phase II reactions.

Phase
II metabolism
Its purpose is to attach small, polar, ionizable endogenous compounds to the functional handles of phase I metabolites or parent compounds that already have suitable existing functional groups to form water-soluble conjugated products.

Phase
I and Phase II reactions complement each other in detoxifying, and facilitating the elimination of drugs and xenobiotics.
Biotransformation is a major mechanism for drug elimination

Results
of biotransformation:
Production of metabolites that are more polar than the parent drug usually terminates the pharmacologic action of the parent drug After phase I reactions, similar or different pharmacologic activity, or toxicological activity.
Results of Biotransformation
Tetrahydrocannabinol (D1-THC) Metabolism

7
CH3
6 5 1 4 2 3
CH2OH OH H3C OH
COOH OH H3C O CH3 C5H11
H3C O CH3 D1-THC C5H11
Phase I
O CH3 7-Hydroxy-D1-THC COOR OR
C5H11
D1-THC-7-oic Acid
COO-
Where R =
H3C O CH3 C5H11 H
O HO
OH OH
Glucuronide conjugate at either COOH or phenolic OH group
-Glucuronyl moiety
The metabolite is polar, ionizable and hydrophilic
Possible consequences of biotransformation:

Inactive
metabolites (most common) Metabolites with increased or decreased potencies Metabolites with qualitatively different pharmacologic actions Toxic metabolites Active metabolites from inactive prodrugs.
Possible consequences of biotransformation:

Metabolites
are often more polar than the parent compounds. increased polarity may lead to:
A more rapid rate of clearance because of possible secretion by acid or base carriers in the kidney
This
It may lead to decreased tubular reabsorption.
Drug Metabolism Reactions
Enzymes catalyzing phase I biotransformation reactions

cytochrome
P-450 aldehyde and alcohol dehydrogenase deaminases esterases amidases epoxide hydrase
Enzymes catalyzing phase II biotransformation reactions

glucuronyl
transferase (glucuronide conjugation) sulfotransferase (sulfate conjugation) transacylases (amino acid conjugation) acetylases ethylases methylases glutathione transferase
General Metabolic Pathways

Hydrolytic Reactions Esters and amides Epoxides and arene oxides by epoxide hydrase Oxidation Aromatic moieties Olefins Benzylic & allylic C atoms and a-C of C=O and C=N At aliphatic and alicyclic C C-Heteroatom system C-N (N-dealkylation, N-oxide formation, Nhydroxylation) C-O (O-dealkylation) C-S (S-dealkylation, Soxidation, desulfuration) Oxidation of alcohols and aldehydes Miscellaneous
Phase II Conjugation
Phase I Functionalization
Drug Metabolism
Glucuronic acid conjugation Sulfate Conjugation Glycine and other AA Glutathion or mercapturic acid Acetylation Methylation
Reduction Aldehydes and ketones Nitro and azo Miscellaneous
Sites of Drug Metabolism

Liver
Major site Well organized with all enzyme systems
Drug Metabolism
Extrahepatic microsomal enzymes (oxidation, conjugation)
Hepatic microsomal enzymes (oxidation, conjugation)
Hepatic non-microsomal enzymes (acetylation, sulfation,GSH, alcohol/aldehyde dehydrogenase, hydrolysis, ox/red)

The first-pass effect Following drugs are metabolized extensively by first-pass effect:
Isoproterenol, Lidocaine Meperidine, Morphine, Pentazocine, Propoxyphene, Propranolol, Nitroglycerin, Salicylamide

Intestinal
Mucosa:
The extra-hepatic metabolism, contains CYP3A4 isozyme

Intestinal
Mucosa:
Isoproterenol exhibit considerable sulfate conjugation in GI tract Levodopa, chlorpromazine and diethylstilbestrol are also reportedly metabolized in GI tract

Intestinal
Mucosa:
Esterases and lipases present in the intestine may be particularly important carrying out hydrolysis of many ester prodrugs

Intestinal
Mucosa:
Bacterial flora present in the intestine and colon reduce many azo and nitro drugs (e.g., sulfasalazine)

Intestinal
Mucosa:
Intestinal b-glucuronidase can hydrolyze glucuronide conjugates excreted in the bile, thereby liberating the free drug or its metabolite for possible reabsorption (enterohepatic circulation or recycling)
Enzymes Involved in Drug Metabolism

CYP450 Hepatic
microsomal flavin containing monooxygenases (MFMO or FMO) Monoamine Oxidase (MAO) Hydrolases

Cytochrome P450 system: Localized in the smooth endoplasmic reticulum. It is a Pigment that, with CO bound to the reduced form, absorbs maximally at 450nm

Cytochrome P450 system: Cytochromes are hemoproteins (heme-thiolate) that function to pass electrons by reversibly changing the oxidation state of the Fe in heme between the 2+ and 3+ state and serves as an electron acceptordonor
Cytochrome P450: Naming

Before
we had a thorough understanding of this enzyme system, the CYP450 enzymes were named based on their catalytic activity toward a specific substrate, e.g., aminopyrine N-demethylase now known as CYP2E1.

CYP
N 1 L N2 N1 - Family (>40% homology) L - subfamily (> 55% homology) N2 - isoform (specific enzyme responsible for a particular reaction)
Drug Metabolism
Nomenclature
Family
CYP2D6
Sub-Family Individual Gene/ Isoform

Major human forms of P450:
Quantitatively, in the liver the percentages of total P450 protein are:
CYP3A4 28% CYP2Cx 20% CYP1A2 12% CYP2E1 6% CYP2A6 4% CYP2D6 4%

By
number of drugs metabolized:
CYP3A4 35% CYP2D6 20% CYP2C8 and CYP2C9 17% CYP2C18 and CYP2C19 - 8% CYP 1A1 and CYP1A2 -10% CYP2E1 4% CYP2B6 3%
ROLE OF CYP ENZYMES IN HEPATIC DRUG METABOLISM

RELATIVE HEPATIC CONTENT OF CYP ENZYMES
CYP2D6 2% CYP2E1 7%
% DRUGS METABOLIZED BY CYP ENZYMES
CYP 2C19 11% CYP 2C9 14%

CYP 2C 17% OTHER 36% CYP 1A2 12% CYP 3A4-5 26%
CYP2D6 23%
CYP 1A2 14% CYP2E1 5%
CYP 3A4-5 33%
Few Important CYP450 Isozymes

CYP family
CYP1 CYP2
Main functions
Xenobiotic metabolism Xenobiotic metabolism, Arachidonic acid metabolism
CYP3
CYP7 CYP11 CYP17 CYP19 CYP21 CYP24 CYP27
Xenobiotic and steroid metabolism

