SYAHRIJUITA

Bagian Biokimia FK UNHAS

TIU
memahami reaksi-reaksi kimia untuk merubah zat-zat
asing berbahaya bagi tubuh menjadi zat-zat yang tidak
berbahaya dan mudah diekskresi.
TIK
1. Mengetahui zat-zat asing untuk tubuh yang berasal
dari bahan aditif makanan, polutan dan obat-obatan.
2. Memahami reaksi-reaksi metabolisme xenobiotik
dalam tubuh.
3. Mengetahui peran enzim-enzim metabolisme
xenobiotik dan faktor-faktor yang mempengaruhi
efektivitasnya.
4. Mengetahui efek biologis xenobiotik.
Pendahuluan

Xenobiotik: bahan kimia asing

(Yunani xenos, “orang asing”)
= senyawa yang asing bagi tubuh.

Tubuh terpajan  diproses pada tingkat sel

(metabolisme xenobiotik)
Kepentingan biomedis

* untuk mempelajari cara kita menghadapi

serangan
bahan kimia tersebut.
* merupakan hal mendasar untuk memahami
secara
rasional;
farmakologi
toksikologi
penatalaksanaan kanker
ketagihan obat
1. Zat-zat exogen/asing untuk tubuh (Xenobiotik)
Asalnya dari:
- zat aditif makanan,
- polutan(bifenil poliklorinasi=PCB.)
- obat-obatan (morfin, fenobarbital dll.),
- karsinogen,
- produk petroleum,
- insektisida.
- > 200.000 bahan kimia sintetis di
lingkungan
2. Metabolisme xenobiotik

Lokasi dan enzim yang berperan.

* terutama terjadi di dalam hati, secara:
oksidasi, reduksi, metilasi, hidrolisis, konyugasi.
* dilakukan oleh oxygen-transferring enzymes
(oxygenases), termasuk sit P450.
Enzim-enzim ini mampu mengoksidasi ribuan
komponen
organik bentuk hidrofobik  hidrofilik sehingga
mudah diekskresi.
Ada beberapa xenobiotik diekskresikan tanpa
mengalam perubahan.
Fase-fase metabolisme xenobiotik
Fase 1.
Reaksi hidroksilasi dikatalisis oleh enzim
monooksigenase atau sitokrom P450, mengakhiri
kerja obat, tetapi tidak selalu terjadi.
Sit P450 juga mengkatalisis reaksi: deaminasi,
dehalogenasi, desulfurasi, epoksidasi,
peroksigenasi dan reduksi.
Reaksi hidrolisis dikatalisis oleh esterase dan
reaksi lain yg tidak dikatalisis oleh P450.
 Fase 2.

Senyawa terhidroksilasi & senyawa lain yang di

produksi dalam fase 1 diubah oleh enzim spesifik
menjadi metabolit polar lewat:
- konyugasi dengan asam glukuronat, sulfat, asetat,
glutation atau asam amino
tertentu.
- metilasi
Tujuan kedua fase: untuk meningkatkan kelarutannya
didalam air (polaritas) sehingga memudahkan
ekskresinya.
Sifat sangat hidrofobiknya harus dirubah menjadi
lebih polar agar tidak tertahan lama di jaringan
adiposa.
Fase 1
Pada kasus
1* senyawa secara biologis inaktif menjadi senyawa biologis aktif,
senyawa tsb disebut prodrug atau prokarsinogen.
Mis. Benzol(a)pyrene dari daging panggang berubah menjadi
berpotensi karsinogen oleh enzim-enzim detoksikasi SER.

