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KATHMANDU UNIVERSITY
DEPARTMENT OF PHARMACY
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within the body.
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transforms absorbed nutrients (food, oxygen,
etc.) into substances required for normal body
f ti
functions.
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INTRODUCTION
| Biotransformation also serves as an important
defense mechanism by converting toxic xenobiotics
and body wastes into less harmful substances and/or
substances that can be excreted from the body.
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| Lipophilic to
toxicants
icants are hard
h d ffor the
th body
b d to
t
eliminate
| Most lipophilic toxicants can be transformed into
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hydrophilic metabolites
| Hydrophilic
y p chemicals are easier for the bodyy to
eliminate.
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INTRODUCTION
| When biotransformation results in metabolites of
lower toxicity, the process is known as
detoxification.
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| In many cases, however, the metabolites are more
t i than
toxic th the
th parentt substance.
bt This
Thi is
i known
k as
bio-activation.
| Occasionally,
Occasionally biotransformation can produce an
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unusually reactive metabolite that may interact
with cellular macromolecules (e.g., DNA).
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concentrations than are usual.
| The body removes xenobiotics by xenobiotic
metabolism. This consists of the deactivation and the
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secretion of xenobiotics, and happens mostly in the
liver.
| Hepatic enzymes are responsible for the metabolism of
xenobiotics by first activating them and then
conjugating the active secondary metabolite followed by
excretion in bile or urine. E.g.hepatic microsomal 5
cytochrome P450.
BIO-CHEMICAL REACTIONS
| Bio-Chemical reactions are continually taking
place in the body……………………..
p y
• They are a normal aspect of life,
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• pparticipating
p g in the buildingg upp of new tissue,,
• tearing down of old tissue,
• conversion of food to energy,
gy,
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• disposal of waste materials,
• and elimination of toxic xenobiotics.
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BIO-CHEMICAL REACTIONS
Enzymes are the catalysts for nearly all
biochemical reactions in the body.y
Without these enzymes, essential
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biotransformation reactions would take place
slowly or not at all, causing major health
problems.
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it is the drug or chemical transforming enzymes
that hold the key to xenobiotic transformation.
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BIOTRANSFORMING ENZYMES
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| When a substrate fits into the enzyme's structure,
an enzyme-substrate complex can be formed.
| This allows the enzyme to react with the substrate
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with
i h the
h result
l that
h two different
diff products
d are
formed.
| If th
the substrate
b t t does
d nott fit into
i t the
th enzyme, no
complex will be formed and thus no reaction can
occur.
occur 8
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BIOTRANSFORMING ENZYMES
SPECIFICITY OF BIOTRANSFORMING
ENZYMES
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CATEGORIZATION OF THE
BIOTRANSFORMATION REACTIONS
| They are usually classified as Phase I and Phase II
reactions.
| Phase I reactions are generally reactions which modify
the chemical by adding a functional structure to "fit"
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into the Phase II enzyme for conjugation (joined
together) with another substance.
| Phase II reactions consist of those enzymatic reactions
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that
h conjugate
j the modified
ifi xenobiotic
i i with
i another
substance.
| The conjugated products are larger molecules than the
substrate and generally polar in nature (water-soluble).
Thus, they can be readily excreted from the body. They
also
l have
h poor ability
bilit to
t cross cell
ll membranes.
b 11
CATEGORIZATION OF THE
BIOTRANSFORMATION REACTIONS
| In some cases, the xenobiotic already has a functional
group that can be conjugated and can be biotransformed
b a Phase
by Ph II reaction
ti without
ith t going
i through
th h a Phase
Ph I
reaction. Examples:-
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| Phenol can be directly conjugated ;
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Phase I – Reactions Nonsynthetic Reactions
•hydroxylations
aromatic, aliphatic, nitrogen
OXIDATION •dealkylations(N-, S-, P)
deaminations
•deaminations
•N-, S-, P- oxidations oxidoreductases
•S-replacements oxidases
•epoxidations monoamine oxidases
mixed function oxidases
•others
th
esterases
•esters
amidases
HYDROLYSIS •amides
peptidases
•hemiacetals,acetal
s, h
hemiketals,
ik t l ketals
k t l lipases
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hydrolases
OXIDATION
| Oxidation is a chemical reaction in which
a substrate loses electrons.
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EXAMPLES OF OXIDIZING REACTIONS.
¾ Alcohol dehydrogenation
¾ Ald h d d
Aldehyde dehydrogenation
h d ti
¾ Alkyl/acyclic hydroxylation
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¾ Aromatic hydroxylation
¾ Deamination
¾ Desulfuration
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¾ N-hydroxylation
¾ N-oxidation
¾ Sulphoxidation
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REDUCTION
| Reduction is a chemical reaction in which the substrate
gains electrons.
| Reductions
R d ti are mostt likely
lik l to
t occur with
ith xenobiotics
bi ti ini
which oxygen content is low.
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| Reductions can occur across nitrogen
nitrogen-nitrogen
nitrogen double
bonds (azo reduction) or on nitro groups (NO2).
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EXAMPLES OF REDUCING REACTIONS.
¾ Azo reduction
¾ Dehalogenation
Disulfide reduction
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¾
¾ Nit o reduction
Nitro ed ction
¾ N-oxide reduction
¾ Sulfoxide reduction
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HYDROLYSIS
| Hydrolysisis a chemical reaction in which the
addition of water splits the toxicant into two
fragments or smaller molecules.
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| Thehydroxyl group (OH-) is incorporated into
one fragment and the hydrogen atom is
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i
incorporated
t d into
i t the
th other.
th
| Larger
L chemicals
h i l suchh as esters,
t amines,
i
hydrazines, and carbamates are generally
biotransformed by hydrolysis.
hydrolysis 18
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HYDROLYSIS
SUMMARY OF
PHASE -I BIOTRANSFORMATION
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biotransformation or converted to an
intermediate metabolite that is ready for Phase
II biotransformation.