Cholesterol 7-hydroxylation Cholesterol side-chain cleavage, Steroid 11 hydroxylation, Aldosterone synthesis Steroid 17-hydroxylation Androgen aromatization Steroid 21-hydroxylation Steroid 24-hydroxylation Steroid 27-hydroxylation
Few Important CYP450 Isozymes

Recommended name
secologanin synthase trans-cinnamate 4-monooxygenase benzoate 4-monooxygenase calcidiol 1-monooxygenase cholestanetriol 26-monooxygenase -monooxygenase
Family/gene
CYP72A1 CYP73 CYP53 CYP27 CYP27 CYP7
flavonoid 3'-monooxygenase
3,9-dihydroxypterocarpan 6a-monooxygenase leukotriene-B4 20-monooxygenase methyltetrahydroprotoberberine 14-monooxygenase
CYP75
CYP93A1 CYP4F CYP93A1
tyrosine N-monooxygenase
CYP79
Role of CYP Monooxygenases in Oxidative Biotransformation

Oxidation of Xenobiotics
RH
+ NADPH + O2 + H+ ROH + NADP+ + H2O
Mixed
function oxidases or monooxygenases
NADP+
Drug CYP eR-Ase
CYP Fe+3
PC
Drug
Drug OH CYP Fe+3 Drug OH e-
NADPH CO
CYP-Fe+2 hu Drug
CO
CYP Fe+2 Drug O2 O2 CYP Fe+2 Drug
H2O 2H+
Electron flow in microsomal drug oxidizing system

Oxidation of Xenobiotics It requires molecular oxygen and the reducing agent NADPH. One atom of Oxygen is introduced into the substrate (RH) and the other atom is incorporated in water.

1.
In the overall reaction:

the drug is oxidized oxygen is reduced to water.
Reducing equivalents are provided by nicotinamide adenine dinucleotide phosphate (NADPH), and generation of this cofactor is coupled to cytochrome

Drug Metabolism
Oxidative Reactions
Arenols OH Arene Oxides O Epoxides O C C
C C
Benzylic, allylic aliphatic C Hydroxylation C OH

C H
R N H Miscellaneous Oxidations
S C S P
R N OH R NH + O CHR R N O
"Activated Oxigen" [FeO]

3+
R N CH2R
R O CH3
R N
O C O P Desulfuration
R OH O O-Dealkylation SH, S CH3 S-Dealkylation and S-Oxidation
S CH3
N-Hydroxylation N-Dealkyaltion and Oxidative Deamination N-Oxide Formation
Aromatic Hydroxylation
R1 CYP450 R1 Spontaneous R1
Mixed function oxidation of arenes to arenols via an epoxide intermediate arene oxide
Major route of metabolism drugs with phenyl ring Occurs primarily at para position Substituents attached to aromatic ring influence the hydroxylation Activated rings (with electron-rich substituents) are more susceptible while deactivated (with electron withdrawing groups, e.g., Cl, N+R3, COOH, SO2NHR) are generally slow or resistant to hydroxylation for
O OH R1 Epoxide hydrolase Epoxide Hydrase Aromatase OH OH R1 Glutathione

OH
R1
OH OH
S Glutathione R1 Macromolecule
OH
Macromolecule
H
H N O O N H
CYP2C19
HO O
H N O N H
N CH3
Phenytoin
O O
p-hydroxyphenytoin
OH C CH
Amphetamine
OH O
H N
CH3 CH3
ONa O
CH3
Warfarin sodium
HO O C OH O
HO
17-a-Ethinylestradiol
Ca+2
Propranolol
O
CH3 H3C HN C O N F
N N O CH3
Phenylbutazone
2
Atorvastatin
Cl H N HN N
H3C H3C
O N S O
O OH
Cl
Clonidine
Probenecid
Antihypertensive drug clonidine undergo little aromatic hydroxylation and the uricosuric agent probenecid has not been reported to undergo any aromatic hydroxylation
CH3 O N
Cl
N S
CH3
CH3 Cl
Preferentially the more electron rich ring is hydroxylated
Diazepam
Chlorpromazine
NIH Shift: Novel Intramolecular Hydride shift named after National Institute of Health where the process was discovered. This is most important detoxification reaction for arene oxides
R Spontaneous Rearrangement R NIH Shift
+ -
R Arenol H
O Arene Oxide
H O
OH
OH
Oxidation of Aromatic Moieties

PCB & TCDD Resistant to aromatic oxidation Metabolic stability coupled to the lipophilicity explains their long persistence in the body once absorbed.
Oxidation of olefinic bonds (also called alkenes)

O Epoxide hydrolase OHOH
Alkene
Epoxide
trans dihydrodiol derivative
The second step may not occur if the epoxide is stable, usually it is more stable than arene oxide May be spontaneous and result in alkylation of endogenous molecules Susceptable to enzymatic hydration by epoxide hydrase to form trans1,2-dihydrodiols (also called 1,2-diols or 1,2-dihydroxy compounds) Terminal alkenes may form alkylating agents following this pathway
O CYP3A4 N O NH 2 O N NH 2 O Epoxide hydrolase N NH2 HO OH
Carbamazepine (Active)
Carbamazepine 10,11 epoxide (Active & Toxic)
Carbamazepine trans 10,11 diol (Inactive)
Oxidation of Olefins
Epoxidation of the olefinic 10,11 double bond Further conversion to 1,2 diols
Protriptyline
(Vivactil) Antipsychotic Cyproheptadine (Periactin) H1 Antagonist
Aflatoxin B1
Carcinogenic
agent Contains olefinic (C2-C3) double bond adjacent to a cyclic ether oxygen
Aflatoxin B1
It
is oxidized to the corresponding 2,3-oxide (extremely reactive) The oxide binds covalently to DNA, RNA and proteins 2,3-dihydro-2-(N 7-guanyl)-3hydroxyaflatoxin B1
Benzylic Carbon Hydroxylation

R2 R1 C H R1 R2 C OH
Hydroxylate a carbon attached to a phenyl group (aromatic ring) R1 and R2 can produce steric hindrance as they get larger and more branched So a methyl group is most likely to hydroxylate Primary alcohol metabolites are often oxidized further to aldehyde and carboxylic acids and secondary alcohols are converted to ketones by soluble alcohol and aldehyde dehydrogenase
Benzylic Carbon Hydroxylation

O S O O N H N H CH3 CYP2C9 H HO
Tolbutamide Metabolism
O S
O O N H N H CH3
H3C
C H
O ONa N CH3
O H3C
Tolmetin sodium
Dicarboxylic acid is the major metabolite
Oxidation at Benzylic Carbon Atom

