Identification of Covalent Inhibitors Targeting NSP13 via Integrated Biochemical and Chemoproteomics Approaches

About this poster

This poster describes an integrated hit-to-lead workflow to discover covalent inhibitors of the SARS-CoV-2 helicase NSP13, a conserved enzyme required for viral RNA synthesis, replication, and transcription. It outlines an HTS campaign using ATPase and helicase unwinding assays, followed by a selection funnel that combines intact-mass confirmation of covalent adduct formation, intrinsic thiol-reactivity profiling, peptide mapping by LC–MS/MS to localize modified cysteines, and chemoproteomics in cellular lysates to assess specificity and site selectivity. By integrating biochemical and proteomics readouts, the workflow identifies covalent NSP13 binders with demonstrated activity and supports structure–activity studies to optimize potency and selectivity. Download the poster to explore the screening funnel and assay cascade in detail.