Mechanism-based inactivation of cytochrome P450 3A by evodol

Abstract

1. Evodol is one of the furanoids isolated from the fruits of Evodia rutaecarpa that has been widely prescribed for the treatment of gastrointestinal diseases in China. The aim of this study was to investigate the inhibitory effect of evodol on CYP3A.

2. A 30-min preincubation of evodol with human liver microsomes raised an obvious left IC₅₀ shift, 3.9-fold for midazolam 1’-hydroxylation and 3.2-fold for testosterone 6β-hydroxylation. Evodol inactivated CYP3A in a time-, concentration- and NADPH-dependent manner, with K_I and k_inact of 5.1 μM and 0.028 min⁻¹ for midazolam 1’-hydroxylation and 3.0 μM and 0.022 min⁻¹ for testosterone 6β-hydroxylation.

3. Co-incubation of ketoconazole attenuated the inactivation while the inclusion of glutathione (GSH) and catalase/superoxide dismutase displayed no such protection.

4. cis-Butene-1, 4-dial (BDA) intermediate derived from evodol were trapped by glutathione and N-acetyl-lysine in microsomes and characterised by HR-MS spectra. The BDA intermediate was believed to play a key role in CYP3A inactivation. CYP3A4 and 2C9 were the primary enzymes contributing to the bioactivation of evodol.

5. To sum up, for the first time evodol was characterised as a mechanism-based inactivator of CYP3A.

Keywords:

• Evodol
• Evodia rutaecarpa
• mechanism-based inactivator
• cytochrome P450 3A
• cis-butene-14-dial

Author contributions

The experiment was designed by: Jie Zhao, Jie Xu; Experiment was performed by: Jie Zhao, Jingyu He; Data were analysed by: Jie Zhao, Jingyu He; the Manuscript was written by: Jie Zhao; the Manuscript was reviewed by: Jie Zhao, Jingyu He, Jie Xu.

Disclosure statement

The authors report no declarations of interest.

Additional information

Funding

This work was not funded.