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Inhibition of the in vitro proteolytic activity of EV-93 3Cpro by rupintrivir. Reverse-phase chromatograms show in red the products of the digestion of the EV-93 2C↓3A peptide (Ac-RHSVGATLEALFQ↓GPPVYREIKIS-NH2) by EV-93 3Cpro (p) without rupintrivir (r) (A) or with rupintrivir at two inhibitor-to-protein molar ratios, 1:3 (B) and 30:3 (C). (D) C147A mutant protein. Chromatograms of the substrate peptide alone are shown in blue in all panels.
