Drug Metabolism and Pharmacokinetics 2009-01-01

Evaluation of induction potency of new drug candidates on CYP1A2 and CYP3A4 using real-time one-step RT-PCR in primary cultures of cryopreserved human hepatocytes.

Masuhiro Nishimura, Shizuo Narimatsu, Shinsaku Naito

Index: Drug Metab. Pharmacokinet. 24(5) , 446-50, (2009)

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Abstract

This study evaluates the induction potency of new drug candidates on mRNA levels of CYP1A2 and CYP3A4 in primary cultures of cryopreserved human hepatocytes. Analysis was performed by quantitative real-time RT-PCR using primers and TaqMan probes. Positive controls for CYP1A2 and CYP3A4 used beta-naphthoflavone (beta-NF) and rifampicin (Rif), respectively. In the first stage of the study, the lot showing the best induction of mRNA expression CYP1A2 and CYP3A4 from among eight lots of hepatocytes was selected. In the second stage, we evaluated the levels of CYP1A2 and CYP3A4 gene expression in hepatocytes after exposure to eight NO-1886 (ibrolipim) derivatives. A combination of real-time one-step RT-PCR and primary culture of cryopreserved human hepatocytes is suitable for evaluating of induction potency of a large number of new drug candidates.