Metabolism

• Mainly by liver enzymes belonging to the

cyctochrome P-450 family
• Almost a dozen variations have been identified in
humans.
– The most important one, CYP3A4, is involved in the
metabolism of approximately half the drugs that
are used today

CYP2C19 next in importance.

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 The CYP- family of enzymes

These enzymes accept compounds with widely

different shapes and sizes of
• Introduce oxygen into the molecule using molecular
oxygen.
• Mechanism

2
 Metabolism
• Introduction of an OH group at a saturated carbon
atom
• Mechanism -1 :
– 1. Abstraction of a hydrogen atom, preferred from a relatively weak C-
H bond, thereby forming a relatively stable carbon centered radical
– 2. Reaction with molecular oxygen to form a hydroperoxy radial
– 3. Abstraction of a hydogen atom from another molecule giving a
hydroperoxide
– 4. Reduction of the hydroperoxide to an alcohol.
.
H .H O HO
C
H . C O2
O O
H -H C H CH2-OH
H C H
H H

carbon centered radical

3
• Introduction of an OH group at a saturated carbon
atom (sp3 hybridized)
• A key part of the active site in the Cytochrome enzymes is the
iron center (Fe+2)
• Reaction with oxygen to form an Fe- hydroperoxy radical
2 3

step 1 Fe + O O Fe

O
O
• Abstraction of a relatively weak C-H bond from the substrate
to form Fe-hydro-peroxide and a carbon centered radical
3 3
+ H R
step 2 Fe Fe +R
O O
O O
H

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 CYP mediated oxidation at sp3 carbon

• Reaction of the carbon centered radical with the iron

peroxide to give an alcohol and Fe(O) species.
3 2 2
step 3
Fe + +R R O + Fe Fe
H
O O
O
H

• Overall
CYP
H R R-O-H
O2

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 Chemistry: C-H bond dissociation energies
• Bond strengths
H
H H Primary C-H bond ~ 98 Kcal/mol
C
H 3C C C H Seconday C-H bond ~ 95 Kcal /mol
H 3C H H
Tertiary C-H bond ~ 91 Kcal mol

C C C
C > C > H
C H H
Tertiary radical > secondary radical > primary radical

• Allylic and benzylic radicals are as or more stable than

tertiary radicals resonance stabilization

H
-H H -H H H
H
H H H
Benzylic radical allylic radical

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 Oxidation at an allylic, or benzylic position is often
preferred. Why?
Possible monohydroxylation products

H H H H H H H OH H OH H H H H H H
C C CYP450 C C C C C
CH 3 CH 3 CH 3 CH 3

C H C H H C H
H H H H H H H OH

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 Demethylation of R2N(CH3) to R2N-H
Demethylation of R-O-CH3 to R-OH

H (+) O
CH 3 H2 H
N P450 C
C H H N
N O H
O2

H (+) H
O
O O H O
O P450 C
CH 3 CH 2 H H
O2

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 Demethylation of R2N(CH3) to R2N-H
Demethylation of R-O-CH3 to R-OH

Why are R2N-CH3 and Ar-O-CH3 demethylations favourable?

H2
. H
.
H
OH
-H N
C H N H
N H

O
. O H
.
C H -H O H
H2 OH
H
H

realtively weak C-H bond resonance stabilized radicals

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 Metabolism: Introduction of OH groups into
aromatic rings
R R
P450

O2 O
H

• Mechanism involves formation of an epoxide

followed by rearrangement
R R R R
H (+)
P-450/ O2
H H

H O H
O O-H
H

• Why a different mechanism that observed in aliphatic

systems?
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 Why a different mechanism?
• Aromatic and alkene C-H bond strength is much
higher than aliphatic C-H bond strength
• in other words,
• aromatic and alkene radicals are much less stable
than alkyl and allyl radicals
• Result:
• abstraction of a hydrogen from an aromatic ring or
attached to a double bond is difficult for the
enzyme.

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 Effect of substituents on aromatic
hydroxylation
• If the aromatic ring acts as a nucleophile
• A substituent on the ring that is electron donating {EDG} will increase its
reactivity
EDG increased reactivity towards electrophiles

Common EDG groups: CH3 and other alkyls

OR

• A substituent that is electron withdrawing {EWG} will decrease its

reactivity
EWG decreased reactivity towards electrophiles

Common EWG groups: Halogens; F, Cl

O ketone, acid, ester

R

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 EWG groups reduce the rate of metabolism
drugs containing a benzene ring.
• In many drugs with aromatic rings have EWG substituents
are introduced in order to slow down their rate of
metabolism and increase their half-life in the body. (Other
effect of F and Cl: increase in lipophilicity [logP])
Celebrex Lipotor
F
O
N
H OH OH O
SO2 NH2
N H
N O
N
F3 C

Ciprofloxacin Valium Prozac

O O
F CO2 H CH3 H
N N O
N N H 3C
H N
N CF3
H

Cl

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 Metabolism at aromatic rings other than
benzene
• Electron poor aromatic rings tend to be metabolically more
stable than a benzene ring
N
Decreasing electron density
N N
Increasing metabolic stability
benzene pyridine pyrimidine

.. .. .. Increasing electron density in the double

S O N bonds
.. .. H Increases the rate of epoxide formation
thiophene furan pyrrole (Increase rate of metabolism
benzene


Explanation .. Electrophile
N N
H H
pyrrole

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 Drug-Drug Interactions
• Many drug- drug interactions are due to
metabolism issures
• Need to be aware of the
– Metabolism (enzyme involved)
– The degree of inhibition and
– Therapeutic Index [TI] of the drug.

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 Therapeutic Index (TI)
• Toxic Dosage for 50% of patients / Therapeutic
Dosage for 50 % of the patients
• Or TD50/ED50
• Ideally a large number: low toxicity relative to
effective dosage
• If less than 5 it gives a narrow window and a small
margin of error for safety
• Digoxin, for coronary heart disease has a rather small
TI
• Acetominiphen has a large TI, a relatively safe drug
• Many cancer drugs have rather small TIs !!! Why?

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 Liver metabolism
• Avoiding liver enzyme metabolism - also
called “first pass metabolism”
• Oral administration: Drugs are absorbed
through the gut, go to the liver;
– Oxidative metabolisms occur mainly in the liver
and gut
– To avoid this have the drug go into the blood
stream and before it can be metabolized by liver
enzymes.

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 Methods of administration
• Oral – most common, reasonable compliance
• Intravenous injection typical of many anti-cancer
drugs, often problems with formulations
• Intramuscular injection –most vaccines, acid
sensitive penillins
• Sublingual - angia drugs –nitoglycerin
• Rapid absorption into the blood, rapid action
(KCN)
• Inhalation- asthma drug –salbutamol “puffer”
• Transdermal- hormones, nicotine patch
• Rectal suppositories –aspirin

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• Examples
– Nitroglycerin = anti-anginal
– Aspirin = analgesic

• Need to adjust dosage –less rapid metabolism

– Avoiding stomach acidity which can destroy acid
sensitive compounds

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