TY - JOUR T1 - Identification of the human cytochrome P450 enzymes involved in the two oxidative steps in the bioactivation of clopidogrel to its pharmacologically active metabolite JF - Drug Metabolism and Disposition JO - Drug Metab Dispos DO - 10.1124/dmd.109.029132 SP - dmd.109.029132 AU - Miho Kazui AU - Yumi Nishiya AU - Tomoko Ishizuka AU - Katsunobu Hagihara AU - Nagy A Farid AU - Osamu Okazaki AU - Toshihiko Ikeda AU - Atsushi Kurihara Y1 - 2009/01/01 UR - http://dmd.aspetjournals.org/content/early/2009/10/07/dmd.109.029132.abstract N2 - The aim of the current study is to identify the human cytochrome P450 (CYP) isoforms involved in the two oxidative steps in the bioactivation of clopidogrel to its pharmacologically active metabolite. In the in vitro experiments using cDNA-expressed human CYP isoforms, clopidogrel was metabolized to 2-oxo-clopidogrel, the immediate precursor of its pharmacologically active metabolite. CYP1A2, CYP2B6 and CYP2C19 catalyzed this reaction. In the same system using 2-oxo-clopidogrel as the substrate, detection of the active metabolite of clopidogrel required the addition of glutathione to the system. CYP2B6, CYP2C9, CYP2C19 and CYP3A4 contributed to the production of the active metabolite. Secondly, the contribution of each CYP involved in both oxidative steps was estimated using enzyme kinetic parameters. The contribution of CYP1A2, CYP2B6 and CYP2C19 to the formation of 2-oxo-clopidogrel was 35.8, 19.4 and 44.9%, respectively. The contribution of CYP2B6, CYP2C9, CYP2C19 and CYP3A4 to the formation of the active metabolite was 32.9, 3.76, 20.6 and 39.8%, respectively. In the inhibition studies with antibodies and selective chemical inhibitors to CYPs, the outcomes obtained by inhibition studies were consistent with the results of CYP contributions in each oxidative step. These studies showed that CYP2C19 contributed substantially to both oxidative steps required in the formation of clopidogrel active metabolite and that CYP3A4 contributed substantially to the second oxidative step. These results help explain the role of genetic polymorphism of CYP2C19 and also the effect of potent CYP3A inhibitors on the pharmacokinetics and pharmacodynamics of clopidogrel in humans, and on clinical outcomes.The American Society for Pharmacology and Experimental Therapeutics ER -