METABOLISME XENOBIOTIK

TOXICANTS BIOTRASFORMATION
Wongwiwat Tassaneeyakul
Department of Toxicology
Khon Kaen University

Ema Qurnianingsih, dr., M.Si

Modif: Nastiti Wijayanti

 Xenobiotik asal kata : xenos (Yunani)
berarti asing

Definisi : senyawa asing yang terdapat di

dalam tubuh (karsinogen kimiawi,
insektisida tertentu, obat, dll)
ratusan ribu jenis
 Biotransformation
The elimination of xenobiotics often depends on their conversion
to water-soluble chemicals through biotransformation, catalyzed
by multiple enzymes primarily in the liver with contributions from
other tissues.
Biotransformation changes the properties of a xenobiotic usually
from a lipophilic form (that favors absorption) to a hydrophilic form
(favoring excretion in the urine or bile).
The main evolutionary goal of biotransformation is to increase the
rate of excretion of xenobiotics or drugs.
Biotransformation can detoxify or bioactivate xenobiotics to
more toxic forms that can cause tumorigenicity or other toxicity.
NST110, Toxicology
Department of Nutritional Sciences and Toxicology
University of California, Berkeley
 XENOBIOTIC METABOLISM

metabolism is what the body does to the

toxicants ..

Toxicants may converted to

1. Less toxic chemicals (metabolite) , or
2. More toxic chemicals (metabolite) , or
3. Chemicals with different type of
effect or toxicity

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 XENOBIOTIC METABOLISM

Toxicants (xenobiotics) catalyze by enzymes to

form metabolite (s) with modified structure
Several routes of metabolism found in vivo
May inactivate or bioactivate action
Liver is the major site of metabolism
Genetically variation with some enzymes
Not constant - can be changed by other
chemicals; basic of many interactions
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Sites of Metabolism

where ever appropriate enzymes occur;

plasma,
kidney,
lung,
gut wall and LIVER

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The Liver

Hepatocytes (liver cells)

portal smooth
venous endoplasmic bile
blood reticulum Feces
microsomes
contain cytochrome P450
(CYP)
systemic
arterial
blood

venous blood Urine,

Expiration
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 Lipophilic toxicant

Biotransformation

Hydrophilic metabolite

Metabolite excreted
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 (Xenobiotics)

Highly lipophillic
compounds

Lipophillic compounds

Phase I biotransformation
Polar compounds
(oxidation, reduction, hydrolysis)

Hydrophillic cpds

Phase II biotransformation
(conjugation)

(Renal excretion, Biliary excretion)

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Types of Biotransformation Reaction

Any structural change in a molecule

Phase I - creates site for phase II
oxidation (adds O) e.g. microsomes;
reduction ;
hydrolysis (e.g. by plasma esterases)
others
Phase II - couples group to existing (or phase I
formed) conjugation site
glucuronide (with glucuronic acid)
sulphate
others
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 Metabolisme xenobiotik 2 fase :
Fase I : katalisator sitokrom P450
(hidroksilasi)

Fase II : konjugasi larut air ekskresi

 Fase I
Reaksi utama yang terjadi : hidroksilasi
Enzim : monooksigenase sitokrom P450

Reaksi :
RH + O2 + NADPH + H+ R-OH + H2O + NADP
P450(r) P450(o)

RH + O2 R-OH + H2O

Prinsip reaksi :
Mengkatalisa pengikatan satu atom O pada substrat, atom O yg lain direduksi
menjadi H2O
 Fase I
Reaksi lain yang dikatalisa enzim ini :
deaminasi, dehalogenasi, desulfurasi,
epoksidasi, peroksigenasi, reduksi

Reaksi utama : hidroksilasi

Enzim : monooksigenase sitokrom P450
 PHASE 1 reactions
Hydroxylation -CH2CH3 -CH2CH2OH

Oxidation -CH2OH -CHO -COOH

N-dealkylation -N(CH3)2 - NHCH3 + CH3OH

Oxidative deamination -CH2CHCH3 -CHCOCH3 + NH3

|
NH2

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Xenobiotic-Metabolizing Enzymes
(XME)

Phase 1
P450s
Flavin-containing monoxygenases (FMO)
Epoxide hydrolases

Phase 2 Transferases
Sulfotransferases (ST)
UDP-glucuronosyltransferases (UGT)
Gluthione S-transferases (GST)

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Phase I Enzymes

Cytochromes P450
Flavin Containing Monooxygenase
Epoxide Hydrolase
Alcohol /Aldehyde Dehydrogenases
Monoamine Oxidases
Xanthine oxidase

