title: Expression and characterization of cynomolgus monkey cytochrome CYP3A4 in a novel human embryonic kidney cell-based mammalian system.
abstract: Cynomolgus monkeys are a commonly used species in preclinical drug discovery, and have high genetic similarity to humans, especially for the drug-metabolizing cytochrome P450s. However, species differences are frequently observed in the metabolism of drugs between cynomolgus monkeys and humans, and delineating these differences requires expressed CYPs. Toward this end, cynomolgus monkey CYP3A4 (c3A4) was cloned and expressed in a novel human embryonic kidney 293-6E cell suspension system. Following the preparation of microsomes, the kinetic profiles of five known human CYP3A4 (h3A4) substrates (midazolam, testosterone, terfenadine, nifedipine, and triazolam) were determined. All five substrates were found to be good substrates of c3A4, although some differences were observed in the Km values. Overall, the data suggest a strong substrate similarity between c3A4 and h3A4. Additionally, c3A4 exhibited no activity against non-h3A4 probe substrates, except for a known human CYP2D6 substrate (bufuralol), which suggests potential metabolism of human cytochrome CYP2D6-substrates by c3A4. Ketoconazole and troleandomycin showed similar inhibitory potencies toward c3A4 and h3A4, whereas non-h3A4 inhibitors did not inhibit c3A4 activity. The availability of a c3A4 preparation, in conjunction with commercially available monkey liver microsomes, will support further characterization of the cynomolgus monkey as a model to assess CYP3A-dependent clearance and drug-drug interactions.
authors: Pharmaceutical Candidate Optimization (P.B., M.S.) and Applied Biotechnology (Si.S., K.G., P.K., S.K., Sa.S.), Biocon Bristol-Myers Squibb Research and Development Center (BBRC), Syngene International Limited, Plot No. 2 & 3, Bommasandra IV Phase, Bangalore, India; Bristol-Myers Squibb, Wallingford, Connecticut (M.W.S.); Bristol-Myers Squibb, Pennington, New Jersey (A.D.R.); and Bristol-Myers Squibb India Ltd. BBRC, Bangalore, India (M.R., S.M.).
authors: Sindhuja
authors: S
authors: Selvakumar
authors: Priyadeep
authors: P
authors: Bhutani
authors: Kaushik
authors: K
authors: Ghosh
authors: Prasad
authors: P
authors: Krishnamurthy
authors: Sanjith
authors: S
authors: Kallipatti
authors: Sabariya
authors: S
authors: Selvam
authors: Manjunath
authors: M
authors: Ramarao
authors: Sandhya
authors: S
authors: Mandlekar
authors: Michael W
authors: MW
authors: Sinz
authors: A David
authors: AD
authors: Rodrigues
authors: Murali
authors: M
authors: Subramanian
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D051544:Cytochrome P-450 CYP3A / Q000235:genetics / Q000378:metabolism*; D065692:Cytochrome P-450 CYP3A Inhibitors; D004347:Drug Interactions; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D008252:Macaca fascicularis; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology; D008954:Models, Biological; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D013045:Species Specificity; D013379:Substrate Specificity; D014162:Transfection
chemical_list: D065692:Cytochrome P-450 CYP3A Inhibitors; D004364:Pharmaceutical Preparations; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human