Celecoxib
Undergoes
benzylic oxidation at its C-5 methyl group to give hydroxycelecoxib as a major metabolite.
Oxidation at Allylic Carbon Atom

1-THC
It
contains 3 allylic carbon centers (C7, C6, C3). Allylic oxidation occurs at C-7 to yield 7-hydroxy-1-THC It is as active or even more active than the parent compound.
Oxidation at Allylic Carbon Atoms

H R1 C C H C H C R4 R3 R1 H C C H C OH C R4 R3
R2 H
R2 H
7 6 5
CH3
1 4 2 3
7CH2OH
CH3 OH HO + HO OH + O CH3 C5H11 H3C
CH3 OH
OH H3C
H3C O CH3 D1-THC C5H11
H3C
O CH3
C5H11
O CH3
C5H11
7-Hydroxy-D1-THC
6a-Hydroxy-D1-THC
6-Hydroxy-D1-THC
H2C
3 2
H2C H H3CO OH HO N
HO H3CO N Quinine
N 3-Hydroxyquinine
Oxidation at Allylic Carbon Atom

Examples:
Hexobarbital Pentazocin Safrole
O-Glucuronide Cojugate O O CH3 O CH3 Hexabarbital

2' 3'
O CH3 O O CH3
CH3 O
O CH3
OH
3'-Hydroxyhexabarbital
3'-Oxohexabarbital
Pentazocin e
Oxidation at Carbon Atoms to Carbonyls and Imines

Alpha
carbon
Carbon adjacent to the carbonyl (C=O) and imino (C=N) functionalities
Hydroxylation at C to C=O and C=N

CH 3 N O
3
CH 3 N Cl N
O OH N-demethylation
H N Cl N
O OH
O R C
H C H R' R
O C
H C OH R' Cl
N
The benzodiazepines are classic examples with both functionalities
Diazepam
(3S) N -Methyloxazepam or 3-Hydroxydiazepam CH 3 N O2 N N
Oxazepam
(CH3 CH 2 )2 NCH2 CH 2 N Cl N
O
3
O
3
Oxidized to their 3-hydroxy metabolites
F Nimetazepam
Flurazepam
The sedative hypnotic glutethimide possesses C a to carbonyl function
3 1
CH2 CH 3 C6 H5
HO
4
CH2 CH 3 C6 H5
N O H Glutethemide
N O H 4-Hydroxyglutethemide
Oxidation at Carbon Atoms to Carbonyls and Imines

Hydroxylation
of the carbon alpha to the carbonyl moieties generally occur at a limited extent in drug metabolism.
Oxidation at Aliphatic or Alicyclic Carbon Atoms
Oxidation
Metabolic oxidation at the terminal methyl group
- 1 Oxidation
Oxidation of the penultimate carbon atom (next-to-the-last carbon)
Aliphatic hydroxylation
H R1 H R1 C H H C H H C H R1 H H C H OH H C H C H H C H H C H H C H OH
Catalyzes hydroxylation of the and -1 carbons in aliphatic chains Generally need three or more unbranched carbons
O N H O H N O N H OH O H
CH3
O
CYP450
O
Pentobarbital Metabolism
O
CH3 O
CH3
Ibuprofen Metabolism
H 3C CH3
OH
CYP450
H3C OH CH3
OH
+
HOOC CH3
OH

Valproic
Acid (Depakene) Undergoes both and - 1 Oxidation to 5- hydroxy and 4hydroxy metabolites respectively.

Amobarbital Pentobarbital Thiamylal Secobarbital Chlorpropramide Meprobamate Glutethimide Ethosuximide Phenylbutazone
Alicyclic (nonaromatic ring) Hydroxylation

Cyclohexyl group is commonly present in many drug molecules The mixed function oxydase tend to hydroxylate at the 3 or 4 position of the ring Due to steric factors if position 4 is substituted it is harder to hydroxylate the molecules
OH O S H3C O O N H O N H O O S H3C O N H O N H
CYP450
Acetohexamide (Dymelor) Metabolism
Oxidation at Alicyclic Carbon Atoms

Glipizide
Oxidized to trans-4 and cis-3hydroxylcyclohexyl metabolite (6:1 ratio)
Oxidation at Alicyclic Carbon Atoms

Phencyclidine
(PCP)
4-hydroxypiperidyl (Aliphatic) 4-hydroxycyclohexyl derivatives (Alicyclic)
Oxidation Involving CarbonHeteroatom Systems

C-N, C-O and occasionally C-S Two basic types of biotransformation processes: 1. Hydroxylation of alpha-C attached directly to the heteroatom (N,O,S). The resulting intermediate is often unstable and decomposes with the cleavage of the C-X bond:
H H R X Ca R X O Ca R XH + O
Usually Unstable
Oxidative N-, O-, and S-dealkylation as well as oxidative deamination reaction fall under this category
Oxidation Involving CarbonHeteroatom Systems

Two basic types of biotransformation processes: 2. Hydroxylation or oxidation of heteroatom (N, S only, e.g., N-hydroxylation, N-oxide formation, sulfoxide and sulfone formation) Metabolism of some N containing compounds are complicated by the fact that C or N hydroxylated products may undergo secondary reactions to form other, more complex metabolic products (e.g., oxime, nitrone, nitroso, imino)
N-Dealkylation (Deamination)
H R1 C N R3
CYP450
R1
OH C N R3
Spontaneous
R1 C R2 O
HN R4
R3
R 2 R4
R2 R4
Deamination and N-dealkylation differ only in the point of reference; If the drug is R1 or R2 then it is a deamination reaction and If the drug is R3 or R4 then it is an N-dealkylation In general, least sterically hindered carbon (a) will be hydroxylated first, then the next, etc. Thus the more substituent on this C, the slower it proceeds; branching on the adjacent carbon slows it down, i.e. R1, R2 = H is fastest. Any group containing an a-H may be removed, e.g., allyl, benzyl. Quaternary carbon cannot be removed as contain no a-H The more substituents placed on the nitrogen the slower it proceeds (steric hindrance) The larger the substituents are the slower it proceeds (e.g. methyl vs. ethyl). In general, small alkyl groups like Me, Et and iPro are rapidly removed; branching on these substituents slows it down even more
OH N N CH3
CYP2C19
CH2
Spontaneous
CH3
CH3
CH3
Imipramine N-Dealkylation
C-N systems
1. Aliphatic (1o, 2o, 3o,) and alicyclic (2o and 3o) amines;
2. Aromatic and compounds;