2* Reaksi tambahan (reaksi hidroksilasi lebih lanjut) mengubah

senyawa aktif menjadi kurang aktif /inaktif sebelum konyugasi

3* Reaksi konjugasi sendiri yang mengubah produk aktif menjadi

bentuK yg kurang/inaktif yg diekskresikan kedalam
urine/empedu.
 Konjugasi sangat jarang meningkatkan aktivitas
biologis suatu xenobiotik

Istilah detoksikasi tidak selalu sesuai, sebab adanya

reaksi metabolisme yang malah meningkatkan
aktivitas biologik dan toksisitasnya.
- Hati mengandung jumlah Sit P450 terbanyak
tetapi juga ditemukan dalam sebagian besar
jaringan.
- Terletak dalam retikulum endoplasmik halus
atau di dalam mitokondria (enzim-enzim
stereogenik)
- Pada beberapa keadaan, produknya bersifat
mutagenik/karsinogenik.
- Sebagian besar memiliki massa molekul 55 kDa.
- Banyak yang bersifat dapat dirangsang.
- Beberapa memperlihatkan polimorfisme, yang
dapat menyebabkan metabolisme obat yang
atipik.
 Table 1. Xenobiotics Metabolized by Cyt. P450

Reaction Examples
Aliphatic hydroxylation Valproic acid, pentobarbital
Aromatic hydroxylation Debrisoquine, acetanilide
Epoxidation Benzene, benzo[α]pyrene
Dealkylation Aminopyrine, phenacetin, 6-
methyl-thiopurine
Oxidative deamination Amphetamine
Nitrogen or sulfur 2-Acetylaminofluorence,
oxidation chlorpromazine
Dehalogenation Halothane
Alcohol oxidation Ethanol
 Type of Enzyme Representative
reaction substrate

Oxidation Cytochrome P450 Toluene

Alcohol dehydrogenase Ethyl alcohol
Flavin containing Dimethylaniline
Monooxygenase

Reduction Ketone reductase Metyrapone

Hydration Epoxide hydrolase Benzo[α]pyrene

-7,8-epoxide
Hydrolisis Esterase Procaine

Conjugation UDP glucuronyl Acetaminophen

transferase
Sulfotransferase β-Napthol
N-acetyltransferase Sulfanilamide
Methyltranseferase Thioracil
Glutathione Acetaminophen
transferase
 Berbagai isoform sit P450 menghidroksilasi
beragam xenobiotik pada fase 1 metabolisme.

Reaksi kimia fase 1.

RH + O2 + NADPH + H+ ROH + H2O + NADP

Cyt P-450 red Cyt P-450 oks

RH + O2 ROH + H2O
RH mewakili xenobiotik dengan keragaman luas.
Satu oksigen memasuki ROH dan satu atom lagi
memasuki molekul air.
Reaksi kimia fase 2.
Reaksi konyugasi ada lima tipe.
A. Glukuronidasi (paling sering terjadi).
Donor glukuronil : UDP-asam glukuronat
Katalisator: glukuronil transferase
(ret.endoplasma,sitosol)
Molekul: 2-asetilaminofluoren(karsinogen),anilin,
asambensoat, meprobamat (tranqulizer),
fenol dan banyak molekul steroid
diekskresikan sebagai glukuronida, yang
dapat
terikat dengan gugus oksigen, nitrogen,
sulfur
substratnya.
B. Sulfasi
Molekul: sebagian alkohol,arilamin dan fenol.
Donor: sulfat
Molekul biologis lain:
- senyawa steroid,
- glikosaminoglikan,
- glikolipid,
- glikoprotein,
- adenosin3’-fosfat-5’-fosfosulfat (PAPS)
disebut sulfat aktif.
C. Konyugasi dengan Glutation.

Glutation = -glutamil-sisteinilglisin (tripeptida) (GSH)

karena gugus sulfhidril pada sistein, merupakan gugus
fungsional.
Xenobiotik elektrofilik potensial beracun (karsinogen
tertentu) (R) akan terkonyugasi ke GSH nukleofilik
Reaksi: R + GSH  R-S-H
glutation S-transferase
(tinggi dalam hati)
Bila R tidak terkonyugasi, bebas terikat kovalen
dengan DNA, RNA atau protein sel  rusak.
GSH: mekanisme pertahanan penting terhadap toksik
obat dan karsinogen.
 Konyugat glutation dimetabolisme lanjut.

