METABOLISME XENOBIOTIK

TOXICANTS
TOXICANTS BIOTRASFORMATION
BIOTRASFORMATION
Introduction

Arifah
Arifah Sri
Sri Wahyuni
Wahyuni
arifah.wahyuni@ums.ac.id
arifah.wahyuni@ums.ac.id
Learning Outcomes
1. Menjelaskan prinsip biotransformasi pada fase
toxicokinetik
2. Memahami biotransformasi pada eliminasi toxicant.
3. Membedakan rekasi Phase I & Phase II pada
biotransformasi.
4. Mengidentifikasi bioactivasi or toxikasi.

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XENOBIOTIK
 Xenobiotik  asal kata : xenos (Yunani)
berarti asing
 Definisi: senyawa asing yang terdapat di
dalam tubuh (karsinogen kimiawi,
insektisida tertentu, obat, dll)

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BIOTRANSFORMASI
 Eliminasi xenobiotik tergantung pada perubahan obat
menjadi metabolit yang larut air melalui biotransformation
 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 goal of biotransformation is to increase the rate of
excretionof xenobiotics or drugs.
 Metabolism - is “changed” so that it can be excreted

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 Biotransformation:
Modification of the chemical structure of
parent compound in organism, catalysed
by enzymes
 Metabolites with different potency
 Most of enzymes bound in hepatocytes
 The liver and the first pass metabolism
 Lipophilic compounds – extensive metabolism
 Polar metabolites – excreted by urine

M. Balíková: The Fate of Poison 6

fundamental concepts in drug
biotransformation
 Xenobiotic yang Lipid solubel  poorly excreted in
the urine, cenderung disimpan dalam fat and/or
circulate sampai diubah (phase I biotransformation)
ke metabolit yang more water soluble atau konjugate
metabolite (phase II biotransformation
 Water soluble drugs are more readily excreted in the
urine. They may be metabolized, but generally not
by the CYP enzyme systems

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Elimination ways:
 Urine – polar compounds
 Feaces – lipophilic compounds
 Lungs – gases, volatiles
 Hair – deposits of basic
compounds above all
 Saliva – hydrophilic/lipophilic
 Sweat – hydrophilic/lipophilic
 Major biotransformation sites
Extrahepatic microsomal
• Liver enzymes
(oxidation, conjugation)
• Other….
– kidneys Hepatic
– muscle microsomal
tissue enzymes
(oxidation,
– intestinal
conjugation)
wall
– lungs Hepatic non-
– skin microsomal enzymes
(acetylation,
– blood sulfation,GSH,
alcohol/aldehyde
dehydrogenase,
hydrolysis, ox/red)
Phase I
– functionalizati
on reactions
Phase II
– conjugation
reactions
 Main ways of biotransformation of drugs
I phase
• Oxydation: diazepam,
pentazocin, sydnocarb,
phenotiazin, phenobarbital,
aspirin, butadion, lidokain,
morphin, codein, ethanol,
rifampicin
• Reduction: hestagens,
metronidazol, nitrazepam,
levomycetin, chlozepid
• Hydrolysis: levomycetin,
novocain, cocain, glycosides,
ditilin, novocainamid, xycain,
fentanyl
II phase
• Conjugation with sulfate:
morphin, paracetamol, isadrin
• Conjugation with glucuronic
acid: teturam, sulfonamides,
levomycetin, morphin
• Conjugation with remains of  -
aminoacids: nicotinic acid,
paracetamol
• Acetylation: sulfonamides,
isoniasid, novocainamid
• Methylation: morphin, unitiol,
ethionamid, noradrenalin
 Phase I biotransformation reactions*

Oxidation (most important).

– Add O, remove H, increase valence.
– Cytochrome P-450, MFO, alcohol dehydrogenase,
– oxidases, others.
Reduction (less important).
– Remove O, add H, decrease valence.
– Reductases.
Hydrolysis.
– Add water.
– Esterases, phosphtases, others

*may result in inactivation, activation, or no change in pharmacological

potency/activity
Metabolic enzymes

1. Microsomal:
1. CYP450 monooxygenases
2. Flavin monooxygenase
2. Non-microsomal
1. Alcohol dehydrogenase
2. Aldehyde dehydrogenase
3. Monoamine and diamine oxidases
3. Both
1. Esterases and Amidases
2. Prostaglandin synthase
3. Peroxidases
 ROLE OF CYP ENZYMES IN HEPATIC
DRUG METABOLISM

RELATIVE HEPATIC CONTENT % DRUGS METABOLIZED

OF CYP ENZYMES BY CYP ENZYMES
CYP 2E1
CYP2D6 7%
2%

CYP 2C19
11%
CYP 2C9
CYP 2C
14%
17% CYP2D6
OTHER 23%
36%
CYP 1A2
CYP 1A2
14%
12%

CYP2E1
CYP 3A4-5 CYP 3A4-5
5%
26% 33%
Participation of the CYP Enzymes in Metabolism of
Some Clinically Important Drugs

