Design of SARS-CoV-2 papain-like protease inhibitor with antiviral efficacy in a mouse model

The emergence of SARS-CoV-2 variants and drug-resistant mutants calls for additional oral antivirals. The SARS-CoV-2 papain-like protease (PL^pro) is a promising but challenging drug target. In this study, we designed and synthesized 85 noncovalent PL^pro inhibitors that bind to the newly discovered Val70^Ub site and the known BL2 groove pocket. Potent compounds inhibited PL^pro with inhibitory constant K_i values from 13.2 to 88.2 nM. The co-crystal structures of PL^pro with eight leads revealed their interaction modes. The in vivo lead Jun12682 inhibited SARS-CoV-2 and its variants, including nirmatrelvir-resistant strains with EC₅₀ from 0.44 to 2.02 μM. Oral treatment with Jun12682 significantly improved survival and reduced lung viral loads and lesions in a SARS-CoV-2 infection mouse model, suggesting PL^pro inhibitors are promising oral SARS-CoV-2 antiviral candidates.