In vivo induction of human cytochrome P450 3A4 by rifabutin in chimeric mice with humanized liver

Abstract

The induction of human cytochrome P450 enzymes (CYPs) often poses a serious problem in clinical practice. The induction of CYP3A leads to a decrease in the pharmacological potency of drugs, since many drugs are substrates of CYP3A. The present study examined the in vivo induction potency of human CYP3A in chimeric mice with humanized liver, recently established in Japan, by a specific inducer of human CYP3A enzyme activity in this experimental condition, rifabutin, which is an analogue of rifampicin. The chimeric mice were treated intraperitoneally daily for 4 days with rifabutin (50 mg kg⁻¹ day⁻¹). The mRNA, protein and enzyme activity in liver of the chimeric mice were measured by reverse-transcriptase polymerase chain reaction, Western blot analysis and high-performance liquid chromatography, respectively. In the chimeric mice, the human CYP3A4 mRNA expression, CYP3A4 protein content, testosterone 6ß-hydroxylase activity and dexamethasone 6-hydroxylase activity were increased 7.4-, 3.0-, 2.4- and 1.9-fold, respectively, by treatment with rifabutin. The mRNA expression of other human CYPs, transporters and nuclear receptors was not significantly changed by rifabutin. On the other hand, rifabutin was demonstrated not to increase the murine Cyp3a enzyme activities in the control mice. It was demonstrated that human CYP3A4 expressed in the chimeric mice with humanized liver was induced by rifabutin, suggesting that human CYP3A4 in the chimeric mice had induction potency. This chimeric mouse model may be a useful animal model to estimate and predict the in vivo induction of CYPs in human.

Kewords: :

• Induction
• chimeric mice with humanized liver
• rifabutin
• cytochrome P450

Acknowledgements

Work was supported by a Research on Advanced Medical Technology, Health, and Labor Sciences Research Grant from the Ministry of Health, Labor, and Welfare of Japan. The authors thank Brent Bell for reviewing the manuscript.