Cytochrome P450 gene families

Human 14+
Molluscs 1

CYP450
Plants 22

Insects 3 Bacteria 18
Yeasts 2 Nematodes 3
Fungi 11

3
 ENZYMES REACTIONS

Phase 1 "oxygenases"
Table 3–1 Xenobiotic Metabolizing Enzymes
Cytochrome P450s (P450 or CYP) C and O oxidation, dealkylation, others

Flavin-containing monooxygenases (FMO) N, S, and P oxidation

Epoxide hydrolases (mEH, sEH) Hydrolysis of epoxides

Phase 2 "transferases"

Sulfotransferases (SULT) Addition of sulfate

UDP-glucuronosyltransferases (UGT) Addition of glucuronic acid

Glutathione-S-transferases (GST) Addition of glutathione

N-acetyltransferases (NAT) Addition of acetyl group

Methyltransferases (MT) Addition of methyl group

Other enzymes

Alcohol dehydrogenases Reduction of alcohols

Aldehyde dehydrogenases Reduction of aldehydes

NADPH-quinone oxidoreductase (NQO) Reduction of quinones

mEH and sEH are microsomal and soluble epoxide hydrolase.

UDP, uridine diphosphate;
NADPH, reduced nicotinamide adenine dinucleotide phosphate.
 CYP Biotransformations

• Chemically diverse small molecules are

converted, generally to more polar compounds
• Reactions include:
– Aliphatic hydroxylation, aromatic hydroxylation
– Dealkylation (N-,O-, S-)
– N-oxidation, S-oxidation
– Deamination
– Dehalogenation
 Cytochrome P450 Isoforms (CYPs) - An Overview

• NADPH + H+ + O2 + Drug → NADP+ + H2O + Oxidized

Drug
• Carbon monoxide binds to the reduced Fe(II) heme and
absorbs at 450 nm (origin of enzyme family name)
• CYP monooxygenase enzyme family is major catalyst of
drug and endogenous compound oxidations in liver,
kidney, G.I. tract, skin, lungs
• Oxidative reactions require the CYP heme protein, the
reductase, NADPH, phosphatidylcholine and molecular
oxygen
• CYPs are in smooth endoplasmic reticulum in close
association with NADPH-CYP reductase in 10/1 ratio
• The reductase serves as the electron source for the
oxidative reaction cycle
NADP+ Drug
CYP Fe+3
CYP e ­

R­Ase Drug
PC Drug OH
NADPH

CO CYP Fe+3
CO
CYP­Fe+2 CYP Fe+2 Drug OH
Drug hυ
Drug
e­
O2
CYP Fe+2 H2O
O2 Drug
2H+

Electron flow in microsomal drug oxidizing system

Table 1. Human CYP families and their main functions. Data adapted from (Gonzalez 1992,
Nelson et al. 1996, White et al. 1997, Nelson 1999, Lund et al. 1999).

CYP family Main functions

CYP1 Xenobiotic metabolism
CYP2 Xenobiotic metabolismArachidonic acid metabolism
CYP3 Xenobiotic and steroid metabolism
CYP4 Fatty acid hydroxylation
CYP5 Thromboxane synthesis
CYP7 Cholesterol 7α-hydroxylation
CYP8 Prostacyclin synthesis
CYP11 Cholesterol side-chain cleavage Steroid 11β -
hydroxylation Aldosterone synthesis
CYP17 Steroid 17α-hydroxylation
CYP19 Androgen aromatization
CYP21 Steroid 21-hydroxylation
CYP24 Steroid 24-hydroxylation
CYP26 Retinoic acid hydroxylation
CYP27 Steroid 27-hydroxylation
CYP39 Unknown
CYP46 Cholesterol 24-hydroxylation
CYP51 Sterol biosynthesis
 What CYPs important in drug
metabolism
• CYP1A2-chromosome 15
• CYP2C9-chromosome 10**
• CYP2C19-chromosome 10**
• CYP2D6- chromosome 22**
• CYP2E1-chromosome 10
• CYP3A (4/5/7)-chromosome 7
** most genetic information
 Cytochrome P450 enzymes
Especially CYP 3A4, CYP 2D6, and CYP 2C9 are involved in
the metabolism of xenobiotics and drugs.

Metabolic Contribution
hepatic only CYP 1A2 other CYP 3A4
CYP 2C9 2% 3% CYP 2D6
10% CYP 2C9
CYP 1A2
other

