The Organic Chemistry of

Drug Design and Drug

Action
Chapter 8
Drug Metabolism
Drug Metabolism

 Foreign organism – elicits antibody response

 Low molecular weight xenobiotics – nonspecific enzymes
convert them into polar molecules for excretion
 Enzymatic biotransformations of drugs – drug metabolism
Principal site of drug metabolism is the
liver; also kidneys, lungs, GI tract
Pathway of Oral Drugs
take via mouth absorbed through small bloodstream
intestine or stomach
liver
(first metabolized)

Drug metabolism by liver enzymes – first-pass effect

 Avoid first-pass effect by changing the
route of administration
 sublingual route (under the tongue) bypasses liver

 rectal route (suppository or enema)

 intravenous (i.v.) injection – rapid response, circulation time

of 15 seconds
Avoid first-pass effect by changing the
route of administration (cont’d)
 intramuscular (i.m.) injection – for large volumes or slow
absorption
 subcutaneous (s.c.) injection – through loose connective
tissue of s.c. layer of skin
 pulmonary absorption – gaseous or highly volatile drugs
 Drug metabolism is desirable once drug has reached site of
action – may produce its effect longer than desired or
become toxic.
 Drug metabolism studies are essential for the safety of
drugs. Metabolites must be isolated and shown to be
nontoxic.
An active metabolite that is less toxic

 Terfenadine is cardiotoxic, since it binds to the hERG

channel
 Fexofenadine has similar antihistamine activity, but no
hERG activity
Synthesis of Radioactive Compounds

 To increase sensitivity for detection of metabolites, radioactivity is

incorporated into the drug candidate.
 Incorporate a commercial radioactive compound near the end of the
synthesis, if possible.
 Usually the radioactive synthesis is different from that of the unlabeled
compound.
 [14C] preferable to [3H] – 3H has shorter t1/2; isotope effect on C-H
cleavage; loss of 3H as 3H2O if C-H cleavage occurs
 Only a trace amount of radioactivity is used (maybe 1 in 106 molecules); the
remainder of the molecules is nonradiolabeled.
Metabolism of erythromycin

If the NMe2 group is labeled with 14C, the

[14C]-CO2 can be measured.
If the drug is a natural product, a biosynthetic
approach to radioactive incorporation is best
If the drug is not a natural product, a
chemical synthesis is needed.
 The radioactive drug is used in metabolism and
bioavailability studies in rats, mice, or guinea pigs, then in
dogs and/or monkeys.
 If >95% of the radioactivity is found in urine and feces,
and is nontoxic, it can be administered to humans.
 Phase I clinical trials on healthy volunteers – radiolabeled
drug administered to humans for human metabolism
studies.
Advances that Made Metabolism
Studies Less Difficult
 More commercially-available radioactive compounds
 High performance liquid chromatography (HPLC); new
column packings; capillary GC; capillary electrophoresis
 New mass spectrometric methods – tandem mass
spectrometry/mass spectrometry; GC/mass spectrometry;
*HPLC/electrospray mass spectrometry
 New nuclear magnetic resonance (NMR) techniques
*HPLC/NMR
*HPLC/NMR/MS
Principal Steps in Drug Metabolism
Studies
1. Isolation (often, this step can be omitted) – extractions,
ion exchange
2. Separations – HPLC, GC
3. Identification – mass spectrometry (MS), NMR
4. Quantification – radioactive labeling, GC, HPLC
LC/MS/MS is a rapid method in which a sample is injected
into the HPLC, then each peak is run into an electrospray
ionization MS for parent ion data, then the parent ion is run
into a second MS for fragmentation data.
Pathways for Drug Deactivation and
Elimination
 Rate and pathway of drug metabolism are affected by species, strain, sex,
age, hormones, pregnancy, and liver diseases.
 Drug metabolism is stereoselective, if not stereospecific.
 Generally, enantiomers act as two different xenobiotics – different
metabolites and pharmacokinetics.
 Sometimes the inactive enantiomer produces toxic metabolites or may inhibit
metabolism of active isomer.
 Metabolism of enantiomers may depend on the route of administration.
 For example, the anti-arrhythmia drug verapamil is 16 times more potent
when administered i.v. than orally.
Verapamil is 16 times more active IV
than orally
 The more active (-) isomer is metabolized faster than the
(+) isomer by the liver
 Inactive (R)-isomer is metabolized to active (S)-isomer
 No need to use a single enantiomer
One enantiomer can be metabolized to the other.
Drug metabolism reactions – two
categories
 Phase I transformations – introduce or unmask a
functional group, e.g., by oxygenation or hydrolysis
 Phase II transformations – generate highly polar
derivatives (called conjugates) for excretion
Phase I Transformations Oxidative
Reactions
Late 1940s, early 1950s
 Metabolism of 4-dimethylaminoazobenzene shown to
require O2 and a reducing system (NADPH). Called a mixed
function oxidase.
 One atom of O from O2 is incorporated into product; a
heme protein is involved.
 Cytochrome P450 – family of heme enzymes that catalyzes
the same reaction on different substrates (isozymes)
Drug-Drug Interactions

