Validation and invalidation of SARS-CoV-2 main protease inhibitors using the Flip-GFP and Protease-Glo luciferase assays  

作　　者：Chunlong Ma Haozhou Tan Juliana Choza Yuyin Wang Jun Wang 

机构地区：[1]Department of Pharmacology and Toxicology,College of Pharmacy,the University of Arizona,Tucson,AZ 85721,USA

出　　处：《Acta Pharmaceutica Sinica B》2022年第4期1636-1651,共16页药学学报（英文版）

基　　金：This research was supported by the National Institute of Allergy and Infectious Diseasess at the National Instiute of Health(NIH,USA,grants AI147325,AI157046,and AI158775);the Arizona Biomedical Research Centre Young Investigator grant(ADHS18-198859,USA)to Jun Wang.

摘　　要：SARS-CoV-2 main protease(M^(pro))is one of the most extensively exploited drug targets for COVID-19.Structurally disparate compounds have been reported as M^(pro) inhibitors,raising the question of their target specificity.To elucidate the target specificity and the cellular target engagement of the claimed M^(pro) inhibitors,we systematically characterize their mechanism of action using the cell-free FRET assay,the thermal shift-binding assay,the cell lysate Protease-Glo luciferase assay,and the cell-based FlipGFP assay.Collectively,our results have shown that majority of the M^(pro) inhibitors identified from drug repurposing including ebselen,carmofur,disulfiram,and shikonin are promiscuous cysteine inhibitors that are not specific to M^(pro),while chloroquine,oxytetracycline,montelukast,candesartan,and dipyridamole do not inhibit M^(pro) in any of the assays tested.Overall,our study highlights the need of stringent hit validation at the early stage of drug discovery.

关 键 词：SARS-CoV-2 ANTIVIRAL Main protease EBSELEN CARMOFUR FlipGFP assay Protease-Glo luciferase assay 

分 类 号：R917[医药卫生—药物分析学]