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| The intermediates from Phase I transformations
mayy be p
pharmacologically
g y more effective and
in many cases more toxic than the parent
xenobiotic.
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SUMMARY OF
PHASE -I BIOTRANSFORMATION
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| The enzymes responsible are oxido-reductases;
called mixed-function oxidases.
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| Mostprominent and important among these is
the
h cytochrome
h P450 system consists
i off Cyt
C P
450 and Cyt P 450 reductase
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STRUCTURE OF CYTOCHROME P450 SYSTEM
cytochrome P 450
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CYP450 Reaction Sequence
OH
DRUG DRUG
CYP450
Fe3+
CYP450 CYP450
incorporation of
Fe3+ Fe3+ oxygen
DRUG DRUG peroxide
NADPH + H+ dioxygen
etc.
OH e-
CYP450
reductase
CYP450
Fe3+ CYP450
NADPH + H+ Fe2+
2
DRUG
DRUG
O H+
e-
H 2O
CYP450 CYP450 O2
Fe2+ Fe2+
DRUG DRUG
O21- O2
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IMPORTANCE & QUANTITY OF CYP450 SYSTEM
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P450 3A
CYTOCHROME
PHASE -II BIOTRANSFORMATION
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((-NH2),
NH2), and carboxyl ((-COOH).
COOH).
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possess sufficient hydrophilicity to permit
elimination from the body.
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o Glucuronide conjugation
o Sulfate conjugation
Others :
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o Acetylation
o Methylation
y
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GLUCURONIDE CONJUGATION
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| Molecules added directly to the toxicant or its
phase I metabolite is glucuronic acid, a
molecule derived from glucose, a common
carbohydrate (sugar) that is the primary source
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of energy for cells.
| The sites of glucuronidation reactions are
substrates having an oxygen, nitrogen, or sulfur
bond.
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GLUCURONIDE CONJUGATION
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| It is a high-capacity
high capacity pathway for xenobiotic
conjugation.
| It usually decreases toxicity, although there are
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some notable exceptions, for example, the
production of carcinogenic substances.
| They are generally quite hydrophilic and are
excreted by the kidney or bile.
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GLUCURONIDE CONJUGATION
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GLUCURONIDE CONJUGATION
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OH UTP
OH
O OH
PPi
OH COOH
HO H CO2H
O OH UDP
HO
OH
glucose OH
HO
HO O
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H OH
HO
H H
glucuronic acid OH
UDP
O
OH O
H CO2H UDP H CO2H OH
COH HO HO C
+
HO HO
HO O HO O
H OH H OH
O
salicylic acid H H H H 31
a glucuronide
UDP-glucuronide
derivative
SULFATE CONJUGATION
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xenobiotics.
xenobiotics
| The highly polar sulfate conjugates are readily
secreted in the urine.
urine
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| In general, sulfation is a low-capacity pathway
for xenobiotic conjugation.
| Often glucuronidation or sulfation can conjugate
the same xenobiotics.
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SULFATE CONJUGATION
sulfate ester formation
NH2
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HO + N N O
N N O SO
O O
Sulfates are carried as O
O S O PO
phosphoadenosine- O
phosphosulfate derivatives O y
The enzymes catalyzing
y g
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O
(PAPS) - a high energy form. this type of reaction are
HO O called sulfotransferases.
O P O
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BIOTRANSFORMATION SITES
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| Within the liver cell, the primary subcellular
components that contain the transforming
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enzymes are the microsomes (small vesicles) of
the endoplasmic
p reticulum and the soluble
fraction of the cytoplasm (cytosol).
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BIOTRANSFORMATION SITES
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Liver SER
smooth
endoplasmic
reticulum
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MODIFIERS OF BIOTRANSFORMATION
| Age
| Genetic variability in biotransforming capability
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MODIFIERS OF BIOTRANSFORMATION
| Poor nutrition
| Enzyme inhibition and enzyme induction
can be caused by prior or simultaneous exposure
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to xenobiotics. In some situations exposure to a
substance will inhibit the biotransformation
capacity for another chemical due to inhibition of
specific
p enzymes
y
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| Examples of enzyme inducers are alcohol,
isoniazid, polycyclic halogenated aromatic
h d
hydrocarbons
b ((e.g., di
dioxin),
i ) phenobarbital,
h b bit l
and cigarette smoke
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MODIFIERS OF BIOTRANSFORMATION
| Dose level
| At low doses,
doses a xenobiotic may follow a
biotransformation pathway that detoxifies the
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substance.
| However, if the amount of xenobiotic exceeds the
specific enzyme capacity, the biotransformation
pathway is "saturated"
saturated . In that case,
case it is possible that
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the level of parent toxin builds up.
| In other cases, the xenobiotic may enter a different
biotransformation pathway that may result in the
production of a toxic metabolite.
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REFERENCES
| National Library of Medicine; Emily Monosson ; 2008
"Biotransformation”.
| Encyclopedia of Earth; Eds. Cutler J. Cleveland, Washington,
D.C.: Environmental Information Coalition, National Council
bibekmahatsingh@gm
for Science and the
Environment<http://www.eoearth.org/article/Biotransformation
>Diaz E (editor). (2008). Microbial
| Biodegradation:
g Genomics and Molecular Biology
gy ((1st ed.
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ed.). Caister Academic Press. ISBN 978-1-904455-17-
2.http://www.horizonpress.com/biod.
| www. wikipedia,
p , the free encyclopedia
y p
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THANK - YOU