BIOTRANSFORMATION

By dr. shah murad

shahmurad65@gmail.com

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 Biotransformation or drug
metabolism
Chemic al altera tio n o f th e drug in
the b ody

• Drugs taken should be changed

from their original form to more
excretable form, by biological
processes of human body
• METABOLISM includes,Anabolism
and Catabolism(breakdown of
some compounds and synthesis of 2
• It is needed to render non
polar(lipid soluble) compounds
POLAR(lipid insoluble) so that
they are not reabsorbed in the
renal tubules and are excreted.
• Most hydrophylic drugs like
streptomycin,neostigmine are
not biotransformed and are
excreted unchanged. 3
• Mechanisms which metabolize
drugs(essentially foreign
substances) have developed to
protect the body from ingested
toxins.

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• The primary site for drug
metabolism is liver; others are –
kidney,intestine,lungs and
plasma.
• Metabolism of drugs may lead
to INACTIVATION…ACTIVE
METABOLITE FROM ACTIVE
DRUG or ACTIVATION OF
INACTIVE DRUG 5
DRUG MAY BE:

• In active form
• In inactive form
• In toxic form

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Drug may be:

• As raw material
• May be synthetic form
• May be semi-synthetic form
• Taken as nutrition only
• Taken as essential body
compounds, like VITAMINS

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Why metabolism of drug is
essential???
• All forms of drug should be changed from
one to another form…….because
2. It may require to be changed from
inactive to active form for its
pharmacodynamic action
3. If it is active, it may require to be
changed chemically for its required
therapeutic action
4. If it is in more active(toxic)form,it is
necessary to be changed chemically for
its therapeutic action with less toxic
effects

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Drug is BASICALLY:

• Recognized as Antigen for the

body tissue

So……it should be changed (by

metabolism) TO BE
RECOGNISED BY BODY
TISSUES for ITS THERAPEUTIC
ACTION
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Biotransformation is divided
in 2 PHASES

• PHASE I REACTIONS(non
synthetic reactions)

• PHASE II REACTIONS(synthetic
or conjugation reactions)
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Phase I reaction

• Phase I reactions usually

convert the parent drug to a
more polar metabolite by
introducing or unmasking a
functional group like –OH,
-NH2 ,-SH

• Often these metabolites are

inactive, although in some
cases activity is only modified 11
• If phase I metabolites are
sufficiently polar, they may be
readily excreted.

• However ,many phase I

products are not eliminated
rapidly and undergo PHASE II
reactions 12
Phase II reaction

• Endogenous substrate like

glucuronic acid, sulfuric acid,
acetic acid, or an amino acid
combines with the newly
incorporated functional group to
form a highly polar conjugate.
• These are known as synthetic
reactions.
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• ABSORPTION >> METABOLISM >> ELIMINATION

DRUG >>>>>phase I≈≈≈phase II >> >> >>Elimination

conjugate>>Elimination
/
Drug{drug metabolites with modified activity inactive drug metabolite>>

Drug >> >> >> >> >> >> >> >> >> >> >> Elimination

Lipophobic material….......…Hydrophobic material >> > >> >> >> >>

Elimination

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 Phase II>>>>Phase
I>>>>>Elimination
• In some cases the parent drug may
already possess a functional group
that may form a conjugate directly.For
example HYDRAZIDE part of
ISONIAZID is known to form an N-
acetyl conjugate in a phase II
reaction.This conjugate is then a
substrate for a >>>>>>>phase I
reaction(hydrolysis) to ISONICOTINIC
ACID 15
 Active metabolite
Active drug
Chloral Trichloroethanol
hydrate>> paracetamol
Oxyphenylbutazone
Phenacetin Oxazepam
Digoxin
Phenylbutazone Desipramine
Nortriptyline
Diazepam Morphine
Canrenone
E 3/74
Digitoxin>>
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Prodrug Acive form
Levodopa Dopamine
Enalapril Enalaprilat
Alfa- Alfa-methyl ne
methyldopa Progl triazine
Proguanil Prednisolone
Prednisone Ampicillin
Becampicillin 5-
Sulfasalazine aminosalicylic
Fluorouracil a 17

Fluorouridine
NONSYNTHETIC(phase
I)reactions

• Oxidation

• Reduction

• Hydrolysis

• Cyclization

• Decyclization
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• The most important single group of
reactions is the oxidations,in
particular those undertaken by the
so-called MIXED
FUNCTION(microsomal)OXIDASES
which are capable of metabolising a
wide variety of compounds.
• The most important enzyme is a
haem,cytochrome P450,which take
part in a process whereby molecular
oxygen is bound and incorporated
into the drug molecule,so forming a
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 Oxidation

