Evaluation of MTBH, a novel hesperetin derivative, on the activity of hepatic cytochrome P450 isoform in vitro and in vivo using a cocktail method by HPLC-MS/MS

Abstract

1. 8-methylene-tert-butylamine-3′,5,7-trihydroxy-4′-methoxyflavanone (MTBH), a novel hesperidin derivative, has potential in the prevention of hepatic disease, however, its effects on cytochrome P450 isoforms (CYP450s) remains unexplored. The purpose was to investigate the effects of MTBH on the mRNA, protein levels, and activities of six CYP450s (1A2, 2C11/9, 2D2/6, 3A1/4, 2C13/19, and 2E1) in vitro and in vivo.

2. In vitro study, rat and human liver microsomes were adopted to elucidate the inhibitory effect of MTBH on six CYP450s using probe drugs. In vivo study, Sprague-Dawley male rats were treated with MTBH (25, 50, or 100 mg/kg for 28 consecutive days), phenobarbital (80 mg/kg for 12 consecutive days), or 0.5% CMC-Na solution (control group) by intragastric administration, then, the mRNA, protein levels and activities of liver CYP450s were analysed by real-time PCR, western blotting and probe-drug incubation systems, respectively.

3. The in vitro study indicated that MTBH inhibits the activities of CYP3A1/4 and CYP2E1 in rat and human liver microsomes. In vivo data showed that MTBH inhibits mRNA, protein levels, and activities of CYP3A1 and CYP2E1 in medium- and high-dose MTBH groups.

4. MTBH has the potential to cause drug-drug interactions when co-administered with drugs that are metabolised by CYP3A1/4 and CYP2E1.

Keywords:

• MTBH
• cytochrome P450 isoforms
• cocktail
• liver microsome
• LC-MS/MS

Acknowledgments

The authors would like to thank Hefei Kaifan Analytical Technology Co., Ltd for their work on LC-MS/MS analysis of substrate metabolites (cocktail assay) for assessing the in vitro inhibition of CYP450 enzymes in RLMs and HLMs. The authors thank Guanjun Chen and Sheng Wang of the Center for Scientific Research of Anhui Medical University for valuable help in our experiment.

Disclosure statement

All the authors have declared that there are no conflicts of interest.

Additional information

Funding

This study was supported by the National Natural Science Foundation of China (No. 81703800), the First Affiliated Hospital of Anhui Medical University for the Cultivation Program of Youth Science Foundation (No. 2792), Grants for Scientific Research of BSKY (No. XJ201625) from Anhui Medical University, the Fund of Anhui Medical University Doctoral Start Research (No. 0601069201) and Innovation and Entrepreneurship Training Program for Undergraduates of Anhui Medical University (No.S202010366011).