Discovery of Chlorofluoroacetamide-Based Covalent Inhibitors for Severe Acute Respiratory Syndrome Coronavirus 2 3CL Protease

Abstract

The coronavirus disease 2019 (COVID-19) pandemic has necessitated the development of antiviral agents against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). 3C-like protease (3CL^pro) is a promising target for COVID-19 treatment. Here, we report a new class of covalent inhibitors of 3CL^pro that possess chlorofluoroacetamide (CFA) as a cysteine-reactive warhead. Based on an aza-peptide scaffold, we synthesized a series of CFA derivatives in enantiopure form and evaluated their biochemical efficiency. The data revealed that <b>8a</b> (<b>YH-6</b>) with the <i>R</i> configuration at the CFA unit strongly blocks SARS-CoV-2 replication in infected cells, and its potency is comparable to that of nirmatrelvir. X-ray structural analysis showed that <b>YH-6</b> formed a covalent bond with Cys145 at the catalytic center of 3CL^pro. The strong antiviral activity and favorable pharmacokinetic properties of <b>YH-6</b> suggest its potential as a lead compound for the treatment of COVID-19.