Gugus glutamil, glisinil milik glutation akan

dikeluarkan oleh enzim spesifik dan gugus asetil
(asal asetil-KoA) ditambahkan kepada gugus
aminosisteinil lainnya dan dihasilkan asam
merkapturat, yaitu konyugat L-asetil-sistein, yang
kemudian diekskresikan ke dalam urine.
D. Asetilasi

Reaksi: X + Asetil-KoA  Asetil-X + KoA

asetil transferase (sitosol)

Donor asetil: Asetil-KoA

Substrat asetilasi: Isoniazid (R/ tbc)
Tipe polimorfik enzim: ada orang disebut asetilator

lambat dan cepat.

Asetilator lambat: orang lebih sering alami efek
toksik tertentu dari isoniazid, karena obat
tersebut bertahan lama pada orang tersebut.
E. Metilasi

Sejumlah kecil xenobiotik akan mengalami

metilasi oleh
enzim metiltransferase, dengan memakai
S-adenosilmetionin sebagai donor metil.
3. Faktor-faktor yang memengaruhi aktivitas enzim yang
memetabolisme xenobiotik:
Genetik: ada perbedaan aktivitas enzim antar individu.

Usia dan jenis kelamin.

Asupan xenobiotik (fenobarbital, PCB, seny.hidrokarbon

tertentu) dapat menginduksi enzim.
Penting diketahui apakah seseorang sudah pernah
terpajanzat penginduksi ini.

Metabolit xenobiotik tertentu dapat menghambat /

mengstimulasi aktivitas enzim, hal ini dapat
memengaruhitakaran sejumlah obat tertentu yang
diberikan pada pasien.

Berbagai penyakit (sirosis hepatis) dapat memengaruhi

aktivitas enzim sehingga perlu penyesuaian dosis
pelbagai obat yang diberikan kepada orang tersebut.
4. Efek biologik xenobiotik
mencakup:

1). Efek farmakologik,

Respon farmakologik, khususnya
farmakogenetika:
beberapa reaksi penting pada obat yang
mencerminkan perbedaan yang ditentukan
secara genetik pada struktur enzim atau protein
antar individu
Enzim atau Protein yang terkena Reaksi atau Akibat

Glukosa-6-fosfat dehidrogenase Anemia hemolitik akibat

(G6PD) (mutasi) (MIM 305900) konsumsi obat seperti primaquin

Saluran pelepasan Ca2+ (reseptor Hipertemia maligna (MIM 145600)

rianodin) dalam retikulum sesudah pemberian obat anestesi
sarkoplasma (MIM 180901) tertentu (misal, halotan)
CYP2D6 (Polimorfisme) (MIM Metabolisme lambat obat-obat
124300) tertentu (misal, debrisouquin),
yang menyebabkan akumulasi
obat tersebut.

CYP2A6 (Polimorfisme) (MIM Gangguan metabolisme nikotin

122720) yeng memberikan perlindungan
agar orang tidak menjadi
perokok yang tergantung pada
tembakau
2. Efek toksik xenobiotik, tiga tipe:

a. Cedera sel (sitotoksisitas)--- kematian sel.

