Kinetic Characterization and Inhibitor Screening for the Proteases Leading to Identification of Drugs against SARS-CoV-2
Journal
Antimicrobial agents and chemotherapy
Journal Volume
65
Journal Issue
4
Date Issued
2021
Author(s)
Kuo, Chih-Jung
Kao, Han-Chieh
Tsai, Ya-Min
Liu, Yi-Kai
Wang, Lily Hui-Ching
Hsieh, Ming-Chang
Liang, Po-Huang
Abstract
Coronavirus (CoV) disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has claimed many lives worldwide and is still spreading since December 2019. The 3C-like protease (3CLpro) and papain-like protease (PLpro) are essential for maturation of viral polyproteins in SARS-CoV-2 life cycle and thus regarded as key drug targets for the disease. In this study, 3CLpro and PLpro assay platforms were established, and their substrate specificities were characterized. The assays were used to screen collections of 1,068 and 2,701 FDA-approved drugs. After excluding the externally used drugs which are too toxic, we totally identified 12 drugs as 3CLpro inhibitors and 36 drugs as PLpro inhibitors active at 10 μM. Among these inhibitors, six drugs were found to suppress SARS-CoV-2 with the half-maximal effective concentration (EC50) below or close to 10 μM. This study enhances our understanding on the proteases and provides FDA-approved drugs for prevention and/or treatment of COVID-19.
Subjects
3CLpro; COVID-19; PLpro; SARS-CoV-2; antivirals; drug repurposing; inhibitors
Other Subjects
anti-SARS-CoV-2 agent; anticoronavirus agent; coronavirus 3C protease; coronavirus papain-like protease; levothyroxine; loperamide; manidipine; maprotiline; peptide; proanthocyanidin; proteinase inhibitor; reserpine; tolcapone; antivirus agent; peptide hydrolase; proteinase inhibitor; antiviral activity; Article; CC50 (cytotoxic concentration); cell viability; concentration response; controlled study; coronavirus disease 2019; drug approval; drug cytotoxicity; drug identification; drug repositioning; drug screening; drug targeting; EC50; enzymatic assay; enzyme activity; enzyme inhibition; enzyme kinetics; enzyme linked immunosorbent assay; enzyme specificity; fluorescence analysis; fluorescence resonance energy transfer; Food and Drug Administration; high performance liquid chromatography; IC50; nonhuman; priority journal; SARS-CoV-2/NTU13/TWN/Human/2020; Severe acute respiratory syndrome coronavirus 2; Vero C1008 cell line; viral plaque assay; virus replication; animal; cell line; Chlorocebus aethiops; drug effect; human; kinetics; metabolism; Vero cell line; Animals; Antiviral Agents; Cell Line; Chlorocebus aethiops; COVID-19; Humans; Kinetics; Peptide Hydrolases; Protease Inhibitors; SARS-CoV-2; Substrate Specificity; Vero Cells
Type
journal article