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Detoxication (detoxification)
Prodrugs
Active Metabolite
Reactive Metabolite
Metabolism
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Xenobiotics
Definition:
Biotransformation of drug is defined as the
conversion from one chemical form to another.
the term is used synonymously with metabolism.
Liver > lungs > Kidney > Intestine > Placenta >
Skin > Brain > Testes > Muscle > Spleen
Non-microsomal
enzymes
Not inducible
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Why Biotransformation?
Most drugs are excreted by the kidneys.
For renal excretion drugs should:
have small molecular mass
be polar in nature
not be fully ionised at body pH
Most drugs are complex and do not have these
properties and thus have to be broken down to
simpler products.
Drugs are lipophilic in nature.
Oxidation
Conjugation
Reduction
Hydrolysis
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Phase I:
A polar functional group is either introduced
or unmasked if already present on the
otherwise lipid soluble Substrate,
E.g. OH, -COOH, -NH2 and SH.
Thus, phase I reactions are called as
functionalization reactions.
Phase I reactions are Non-synthetic in nature.
The majority of Phase I metabolites are
generated by a common hydroxylating
enzyme
03-12-2010
system known as
KLECOP, Nipani
cytochrome P450. 15
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Oxidative reaction:
1) Oxidation of aromatic carbon atoms
B. Carbon-Sulfur system
S- Dealkylation
Desulfuration
S-oxidation
2) Hydrolysis of Amides.
4) Hydrolytic Dehalogination
OH
Arenol (major)
R R R OH
H2O
O
epoxide hydrase
OH
Arene oxide
Arene (highly reactive electrophile) Dihyrdrodiol
GSH
5-epoxide transferase
R
R OH OH
SG OH
Tissue toxicity in instances
when glutathione is depleted. Glutathione conjugate Catechol(min. Pro.)
(min.pro.)
E.g. Epoxides of Bromobenzene and Benzopyrene.
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Oxidation of olefins (C=C bonds):
Oxidation of non-aromatic C=C bonds is
analogous to aromatic hydroxylation. i.e. it
proceeds via formation of epoxides to yield 1,2-
dihydrodiols.
O HO OH
H2O
N
N epoxide hydrase N
CONH2 CONH2 CONH2
SO2NHCONHC4H9 SO2NHCONHC4H9
Corresponding Corresponding
aldehyde carboxylic acid.
Tolbutamide Prmary carbinol
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Oxidation of Allylic carbon Atoms:
Carbon atoms adjacent to Olefinic double bonds (are
allylic carbon atoms) also undergo hydroxylation in a
manner similar to Benzylic Carbons.
E.g. Hydroxylation of Hexobarbital to 3`-hydroxy
Hexobarbital.
Allylic carbon atom
OH
3'
2'
H3C H3C
O O O O
HN N HN N
CH3 CH3
O O
E.g. Diazepam
O
N N
3 OH
N N
IC IC
H CH3 OH CH3
O N N 4' O N N OH
H2N
H2N
H H
Carbonyl
N-Dealkylated
Carbinolamine metabolite
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Intermediate
KLECOP, Nipani 29
A tertiary nitrogen attached to different alkyl groups
undergoes dealkylation by removal of smaller alkyl
group first.
Example:
E.g. Lidocaine
CH3
CH3
O C2H5
O C2H5
N C C N
N C C N H2
H H2
OH C2H5
C2H5
CH3
CH3
N- Hydroxy Lidocaine
Lidocaine
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Oxidation of Carbon-Sulfur Systems:
S-Dealkylation:
The mechanism of S-Dealkylation of
thioethers is analogous to N-dealkylation .IT
proceed via -carbon hydroxylation.
The C-S bond cleavage results in formation of
a thiol and a carbonyl product.
E.g. 6-Methyl mercaptopurine.
SCH3 SCH2OH SH
N N N
N N N
+ HCHO
N N N
N N N
H H H
6-Methyl Hydroxylated
intermediate 6-Mercaptopuri9ne
Mercaptopurine
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Desulfuration:
This reaction also involves cleavage of carbon-sulfur bond
(C=S).
