Discovery of a novel SARS-CoV-2 main protease inhibitor by a simple and optimized colorimetric screening assay
10.16438/j.0513-4870.2022-0365
- VernacularTitle:基于比色法原理的新冠病毒主蛋白酶小分子抑制剂高通量筛选模型的优化与应用
- Author:
Gan-gan YAN
1
;
Hao-hao YAN
1
;
Zhi-cheng LIU
1
;
Hai-yan QI
1
;
Xiao-li LIU
1
;
Xiao-ping LIU
1
;
Jing ZHANG
2
;
Yun-yu CHEN
1
Author Information
1. Institute for Drug Screening and Evaluation, Wannan Medical College, Wuhu 241002, China
2. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
- Publication Type:Research Article
- Keywords:
SARS-CoV-2;
main protease inhibitor;
colorimetric screening assay;
fluorescence resonance energy transfer;
fluorescence polarization;
ginkgolic acid C13:0
- From:
Acta Pharmaceutica Sinica
2022;57(10):3019-3026
- CountryChina
- Language:Chinese
-
Abstract:
For rapid discovery of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) main protease (Mpro) inhibitors from a natural product library, a novel colorimetric screening assay was developed. According to the colorimetric principle, the synthetic peptide TSAVLQ-para-nitroanilide (pNA) was used as the Mpro hydrolysis substrate. Subsequently, the working concentration of pNA substrate, Mpro working concentration, hydrolysis time and DMSO tolerance were optimized for the development of a simple and robust colorimetric screening assay. Through these systematic optimizations, we selected 0.4 μmol·L-1 Mpro and 100 μmol·L-1 pNA substrate as the optimal working concentrations in this colorimetric screening assay, and a high Z' factor of 0.9 was achieved. Using this screening assay, natural product ginkgolic acid C13:0 (GA13:0) was identified as a novel competitive Mpro inhibitor in vitro. Taken together, we have successfully developed a simple and optimized colorimetric screening assay, which will be vital for the discovery of novel SARS-CoV-2 Mpro inhibitors.
