Effect of donepezil combined with hypoxia on CYP3A4 and its safety-evaluation
- VernacularTitle:多奈哌齐协同缺氧对CYP3A4酶的影响及安全性评价
- Author:
Xiao-xia HAN
1
;
Yue-xin LI
;
Wei TENG
;
Fang WANG
;
Hai-ying HONG
;
Ze-shuai YI
;
Ying SONG
;
Yu-yan ZHOU
;
Bao-xin LI
;
Pan FAN
Author Information
- Publication Type:Journal Article
- Keywords: donepezil; CYP3A4; hypoxia; long QT syndrome; drug metabolism; cardiac toxicity
- From: Chinese Pharmacological Bulletin 2025;41(12):2354-2361
- CountryChina
- Language:Chinese
- Abstract: Aim To investigate the regulatory mecha-nisms of donepezil on the expression and enzymatic ac-tivity of cytochrome P450 3A4(CYP3A4),elucidate the synergistic impact of hypoxia on CYP3A4 function,and reveal its potential association with drug-induced cardiotoxicity,particularly QT interval prolongation.Methods Western blot,co-immunoprecipitation,and gene knockdown techniques were employed to evaluate the effects of donepezil and hypoxia on CYP3A4 pro-tein expression.CYP3A4 enzymatic activity was as-sessed using an in vitro incubation system with rat liver microsomes combined with high-performance liquid chromatography(HPLC),and the half-maximal inhib-itory concentration(IC50)was determined.Results Donepezil(10 μmol·L-1)and hypoxia reduced CYP3A4 protein expression to 31.75％and 45.90％of the control levels,respectively.Both interventions activated the gp78-mediated ubiquitin-proteasome path-way,significantly increasing CYP3A4 ubiquitination levels by 2.1-fold compared to the control group,thereby promoting proteasomal degradation.Donepezil inhibited CYP3A4 enzyme activity with an IC50 of 83.4μmol·L-1,and hypoxia synergistically enhanced this inhibitory effect,reducing the IC50 to 20.79 μmol·L-1.Conclusion Donepezil downregulates CYP3A4 function through dual mechanisms involving ubiquitin-mediated proteasomal degradation and direct enzymatic inhibition.Hypoxia potentiates this effect,leading to impaired metabolism of CYP3A4 substrate drugs,ele-vated plasma drug concentrations(1.6-2.3-fold in-crease compared to normal metabolic conditions),and an increased risk of QT interval prolongation and other forms of cardiotoxicity.
