Sistem Enzim

Sitokrom - P450
Sistem Enzim Sitokrom-P450

adalah suatu sistem enzim

(gabungan beberapa enzim atau
enzim dan kofaktor) yang
terdapat pada membran lipofilik
retikulum endoplasmik
/mikrosom halus sel hati
 Sistem Sitokrom P450
 Disebut juga MFOs (mixed Function
Oxidases) karena fungsinya yang banyak
dalam mengkatalisis berbagai reaksi
 Monooksigenase  membutuhkan satu
molekul oksigen per molekul substrat dan
satu atom di antaranya akan terikat dalam
molekul substrat tersebut.
 Berfungsi sebagai sistem transpor elektron
multikomponen yang bertanggung jawab
terhadap metabolisme berbagai substrat
endogen dan eksogen
 Reaksi katalitik CY P450
(Monooksigenase)

RH + O2 + 2 H+ + 2 e- ROH + H2O
 Sistem Sitokrom P450 melibatkan
2 komponen enzim utama
 Sitokrom-P450 yaitu suatu protein hem yang
berperan sebagai akseptor elektron dan
oksidase terminal
 NADPH sitokrom P450 reduktase (suatu
flavoprotein) yang berfungsi sebagai
pembawa elektron dan meloncatkan elektron
dari NADPH ke sitokrom P450. Setiap mol
enzim ini mengandung 1 mol FMN dan 1
mol FAD
 Sistem Sitokrom P450
dalam reaksinya membutuhkan
 Sitokrom P450
 NADPH Sitokrom P 450 reduktase (FMN
& FAD)
 NADPH
 Molekul oksigen
Sitokrom P450
Merupakan suatu
hemoprotein, yaitu
protein yang
mempunyai gugus
hem/logam Fe
yang bisa bertindak
sebagai oksidator
atau reduktor
 CY P450
 Cytochromes P450 enzymes constitute a large
superfamily of haem-thiolate proteins involved
in the metabolism of a wide variety of both
exogenous and endogenous compounds.
 They were first discovered in 1955 in rat liver
microsomes
 They are characterized by an intense
absorption band at 450 nm in the presence of
carbon monoxide.
Kurva Absorpsi Kompleks
CY P450 - CO
 Sitokrom P450

Penentu kecepatan reaksi dari banyak

reaksi oksidasi in vivo  terdapat di
berbagai jaringan
NADPH Sitokrom P450 Reduktase
 Reaksi katalitik Sistem CY P450
(Monooksigenase)

RH + O2 + 2 H+ + 2 e- ROH + H2O
Catalytic
cycle of
CY P450

The intermediate states enclosed in a dashed box

have not been directly observed and are hypothetical
 Catalytic cycle of CY P450
1. Substrate binding: The binding of a substrate to a
P450 causes a lowering of the redox potential by
approximately 100mV, which makes the transfer of
an electron favourable from its redox partner,
NADH or NADPH. This is accompanied by a
change in the spin state of the haem iron at the
active site. It has also been suggested that the
binding of the substrate brings about a
conformational change in the enzyme which
triggers an interaction with the redox component
 Catalytic cycle of CY P450
2. The first reduction: The next stage in the
cycle is the reduction of the ion by an
electron transfered from NAD(P)H via an
electron transfer chain.
3. Oxygen binding: An O2 molecule binds
rapidly to the Fe2+ ion forming Fe2+-O2 .
There is evidence to suggest that this
complex then undergoes a slow conversion
to a more stable complex Fe3+-O2-
 Catalytic cycle of CY P450