Thiopental
KLECOP, Nipani Pentobarbital 33
S-Oxidation:
Alcohol Aldehyde/ketone
Hemiacetal
Ether
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Oxidation of Alcohol, Carbonyl and
Carboxylic Acid
Aldehyde
alcohol
CH3CHO CH3COOH
CH3CH2OH
dehydrogenase
dehydrogenase
Ethanol Acetaldehyde Aceticacid
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Miscellaneous oxidative reactions:
Oxidative aromatization /dehydrogenation:
COOCH3 COOH
NO2 NO2 CH2OH
CH3
NH N
CH3 CH3
COOCH3 COOCH3
Cl O
oxi. Covalent binding to tissues.
Cl Cl Cl Cl
-HCL
H
Chloroform Phosgene
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Reductive reaction:
Bioreductions are also capable of generating polar
functional group such as hydroxy and amino which can
undergo further biotransformation or conjugation.
Reduction of carbonyls:
Aliphatic aldehydes :
Cl3C-CHOH2O Cl3C-CH2OH
O OH
C 2 H5 C 2 H5
H2 C H CH3 H2 C H CH3
C N C N
H3 C CH3 H3 C CH3
Methadone Methadol
O (CH2)3 N(CH3)2 OH
H2 N N
O2N N
O O
R-C-OR' R-C-OH + R'-OH
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Organic acid esters:
Esters with a large acidic group : E.g. Clofibrate
COOC2H5 COOH
Cl O CH3 Cl O CH3 C 2H5OH
CH3 CH3
+
OH C2 H5 OH C2 H5
O P O O P O HO OH + 2 H3PO4
OH C2 H5 OH C 2 H5
Procanamide PABA
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Secondary amide with aromatic Substituent on N-atom:
E.g. Lidocaine
CH3 CH3
O C 2H5 C 2H5
N C N NH2 + HOOC C N
H H2 C 2H5 H2 C 2H5
CH3 CH3
N N
CONH2 H
Carbamazepine Iminostilbene
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Hydrolytic cleavage of non
aromatic heterocyclics:
Four membered lactams: E.g. Penicillins
O O
H H
C N S CH3 C N
H2 S CH3
H2
N CH3 HO N CH3
O
COOH O
COOH
O
O N O COOH
CH3 NH2
DDT DDE
Synthetic Conjugation
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Phase II
Phase II - combines functional group of compound
with endogenous substance
E.g. Glucuronicacid, Sulfuric acid, Amino Acid, Acetyl.
Products usually very hydrophilic
The final compounds have a larger molecular weight.
UDPG - Dehydrogenase
UDPG +2NAD + H2O UDPGA
+2NADH + 2H+
UDPGA + RXH RX
Glucuronic Acid +UDP
Where,
X = O, COO, NH or S
OH OH
OH UDP OH
OH
OH
UDPGA Benzoic Acid Glucuronide Benzoic acid
Ex.
Chloramphenicol, Morphine, Salicylic Acid, Paracetamol
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Common, minor pathway.
Methyltransferases
-CH3 transfer to O, N, S, C
R NH2 R NH COCH3
CH3COS-
COA NHCOCH3
NH 2
NH 2
N- Acetylated PAS
Paraaminosalicyclic Acid Acetyl Co enzyme
Ex.
Histamine, PAS, PABA
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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
ATP Sulfurylase/Mg++
ATP + SO42- APS + Ppi
APS Phosphokinase/Mg++
APS + ATP PAPS + ADP
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67
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).
X= O,NH
Acetyl Synthetase
RCOOH + ATP RCOAMP + H2O + Ppi
Acyl CoA Transferase
RCOAMP + CoA-SH RCSCoA + AMP
-Glutamyl
transferase
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Glycine Cysteinyl glycinase
N- acetylase
Cyanide Conjugation
Conjugation of cyanide ion involves transfer of sulfur
atom from thiosulfate to the cyanide ion in presence of
enzyme rhodanese to form inactive thiocyanate.
2- - rhodanese - 2-
S2O3 + CN SCN + SO3
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Biotransformation-Conclusion
Change the Xenobiotics to a form that can be
eliminated from the body
Change the Xenobiotics to a less biologically active
form
Bioactivation to more toxic forms can also occur
Synthetic Phase II reactions are carried out by other
enzymes.