Improved fluorescence-based assay for rapid screening and evaluation of SARS-CoV-2 main protease inhibitors.

Publication date: Mar 01, 2024

The outbreak of coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a global threat to human health. In parallel with vaccines, efficacious antivirals are urgently needed. SARS-CoV-2 main protease (Mpro) is an attractive drug target for antiviral development owing to its key roles in virus replication and host immune evasion. Due to the limitations of currently available methods, the development of novel high-throughput screening assays is of the highest importance for the discovery of Mpro inhibitors. In this study, we first developed an improved fluorescence-based assay for rapid screening of Mpro inhibitors from an anti-infection compound library using a versatile dimerization-dependent red fluorescent protein (ddRFP) biosensor. Utilizing this assay, we identified MG-101 as a competitive Mpro inhibitor in vitro. Moreover, our results revealed that ensitrelvir is a potent Mpro inhibitor, but baicalein, chloroquine, ebselen, echinatin, and silibinin are not. Therefore, this robust ddRFP assay provides a faithful avenue for rapid screening and evaluation of Mpro inhibitors to fight against COVID-19.

Concepts Keywords
Competitive 3C-like proteinase, SARS-CoV-2
Coronavirus Antiviral Agents
Library Antiviral Agents
Pandemic Coronavirus 3C Proteases
Coronavirus 3C Proteases
high-throughput screening
main protease inhibitor
Protease Inhibitors
Protease Inhibitors
red fluorescent protein


Type Source Name
disease IDO assay
disease MESH coronavirus disease 2019
disease VO Severe acute respiratory syndrome coronavirus 2
disease IDO replication
disease IDO host
disease MESH infection
drug DRUGBANK Chloroquine
drug DRUGBANK Ebselen

Original Article

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