4. The second reduction: A second reduction is required

by the stoichiometry of the reaction. This has been
determined to be the rate-determining step of the
reaction. A comparison between the bond energies of
O2, O2- and O22- suggest that the complex Fe3+-O22- is
the most favourable starting point for the next stage of
the reaction to occur. However, evidence from
resonance Raman spectroscopy indicates the presence
of a superoxide (O2- ) complex.
 Catalytic cycle of CY P450
5. O2 cleavage: The O22- reacts with two protons
from the surrounding solvent, breaking the O-
O bond, forming water and leaving an (Fe-O) 3+
complex.
6. Product formation: The Fe-ligated O atom is
transferred to the substrate forming an
hydroxylated form of the substrate.
7. Product release: The product is released from
the active site of the enzyme which returns to
its initial state.
Beberapa contoh reaksi oksidasi
yang dikatalisis CY P450
 Hidroksilasi aromatik: Lignokain,
Asetanilida
 Hidroksilasi alifatik: Pentobarbital,
Tolbutamid
 Epoksidasi: Benzo (a) piren
 Dealkilasi-N: Diazepam, Imipramin
 Dealkilasi-O: Kodein, Fenasetin
 Dealkilasi-S: 6-metil tiopurin, Klorpromazin
Beberapa contoh reaksi oksidasi
yang dikatalisis CY P450
 Oksidasi-S: Klorpromazin
 Oksidasi-N: 3-metilpiridin, 2-
asetilaminofluoren, Trimetilamin
 Oksidasi Fosfotionat: Paration
 Deaminasi oksidatif: Amfetamin
 Dehalogenasi oksidatif: Halotan
Reaksi Reduksi
Yang dikatalisis CY P450
 Reduksi Azo:
Prontosil merah > Sulfanilamida
 Reduksi Nitro:
Kloramfenikol > Arilamina
 Reduksi Karbonil:
 Dehalogenasi reduktif: Reduksi
Halotan dalam keadaan anaerob
 Konversi epoksida kembali menjadi
senyawa asal
 Beberapa reaksi Hidroksilasi
yang dikatalisis CY P450

 Hidroksilasi aromatik: Lignokain,

Asetanilid
 Hidroksilasi alifatik: Pentobarbiton,
Tolbutamid
Isoform CY P450
 Kurang lebih 150 isoform, 18 famili, 43
subfamili
 Pembagian ke dalam kelompok (famili
atau subfamili) didasarkan pada kesamaan
sekuens asam aminonya
– Dalam famili, kesamaan > 40%
– Dalam subfamili, kesamaan > 55%
– Dalam 1 individu enzim, kesamaan > 97%
18 Famili CY P450

1. CYP1 10. CYP19

2. CYP2 11. CYP20
3. CYP3 12. CYP21
4. CYP4 13. CYP24
5. CYP5 14. CYP26
6. CYP7 15. CYP27
7. CYP8 16. CYP39
8. CYP11 17. CYP46
9. CYP17 18. CYP51
 Human CYP450

 At least 12 CY P-450 gene families

have been identified in humans
 3 families are involved in the majority
of the drug biotransformations; these
are CYP1, CYP2 and CYP3
1. CYP1: drug metabolism (3 subfamilies)  CYP1A1, CYP1A2,
CYP1B1
2. CYP2: drug and steroid metabolism (13 subfamilies) 
CYP2A6, CYP2A7, CYP2A13, CYP2B6, CYP2B7, CYP2C8,
CYP2C9, CYP2C11, CYP2C18, CYP2C19, CYP2D6, CYP2E1,
CYP2F1, CYP2G1P, CYP2G2P, CYP2J2, CYP2R1, CYP2S1,
CYP2U1, CYP2W1
3. CYP3: drug metabolism (1 subfamily)  CYP3A4, CYP3A5,
CYP3A7, CYP3A43
4. CYP4: arachidonic acid or fatty acid metabolism (6 subfamili)
 CYP4A11, CYP4A22, CYP4B1, CYP4F2, CYP4F3,
CYP4F8, CYP4F11, CYP4F12, CYP4V2, CYP4X1, CYP4Z1
5. CYP5: thromboxane A2 synthase (1 subfamily)  CYP5A1
6. CYP7: bile acid biosynthesis 7-alpha hydroxylase of steroid
nucleus (2 subfamilies)  CYP7A1, CYP7B1
7. CYP8 (2 subfamilies)  CYP8A1
(prostacyclin synthase), CYP8B1 (bile acid
biosynthesis)
8. CYP11: steroid biosynthesis (2 subfamilies)
 CYP11A1, CYP11B1, CYP11B2
9. CYP17: steroid biosynthesis (1 subfamily)
17-alpha hydroxylase  CYP17A1
10. CYP19: steroid biosynthesis (1 subfamily):
aromatase synthesizes estrogen  CYP19A1
11. CYP20: unknown function (1 subfamily) 
CYP20A1
12. CYP21: steroid biosynthesis (2 subfamilies)
CYP21A2
13. CYP24: vitamin D degradation (1 subfamily, 1
gene) CYP24A1
14. CYP26: retinoic acid hydroxylase (3 subfamilies, 3
genes) CYP26A1, CYP26B1, CYP26C1
15. CYP27: (3 subfamilies, 3 genes) CYP27A1 (bile
acid biosynthesis), CYP27B1 (vitamin D3 1-alpha
hydroxylase, activates vitamin D3), CYP27C1
(unknown function)
16. CYP39: 7-alpha hydroxylation of 24-
hydroxycholesterol (1 subfamily, 1 gene) CYP39A1
17. CYP46: cholesterol 24-hydroxylase (1 subfamily, 1
gene) CYP46A1
18. CYP51: cholesterol biosynthesis (1 subfamily, 1
gene, 3 pseudogenes) CYP51A1 (lanosterol 14-
alpha demethylase)
 Why is this detailed information
important or useful?