• It involves addition of oxygen/-ve

charged radical or removal of H/+ve

charged radical.
• Most important reactions for drug
biotransformation>>>>>hydroxylation,
oxygenation at C,N or S atoms,N or O-
dealkylation,oxidative 20
• In many cases the initial insertion of O2

atom into the drug molecule produces

short lived highly reactive

quinone/epoxide/superoxide intermediates

which then convert to more stable

compounds.
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• Oxidative reactions are mostly carried out
by a group of mono-oxygenases in the
LIVER,which in the final step involve a
cytochrome P-450
haemoprotein,NADPH,Cytochrome P-450
reductase and O2.
• More than 100 cytochrome P-450 isoenzymes
differing in their affinity for various
substrates(drugs),have been identified.
• An isoenzyme is one of a group of enzymes
that catalyse the same reaction but differ
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in
protein structure.
Types of CYTOCHROME P-
450(CYP)
• CYP isoenzymes are grouped as
numerals like 1,2,3,4………
• Subfamilies are designed by
capital letters like A,B,C………,
while individual isoenzymes are
again alloted numerals like
1,2,3……like
CYP3A4,CYP3A5,CYP2D6,CYP2
C8,CYP2C19,CYP1A2,CYP2E1…23
• The relative amount of different
cytochrome P-450s differs
among species and among
individuals of the same species.
• These differences largely
account for marked
interspecies and interindividual
differences in rate of
metabolism of drugs. 24
• CYP2C8/9: are important in the
biotransformation of >15 commonly
used drugs including
• phenytoin and
• warfarin which are narrow safety
margin drugs.

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• CYP2C19: metabolizes >12 frequently
used drugs including
OMEPERAZOLE,lansoprazole.

• CYP3A4/5: is responsible for

metabolism of largest
number(>50%)of drugs.in addition to
liver,these isoforms are present in 26
st
• CYP2D6: this is the next most
important CYP isoform which
metabolizes nearly 20% drugs
including tri-cyclic
antidepressants,neuroleptics,antiarrh
ythmics,beta blockers and
opiates.INHIBITION OF THIS ENZYME
BY quinidine RESULTS IN FAILURE OF
CONVERSION OF CODEINE TO
MORPHINE<<<<>>>>>analgesic effect
of codeine is lost.
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• CYP2E1:it catalyses formation
of minor metabolites of few
drugs,notably the hepatotoxic
N-acetyl benzoquinoneimine
from PARACETAMOL….chronic
alcoholism induces this
isoenzyme.It also catalyses a
reaction involved in the
metabolism of 28
alcohol,oestradiol,ethynyloestra
• BARBITURATES,phenothiazines,steroi
ds,benzodiazepines,theophylline and
many other drugs are oxidized in this
way.

• Some other drugs eg;

adrenaline,alcohol,mercaptopurine
are oxidized by mitochondrial or
cytoplasmic enzymes. 29
 Reduction
• Reductive reactions are much less common
than oxidations but some are important.for
example WARFARIN is inactivated by
conversion of a ketone to a hydroxyl group
by CYP2A6.
• This reaction is the conversion of oxidation
and involves CYP enzymes working in the
opposite direction.

• Drugs primarily reduced are

chloralhydrate,chloramphenicol,halothene.
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 Hydrolysis

• This is cleavage of drug molecule by taking

up a molecule of water.
ESTER+H2O →(esterase)→ Acid + Alcohol
Similarly amides and polypeptides are
hydrolysed by amidases and peptidases.
• Hydrolysis occurs in liver,intestine,plasma
and other tissues.EXAMPLES are>>>choline
esters,procaine,lidocaine,procainamide,pet
hidine,oxytocin.
• These reactions do not involve hepatic
microsomal enzymes.
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Cyclization

• This is formation of RING

STRUCTURE,from a straight
chain compound;eg.proguanil.
• Main site of biotransformation is
liver and plasma.

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 Decyclization
• This is opening up of ring structure of
the cyclic drug molecule,eg.
Barbiturates,phenytoin.

• This is generally a minor pathway of

metabolism. 33
 Some Phase I reactions
• OXI DATI VE REACTI ONS:
Hydroxylation(phenytoin,tolbutamide,ph
enobarbital,cortisol,phenylbutazone)Al
kylation(imipramine,codeine,6-
methylthiopurine)Deamination(amphet
amine,diazepam)Desulphuration(parat
hion)Sulphoxidation(chlorpromazine)N
on-microsomal oxidation(ethyl
alcohol)
N-oxidation(acetaminophen,nicotine)5-
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 P-450 independent
OXIDATIONS
• Amine oxidation >>> epinephrine

• Dehydrogenation >>> ethanol,chloral

hydrate 35
• REDU CTIVE REACTI ONS:
Chloramphenicol,clonazepam,dan
trolene,naloxone

