title: Evaluation of 89 compounds for identification of substrates for cynomolgus monkey CYP2C76, a new bupropion/nifedipine oxidase.
abstract: Cynomolgus monkeys are widely used in preclinical studies during drug development because of their evolutionary closeness to humans, including their cytochrome P450s (P450s). Most cynomolgus monkey P450s are almost identical (≥90%) to human P450s; however, CYP2C76 has low sequence identity (approximately 80%) to any human CYP2Cs. Although CYP2C76 has no ortholog in humans and is partly responsible for species differences in drug metabolism between cynomolgus monkeys and humans, a broad evaluation of potential substrates for CYP2C76 has not yet been conducted. In this study, a screening of 89 marketed compounds, including human CYP2C and non-CYP2C substrates or inhibitors, was conducted to find potential CYP2C76 substrates. Among the compounds screened, 19 chemicals were identified as substrates for CYP2C76, including substrates for human CYP1A2 (7-ethoxyresorufin), CYP2B6 (bupropion), CYP2D6 (dextromethorphan), and CYP3A4/5 (dextromethorphan and nifedipine), and inhibitors for CYP2B6 (sertraline, clopidogrel, and ticlopidine), CYP2C8 (quercetin), CYP2C19 (ticlopidine and nootkatone), and CYP3A4/5 (troleandomycin). CYP2C76 metabolized a wide variety of the compounds with diverse structures. Among them, bupropion and nifedipine showed high selectivity to CYP2C76. As for nifedipine, CYP2C76 formed methylhydroxylated nifedipine, which was not produced by monkey CYP2C9, CYP2C19, or CYP3A4, as identified by mass spectrometry and estimated by a molecular docking simulation. This unique oxidative metabolite formation of nifedipine could be one of the selective marker reactions of CYP2C76 among the major CYP2Cs and CYP3As tested. These results suggest that monkey CYP2C76 contributes to bupropion hydroxylation and formation of different nifedipine oxidative metabolites as a result of its relatively large substrate cavity.
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Tokyo, Japan (S.H., N.M., M.Shi., S.U., S.I., H.Y.); Pharmacokinetics and Safety Research Department, Kaken Pharmaceutical Co., Ltd., Shizuoka, Japan (S.H., M.Sa.); School of Pharmacy, Hyogo University of Health Sciences, Kobe, Japan (H.F.); Novartis Pharma K.K., Tokyo, Japan (S.I.); and Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Japan (K.I., Y.U.).
authors.forename: Shinya
authors.initials: S
authors.lastname: Hosaka
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Tokyo, Japan (S.H., N.M., M.Shi., S.U., S.I., H.Y.); Pharmacokinetics and Safety Research Department, Kaken Pharmaceutical Co., Ltd., Shizuoka, Japan (S.H., M.Sa.); School of Pharmacy, Hyogo University of Health Sciences, Kobe, Japan (H.F.); Novartis Pharma K.K., Tokyo, Japan (S.I.); and Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Japan (K.I., Y.U.).
authors.forename: Norie
authors.initials: N
authors.lastname: Murayama
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Tokyo, Japan (S.H., N.M., M.Shi., S.U., S.I., H.Y.); Pharmacokinetics and Safety Research Department, Kaken Pharmaceutical Co., Ltd., Shizuoka, Japan (S.H., M.Sa.); School of Pharmacy, Hyogo University of Health Sciences, Kobe, Japan (H.F.); Novartis Pharma K.K., Tokyo, Japan (S.I.); and Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Japan (K.I., Y.U.).
authors.forename: Masahiro
authors.initials: M
authors.lastname: Satsukawa
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Tokyo, Japan (S.H., N.M., M.Shi., S.U., S.I., H.Y.); Pharmacokinetics and Safety Research Department, Kaken Pharmaceutical Co., Ltd., Shizuoka, Japan (S.H., M.Sa.); School of Pharmacy, Hyogo University of Health Sciences, Kobe, Japan (H.F.); Novartis Pharma K.K., Tokyo, Japan (S.I.); and Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Japan (K.I., Y.U.).
authors.forename: Makiko
authors.initials: M
authors.lastname: Shimizu
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Tokyo, Japan (S.H., N.M., M.Shi., S.U., S.I., H.Y.); Pharmacokinetics and Safety Research Department, Kaken Pharmaceutical Co., Ltd., Shizuoka, Japan (S.H., M.Sa.); School of Pharmacy, Hyogo University of Health Sciences, Kobe, Japan (H.F.); Novartis Pharma K.K., Tokyo, Japan (S.I.); and Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Japan (K.I., Y.U.).
authors.forename: Shotaro
authors.initials: S
authors.lastname: Uehara
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Tokyo, Japan (S.H., N.M., M.Shi., S.U., S.I., H.Y.); Pharmacokinetics and Safety Research Department, Kaken Pharmaceutical Co., Ltd., Shizuoka, Japan (S.H., M.Sa.); School of Pharmacy, Hyogo University of Health Sciences, Kobe, Japan (H.F.); Novartis Pharma K.K., Tokyo, Japan (S.I.); and Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Japan (K.I., Y.U.).
authors.forename: Hideki
authors.initials: H
authors.lastname: Fujino
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Tokyo, Japan (S.H., N.M., M.Shi., S.U., S.I., H.Y.); Pharmacokinetics and Safety Research Department, Kaken Pharmaceutical Co., Ltd., Shizuoka, Japan (S.H., M.Sa.); School of Pharmacy, Hyogo University of Health Sciences, Kobe, Japan (H.F.); Novartis Pharma K.K., Tokyo, Japan (S.I.); and Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Japan (K.I., Y.U.).
authors.forename: Kazuhide
authors.initials: K
authors.lastname: Iwasaki
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Tokyo, Japan (S.H., N.M., M.Shi., S.U., S.I., H.Y.); Pharmacokinetics and Safety Research Department, Kaken Pharmaceutical Co., Ltd., Shizuoka, Japan (S.H., M.Sa.); School of Pharmacy, Hyogo University of Health Sciences, Kobe, Japan (H.F.); Novartis Pharma K.K., Tokyo, Japan (S.I.); and Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Japan (K.I., Y.U.).
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Iwano
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Tokyo, Japan (S.H., N.M., M.Shi., S.U., S.I., H.Y.); Pharmacokinetics and Safety Research Department, Kaken Pharmaceutical Co., Ltd., Shizuoka, Japan (S.H., M.Sa.); School of Pharmacy, Hyogo University of Health Sciences, Kobe, Japan (H.F.); Novartis Pharma K.K., Tokyo, Japan (S.I.); and Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Japan (K.I., Y.U.).
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Uno
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Tokyo, Japan (S.H., N.M., M.Shi., S.U., S.I., H.Y.); Pharmacokinetics and Safety Research Department, Kaken Pharmaceutical Co., Ltd., Shizuoka, Japan (S.H., M.Sa.); School of Pharmacy, Hyogo University of Health Sciences, Kobe, Japan (H.F.); Novartis Pharma K.K., Tokyo, Japan (S.I.); and Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Japan (K.I., Y.U.) hyamazak@ac.shoyaku.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yamazaki
mesh_terms: D000818:Animals; D016642:Bupropion / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D006801:Humans; D006900:Hydroxylation / Q000502:physiology; D008252:Macaca fascicularis / Q000378:metabolism*; D062105:Molecular Docking Simulation / Q000379:methods; D009543:Nifedipine / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism*
chemical_list: D016642:Bupropion; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D009543:Nifedipine