Impact of Single Halogen Atom Substitutions on Antiviral Profile of Inhibitors Targeting SARS-CoV-2 Main Protease
Haydar Bulut, Nobuyo Higashi‐Kuwata, Hiromi Ogata-Aoki, Hironori Hayashi, Nobutoki Takamune, Naoki Kishimoto, Debananda Das, Mi Li, Alexander Wlodawer, Shogo Misumi, Hirokazu Tamamura, Hiroaki Mitsuya · ACS Omega · 2026
High Resolution Image Download MS PowerPoint Slide The SARS-CoV-2 main protease (M pro ) remains a prime antiviral target because its inhibition halts viral replication. To probe how subtle atomic changes influence drug performance, we carried out a systematic halogen scan on a potent ketoamide scaffold, replacing a single fluorine with chlorine, bromine, or iodine. Enzymatic assays revealed that the F- and Cl-substituted analogues inhibit M pro at nanomolar levels, whereas Br and I variants are 10- to 20-fold weaker. Cell-based antiviral tests mirrored this trend, yet uptake studies showed the opposite: iodine markedly enhances intracellular accumulation. High-resolution X-ray structures (1.6–1.8 Å) explain the dichotomy: small halogens fit snugly in the S1′ σ-hole pocket, maximizing hydrogen-bond geometry, while bulkier atoms distort binding but create a lipophilic patch that boosts permeability. These data yield the first fluorine-to-iodine structure–activity map for SARS-CoV-2 M pro inhibitors. These findings highlight the critical role of halogen selection in antiviral inhibitor design.