Xenobiotik reaktif melalui pengikatan kovalen
dengan makromolekul sel (target DNA, RNA dan
protein).
Terlibat dalam fungsi selular sel (protein atau
enzim fosforilasi oksidatif, pengaturan
permeabilitas membran)
menyangkut kelangsungan hidup jangka
pendek sel akan terlihat nyata parahnya fungsi sel.
b. Efek imunologis
Xenobiotik reaktif terikat dengan protein,
memodifikasi dan mengubah sifat antigenesitas
spesies tersebut.
Xenobiotik ini bekerja sebagai hapten, yaitu
molekul kecil yang tidak dengan sendirinya
merangsang sintesis antibodi tetapi akan
bergabung dengan antibodi begitu unsur ini
terbentuk. Antibodi ini merusak sel melalui
mekanisme imunologi, yang mengganggu proses
biokimia seluler.
c. Efek karsinogenik
Reaksi antara spesies karsinogen kimiawi yang
aktif dengan DNA bermakna penting dalam
peristiwa karsinogenesis kimiawi.
Zat kimia lain (zat alkilasi) dapat bereaksi
langsung dengan DNA (karsinogen direk), tanpa
mengalami aktivasi kimiawi intra sel
Produk enzim monooksigenase tertentu pada
sebagian substrat prokarsinogen adalah epoksida
sangat reaktif dan mutagenik oleh enzim epoksida
hidroksilase (ret.endoplasma) akan mengubah
menjadi dihidroliol yang jauh kurang reaktif.
 GSH S-transfrase/
epoksida hidrolase
Sit P450
Zenobiotik Metabolik reaktif Metabolit nontoksik

Pengikatan kovalen
pada makromolekul

Cedera sel Hapten Mutasi

Produksi antibodi Kanker

Cedera sel
Gambar 1. Efek Metabolisme xenobiotik
Kesimpulan.

Telah dibahas:
1. Arti dan macam-macam xenobiotik
2. Reaksi-reaksi dalam metabolisme xenobiotik
untuk kepentingan biomedis, termasuk faktor-
faktor yang mempengaruhi aktivitas enzim dan
efek biologik xenobiotik.
Introduction.
Oxygen:- essensial for the life of aerobic organism
O2 usage is commited to;
- 90% Oxidative phosphorylation.
- 5-10% Enzymes for hydroxylation,
oxygenation.
- toxic; when O2 accepts single electrons,
- 3-5% transformed into highly reactive
oxygen radicals (ROS) that damage cellular
lipids, proteins, and DNA, contribute to
cellular death and degeneration in a wide
range of diseases.
  Cerebrovascular disorder
 Atherogenesis  Ischemia/reperfusion injury
 Emphysema bronchitis  Neurodegenerative disorder
 Duchene-type muscular dystrophy Amyotropic lateral sclerosis
 Pregnancy/pre-eclampsia (Lou Gehrig’s disease)
 Retrolental fibroplasia
 Cervical cancer
 Alzheimer’s disease
 Down syndrome
 Alcohol-induced liver disease  Ischemia-reperfusion injury
 Hemodyalisis following stroke
 OXPHOS diseases
 Diabetes (mitochondrial DNA disorders)
 Acute renal failure  Multiple sclerosis
 Aging
 Parkinson’s disease
Radicals
are compounds that contain a single electron,
usually in an outside orbital.
Oxygen is a biradical, a molecule that has two
unpaired electrons in separate orbitals.
Through a number of enzymatic and
nonenzymatic processes that routinely occur in
cells, O2 accepts single electrons to form
reactive oxygen species (ROS)
ROS
-are highly reactive oxygen radicals or
compounds that are readily converted in cells to
these reactive radicals.
-are important in metabolism some-enzyme
use H2O2 as a substrate.
-may be generated nonenzymatically or
enzymatically as accidental by products or
major products of reactions.
-The ROS formed by reduction of O2 are
*the radical superoxide
*the non radical hydrogen peroksida
*the hydroxyl radical
 Superoxide.
 -may be generated nonenzymatically from CoQ,

or from metal containing enzymes (e.g.,

cyt.P450, xanthine oxidase, and NADPH
oxidase).
 -The highly toxic hydroxyl radical is formed

nonenzymatically from superoxide in the

presence of Fe2+ or Cu+ by the Fenton reaction,
and from hydrogen peroxide in the Haber-Weiss
reaction.
 Oxygen radicals and their derivatives can be
deadly to cells.
 The hydroxyl radicals causes oxidative damage

to proteins, and DNA. It also forms lipid

peroxides and malondialdehyde from
membran lipids containing polyunsaturated
fatty acids (PUFA).
 -also a source of chronic damage to tissue
biomolecules.
 In some cases, free-radical damage is the

direct cause of a disease state (e.g., tissue

damage initiated by exposure to ionizing
radiation).
 In neurodegenerative diseases, such as