1. Information on which human P450 enzyme

metabolizes a drug can help predict or explain
drug interactions.
2. Knowledge of genetic differences in the
activity of specific cytochrome P450 enzymes
can help predict who will be poor or rapid
metabolizers of specific drugs.
 CYP3A
 Enzim-enzim subfamili CYP3A merupakan enzim CYP
yang paling banyak pada manusia: 30% dari CYP hati &
70% dari CYP usus
 CYP3A4 : isoform CYP utama pada hati orang dewasa
dan berperan dalam metabolisme sebagian besar obat
 CYP3A4 & CYP3A3: isoform CYP utama di usus halus,
97% substratnya identik
 CYP3A5 : isoform CYP utama pada lambung
 CYP3A5 is present in only 20%-30% of Caucasians, but
being deficient in CYP3A5 poses no problem because the
CYP3A4 enzyme is available to assume its functions.
Important CYP Isoforms

1. CYP3A4: 50%
2. CYP1A2: carcinogenesis
3. CYP2C9: polymorphism
4. CYP2C19: polymorphism
5. CYP2D6: polymorphism
6. CYP2E1: carcinogenesis toxic effect
 Substrat CYP3A4
Analgesics Acetaminophen, alfentanil HCl,
codeine, dextromethorphan
Antiarrhitmics Disopyramide, lidocaine HCl,
quinidine
Anticonvulsants Carbamazepine, ethosuximide

Antidepressants Citalopram, desipramine,

nefazodone HCl, sertraline HCl,
trazodone
 Substrat CYP3A4

Antifungal Itraconazole, ketoconazole

Antihistamines Loratadine

Benzodiazepines Alprazolam, clonazepam,

midazolam HCl, triazolam
Calcium channel Amlodipine, felodipine,
blockers isradipine, mibefradil,
verapamil HCl
 Substrat CYP3A4
Chemotherapeutics Busulfan, doxorubicin HCl,
etoposide, paclitaxel, tamoxifen
citrate, vinblastine sulfate, vincristine
sulfate
Cholesterol-lowering Atorvastatin calcium, fluvastatin
drugs sodium, lovastatin, pravastatin
sodium, simvastatin
Immunosuppressants Cyclosporine, tacrolimus
Macrolide Antibiotics Clarithromycin, erythromycin,
troleandomycin
 Substrat CYP3A4
Steroids Estradiol, cortisol,
methylprednisolone, prednisone,
testosterone
Others Cisapride, rifampin, R-warfarin
 Inducer CY3A4

 rifampicin, rifabutin, glucocorticoids,

carbamazepine, phenobarbitone, phenytoin,
artemisinin, nevirapine

 Some xenobiotics stimulate CYP3A4 mediated

degradation of other substrates ("positive
cooperativity"), or even themselves.
[flavonoids and some steroids]
 Inhibitor CYP3A4
Antiarrhithmics Amiodarone

Antibiotics/ Clarithromycin, erythromycin,

Antibacterials metronidazole, norfloxacin,
troleandomycin
Antidepressants Fluoxetine, fluvoxamine, mirtazapine,
nefazodone HCl, paroxetine, sertraline
Antifungal Fluconazole, ketoconazole, itraconazole