• HYD ROLYTIC REACTI ONS:

Enalapril>>>>>enalaprelat
Procaine>>>>>procainamide
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 Non-hepatic sites for drug
metabolism

• Suxamethonium,procaine >> in

PLASMA

• Prostanoids >> in LUNGS

• Tyramine >> in INTESTINE

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DETERMINANTS of
Biotranformation
• The rate of biotransformation of a drug
may vary markedly among different
individuals>>>>this variation is most often
due to genetic or drug-induced
differences.for a few drugs,age or disease-
related differences in drug metabolism are
significant.
• Gender is important for only a few
drugs(eg; ethanol). First-pass metabolism
of alcohol is lower in women than in men.

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• Because the rate of metabolism is
often the primary determinant of
clearance,variation in drug
metabolism must be considered
carefully when designing a dosage
regimen. 39
 Genetic factors which
influence drug metabolism
• Hydrolysis of esters----- succinylcholine is an ester
that is metabolized by plasma cholinesterase
(pseudo-cholinesterase OR butyrylcholinestrase).
• In most individuals ,this process occurs very
rapidly,and a single dose of this neuromuscular
blocking drug has a duration of action of about 5
minutes.
• Approximately 1 person in 2500 has an abnormal
form of this enzyme that metabolizes
succinylcholine and similar esters much more
slowly.In such persons,the neuromuscular
paralysis produced by a single dose of
succinylcholine may last in many hours.
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Acetylation of amines
(SLOW & FAST ACETYLATORS)
• Isiniazid & some other amines such as
hydralazine and procainamide are
metabolized by N-acetylation.Persons
defficient in acetylation capacity,called
”SLOW ACETYLATORS” may have prolonged
or toxic responses to normal doses of these
drugs.
• Slow acetylators constitute about 50% of
white and african-american persons in the
US and a much smaller fraction of asian and
in Eskimos populations.
• The slow acetylation trait is inherited as an
AUTOSOMAL RECESSIVE gene. 41
Phase II reactions
• If a drug molecule has a suitable “handle”
(eg; a hydroxyl,thiol or amino group),either
in the parent molecule or in a product
resulting from phase I metabolism,it is
susceptible to conjugation,ie. Attachment
of a substituent group.
• The resulting conjugate is almost always
pharmacologically inactive and less lipid
soluble than its precursor and is excreted
in urine or bile.

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• The groups most often involved in
conjugate formation are
glucuronyl,sulfate,methyl,acetyl,glycyl and
glutathione.
• Glucuronide formation involves the
formation of a high-energy phosphate
compound,uridine diphosphate (UDP)
glucuronic acid(UDPGA),from which
glucuronic acid is transferred to an
electron-rich atom (N, O,or S) on the
substrate ,forming an amide,ester or thiol
bond.
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• Uridine diphosphate(UDP) glucuronyl
transferase,which catalyses phase 2
reactions,has very broad substrate
specificity embracing many drugs and
other foreign molecules.
• Several important endogenous
substances,such as bilirubin and
adrenal corticosteroids,are also
conjugated by the same system. 44
• Acetylation and methylation
reactions occur with acetyl-CoA
and S-adenosyl
methionine,respectively,acting
as the donor compounds.

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• In the liver,the enzymes of phase II
reaction are located intracellularly
and mostly attached to the smooth
endoplasmic reticulum and are often
called microsomal enzymes >>>>>in
order to reach these enzymes drugs
must cross the hepatocyte membrane.

• Non-ionized drug is therefore resistant

to hepatic degradation(unless there is
a specific active transport 46
• Microsomal enzymes constitute a
mixed function oxidase system and
require NADPH and oxygen.

• It involves an electron—transfer chain

consisting of three
components>>>>They are a flavo-
protein –the NADPH-cytochrome-C(P-
450) reductase, a hemoprotein-the
cytochrome P-450,and a lipid
component-the phosphotidylcholine.
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• After xenobiotics undergo enzymatic
phase I metabolism
reactions>>>>some reactive groups
may still persist and are now
amenable to the process of
conjugation(phase II reaction)
involving the chemical combination of
this reactive group with a molecule
eg,glucuronic acid
,glycine,sulphate,acetate
• Such a conjugate is essentially
pharmacologically inert and less lipid
soluble (more water soluble) as 48
 Examples of phase II
•
reaction >>> acetyl salicylic acid >>> salicylic
GLYCIN E CON JUG ATIO N
acid

• Sulphate conjugation > >> acetaminophen >>>

acetaminophen sulphate

• Ace tylat ing >>> isoniazid >>> acetylated

isoniazid

• Met hylat ion >>> nor-epinephrine >>> nor-

metanephrine

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