1. Immunochemical detection of cytochrome P450 enzymes in liver microsomes of 27 cynomolgus monkeys.
- Author
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Uehara S, Murayama N, Nakanishi Y, Zeldin DC, Yamazaki H, and Uno Y
- Subjects
- Animals, Anticoagulants metabolism, Aryl Hydrocarbon Hydroxylases immunology, Aryl Hydrocarbon Hydroxylases metabolism, Coumarins metabolism, Cytochrome P-450 CYP3A immunology, Cytochrome P-450 CYP3A metabolism, Cytochrome P-450 Enzyme System biosynthesis, Cytochrome P-450 Enzyme System genetics, Cytochrome P-450 Enzyme System immunology, Enzyme Assays, Female, Humans, Hydroxylation, Immunoblotting, Inactivation, Metabolic, Macaca fascicularis, Male, Microsomes, Liver metabolism, Oxazines metabolism, Sensitivity and Specificity, Steroid Hydroxylases immunology, Steroid Hydroxylases metabolism, Cytochrome P-450 Enzyme System metabolism, Microsomes, Liver enzymology
- Abstract
The cynomolgus monkey is widely used as a primate model in preclinical studies because of its evolutionary closeness to humans. Despite their importance in drug metabolism, the content of each cytochrome P450 (P450) enzyme has not been systematically determined in cynomolgus monkey livers. In this study, liver microsomes of 27 cynomolgus monkeys were analyzed by immunoblotting using selective P450 antibodies. The specificity of each antibody was confirmed by analyzing the cross-reactivity against 19 CYP1-3 subfamily enzymes using recombinant proteins. CYP2A, CYP2B6, CYP2C9/19, CYP2C76, CYP2D, CYP2E, CYP3A4, and CYP3A5 were detected in all 27 animals. In contrast, CYP1A, CYP1D, and CYP2J were below detectable levels in all liver samples. The average content of each P450 showed that among the P450s analyzed CYP3A (3A4 and 3A5) was the most abundant (40% of total immunoquantified P450), followed by CYP2A (25%), CYP2C (14%), CYP2B6 (13%), CYP2E1 (11%), and CYP2D (3%). No apparent sex differences were found for any P450. Interanimal variations ranged from 2.6-fold (CYP3A) to 11-fold (CYP2C9/19), and most P450s (CYP2A, CYP2D, CYP2E, CYP3A4, and CYP3A5) varied 3- to 4-fold. To examine the correlations of P450 content with enzyme activities, metabolic assays were performed in 27 cynomolgus monkey livers using 7-ethoxyresorufin, coumarin, pentoxyresorufin, flurbiprofen, bufuralol, dextromethorphan, and midazolam. CYP2D and CYP3A4 contents were significantly correlated with typical reactions of human CYP2D (bufuralol 1'-hydroxylation and dextromethorphan O-deethylation) and CYP3A (midazolam 1'-hydroxylation and 4-hydroxylation). The results presented in this study provide useful information for drug metabolism studies using cynomolgus monkeys.
- Published
- 2011
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