中医药来源2019新型冠状病毒3CL蛋白酶抑制剂的筛选

Discovery of SARS-CoV-23CL protease inhibitors from pre-fractionated traditional Chinese medicine

目的:构建2019新型冠状病毒(SARS-CoV-2)3CL蛋白酶的高通量筛选体系,并利用该体系对180种中药提取物进行筛选。方法:针对SARS-CoV-23CL蛋白酶进行原核表达和纯化,进而建立基于荧光能量共振转移技术的SARS-CoV-23CL抑制剂高通量筛选体系。利用该体系对180种中药提取物进行筛选,并验证活性组分的量效关系。结果:本研究成功构建了SARS-CoV-23CL蛋白酶的高通量筛选体系,并筛选得到5种中药提取物(半枝莲乙酸乙酯部位、防风石油醚部位、紫苏叶环己烷部位、紫苏梗环己烷部位以及紫苏梗乙酸乙酯部位)具有3CL蛋白酶的抑制活性,其半数抑制浓度分别为30.03、20.00、18.34、25.04、18.48μg/mL。结论:该体系可以应用于3CL蛋白酶抑制剂的筛选,为新药研发提供了有力的工具。同时,5种活性提取物为筛选抗SARS-CoV-2活性成分提供了依据。

Objective:To construct of Severe Acute Respiratory Syndrome Coronavirus 2(SARS-CoV-2)3 CL protease high throughput screening system and screen perfectionated traditional Chinese medicines using this system.Methods:Novel coronavirus SARS-CoV-23 CL protease was constructed,expressed and purified,and then the high throughput screening system for 3 CL inhibitors was established based on fluorescence resonance energy transfer technology.A total of 180 kinds of Chinese herbal extracts were screened by this system.The dose-response relationship of the 5 kinds of extracts with good activities were verified.Results:A high throughput screening of SARS-CoV-23 CL protease was successfully constructed,and 5 kinds of extracts(ethyl acetate fraction of Herba Scutellariae Barbatae,petroleum ether fraction of Radix Saposhnikoviae,cyclohexane fraction of Folium Perillae,cyclohexane fraction of stem of Caulis Perillae and ethyl acetate fraction of Caulis Perillae)had inhibitory activity against 3 CL protease.Their 50%inhibitory concentrations are 30.03,20.00,18.34,25.04,18.48μg/mL,respectively.Conclusion:This system could be applied to screen 3 CL protease inhibitors,which provides a powerful tool for the development of new drugs.Meanwhile,the discovery of the 5 active extracts provides a basis for screening new active ingredients from Chinese medicine.