3. Amides Enzymes:
heterocyclic
nitrogen
1. CYP mixed-function oxidases: hydroxylation and N-oxidation
a-C
2. Amine oxidases or N-oxidases (non-CYP, NADPH dependent flavoprotein and require O): N-oxidation
C-N systems
3o Aliphatic and alicyclic amines are metabolized by oxidative N-dealkylation (CYP) Aliphatic 1o, 2o amines are susceptible to oxidative deamination, N-dealkylation and N-oxidation reactions Aromatic amines undergoes similar group of reactions as aliphatic amines, i.e., both N-dealkylation and Noxidation H
H R1
o
O R1 N R2 Ca R1 NH R2 2o or 1o amine +
N R2
o
Ca
3 or 2 amine
Carbinolamine H
H Ca NH2 1o amine
O Ca NH2 Carbinolamine
O + NH3
Carbonyl
Ammonia
Tertiary Aliphatic and Alicyclic Amines

The oxidative removal of alkyl groups from tertiary aliphatic and alicyclic amines are carried out by CYP450 mixed function oxidase enzyme The initial step involves - carbon hydroxylation to form a carbinolamine intermediate, which is unstable and undergoes spontaneous heterocyclic cleavage of the C-N bond to give a secondary amine and a carbonyl moiety (aldehyde or ketone).
Tertiary Aliphatic and Alicyclic Amines

Imipramine is monodemethylated desmethylimipramine (major metabolite). Very little detected. of the bisdemethylated metabolite to is
OH N N CH3
CYP2C19
CH2
Spontaneous
CH3
CH3
CH3
Imipramine N-Dealkylation
3oAmine
CH3 H N O CH3
drugs
CH3 H3C N C O N CH3 CH3
CH3 H3C N O
CH3
CH3 CH3
NH2
Lidocaine
Disopyramide
Tamoxifen
CH3 N
CH3 N CH3
CH3
S
CH3
CH3
CH3
CH3 Cl
Br
Diphenhydramine Chlorpromazine Alicyclic Amine drugs

CH3 N
Benzphetamine
CH3 N
Brompheniramine
CH3 N H
O O
CH3
HO O OH
O CH3
Meperidine
Morphine
Dextromethorphan
Alicyclic Amines Often Generate Lactams
N N N CH3 N CH3
OH
N N CH3
Nicotine
Carbinolamine
Cotinine
Nicotine is hydroxylated initially at the carbon atom alpha to the nitrogen to yield a carbinolamine intermediate.
Enzymatic oxidation generates the amide) metabolite Cotinine. lactam (cyclic
Alicyclic Amines Often Generate Lactams
N CH3
N CH3
Cyproheptadine
1
Lactum metabolite
C6 H 5 H 3C
2 3
C6 H 5 H 3C
C6 H 5 H 3C
N H Phenmetrazine
N OH H Carbinolamine intermediate COOH
N O H 3-Oxophenmetrazine
COOCH 3
Hydrolysis
COOH
HN Methylphenidate
HN Ritalinic Acid
HN O 6-Oxoritalinic Acid
2o & 1o Amines
O
CH3 HN CH3
CH2
CH3 NH 2 Ampetamine
NH3 O
CH3
Methampetamine
Phenylacetone
Cl NHCH 3 O
Cl NH 2 O
Ketamine
Norketamine
Generally, dealkylation of secondary amines occurs before deamination. The rate of deamination is easily influenced by steric factors both on the a-C and on the N; so it is easier to deaminate a primary amine but much harder for a tertiary amine.
Exceptions: Some 2o and 3o amines can undergo deamination directly without dealkylation.
OH O HN Propranolol OH O HN O H CH 3 CH 3
H3C
OH O
Direct Oxidative CH 3 Deamination
OH O H O
H2 N
HN Carbinolamine
O
CH 3 CH 3
NH3
CH 3
Aldehyde Metabolite
Oxidative Deamination Through Primary Amine
OH O
CH3
NH 2 Primary Amine Metabolite (Desisopropyl Propranolol)
N-Oxidation
H N H H N OH N O
Aromatic amines
1 aromatic amine
H H N H H R C H
Hydroxylamine
H N OH R H C H Nitroso N
Nitroso
H O R C H Nitro N O O
1 amines
C H
1 amine
Hydroxylamine
H N CH3 H R C H N
CH3 R OH
H C H Nitrone N
2 amines
C H
CH2 O
2 amine
H CH3 CH3
Hydroxylamine
H N R C H CH3 N O CH3
3 amines
C H
3 amine
N-Oxide
The attack is on the unbonded electrons so 3o amines can be oxidized

Generally, only occurs if nothing else can happen, so it is a rare reaction Performed by both amine oxidases and hepatic MFOs Good examples would include amines attached to quaternary carbons since they cannot be deaminated
H3C H N Cl CH3 H H3C H N Cl CH3 OH
CYP450
Chlorphentermine N-Hydroxylation
H3C H N CH3 H
NH2
Hydroxylamine
Nitroso
Nitro Amantadine
Phentermine
Amides
C-N bond cleavage via a-C hydroxylation (formation of carbinolamide) and Nhydroxylation reactions
Oxidation involving C-O System (O-Dealkylation)
H R1 C R2 O R3
CYP450
R1
OH C R2 O R3
Spontaneous
R1 C R2 O
HO
R3
Converts an ether to an alcohol plus a ketone or aldehyde
Oxidation involving C-O System (O-Dealkylation)

