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SLAS Discov . High-Throughput Screening Identifies Non-nucleoside Inhibitors of the SARS-CoV-2 Polymerase with Novel Mechanisms

tetano

Editor, Senior Moderator
SLAS Discov


. 2025 Nov 20:100289.
doi: 10.1016/j.slasd.2025.100289. Online ahead of print. High-Throughput Screening Identifies Non-nucleoside Inhibitors of the SARS-CoV-2 Polymerase with Novel Mechanisms

Colin R Woodford[SUP] 1 [/SUP], Kristine E Frank[SUP] 2 [/SUP], Haizhong Zhu[SUP] 2 [/SUP], Gilman Dionne[SUP] 2 [/SUP], Michael R Schrimpf[SUP] 2 [/SUP], Sujatha M Gopalakrishnan[SUP] 2 [/SUP], Nathaniel L Elsen[SUP] 3 [/SUP]



Affiliations
Free article Abstract

The RNA-dependent RNA polymerase (RdRp) of coronaviruses, comprising highly conserved non-structural proteins, is a critical player in the viral lifecycle and represents a promising target for developing pan-coronavirus antivirals. Despite substantial efforts to identify RdRp inhibitors through drug repurposing and novel compound discovery campaigns, potent in vitro non-nucleoside inhibitors remain elusive. In this study, we detail the development of a robust PicoGreen assay, which facilitated the screening of AbbVie's extensive chemical library, encompassing over 900,000 small molecules, against the SARS-CoV-2 RdRp. Through a combination of biochemical and biophysical assays, we identified two potent non-nucleoside compounds with activity against our PicoGreen beta-coronavirus panel. Mechanism of action investigations revealed these compounds bind exclusively to the nsp12-8 complex, unveiling a potentially unique inhibitory mechanism. These compounds serve as valuable starting points for structure-activity relationship (SAR) explorations and potential therapeutic leads.

Keywords: RNA-dependent-RNA polymerase; SARS-CoV-2; drug discovery; enzymology; high-throughput screen.

 
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