Others Other Grapefruit juice, quinine

 Inhibitor CYP3A4
 clotrimazole, ketoconazole, itraconazole > fluconazole,
miconazole
 HIV PI's: ritonavir > indinavir ~= saquinavir > nelfinavir;
 Ca-channel blockers; cyclosporine;
 troleandomycin, erythromycin > clarithromycin >> azithromycin
 SSRIs (sertraline, fluoxetine, ..),
 propofol,
 vinblastine, vincristine, ergotamine, progesterone,
dexamethasone, bromocriptine, cimetidine { >>> ranitidine },
 quinidine;
 irreversible inhibition occurs with some 17-alpha ethinyl
substituted steroids (ethinyl-estradiol, ?mifepristone), and
grapefruit juice!
 Substrat CYP1A2
Antidepressants Amitriptyline HCl, clomipramine
HCl, desipramine HCl,
imipramine HCl
Antipsychotics Clozapine, haloperidol

Benzodiazepines Chlordiazepoxide, diazepam

Others Caffeine, propranolol, tacrine

HCl, theophylline, R-warfarin
 Substrat CYP2C9
Sulfonylurea Glibenclamide, glimepiride, glipizide
(antidiabetic) tolbutamide

NSAIDs
Celecoxib, lornoxicam, diclofenac,
(analgesic, ibuprofen, naproxen, piroxicam, meloxicam
antipyretic, anti-
inflammatory)
Antidepressants Amitriptyline, clomipramine, imipramine

Others Diazepam, losartan, omeprazole, phenytoin

(antiepileptic), S-warfarin (anticoagulant),
sildenafil (in erectile dysfunction),
cannabinoids (psychoactive)
 Inhibitor CYP2C9

Antidepressants Fluoxetine,* fluvoxamine,

paroxetine,* sertraline*

Others Amiodarone HCl,

chloramphenicol, fluconazole,
omeprazole
 Substrat CYP2C19
Antidepressants Amitriptyline, citalopram HBr, clomipramine,
imipramine, moclobemide
Antiepileptics nordazepam, diazepam, phenytoin, phenobarbital,
primidone
Proton pump Esomeprazole, lansoprazole, omeprazole,
inhibitors pantoprazole, rabeprazole

Others propranolol, Cyclophosphamide (Alkylating

antineoplastic agent) , glicazide (sulfonylurea),
clopidogrel (antiplatelet drug), diclofenac
(NSAID), indomethacin (NSAID),
propranolol (beta blocker), nelfinavir
(antiretroviral), progesterone (sex hormone)
warfarin (anticoagulant), voriconazole (antifungal)
 Inhibitor CYP2C19

Antidepressants Fluoxetine, fluvoxamine,

paroxetine,* sertraline*

Others Fluconazole, omeprazole

 Substrat CYP2D6
Analgesic Codeine, dextromethorphan, fentanyl,
hydrocodone, meperidine HCl, methadone HCl,
morphine sulfate, oxycodone HCl
Antiarrhitmics Flecainide acetate,* mexiletine, propafenone
HCl*

Antidepressants Fluoxetine HCl, fluvoxamine maleate,

hydroxybupropion,* paroxetine HCl, trazodone
HCl, venlafaxine, tricyclic antidepressants*
Antipsychotics Chlorpromazine HCl,* haloperidol,*
perphenazine,* risperidone,* thioridazine HCl*

Beta blockers Bisoprolol fumarate, labetalol HCl, metoprolol,

pindolol, propranolol, timolol maleate
 Inhibitor CYP2D6
Antiarrhithmics Amiodarone, propafenone HCl,
quinidine
Antidepressants Clomipramine HCl, desipramine
HCl, desmethylcitalopram,*
fluoxetine, S-norfluoxetine,
fluvoxamine,* mirtazapine,*
paroxetine, sertraline, venlafaxine*
Antipsychotics Fluphenazine, haloperidol
Others Cimetidine
 Substrat CYP2E1

 paracetamol (acetaminophen)
 Many volatile anaesthetics: isoflurane,
sevoflurane, enflurane.
 ethanol
 Pentobarbitone
 Tolbutamide
 Propranolol
 rifampicin
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