Effects of Avitinib on CYP450 Enzyme Activity in vitro and in vivo in Rats

Abstract

Purpose

Avitinib is the first third-generation epithelial growth factor receptor (EGFR) inhibitor independently developed in China and is mainly used for treating non-small cell lung cancer. However, pharmacokinetic details are limited. This study explored the in vivo and in vitro effects of avitinib on cytochrome CYP450 enzymes metabolic activity.

Methods

A rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for determining six probe substrates and their metabolites. Avitinib influence on activity levels of CYP isozymes was examined in vitro using human and rat liver microsomes (HLMs/RLMs). For in vivo studies, rats were pretreated with 30 mg/kg avitinib once daily for 7 days (avitinib multiple-doses group), 30 mg/kg avitinib on day 7 (avitinib single-dose group), or an equivalent amount of CMC-Na once daily for 7 days (control group), followed by intragastrical administration of the probe substrates (1 mg/kg tolbutamide and 10 mg/kg phenacetin, bupropion, chlorzoxazone, dextromethorphan, and midazolam). Plasma pharmacokinetics and IC₅₀ values of the probe substrates were then compared. Pharmacokinetic parameters were determined using non-compartmental analysis implemented in a pharmacokinetic program.

Results

In vitro experiments revealed different inhibitory effects of avitinib on the six probe substrates with various IC₅₀ values (bupropion, 6.39/22.64 μM; phenacetin, 15.79/48.36 μM; chlorzoxazone, 23.15/57.09 μM; midazolam, 27.64/59.6 μM; tolbutamide, 42.18/6.91 μM; dextromethorphan, 44.39/56.57 μM, in RLMs and HLMs respectively). In vivo analysis revealed significant differences (P <0.05) in distinct pharmacokinetic parameters (AUC_(0-t), AUC _(0-∞), C_max, MRT_(0-t), MRT _(0-∞), and CLz/F) for the six probe substrates after avitinib pretreatment.

Conclusion

A sensitive and reliable UPLC-MS/MS method was established to determine the concentration of six probe substrates in rat plasma. Avitinib had inhibitory effects on CYP450 enzymes, especially cyp2b1, cyp1a2 in RLMs, CYP2C9 in HLMs, and cyp1a2, cyp2b1, cyp2d1, and cyp2e1 in vivo. Our data recommend caution when avitinib was taken simultaneously with drugs metabolized by CYP450 enzymes.

Keywords:

• cytochrome
• EGFR inhibitor
• UPLC-MS/MS
• rat liver microsome
• drug-drug interaction

Acknowledgments

This work was supported by grants funded by Public Welfare Technology Research Funding Project of Zhejiang (LGF21H310002), Key Research and Development Project of Lishui (2020ZDYF12, 2021ZDYF13 & 2021ZDYF15), Public Welfare Technology Research Funding Project of Lishui (2020GYX18 & 2020GYX23), Ningbo Science & Technology Bureau (2016A610153), Taizhou Science and Technology Project (1802ky29), Medical and Health Science and Technology Plan Project of Zhejiang Province (2021KY411 & 2019KY804).

Disclosure

The authors report no conflicts of interest and no disclosures to declare.