CYP 3A4
CYP 2D6 55%
30%

also small intestine

Table 3–3 Indications and Unwanted Side Effects of Drugs Metabolized by N-Acetyltransferases
DRUG INDICATION MAJOR SIDE EFFECTS
Acebutolol Arrhythmias, hypertension Drowsiness, weakness, insomnia
Amantadine Influenza A, parkinsonism Appetite loss, dizziness, headache, nightmares
Aminobenzoic acid Skin disorders, sunscreens Stomach upset, contact sensitization
Aminoglutethimide Adrenal cortex carcinoma, breast cancer Clumsiness, nausea, dizziness, agranulocytosis
Aminosalicylic acid Ulcerative colitis Allergic fever, itching, leukopenia
Amonafide Prostate cancer Myelosuppression
Amrinone Advanced heart failure Thrombocytopenia, arrhythmias
Benzocaine Local anesthesia Dermatitis, itching, rash, methemoglobinemia
Caffeine Neonatal respiratory distress syndrome Dizziness, insomnia, tachycardia
Clonazepam Epilepsy Ataxia, dizziness, slurred speech
Dapsone Dermatitis, leprosy, AIDS-related complex Nausea, vomiting, hyperexcitability, methemoglobinemia, dermatitis
Dipyrone, metamizole Analgesic Agranulocytosis
Hydralazine Hypertension Hypotension, tachycardia, flush, headache
Isoniazid Tuberculosis Peripheral neuritis, hepatotoxicity
Nitrazepam Insomnia Dizziness, somnolence
Phenelzine Depression CNS excitation, insomnia, orthostatic hypotension, hepatotoxicity
Procainamide Ventricular tachyarrhythmia Hypotension, systemic lupus erythematosus
Sulfonamides Antibacterial agents Hypersensitivity, hemolytic anemia, fever, lupus-like syndromes
Factors Influencing Activity and Level of CYP Enzymes 
Nutrition 1A1;1A2;2E1; 3A3; 3A4,5

Smoking 1A1;1A2

Alcohol 2E1
1A1,1A2; 2A6; 2B6; 2C;
Drugs
2D6; 3A3, 3A4,5
1A1,1A2; 2A6; 1B; 2E1;
Environment
3A3, 3A4,5
Genetic 1A; 2A6; 2C9,19; 2D6;
Polymorphism 2E1

Red indicates enzymes important in drug metabolism

S. Rendic & F. J. Di Carlo Drug Metab Rev 29: 413­580, 1997
M. Del Zompo, giugno 2001
 Table 3–4 Nuclear Receptors That Induce Drug Metabolism

RECEPTOR LIGANDS
Aryl hydrocarbon receptor (AHR) Omeprazole
Constitutive androstane receptor (CAR) Phenobarbital
Pregnane X receptor (PXR) Rifampin
Farnesoid X receptor (FXR) Bile acids
Vitamin D receptor Vitamin D
Peroxisome proliferator activated receptor Fibrates
(PPAR)
Retinoic acid receptor (RAR) all-trans-Retinoic acid

Retinoid X receptor (RXR) 9-cis-Retinoic acid

METABOLISMO Principio attivo

Reazioni di fase I
OSSIDAZIONE
RIDUZIONE
IDROLISI

Metaboliti di fase I
-OH
-COOH
-NH2
-SH

Reazioni di fase II
CONIUGAZIONE

Metaboliti coniugati

ELIMINAZIONE
Physiologic Role of P-glycoprotein
Il Nefrone
Struttura dei segmenti tubulari
Tubulo contorto distale

Dotto collettore corticale

Tubulo contorto
prossimale
Capsula glomerulare o di
Bowman

Ansa discendente o di Ansa ascendente o di

Henle (segmento sottile) Henle (segmento sottile e
spesso)
Dotto collettore midollare
Effetti della funzione renale sulla eliminazione urinaria
dei farmaci
 Attivo Composti endogeni
(vitamine, zuccheri, aminoacidi)

Riassorbimento

Passivo Farmaci

Farmaci anionici
(penicillina/probenecid)
Secrezione (meccanismo attivo)
Farmaci coniugati
 ELIMINAZIONE PER VIA RENALE

• I farmaci liposolubili tendono ad essere escreti a concentrazioni

simili a quelle presenti nel plasma. La loro concentrazione dipende
soprattutto dal volume delle urine

2) I farmaci polari tendono ad essere escreti nelle urine a concentrazioni

superiori a quelle presenti nel plasma , quindi la loro escrezione
dipende più dal volume del filtrato glomerulare che dal volume
delle urine

3) I farmaci coniugati si comportano in maniera simile alle sostanze

polari, ma possono essere escreti in misura maggiore perché
soggetti a meccanismi di secrezione attiva

4) I farmaci che si ionizzano facilmente, cioè acidi e basi, vengono

escreti in maniera pH dipendente
 U = Concentrazione del
CLEARANCE (ml/min) = U x V farmaco nell’urina
P V = Volume urina in 1 min.
P = Concentrazione del
farmaco nel plasma

Quantità di plasma che in un minuto viene depurata dalla sostanza

Cl = 0 - Viene completamente riassorbito (glucosio)

Cl = flusso plasmatico renale (PAI)
Per filtrazione glomerulare e per secrezione attiva tutto il
plasma che attraversa i capillari, sia glomerulari che tubulari,
viene depurato
Cl = volume di plasma ultrafiltrato (inulina)
Non si lega alle proteine, non subisce riassorbimento né
secrezione
Cl < volume di plasma ultrafiltrato - Viene in parte riassorbito
Cl > volume di plasma ultrafiltrato - Viene in parte secreto
PRINCIPALI PARAMETRI FARMACOCINETICI
Cmax: concentrazione massima

Tmax: tempo per raggiungere la Cmax

AUC: area sotto la curva

F%: biodisponibilità

T½: tempo necessario perché la concentrazione

plasmatica si riduca della metà

Vd: volume di distribuzione

Cl: clearance (quantità di farmaco eliminata nell’unità

di tempo)