Comparison of P450 Enzymes Between Cynomolgus Monkeys and Humans: P450 Identities, Protein Contents, Kinetic Parameters, and Potential for Inhibitory Profiles

Chie      Emoto; Noriaki      Yoda; Yasuhiro      Uno; Kazuhide      Iwasaki; Ken      Umehara; Eiji      Kashiyama; Hiroshi      Yamazaki

Abstract

Cynomolgus monkeys are used to predict human pharmacokinetic and/or toxic profiles in the drug developmental stage. Cynomolgus P450s exhibit a high degree of identity (more than 90%) in both cDNA and amino acid sequences with corresponding human P450s. CYP3A protein predominantly exists in cynomolgus monkey liver microsomes, followed by CYP2A, CYP2C, CYP2B6, CYP2E1, and CYP2D. There are many similarities of metabolic properties in cytochrome P450s between cynomolgus monkeys and humans, but the species differences between cynomolgus monkey and human P450s are clearly present in substrate specificity and inhibitor selectivity. Diclofenac 4’-hydroxylation (DFOH) in monkey liver and intestinal microsomes shows much lower activities compared with those in human liver and intestinal microsomes. Sulfaphenazole strongly inhibits DFOH in human liver microsomes, but does not effectively inhibit DFOH in monkey liver and intestinal microsomes. Cynomolgus CYP2C19 exhibits higher activity for DFOH than cynomolgus CYP2C9 although this reaction is a marker reaction of human CYP2C9. On the other hand, cynomolgus CYP2C76 orthologue is not expressed in humans and shows 70-72% identity in amino acid sequences of human CYP2C subfamilies. Cynomolgus CYP2C76 metabolizes non-CYP2C substrates, 7-ethoxyresorufin (human CYP1A substrate) and bufuralol (human CYP2D6 substrate). In addition, cynomolgus CYP3A4 and CYP3A5 also exhibits wider substrate selectivity toward human CYP2D6 and CYP2E1 substrates. These enzymes may be responsible for species difference in drug metabolism between cynomolgus monkeys and humans. The comparative data presented here can be helpful for designing in vivo metabolic assays using cynomolgus monkeys in terms of substrate specificity and inhibitor selectivity.

Keywords: P450, metabolism, liver, intestine, cynomolgus monkey, human, CYP P450, CYP2A, DFOH, Sulfaphenazole, CYP2C9, QGut, extrahepatic tissues, Orthologous P450s