Parkinson’s disease, or in ishemia reperfusion

injury, ROS may perpetuate the cellular
damage caused by another process.
 Oxygen radicals are joined in their destructive
damage by the free radical nitric oxide (NO) and
the reactive oxygen species hypochlorous acid
(HOCl).
 NO combines with O2 or superoxide to form

reactive nitrogen-oxygen species (RNOS), such

as the non radical peroxynitrite or the radical
nitrogen dioxide. RNOS are present in the
environment (e.g., cigarette smoke) and
generated in cells.
 -One of the risks of harnessing O2 as a
substrate for energy metabolism is that the
way, and do, get burned. For these reason we
have a range of antioxidant defenses that
protect us against ROS.
 Cells protect themselves against damage by

ROS and RNOS (other radicals) through repair

processes, compartmentalization of free-
radical production, defense enzymes, and
endogenous and exogenous antioxidants
(free radical scavengers).
 Source and charasteristic of ReactiveOxygen
Species (ROS).
 O2ˉ Superoxide anion.
Produced by the ETC and at other sites.
Cannot diffuse far from the site of origin.
Generates other ROS.
 H2O2 Hydrogen peroxide.
Not a free radical, but can generate free radicals
by reaction with a transition metal (e.g., Fe2+).
Can diffuse into and through cell membranes.
 OH· Hydroxyl radical.
The most reactive species in attacking biologic
molecules.
Produced from H2O2 in the Fenton reaction in the
presence of Fe2+ or Cu+.
RO· , R· , R-S Organic radicals.
Organic free radicals (R denotes the remainder
of the compound).
Produced from ROH, RH (e.e., at the carbon of a
double bond in a fatty acid), or RSH OH· attack.
. RCOO· Peroxyl radical.
An organic peroxyl radical, such as occurs during
lipid degradation (also denoted LOO·)
. HOCl Hypochlorous acid.
Produced in neutrophils during the respiratory
burst to destroy invading organisms.
Toxicity is through halogenation and oxygenation
reactions. Attacking species is OCLˉ .
 O2ˆ ˇ Singlet oxygen.
 Oxygen with anti parallel spins.
 Produced at high oxygen tensions from

absorption of UV light.
 Decays so fast that it is probably not a

significant in vivo source of toxicity.

Source and characteristic of Reactive
Nitrogen-Oxygen Species (RNOS).
. NO Nitric oxide.
RNOS. A free radical produced endogenously
by nitric oxide synthase. Binds to metal ions.
Combines with O2 or other oxygen-
containing radicals to produce additional
RNOS.
. ONOOˉ Peroxynitrite.
RNOS. A strong oxidizing agent that is not a
free radical. It can generate NO2 (nitrogen
dioxide), which is a radical.
 Major sources of Primary ROS in the cell.
 -are constantly being formed in the cell;
approximately 3 to 5% of the oxygen we consume
is converted to oxygen free radicals.
 Some are produced as accidental by-products of
normal enzymatic reactions that escape from the
active site of metal-containing enzymes during
oxidation reactions.
 Others, such as hydrogen peroxide, are
physiologic products of oxidases in peroxisomes.
 Deliberate production of toxic free radicals
occurs during the inflammatory response.
 Natural radiation (UV), air pollutants (O3, NO2),
drugs, and other chemicals (Xenobiotics) also can
increase formation of free radicals in cells.
 Antioxidant Scavenging Enzymes
 The enzymatic defense against ROS includes SOD,
catalase, and glutathione peroxidase.
 SOD
 Conversion of superoxide anion to H2O2 and O2
(dismutation) by SOD is often called the primary
defense against oxidative stress because
superoxide is such a strong initiator of chain
reaction. SOD exists as three isoenzyme forms, a
Cu+ -Zn2+ form present in the cytosol, a Mn2+
form present in mitochondria, and a Cu+ -Zn2+
form found extracellularly. The activity of Cu+
-Zn2+ sOD is increased by chemicals or
conditions (such as hyperbaric oxygen) that
increase the production of superoxide.
 Fig 24-14. SOD converts superoxide to H2O2
 Non enzymatic Antioxidants (Free-Radical
Scavengers)
 Free-radical scavengers convert free radicals to a
nonradical, nontoxic form in nonenzymatic
reactions. Most free-radical scavengers are
antioxidants, compounds that neutralize free
radicals by donating a hydrogen atom (with its
one electron) to the radical. Antioxidants,
therefore,, reduce free radicals and are
themselves oxidized in the reaction. Dietary free-
radical scavengers (e.g., vit E, vit C, carotenoids,
and flavonoids), as well as endogenously
produced free-radical scavengers (e.g., urate and
melatonin), have a common structural feature, a
conjugated double bond system that may be an
aromatic ring.
 Vitamin E (α-tocopherol)
 Vit E, the most widely distributed antioxidant

