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Potent broad-spectrum anti-coronaviral frameshift inhibitors from virtual screen of RNA binding

Neupane, K.; Sekar, R. V.; Ileperuma, S. M.; Reklow, E.; Munshi, S.; Omar, S. I.; Arbabimoghadam, S.; Peterson, J. A.; Tuszynski, J. A.; Hobman, T. C.; Woodside, M. T.

2025-09-18 biophysics
10.1101/2025.09.17.676934 bioRxiv
Show abstract

Coronavirus genomes contain an RNA pseudoknot that directs -1 programmed ribosomal frame-shifting (-1 PRF) to control expression of viral proteins crucial for replication. Ligands that inhibit -1 PRF can thus attenuate viral propagation and have potential as drugs for limiting corona-virus infections. To search for novel small-molecule frameshift inhibitors with anticoronaviral activity, we computationally screened over 14 million compounds for binding to the SARS-CoV-2 pseudoknot, followed by experimental validation of the top hits for inhibition of -1 PRF and viral replication. We identified multiple potent -1 PRF inhibitors, effective at nM concentrations, some of which significantly suppressed SARS-CoV-2 replication in cell culture. Several compounds also inhibited -1 PRF in multiple representative bat coronaviruses, indicating broad-spectrum activity. These results showcase the promise of viral RNA structures like frameshift-stimulatory pseudoknots as targets for broad-spectrum antiviral drugs.

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