OH O H3C H 3C O CH2 N N NH2
us eo tan on Sp
O H 3C H3C O
CH3 N N NH2 NH2 H 3C H 3C O
CY P4 50
NH2
OH N N NH2 NH2
Trimethoprim O-Dealkylation
CH3 N
H N O CH3
H3C
O OH N O CH3
O CH3
OH
CH3
Cl
Codeine
H3C H3C O N N NH2 N
Phenacetin
O N O
Indomethacin
OH O H3C
H N CH3 CH3
Prazosin
Metoprolol
One exception that appears to be a form of Odealkylation is the oxidation of ethanol by CYP2E1 In this case R3 is hydrogen instead of carbon to form the terminal alcohol rather than an ether The enzyme involved is CYP2E1 and has been historically referred to as the Microsomal Ethanol Oxidizing System (MEOS)
H H3C C H OH
CYP450
OH H3C C H OH
Spontaneous
H3C C H O
Oxidation involving C-S System
S-Dealkylation
H R1 C R2 S R3
CYP450
R1
OH C R2 S R3
Spontaneous
R1
C R2
HS
R3
Desulfuration
S R1 C R2 R1 O C R2
S-Oxidation
O R1 S R2 R1 S R2 Sulfoxide O R1 S R2 O Sulfone
S-Dealkylation
S N CH3 N N N S CH2 OH N N H
O CH2
SH N N N N H
N N H 6-(Methylthio)-purine
6-Mercaptopurine
S-Dealkylation
Desulfuration
Desulfuration
S-Oxidation
N O S N CH3 CH3 N O S O N CH3 CH3
N S S Thioridazine
N CH3 CH3
S Ring Sulfoxide
S Ring Sulfone
N S
N CH3
N S S
N CH3 CH3
S CH3 O Mesoridazine
O O Sulforidazine
Other Oxidative Biotransformation Pathway
Oxidative Dehalogenation Hepatic Microsomal Flavin Containing Monooxygenases (MFMO or FMO) Non-Microsomal Oxidation Reactions
Oxidative Dehalogenation
H R C Cl Cl CYP450 R OH C Cl Cl O Spontaneous O R C R C +H2O Cl OH + + H Cl H Cl
Requires two halogens on carbon With three there is no hydrogen available to replace With one, the reaction generally wont proceed The intermediate acyl halide is very reactive
OH OH NHCOCHCl2 Chloramphenicol OH OH NHCOCCl2 OH OH
HCl
OH OH NHCOCCl O Oxamyl Chloride Derivative

Tissue Nucleophiles
O2 N
O2N
O2N
Q. What is Gray Baby Syndrome?
O2N
OH NHCOC OH O Oxamic Acid Derivative
Covalent Binding (Toxicity)
Halothane Metabolized to trifluoroacetic acid Halothane trifluoroacetyl chloride trifluoroacetic acid The metabolite covalently binds in liver microsomal proteins
Chloroform It yields the chemically reactive PHOSGENE (causes hepato- and nephrotoxicity).
Non-Microsomal Oxidation Reactions Monoamine oxidase (outer membrane of mitochondria, flavin containing enzyme )
Dehydrogenases (cytoplasm)
Purine oxidation (Xanthene oxidase)
Monoamine oxidase
H R1 C N H R1 C R2 O
N R3
R2 R3
Non-Microsomal Oxidation Reactions

Two MAOs have been identified: MAOA and MAOB. Equal amounts are found in the liver, but the brain contains primarily MAOB; MAOA is found in the adrenergic nerve endings MAOA shows preference for serotonin, catecholamines, and other monoamines with phenolic aromatic rings and MAOB prefers nonphenolic amines Metabolizes 1 and 2 amines; N must be attached to -carbon; both C & N must have at least one replaceable H atom. 2 amines are metabolized by MAO if the substituent is a methyl group Phenylisopropylamines such as amphetamine and ephedrine are not metabolized by MAOs but are potent inhibitors of MAOs
Alcohol dehydrogenase
R2 R1 C H OH R1 R2 C O
Aldehyde dehydrogenase
R1 C H O R1 C OH O
Metabolizes 1 and 2 alcohols and aldehydes containing at least one H attached to a-C; 1 alcohols typically go to the aldehyde then acid; 2 alcohols are converted to ketone, which cannot be further converted to the acid. The aldehyde is converted back to an alcohol by alcohol (keto) reductases (reversible), however, it goes forward as the aldehyde is converted to carboxylic acid; 3 alcohols and phenolic alcohols cannot be oxidized by this enzyme; No H attached to adjacent carbon
H3C
H2 C
Alcohol Dehydrogenase
OH H3C
H C O
Aldehyde Dehydrogenase
H3C
OH C O
Ethanol Metabolism
Purine oxidation
O HN N N N H Xanthine oxidase O HN N H Xanthine O N N H Xanthine oxidase O HN N H O N OH N H O N H N H HN O H N O
Hypoxanthine
Uric acid (hydroxy tautomer)
Uric acid (keto tautomer)

Drug Metabolism
Reductive Reactions
Bioreduction of C=O (aldehyde and keton) generates alcohol (aldehyde 1o alcohol; ketone 2o alcohol)
Nitro and azo reductions lead to amino derivatives
Less Common Reactions: Reduction of N-oxides to their corresponding 3o amines and reduction of sulfoxides to sulfides are less frequent Reductive cleavage of disulfide (-S-S-) linkages and reduction of C=C are minor pathways in drug metabolism
Reductive dehalogenation is a minor reaction primarily differ from oxidative dehalogenation is that the adjacent carbon does not have to have a replaceable hydrogen and generally removes one halogen from a group of two or three
Reduction of Aldehydes & Ketones

H R C O R C OH
R C O R1 R2 Ketone H C OH
H Aldehyde
H 1 alcohol
R2 2 alcohol
C=O moiety, esp. the ketone, is frequently encountered in drugs and additionally, ketones and aldehydes arise from deamination Ketones tend to be converted to alcohols which can then be glucuronidated. Aldehydes can also be converted to alcohols, but have the additional pathway of oxidation to carboxylic acids.
Reduction of Aldehydes & Ketones

Reduction of ketones often leads to the creation of an asymmetric center and thus two stereoisomeric alcohols are possible Reduction of a, unsaturated ketones found in steroidal drugs results not only in the reduction of the ketone but also of the C=C Aldoketo oxidoreductases carry bioreductions of aldehydes and ketones. out
Alcohol dehydrogenase is a NAD+ dependent oxidoreductase that oxidizes alcohols but in the presence of NADH or NADPH, the same enzyme can reduce carbonyl compounds to alcohols
Reduction of Aldehydes and Ketones

Chloral Hydrate Can undergo enzymatic reduction to form trichloroethanol as a major metabolite (pharmacologically active) Further glucoronidation lead to an inactive conjugated product that is easily excreted in the urine.

Propranolol It is converted to an intermediate aldehyde by N-dealkylation or oxidative deamination. The aldehyde is oxidized to carboxylic acid (naphthoxylactic acid) but a small fraction is reduced to the alcohol derivative (propranolol glycol)

Bioreduction
of ketones leads to the creation of asymmetric center , thereby there are two possible sterioisomeric alcohol.

Product Sterioselectivity The preferential formation of one isomer over the other
O H O R1 C
+
O H2N R2
H HO N R1 C H R2
H2N + N
+
R Chiral Alcohol Ox Nicotinamide moiety + + of NADP or NAD
Ketone
R Red Nicotinamide moiety of NADPH or NADH
Ketone reduction involves a hydride transfer from the reduced Nicotinamide moiety of the cofactor NADPH or NADH to the carbonyl carbon atom of the ketone.