 Changes in the pharmacokinetics and metabolism of drugs

when multiple drugs are taken together.
 One drug may inhibit a cytochrome P450, blocking
metabolism of another drug.
 One drug may induce a cytochrome P450, which increases
metabolism of other drugs.
Hyperforin is found in St. John’s Wort

 Active constituent of St. John’s wort (hyperforin, 8.11)

activates the pregnane X receptor, which regulates P450
3A4 transcription, resulting in more active drug
metabolism
Reactions Catalyzed by Cytochrome
P450
Site of Reactions Catalyzed by P450

 Part of molecule undergoing reaction is determined by:

1. topography of the active site of the isozyme
2. degree of steric hindrance of the heme iron-oxo species to
the site of reaction
3. ease of H atom abstraction or electron transfer from the
compound
CYP450 activity is variable in the
population
 CYP450 is found in liver, kidney and lungs.
 There are a number of different P450 families, which differ in
their substrate and reaction specificity.
 57 human genes for P450 have been indentified. Individuals
also vary in the properties of their P450s.
 CYP450 2C9 and 2D6 are responsible for metabolism of about
half of all drugs.
 Variations in P450s are racially and ethnically distributed.
 Pharmacogenomics—how the genetic characteristics of a
person influences their response to drugs.
Individual variation in CYP450 2C9

 CYP450 2C9 metabolizes phenytoin, S-warfarin,

tolbutamide, losartan, and many nonsteroidal
antiinflammatory agents (NSAIDs).
 At least 33 alleles of CYP450 2C9 have been discovered.
 Most of the mutant alleles of CYP450 2C9 have low or no
enzymatic activity.
CYP450 2C9 and tolbutamide
metabolism
 Tolbutamide is a sulfonylurea antidiabetes drug.
 CYP450 2D9 hydroxylates the aromatic methyl to give a
much lower activity metabolite.
 Individuals with mutant CYP450 2C9 alleles have higher
concentrations of tolbutamide in the blood, longer
duration of action, and lower blood glucose, so they are
more likely to get hypoglycemia.
CYP450 2C9 and warfarin metabolism
 Warfarin is an anticoagulant drug which inhibits vitamin K
2,3-epoxide reductase.
 (S)-Warfarin is hydroxylated at C-6 and C-7 by CYP450 2C9
to give inactive metabolites.
 Mutant alleles of CYP450 2C9 have less activity for
hydroxylation of warfarin, so patients with mutant alleles
need to have lower doses.
 The therapeutic index for warfarin is small even for wild-
type patients.
Individual variation in CYP450 2D6

 P450 2D6 metabolizes opiates, antiarrhytmics, tamoxifen

and b-blockers, among others.
 More than 60 alleles of 2D6 have been discovered. Some of
the alleles of 2D6 have low or no enzymatic activity (PM).
 Some of the alleles of 2D6 have intermediate activity (IM).
 Some of the alleles of 2D6 have somewhat higher activity
(EM).
 Some of the alleles of 2D6 have much higher activity than
wild-type (UM).
CYP450 2D6 and opiate metabolism

 Codeine is O-demethylated to morphine, the active metabolite

in analgesia.
 PMs can’t convert codeine to morphine, so don’t get analgesia.
 UMs convert codeine to morphine very rapidly, so may
experience toxicity.
 Infants have been poisoned by breast milk from UM mothers
taking codeine.
CYP450 2D6 and tamoxifen metabolism

 Tamixofen is an antiestrogen used to treat breast cancer.

 The metabolite, 4-hydroxytamoxifen, binds about 100-fold
more strongly to estrogen receptors.
 2D6 PMs respond poorly to tamoxifen treatment.
Reactions of Flavin Monooxygenase
Flavin monooxygenase is often more stereoselective
than CyP450

CyP450 CyP450
Aromatic Hydroxylation
 Intermediate in aromatic hydroxylation
 Jerina, Daly and Witkop
 1968 National Institutes of Health (NIH)

arene oxide isolated

Mechanism for Arene Oxide Formation
and Aromatic Hydroxylation
Reactions of Arene Oxides
 Rearrangement of Arene Oxide to
Arenol Called the NIH shift
 Competing with the NIH Shift

The more stabilized the carbocation intermediate, the less

favored for hydride shift - more deprotonation.
Species Specificity

Major hydroxylation metabolites

in dogs

in humans
 Toxicity from Arene Oxides

Relationship between soot and

cancer noted in 1775 chimney
sweeps frequently developed
skin cancer
Alkene Epoxidation
anticonvulsant

anticonvulsant
Oxidation of Carbons Adjacent to sp2
Centers
Oxygenation next to aromatic sp2 carbon
 Allylic hydroxylation
 Oxidation Next to a Carbonyl Group
 hydroxylation here for (-)-isomer
hydroxylation
here for (+)- hydroxylation here
isomer for (-)-isomer
N-Oxidation-Hydroxylation of Nitrogen

Basic amines (pKa 8-11) are oxidized

by flavoenzymes. Nonbasic
compounds, such as amides, are
oxidized by P450. Compounds of
intermediate basicity, such as
aromatic amides, are oxidized by both.
 Metabolism of 2° Amines and Amides