A Gaussia luciferase reporter assay for the evaluation of coronavirus Nsp5/3CLpro activity

Scientific Reports, Journal Year: 2024, Volume and Issue: 14(1)

Published: Sept. 5, 2024

Language: Английский

Potent small molecule inhibitors against the 3C protease of foot-and-mouth disease virus

Microbiology Spectrum, Journal Year: 2024, Volume and Issue: 12(4)

Published: March 11, 2024

ABSTRACT Foot-and-mouth disease (FMD) is one of the most devastating diseases livestock which can cause significant economic losses, especially when introduced to FMD-free countries. FMD virus (FMDV) belongs family Picornaviridae and antigenically heterogeneous with seven established serotypes. The prevailing preventive control strategies are limited restriction animal movement elimination infected or exposed animals, be potentially combined vaccination. However, vaccination has limitations including delayed protection lack cross-protection against different Recently, antiviral drug use for outbreaks increasingly been recognized as a potential tool augment existing early response strategies, but research reported on compounds FMDV. FMDV 3C protease (3Cpro) cleaves viral-encoded polyprotein into mature functional proteins during viral replication. essential role 3Cpro in replication high conservation among serotypes make it an excellent target development. We have previously multiple series inhibitors picornavirus 3C-like proteases (3CLpros) encoded by coronaviruses caliciviruses. In this study, we conducted structure-activity relationship studies our in-house focused compound library containing 3CLpro using enzyme cell-based assays. Herein, report discovery aldehyde α-ketoamide potency. These data inform future preclinical that related advancement these further along development pathway. IMPORTANCE Food-and-mouth causes cloven-hoofed animals impact. Emergency limit spread critical, antivirals may overcome measures providing immediate susceptible animals. encodes (3Cpro), attractive discovery. Here, study identified potent 3Cpro. Our results suggest antivirals.

Language: Английский

A Gaussia luciferase reporter assay for the evaluation of coronavirus Nsp5/3CLpro inhibitors

Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown

Published: May 23, 2024

Human coronaviruses (hCoVs) infect millions of people every year. Among these, MERS, SARS-CoV-1, and SARS-CoV-2 caused significant morbidity mortality their emergence highlights the risks associated with possible future coronavirus outbreaks. Therefore, broadly-active anti-coronavirus drugs are needed. Pharmacological inhibition hCoV protease 3CLpro (Nsp5) in COVID-19 patients is clinically beneficial as shown by wide effective use Paxlovid (nirmaltrevir, ritonavir). However, further treatment options required due to drug resistance some strains. To facilitate inhibitor discovery evaluation, we developed an assay allowing rapid reliable quantification activity under biosafety level 1 conditions. It based on ACE2 receptor - Gal4 transcription factor fusion protein separated a recognition site. Cleavage releases factor, which then induces expression Gaussia luciferase. Our compatible proteases from all hCoVs, allows simultaneous measurement inhibitory cytotoxic effects tested compounds. Proof-of-concept IC₅₀ measurements confirmed that nirmaltrevir, GC376 lopinavir inhibit function without inducing cytotoxicity. Overall, luciferase-based reporter suitable for evaluating viral screening potential inhibitors.

Language: Английский