in nature, is a lipid-soluble antioxidant

vitamin that functions principally to protect
against lipid peroxidation in membranes. Vit E
comprises a number of tocopherols that differ
in their methylation pattern. Among these, α-
tocopherol is the most potent antioxidant and
is present in the largest amounts in our diet.
 Vitamin C (Ascorbic Acid)
 Although vit C is an oxidation-reduction
coenzyme that functions in collagen synthesis
and other reactions, it also plays a role in
free-radical defense. Reduced ascorbate can
regenerate the reduced form of vit E by
donating electrons in a redox cycle. It is
water-soluble and circulates unbound in
blood and extracellular fluid, where it has
access to the lipid-soluble vit E present in
membranes and lipoprotein particles.
 In addition, it has a number of non enzymic
effects as a result of its action as a reducing
agent and oxygen radical quencer.
 Carotenoids
 Carotenoids is a term applied to β-carotene (the
precursor of vitamin A) and similar compounds with
functional oxygen-containing substituents on the
rings, such as zeaxanthin and lutein. These
compounds can exert antioxidant effects, as well as
quench singlet O2 (singlet oxygen is a highly
reactive oxygen species in which there are no
unpaired electrons in the outer orbitals, but there is
one orbital that is completely empty).Epidemiologic
studies have shown a correlation between diets that
are high in fruits and vegetables and health
benefits, leading to the hypothesis that carotenoids
might slow the progression of cancer,
atherosclerosis, and other degenerative diseases by
acting as chain-breaking antioxidants.
 Fig 24-18
 Other dietary antioxidants
 Flavonoids are a group of structurally similar

compounds that contain two spatiallyseparate

aromatic rings and are found in red wine,
green tea, chocolate, and other plant derived
food.
 Kepustakaan

 Devlin TM, Text Book of Biochemistry (1993), John Wiley &

Sons (USA), pp.996. Cytochromes P450 Oxidize Exogenous
Lipophylic Substrates.
 Karp G, Cell and Molecular Biology. Concepts and
experiments (2008), John Wiley & Sons (Asia), pp.284.
Smooth Reticulum Endothelial.
 Murray RK, Granner DK, Mayes PA, Rodwel VW, Harper’s
Biochemistry (1996), 24 th ed, Prentice Hall
International(USA), pp.750-755. Metabolism of Xenobiotics.
 Price SA, Wilson LM, Pathophysiology. Clinical Concepts of
Diseases Processes (1992). Terjemahan Edisi 4, Buku 1
Patofisiologi.Konsep Klinis Proses-Proses Penyakit(1995),
(Jakarta), hal.429-430. Fungsi Hati.