细辛挥发油成分与CYP1A2酶的分子对接分析

目的:研究细辛中黄樟醚、肉豆蔻醚、甲基丁香酚、细辛脑四种挥发油成分以及黄樟醚活性中间体和肉豆蔻醚活性中间体同CYP1A2酶的作用方式。方法:通过"Cocktail"探针底物法筛选细辛挥发油成分对CYP1A2,CYP2D6,CYP2E1,CYP3A4和CYP2C19 5种人肝微粒体酶的抑制作用。采用半柔性分子对接的方式研究挥发油成分及中间体与CYP1A2酶的结合能力。结果:结果表明挥发油成分对CYP1A2具有较强的抑制作用。分子对接评分结果分别为3. 048 7 kcal·mol~(-1)(黄樟醚),6. 016 4 kcal·mol~(-1)(肉豆蔻醚),16. 969 2 kcal·mol~(-1)(甲基丁香酚),16. 013 8 kcal·mol~(-1)(细辛脑),23. 923 3 kcal·mol~(-1)(黄樟醚活性中间体)和25. 594 3 kcal·mol~(-1)(肉豆蔻醚活性中间体)。结论:分子对接结果表明黄樟醚中间体和肉豆蔻醚中间体与CYP1A2酶的结合能力最强。进一步确定了黄樟醚和肉豆蔻醚是CYP1A2酶的基于机制性抑制剂,与本课题组前期的IC_(50)-shift及谷胱甘肽捕获实验的结果一致。

Molecular Docking of Volatile Oily Constituents of Chinese Herbal Medicine Asari Radix et Rhizoma and CYP1A2 Enzyme

Objective: To study the mechanisms of action of four volatile oil components (safrole, myristicin, methyleugenol and asarone) and the reactive metabolites of safrole and myristicin with CYP1A2. Method: The inhibitory effects of the volatile oil components of Asari Radix et Rhizoma on the human liver microsomal enzymes CYP1A2, CYP2D6, CYP2E1, CYP3A4 and CYP2C19 were screened by the "Cocktail" probe substrate method. The ability of the volatile oil components and intermediates in binding to CYP1A2 enzyme was studied by means of semi-flexible molecular docking. Result: The screening results showed that the components had a strong inhibitory effect on CYP1A2.Molecular docking scores were 3.048 7 kcal·mol^-1 (safrole), 6.016 4 kcal·mol^-1 (myristicin), 16.969 2 kcal·mol^-1 (methyleugenol), 16.013 8 kcal·mol^-1 (asarone), 23.923 3 kcal·mol^-1 (safrole reactive metabolites) and 25.594 3 kcal·mol^-1 (myristicin reactive metabolites). Conclusion: Molecular docking results indicate that safrole metabolic intermediate and myristicin metabolic intermediate have the strongest ability in binding to CYP1A2 enzyme. This study further confirms that safrole and myristicin are the mechanism-based inhibitors of CYP1A2 enzyme, which is consistent with the results of previous IC₅₀-shift and glutathione capture experiments.