CYP Examples of substrates

Enzyme
1A1 Caffeine, Testosterone, R-Warfarin
1A2 Acetaminophen, Caffeine, Phenacetin, R-Warfarin
2A6 17-Estradiol, Testosterone
2B6 Cyclophosphamide, Erythromycin, Testosterone
2C-family Acetaminophen, Tolbutamide (2C9); Hexobarbital, S-
Warfarin (2C9,19); Phenytoin, Testosterone, R- Warfarin,
Zidovudine (2C8,9,19);
2E1 Acetaminophen, Caffeine, Chlorzoxazone, Halothane
2D6 Acetaminophen, Codeine, Debrisoquine
3A4 Acetaminophen, Caffeine, Carbamazepine, Codeine,
Cortisol, Erythromycin, Cyclophosphamide, S- and R-
Warfarin, Phenytoin, Testosterone, Halothane, Zidovudine

Adapted from: S. Rendic Drug Metab Rev 34: 83-448, 2002

 Enzymes of Biotransformation:
Phase II Enzymes
• Conjugation reactions.
• Enzymes (tranferases) + cofactor.
 Enzyme catalyzes.
 Cofactor donates group.
 Glucuronic acid, glutathione, sulfate, acetyl
group, methyl group.
 Tends to increase molecular size and polarity
for excretion.
 Additional Effects on Drug Metabolism

Species Differences

Phenylbutazone t 1/2
Rabbit 3 jam
Rat 6 jam
Guinea Pig 6 jam
Dog 6 jam
Human 3 hari
• Induction
– Two major categories of CYP inducers
• Phenobarbital is prototype of one group - enhances
metabolism of wide variety of substrates by causing
proliferation of SER and CYP in liver cells.
• Polycylic aromatic hydrocarbons are second type of
inducer (ex: benzo[a]pyrene).
– Orphan Nuclear Receptors (PXR, CAR) are
regulators of drug metabolizing gene expression
 Why to care about biotransformation

 Understanding of drug
effects
 Development of a
toxicological method
 Interpretation of
toxicological findings
 Correct and effective
therapy, reduction of
adverse drug effects
M. Balíková: The Fate of Poison 28
 Acetaminophen and p-Aminophenols
COCH3 COCH3
HN HN
NH2

Acetanilide, 1886 OC2H5

OC 2H5
(accidental discovery of
Phenacetin or
antipyretic activity; high toxicity)
acetophenetidin, 1887
75-80% (nephrotoxic,
70-90%
methemoglobinemia)
COCH3
NH2
HN
Metabolic pathway quantified;
(Brodie &Axelrod, 1948)
popular in US since 1955

OH
Acetaminophen, 1893
 Acetominophen Metabolism
COCH3
HN

~60% ~35%
COCH3 OH
HN
COCH3
CYP2E1*
HN
CYP1A2
CYP3A11
O CO2H
O
COCH3
OH N O
HO SO3H
OH *induced by ethanol, isoniazi

Protein adducts, O
Oxidative stress NAPQI
Toxicity N-acetyl-p-benzoquinone imine
 Acetaminophen Protein Adducts
COCH3 COCH3
HN N
CYPs
HS-Protein
OH O
H2N-Protein

COCH3 COCH3 COCH3

Protein S N HN HN

S Protein NH Protein
O OH OH

S.D. Nelson, Drug Metab. Rev. 27: 147-177 (1995)

K.D. Welch et al., Chem Res Toxicol 18:924-33 (2005)
 Acetaminophen toxicity
mechanism
• The American Liver Foundation reports that 35%
of cases of severe liver failure are caused by
acetaminophen poisoning which may require
organ transplantation.
• CAR and PXR modulate acetaminophen toxicity
(2002, 2004)
• Activation of PXR induces CYP3A11 and markedly
enhances acetaminophen toxicity in wild type
mice
• CAR transcription co-activator KO blocks toxicity
(2005)
 Acetaminophen Toxicity

•N-acetyl cysteine is an effective antidote, especially if

administered within 10 h of ingestion [NEJM 319:1557-
1562, 1988]
•N-acetyl cysteine is an effective agent to
block GSH depletion and rescue from liver
damaging toxicity
 NAPQI toxicity linked to PXR activation
G. Guo et al. 2004, Toxicol Sci 82(2):374-80

COCH3 COCH3
HN N
CAR CYP2E1*
toxicity
PXR CYP3A11
OH O
Xenobiotics

COCH3 SH
HN
glu-cys-gly
oxidative stress
GLY mechanism ?
S CYS toxicity
OH GLU
 Contoh Kasus
Pasien 12 th/pria, kulit putih diobati dengan
imipramine 'dosis rendah' untuk masalah perilaku dan
enuresis. Obat diresepkan selama 1 bulan.
Pada tiga minggu Donny mengeluhkan kelelahan,
kelemahan.
Pada akhir bulan pengobatan, pasien merasa kurang
sehat; kemudian dia mandi air hangat; namun setelah
itu malah ambruk dan mati

Pada pemeriksaan otopsi ditemukan Tingkat plasma

Imipramine >> 1 mg / L (terapeutik 0,1-0,3 mg / L).
Pasien juga termasuk genetik abnormal CYP2D6; 3-
10% of whites, autosomal recessive
Graphical representation of what
happened to Px
 Genetic polymorphism
of
drug metabolizing
enzymes
• Abnormal CYP 2D6 is also known as
‘debrisoquine polymorphism’ - such
patients are “slow metabolizers”,
also called “poor metabolizers” of
debrisoquine

• Some ‘drug list’ items metabolized

by 2D6: carvedilol, cevimeline,
chlorpheniramine, efavirenz,
imipramine, metoprolol,
promethazine and timolol)
CYP2D6 deficiency also
affects
metabolism of codeine
Biotransformation of heroin, codeine

M. Balíková: The Fate of 39

Poison