title: CYP2C19 polymorphisms account for inter-individual variability of drug metabolism in cynomolgus macaques.
abstract: CYP2C19 (formerly known as CYP2C75), highly homologous to human CYP2C19, has been identified in cynomolgus and rhesus macaques, non-human primate species widely used in drug metabolism studies. CYP2C19 is predominantly expressed in liver and encodes a functional drug-metabolizing enzyme. Genetic variants in human CYP2C genes account for the inter-individual variability in drug metabolism; however, genetic variants have not been investigated in macaque CYP2C19. In the present study, re-sequencing of CYP2C19 in 78 cynomolgus and 36 rhesus macaques identified 34 non-synonymous variants. Among these, 6 were located in substrate recognition sites, the domains important for protein function. Eighteen and 6 variants were unique to cynomolgus and rhesus macaques, respectively. Four variants were characterized by site-directed mutagenesis and metabolic assays, and 3 variants (p.Phe100Asn, p.Ala103Val, and p.Ile112Leu) showed substantially reduced activity as compared with wild type in flurbiprofen 4'-hydroxylation, omeprazole 5-hydroxylation, and R-/S-warfarin 7-hydroxylation. These variants, co-segregating in the animals analyzed, influenced metabolic activities because the homozygotes and/or heterozygotes showed significantly reduced catalytic activities in liver toward flurbiprofen 4'-hydroxylation and omeprazole 5-hydroxylation as compared with wild type. Kinetic analysis for R-warfarin 7-hydroxylation and docking simulation indicated that CYP2C19 Ala103Val would change the function and conformation of this enzyme. Ala103Val variation diminished homotropic cooperativity of CYP2C19 with R-warfarin yielding low metabolic capacity. These results indicated that the interindividual variability of CYP2C-dependent drug metabolism is at least partly accounted for by CYP2C19 variants in cynomolgus macaques.
authors.affiliation: Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan 642-0017, Wakayama, Japan. Electronic address: uno-yasuhiro@snbl.co.jp.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Uno
authors.affiliation: Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan 642-0017, Wakayama, Japan; Drug Safety Research Center, Shin Nippon Biomedical Laboratories, Ltd., Kagoshima 891-1394, Kagoshima, Japan.
authors.forename: Akinori
authors.initials: A
authors.lastname: Matsushita
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida 194-8543, Tokyo, Japan.
authors.forename: Mitsunori
authors.initials: M
authors.lastname: Shukuya
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida 194-8543, Tokyo, Japan.
authors.forename: Yasuka
authors.initials: Y
authors.lastname: Matsumoto
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida 194-8543, Tokyo, Japan.
authors.forename: Norie
authors.initials: N
authors.lastname: Murayama
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida 194-8543, Tokyo, Japan. Electronic address: hyamazak@ac.shoyaku.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yamazaki
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism*; D000925:Anticoagulants / Q000378:metabolism*; D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics / Q000378:metabolism*; D005480:Flurbiprofen / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D008252:Macaca fascicularis / Q000235:genetics / Q000378:metabolism*; D008253:Macaca mulatta; D008862:Microsomes, Liver / Q000378:metabolism; D008958:Models, Molecular; D009853:Omeprazole / Q000378:metabolism; D011110:Polymorphism, Genetic; D011487:Protein Conformation; D054328:Proton Pump Inhibitors / Q000378:metabolism*; D014859:Warfarin / Q000378:metabolism
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D000925:Anticoagulants; D054328:Proton Pump Inhibitors; D005480:Flurbiprofen; D014859:Warfarin; D001189:Aryl Hydrocarbon Hydroxylases; D009853:Omeprazole
keywords: Cynomolgus monkey; Cytochrome P450; Genetic polymorphisms; Liver