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Cytochromes P450 (CYP)
Most importance enzyme in xenobiotic metabolism
Berperanan pada bermacam xenobiotik (>50 % obat
menurunkan, meningkatkan, tidak berpengaruh
pada aktivitas obat), spesifitas substrat luas
Juga berperanan pada metabolisme senyawa endogen
(mis. Steroid)
Merupakan hemoprotein
Membrane bound enzyme : locate in smooth
endoplasmic reticulum membrane
All require NADPH and O2
Divided to Family : Subfamily : Isoform
CYP1, CYP2, CYP3 : involved in the metabolism of
xenobiotic W. Tassaneeyakul
 Katalisator serbaguna 60 tipe
reaksi
Produk : lebih larut air ekskresi
Dapat ditemukan di berbagai jaringan,
terutama liver (retikulum endoplasasma
halus, mitokondria)
Kadang, dapat menghasilkan produk
karsinogenik
 Memiliki massa : 55 kDa
Dapat diinduksi interaksi obat (next slide)
Dapat dihambat oleh obat tertentu atau
metabolitnya interaksi obat
Memiliki polimorfisme genetik
metabolisme obat atipikal
Bila terjadi kerusakan jaringan (mis.
Sirosis) mempengaruhi aktivitas
metabolisme obat terganggu
Genotyping profil P450 (masa depan)
individualisasi terapi
Inducibility of CYP

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W. Tassaneeyakul
Content of CYP in human liver

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 Cytochrome P450 dependent Mixed
Function Oxidases

DRUG METABOLITE

=DRUG+O

O2
Liver
microsomes
NADPH NADP+

H+ WATER

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Other (non-microsomal) Phase I reactions

Hydrolysis in plasma by esterases (suxamethonium by

cholinesterase)
Alcohol and aldehyde dehydrogenase in liver cytosolic
(ethanol)
Monoamine oxidase in mitochondria (tyramine,
noradrenaline, dopamine, amines)
Xanthine oxidase (6-mercaptopurine, uric acid
production)
Enzymes for particular substrates (tyrosine
hydroxylase, dopa-decarboxylase etc.)

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 Fase II
Terjadi konversi produk fase I (oleh enzim
spesifik) metabolit polar
Proses :
Konjugasi (asam glukoronat, sulfat, asetat,
glutation, asam amino tertentu)
Asetilasi dan Metilasi
Hasil : metabolit yang lebih larut air
ekskresi di urin atau empedu
 5 tipe reaksi fase II :
(1). Glukoronidasi
Enzim : glucuroniltransferase (retikulum
endoplasma dan sitosol)
Substrat : 2-asetilaminofluoren, anilin, asam
benzoat, meprobamat, fenol, steroid

(2). Sulfasi
Donor sulfat : adenosine 3-fosfat-5-fosfosulfat
(PAPS)
Substrat : alkohol, arylamin, fenol
(3). Konjugasi dengan glutation
Glutation : tripeptida (asam glutamat, sistein,
glisin GSH
Substrat : xenobiotik elektrofilik (karsinogen
tertentu)
Reaksi : R + GSH R-S-G
Enzim : glutation S-transferase
Terdapat banyak pada liver (sitosol)
Pasca konjugasi substrat terkonjugasi
mengalami metabolisme lebih lanjut
(pembuangan gugus glisinil dan glutamil dan
penambahan gugus asetil pada gugs sisteinil) -
-. Asam merkapturat ekskresi
 Fungsi lain glutation :
Berperan pada dekomposisi H2O2 (produk reaksi
oleh glutation peroksidase)
Sebagai reduktan intrasesuler
Sebagai pengangkut beberapa asam amino
melintasi membran pada ginjal
(4). Asetilasi
Reaksi : X + asetil KoA asetil-X + KoA
Enzim : asetiltransferase (sitosol pada sel
liver, terutama)
Substrat : isoniazid

(5). Metilasi
Enzim : metiltransferase
Donor metil : S-adenosilmetionin
Factors Affecting Metabolism
Aktifitas enzim yang terlibat pada
metabolisme xenobiotik dipengaruhi
oleh

Age (reduced in aged & children)

Sex (women more sensitive to ethanol)
Species (phenylbutazone 3h rabbit, 6h horse, 8h monkey,
18h mouse, 36h man)
Race (fast and slow isoniazid acetylators, fast = 95% Eskimo,
50% Brits, 13% Finns 13% Egyptians)
Clinical or physiological conditions

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 AGE
Fetus & Neonate :
Lower drug metabolizing capacity compare to adult
Sensitive to drug,
Long duration of action

GREY BABY SYNDROME

(toxicity of chloramphenicol
in newborn baby)
Volmiting
irregular & rapid respiration
Cyanosis

Birth Adult
 SPECIES

Rate of Hexobarbital metabolism

Species SleepingTime (min)
Mice 12
Rat 90
Dog 315

Therefore animal used for toxicity test must has the

drug metabolism process similar to human
 GENETICS
Activity of xenobiotic metabolizing enzymes
can be vary between individual

Population can be divided to RAPID

METABOLIZER and SLOW METABOLIZERS

Variation in toxicant metabolism

Variation of toxicant level in the body

Variation of toxicant response/toxicity

 DISEASES
Liver diseases : reduced drug metabolism