Acetohexamide Is the metabolized in the liver to give primarily (S)(-)-hydroxyhexamide. The metabolite is as active as the parent compound. It is eliminated through the kidneys.

Acetohexamide It is not recommended to diabetic patients with kidney failure because of the possible accumulation of hydroxyhexamide.
O OH H2C H O O O CH3 C6H 5
HO OH H 2C H O
H CH3 C 6H5 + O
H OH H2C H O
OH CH3 C6H5
R (+)-Warfarin
R,S (+)-Warfarin
R,R (+)-Warfarin
Warfarin Undergoes extensive reduction of its side chain keto group. R,S (+) is the major metabolite
Naltrexone
Reduction of the 6-keto functionality can lead to either 6- or 6- hydroxy metabolites depending on the species.
Humans and rabbits highly sterioselective (6- hydroxy

metabolites ) Chickens - 6- hydroxy metabolite Monkeys and guinea pigs both but predominantly
N OH CH2
HO
O O
Naltrexone
Reduction of Nitro & Azo Compounds

H R C H Nitro N O H R C H Nitroso N O R H C H N OH H R C H 1 amine N
H H
Hydroxylamine
R1
R2
R1
H N
H N
R2
R1
NH2
H2N
R2
Azo
Hydrazo
Two 1 amines
NH
NH2 + N N2
Azido
Amine
R1 and R2 are almost always aromatic
Usually only seen when the NO2 functional group is attached directly to an aromatic ring and are rare
Nitro reduction is carried out by NADPH-dependent microsomal and soluble nitroreductases (hepatic)
NADPH dependent multicomponent hepatic microsomal reductase system reduces the azo
Bacterial reductases in intestine can reduce both nitro and azo
O H2N S O O N H2 H2N S O N H2 H2N
N N NH2
H N O
+
NH2
NH2
Prontosil
Sulfanilamide
1,2,3-Triaminobenzene
O
O2 N N Cl
O S N H N
O2N O N N O
O NNa
HO O OH
N N
Clonazepam
Sulfasalazine
Dantrolene
Reduction of Sulfur Containing Compounds

Sulfoxide reduction (Cannot reduce a sulfone) Disulfide reduction R1
S S R2
O R1 S R2 R1 S R2
R1
S O
R2
Sulfoxide
R1 SH + HS R2
Sulfone
H3C S H3C N S S S N
CH3 CH3
H3C S H3C N SH
Disulfiram
F O
N,N-Diethylthiocarbamic Acid
OH CH3 H H 3C S O
Sulindac

Drug Metabolism
Hydrolytic Reactions
Hydrolyzes (adds water to) esters and amides and their isosteres; the OH from water ends up on the carboxylic acid (or its isostere) and the H in the hydroxy or amine
Enzymes: Non-microsomal hydrolases; however, amide hydrolysis appears to be mediated by liver microsomal amidases, esterases, and deacylases Electrophilicity of the carbonyl carbon, Nature of the heteroatom, substituents on the carbonyl carbon, and substituents on the heteroatom influence the rate of hydrolysis In addition, Nucleophilicity of attacking species, Electronic charge, and Nature of nucleophile and its steric factors also influence the rate of hydrolysis
Table: Naming carbonyl - heteroatom groups

R1 C
O R1 C R2 +
R2 O S O N N N
Name Ester Thioester Carbonate Amide Carbamate Ureide
Susceptibility to Hydrolysis Highest
C O C O N
Lowest
The Reactions
Ester hydrolysis
O R1 C O R2 R1
O C OH HO R2
Amide hydrolysis (slower)

O R1 C H N R2 R1 O C OH H 2N R2
Carbonate hydrolysis
O R1 O C O R2 R1 OH
+
O HO C O R2 HO R2
+
O HO C OH O C O
+
H O H
Carbonate
Carbonic acid derivative
Carbonic acid
The Reactions Carbamate hydrolysis

O R1 O C N O R2 R3 Carbamate R1 OH
+
R2 N HN
R2
+
O HO C OH O C O
+
H O H
HO
R3 Carbamic acid derivative
R3
Carbonic acid
Urea hydrolysis
R1 R2 O N C N R3 R1 R2 NH
+
O HO C N
R3 HN
R2
+
O HO C OH O C O
+
H O H
R4 Urea derivative
R4 Carbamic acid derivative
R3
Carbonic acid
Hydrazide hydrolysis
O R1 C H N N O R2 R3 R1 C OH + H2N N R2 R3
Hydrazide
Hydrazine
Drug Examples
OH OH O O H3C O OH O OH
H3C
O OH N O CH3
O O
+
H3C O
Aspirin
H3C O O O CH3 N O
Salicylic Acid
HO O CH3 N H3C O O CH3 N HO
Cl
Indomethacin
H2N H N O N
Cocaine
CH3 CH3
Benzoylecgonine
H3C O N N NH2
H2N
Methylecgonine
N N O O
Slow Hydrolysis
H3C
Procainamide
CH3
H2N O O CH3 OH
Prazosin
N CH3 CH3
H N O CH3
CH3
Rapid Hydrolysis
Procaine
Lidocaine
Stereoselectivity of Hydrolysis
Etomidate (Amidate, hypnotic): R-(+)-isomer is more rapidly hydrolyzed, but S-(-)-isomer is more rapidly hydroxylated.
Phase 2 Reactions
Synthetic Conjugation
Phase II
Phase II - combines functional group of compound with endogenous substance E.g.Glucuronic acid, Sulfuric acid, Amino Acid, Acetyl. Products usually very hydrophilic The final compounds have a larger molecular weight.
How We Get To Phase 2

Most of the drugs do not become polar upon phase 1 reactions. The Body is left with a plan to further metabolize the Drugs Goal of Phase 2 : Make substances more soluble that couldnt be done in the Phase 1 reactions.
Synthetic Reactions / Phase II

These reactions usually involves covalent attachments of small polar endogenous molecules such as Glucoronic acid, Sulfate, Glycine to either unchanged drugs or Phase I product having suitable functional groups as COOH,-OH,-NH2,- SH. Thus is called as Conjugation reactions. Since the product formed is having high molecular weight so called as synthetic reactions. The product formed is hydrophilic in nature with total loss of pharmacologic activity so called as a true detoxification reaction
Glucuronic Acid Conjugation

Glucuronidation is conjugation pathway the most common
The coenzyme, UDP glucuronic acid is synthesized from the corresponding phosphate Glucuronides are highly hydrophilic and water soluble UDP glucuronosyltransferase is closely associated with Cyp450 so that Phase I products of drugs are efficiently conjugated
Glucuronic Acid Conjugation

