细胞水平新型冠状病毒SARS-CoV-23CL蛋白酶抑制剂筛选模型的建立  

Establishment of a cell-based screening assay for inhibitors of SARS-CoV-23CL protease

作　　者：马铃[1] 赵建元[1] 郭赛赛 谢永丽[1] 岑山[1] MA Ling;ZHAO Jian-yuan;GUO Sai-sai;XIE Yong-li;CEN Shan(Institute of Medicinal Biotechnology,Chinese Academy of Medical Sciences,Beijing 100050,China)

机构地区：[1]中国医学科学院医药生物技术研究所,北京100050

出　　处：《药学学报》2020年第9期2122-2126,共5页Acta Pharmaceutica Sinica

基　　金：中国医学科学院医学与健康科技创新工程项目(2018-I2M-3-004);国家自然科学基金资助项目(81903679)。

摘　　要：新型冠状病毒(SARS-CoV-2)是导致全球新冠病毒肺炎暴发的病原体。SARS-CoV-2的3C样蛋白酶(3CL)在病毒复制中起关键作用,成为抗病毒药物设计的理想靶标。本文利用生物发光共振能量转移(BRET)的技术建立了细胞水平SARS-CoV-2病毒3CL蛋白酶的筛选模型,并初步应用和评价了该筛选模型。结果显示,该方法重复性好(Z’因子为0.59),可以反映3CL蛋白酶底物剪切效率,与细胞水平抗病毒活性分析具有较好的一致性。本工作表明,该方法可以应用于3CL蛋白酶抑制剂的筛选与评价,为新药研发提供了有力工具。Severe acute respiratory syndrome coronavirus 2(SARS-Co V-2)is the pathogen that caused the global COVID-19 outbreak.The 3C-like protease(3CLpro)of SARS-Co V-2 plays a key role in virus replication and has become an ideal target for antiviral drug design.In this paper,we report the validation and use of biolumines‐cence resonance energy transfer(BRET)technology to establish a cell-based assay for screening for SARS-Co V-2virus 3CL protease inhibitors.The results show that the method is able to monitor the cleavage efficiency of 3CL protease with good reproducibility(Z’factor is 0.59),and is consistent with antiviral activity analysis in cell culture.This work demonstrates that this method can be applied to the screening and evaluation of 3CL protease inhibitors,providing a powerful tool for the development of new drugs.

关 键 词：新型冠状病毒 抗新型冠状病毒药物 筛选模型 蛋白酶 

分 类 号：R373.1[医药卫生—病原生物学]