Activity and inhibition of the SARS-CoV-2 Omicron nsp13 R392C variant using RNA duplex unwinding assays

• Published in: SLAS discovery 2024-04, Vol.29 (3), p.100145-100145, Article 100145
• Main Authors: Inniss, Nicole L., Rzhetskaya, Margarita, Ling-Hu, Ted, Lorenzo-Redondo, Ramon, Bachta, Kelly E., Satchell, Karla J.F., Hultquist, Judd F.
• Format: Article
• Language: English
• Subjects: Antiviral Agents - pharmacology; Antiviral drug discovery; COVID-19; COVID-19 - virology; Direct-acting antiviral; Helicase; Humans; Methyltransferases; Mutation - genetics; Nsp13; R392C; RNA Helicases - antagonists & inhibitors; RNA Helicases - genetics; RNA Helicases - metabolism; RNA, Double-Stranded - genetics; RNA, Double-Stranded - metabolism; RNA, Viral - genetics; SARS-CoV-2; SARS-CoV-2 - drug effects; SARS-CoV-2 - genetics; Unwinding assay; Viral Nonstructural Proteins - antagonists & inhibitors; Viral Nonstructural Proteins - genetics; Viral Nonstructural Proteins - metabolism; Virus Replication - drug effects; Virus Replication - genetics
• ISSN: 2472-5552; 2472-5560; 2472-5560
• DOI: 10.1016/j.slasd.2024.01.006

SARS-CoV-2 nsp13 helicase is an essential enzyme for viral replication and a promising target for antiviral drug development. This study compares the double-stranded RNA (dsRNA) unwinding activity of nsp13 and the Omicron nsp13R392C variant, which is predominant in currently circulating lineages. Using in vitro gel- and fluorescence-based assays, we found that both nsp13 and nsp13R392C have dsRNA unwinding activity with equivalent kinetics. Furthermore, the R392C mutation had no effect on the efficiency of the nsp13-specific helicase inhibitor SSYA10-001. We additionally confirmed the activity of several other helicase inhibitors against nsp13, including punicalagin that inhibited dsRNA unwinding at nanomolar concentrations. Overall, this study reveals the utility of using dsRNA unwinding assays to screen small molecules for antiviral activity against nsp13 and the Omicron nsp13R392C variant. Continual monitoring of newly emergent variants will be essential for considering resistance profiles of lead compounds as they are advanced towards next-generation therapeutic development. [Display omitted]