Four general classes of glucuronides: O-, N-, S-, and C Neonates have undeveloped liver UDPglucuronosyltransferase activity, and may exhibit metabolic problem. For example, chloramphenicol (Chloroptic) leads neonates to gray baby syndrome Neonatal jaundice may be attributable to their inability to conjugate bilirubin with glucuronic acid
Glucuronide formation occurs in 2 steps:-
1. Synthesis of an activated coenzyme uridine-5- diphospho -alphaD- Glucuronic acid (UDPGA) from UDP- glucose (UDPG). a-D-Glucose-1-phosphate + UTP
Pyrophosphorylase
UDPG + Ppi
UDPG - Dehydrogenase
UDPG +2NAD + H2O
UDPGA +2NADH + 2H+
2. Transfer of the glucuronyl moiety from UDPGA to the substrate RXH in presence of enzyme UDPglucuronyl transferase to form the conjugate.
UDP-Glucuronyl transferase
UDPGA + RXH
Where, X = O, COO, NH or S
RX Glucuronic Acid +UDP
03-12-2010
KLECOP, Nipani
143
Formation of Glucuronide Conjugate

HO HO HO HO
UTP PPi
O
Phosphorylase
HO HO HO HO
O O OO OO O P O P O
O NH N O
2NAD
+
OPO 32a-D-Glucose-1phosphate
UDPG
HO
U D OH PG
2N de hy dr
DH
og
en
HOOC HO O HO O O HO O P O P O OUDP-Glucuronyl-transferase (microsomal)
as e
O NH O N O
O-
HO HO HO
O O XR
UDP
RXH
OH Uridine-5'-diphosphoa-D-Glucose (UDPG)
HO
HO -D-Glucuronide
Types of Compounds Forming Glucuronides

TYPE
O H N CH3
EXAMPLES
CH3 N
O-Glucuronide
OH
HO O OH
Phenols
Acetaminophen
OH H N O OH Cl Cl
morphine
OH O H N CH3 CH3
O2 N
Alcohols
Chloramphenicol
OH
Propranolol
Enols
H2N
Hydroxycoumarine
S O2 NHOH
N-hydroxyamines/amides
CH3 N OH
N-hydroxydapsone N-Hydroxy-2-acetylaminoflourene
COOH
Aryl acids
OH
Salicylic acid
CH3 O OH
Arylalkyl acids N-Glucuronides Arylamines

O2N NH2
Fenoprofe n
O O S N H O N CH3 CH3
H N N 7-Amino-5-
nitroindazole
Sulfonamides H N
2
Sulfisoxazole
Alkylamines
Desipramine
O NH2 H3C O O H3 C O
CH3
3o Amines
CH3
Amides
Meprobamate
NH2
Cyproheptadine
Sulfate Conjugation
Occurs less frequently than does glucuronidation presumably due to fewer number of inorganic sulfates in mammals and fewer number of functional groups (phenols, alcohols, arylamines and N-hydroxy compounds)
Sulfotransferases are widely-distributed enzymes Cofactor is 3-phosphoadenosine-5-phosphosulfate (PAPS) Produce highly water-soluble sulfate esters, eliminated in urine, bile R OH R O SO3
1. Synthesis of an activated coenzyme 3-phosphoadenosine-5phosphosulfate (PAPS) which acts as a donor of sulfate to the substrate. This also occurs in two steps- an initial interaction between the sulfate and the adenosine triphosphate (ATP) to yield adenosine-5-phosphosulfate (APS) followed by activation of latter to PAPS.
ATP Sulfurylase/Mg++ ATP + SO42APS Phosphokinase/Mg++
APS + Ppi
APS + ATP
PAPS + ADP
2. Transfer of sulfate group from PAPS to the substrate RXH in presence of enzyme Sulfotransferase and subsequent liberation of 3- phosphoadenosine-5-phosphate(PAP). PAPS + RxH Rx-SO3 + PAP
Sulfotransferase
X= O,NH
Examples of compounds undergoing sulfation are: Phenol Paracetamol , Salbutamol Alcohols Aliphatics C-1 to C-5 Arylamines Aniline
Sulfation of Drugs
Phenolic sulfation predominates Phenolic O-glucuonidation competes favorably with sulfation due to limited sulfate availability Sulfate conjugates can be hydrolyzed back to the parent compound by various sulfatases Sulfoconjugation plays an important role in the hepatotoxicity and carcinogenecity of N-hydroxyarylamides
In infants and young children where glucuronyltransferase activity is not well developed, have predominating O-sulfate conjugation
Examples include: a-methyldopa, acetaminophen, phenacetin
H HO H3C HO H COOH N
albuterol,
terbutaline,
OH HO HO
H N CH3 CH3 CH3

HO
OH
H N CH3 CH3 CH3
a-Methyldopa
Albuterol
OH
Terbutaline
Sulfation of Drugs Acetaminophen O-sulfate conjugate is the main urinary metabolite in infants and young children.
Amino Acid Conjugation

The first mammalian drug metabolite isolated, hippuric acid, was the product of glycine conjugation of benzoic acid
R O COH R O O CONHCH2COH
Benzoic Acid, R = H Salicylic Acid, R = OH
Hippuric Acid, R = H Salicyluric Acid, R = OH
Amino acid conjugation of a variety of caroxylic acids, such as aromatic, arylacetic, and heterocyclic carboxylic acids leads to amide bond formation Glycine conjugates are the most common Taurine, arginine, asparagine, histidine, lysine, glutamate, aspartate, alanine, and serine conjugates have also been found
Alternative to glucuronidation Two principle pathways -COOH group of substrate conjugated with -NH2 of Glycine, Serine, Glutamine, requiring CoA activation E.g. conjugation of benzoic acid with Glycine to form Hippuric acid Aromatic -NH2 or NHOH conjugated with -COOH of Serine, Proline, requiring ATP activation
1. Activation of carboxylic acid drug substrate with ATP and coenzyme A (CoA) to form an acyl CoA intermediate. Thus, the reaction is a contrast of glucuronidation and sulfation where the donor coenzyme is activated and not the substrate.
Acetyl Synthetase RCOOH + ATP RCOAMP + H2O + Ppi
Acyl CoA Transferase
RCOAMP + CoA-SH
RCSCoA + AMP
2. Acylation of the alpha- amino acid by the acyl CoA in presence of enzyme N-acyl transferase. RCSCoA + NH2-R-COOH N-Acetyl transferase RCONH-RCOOH + CoA- SH
Brompheniramine Metabolism
CH3 N CH3
P450
CH3 NH
P450
NH2
P450
CHO
COOH
Aldehyde dehydrogenase Br Br Carboxylic Acid metabolite Glycine N-acyltransferase COOH
Br Brompheniramine
Br
Br
CH3 N CH3
N O
H N
Br Brompheniramine N-oxide
Br Glycine conjugate
Glutathione-S-transferase catalyzes conjugation with glutathione Glutathione is tripeptide of Glycine, Cysteine, Glutamic
acid
Glutathione Conjugation
NH 2 HO O HO HS H N O O N H O OH NH 2 HO NH 2 Glutathione reduced form (GSH) N H O O O O O H N O N H S S H N OH O O OH
Glutathione oxidized form (GSSG)
Glutathione is a tripeptide (Glu-Cys-Gly) found virtually in all mammalian tissues Its thiol functions as scavenger of harmful electrophilic parent drugs or their metabolites
Mercapturic Acid Conjugates

