Time-dependent inhibition of CYP3A4 by gallic acid in human liver microsomes and recombinant systems

Abstract

1. Gallic acid is a main polyphenol in various fruits and plants. Inhibitory characteristics of gallic acid on CYP3A4 were still unclear. The objective of this work is hence to investigate inhibitory characteristics of gallic acid on CYP3A4 using testosterone as the probe substrate in human liver microsomes (HLMs) and recombinant CYP3A4 (rCYP3A4) systems.

2. Gallic acid caused concentration-dependent loss of CYP3A4 activity with IC₅₀ values of 615.2 μM and 669.5 μM in HLM and rCYP3A4 systems, respectively. IC₅₀-shift experiments showed that pre-incubation with gallic acid in the absence of NADPH contributed to 12- or 14-fold reduction of IC₅₀ in HLM and rCYP3A4 systems, respectively, supporting a time-dependent inhibition. In HLM, time-dependent inactivation variables K_I and K_inact were 485.8 μM and 0.05 min^–1, respectively.

3. Compared with the presence of NADPH, pre-incubation of gallic acid in the absence of NADPH markedly increased its inhibitory effects in HLM and rCYP3A4 systems. Those results indicate that CYP3A4 inactivation by gallic acid was independent on NADPH and was mainly mediated its oxidative products.

4. In conclusion, we showed that gallic acid weakly and time-dependently inactivated CYP3A4 via its oxidative products.

• 6β-hydroxylase
• CYP3A4
• gallic acid
• testosterone
• time-dependent inhibition

Declaration of interest

The authors report no declaration of interest.

This work was supported by the National Nature Science Foundation of China (no. 81370403) and Specialized Research Fund for the Doctoral Program of Higher Education (no. 20125503110008).