Drug O HO S H N O N H O O Amino Acid -Glutamyl-AA (AA) OH NH 2 -Glutamyl transpeptidase HO Drug O S H N O NH 2
Glutathione Conjugate Drug Glycine Cysteinyl Glycinase HO S NH 2
Acetyl CoA
CoASH
Drug S H 2N N H O CH 3
O S-substituted Cysteine Derivative
O Mercapturic acid conjugate
Acetyl Conjugation
Metabolism for drugs containing a primary amino group, (aliphatic and aromatic amines), amino acids, sulfonamides, hydrazines, and hydrazides The function of acetylation is to deactivate the drug, although Nacetylprocainamide is as potent as the parent antiarrhythmic drug procainamide (Procanbid) or more toxic than the parent drug, e.g., N-acetylisoniazid Acetylation is two-step, covalent catalytic process involving N-acetyl transferase
O H3 C XSCoA
CoASH
O H3 C X
H2N R
O H3 C NHR X-
N -Acetylation of amines
Genetic polymorphism in N-acetyltransferase activity Multiple NAT2 alleles (NAT2*5, *6, *7, and *14) have substantially decreased acetylation activity and are common in Caucasians and populations of African descent. In these groups, most individuals carry at least one copy of a slow acetylator allele, and less than 10% are homozygous for the wild type (fast acetylator) trait. The ratio of NAT2 activity is 7 in Caucasians to 18 in the Chinese population.
Example of Acetylated Drugs

O HO NH O CH3 CH3 S NH2 O OH
Cilastatin
HO H3C O COOH N S
H N
NH
Imipenem
Methyl Conjugation
Minor conjugation pathway, important in biosynthesis of epinephrine and melatonin; in the catabolism of norepinephrine, dopamine, serotonin, and histamine; and in modulating the activities of macromolecules (proteins and nucleic acids)
Except for the formation of quarternary ammonium salts, methylation of an amine reduces the polarity and hydrophilicity of the substrates
A variety of methyl transferase, such as COMT (catechol O-methyl transferase), phenol-O-methyltransferase, N-methyl transferase, Smethyltransferase etc are responsible for catalyzing the transfer of methyl group from SAM to RXH
COOH H2N
H2 N
ATP
PPi + Pi
COOH
H2 N
COOH
Methyltransferase
H3CS
Methionine adenosyltransferase
Mthetionine
S+ Ad O
CH3
HX-R
CH3 -X-R
+
S O Ad
HO
OH
HO
OH
S-Adenosylmethionine Mechanism of methyl conjugation
Age Differences Species and Strain Differences Hereditary or genetic factor Sex differences Enzyme induction/inhibition
End of Presentation
Thank you for listening!

3 Metabolism

Uploaded by

Document Information

Original Description:

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

3 Metabolism

Uploaded by

Copyright:

Available Formats

Metabolic Changes of Drugs and Related Organic Compounds

Kristine Mae F. Gante, RPh

What Roles are Played by Drug Metabolism?

What Roles are Played by Drug Metabolism?

physically removes drugs from the body:

What Roles are Played by Drug Metabolism?

What Roles are Played by Drug Metabolism?

General Pathways of Drug Metabolism

General Pathways of Drug Metabolism

General Pathways of Drug Metabolism

General Pathways of Drug Metabolism

General Pathways of Drug Metabolism

I metabolism can be achieved

General Pathways of Drug Metabolism

General Pathways of Drug Metabolism

General Pathways of Drug Metabolism

Biotransformation is a major mechanism for drug elimination

Tetrahydrocannabinol (D1-THC) Metabolism

COOH OH H3C O CH3 C5H11

H3C O CH3 D1-THC C5H11

Glucuronide conjugate at either COOH or phenolic OH group

The metabolite is polar, ionizable and hydrophilic

Possible consequences of biotransformation:

Possible consequences of biotransformation:

It may lead to decreased tubular reabsorption.

Drug Metabolism Reactions

Enzymes catalyzing phase I biotransformation reactions

Enzymes catalyzing phase II biotransformation reactions

General Metabolic Pathways

Reduction Aldehydes and ketones Nitro and azo Miscellaneous

Sites of Drug Metabolism

Major site Well organized with all enzyme systems

Extrahepatic microsomal enzymes (oxidation, conjugation)

Hepatic microsomal enzymes (oxidation, conjugation)

Hepatic non-microsomal enzymes (acetylation, sulfation,GSH, alcohol/aldehyde dehydrogenase, hydrolysis, ox/red)

Sites of Drug Metabolism

Sites of Drug Metabolism

The extra-hepatic metabolism, contains CYP3A4 isozyme

Sites of Drug Metabolism

Sites of Drug Metabolism

Sites of Drug Metabolism

Sites of Drug Metabolism

Enzymes Involved in Drug Metabolism

Enzymes Involved in Drug Metabolism

Enzymes Involved in Drug Metabolism

Cytochrome P450: Naming

Cytochrome P450: Naming

Sub-Family Individual Gene/ Isoform

Cytochrome P450: Naming

Quantitatively, in the liver the percentages of total P450 protein are:

CYP3A4 28% CYP2Cx 20% CYP1A2 12% CYP2E1 6% CYP2A6 4% CYP2D6 4%

Cytochrome P450: Naming

number of drugs metabolized:

ROLE OF CYP ENZYMES IN HEPATIC DRUG METABOLISM

% DRUGS METABOLIZED BY CYP ENZYMES

CYP 2C19 11% CYP 2C9 14%

CYP 1A2 14% CYP2E1 5%

CYP 3A4-5 33%

Few Important CYP450 Isozymes

Xenobiotic and steroid metabolism

Few Important CYP450 Isozymes

Role of CYP Monooxygenases in Oxidative Biotransformation

+ NADPH + O2 + H+ ROH + NADP+ + H2O