1,2,4-Thiadiazolidin-3,5-Diones as Inhibitors of Cysteine Proteases

A focused library of 1,2,4-thiadiazolidin-3,5-diones (<b>THIA-1</b>-<b>10</b>), previously characterized as hydrogen sulfide (H₂S) donors, was evaluated for inhibitory activity against cysteine proteases. We included two key cysteine proteases aiming at antiviral drug development-SARS-CoV-2 3CLpro (Mpro) and PLpro-alongside reference enzymes Papain and Cathepsin L. The compounds exhibited distinct selectivity profiles and inhibition mechanisms. The ability to act as covalent inhibitors of 3CLpro in the nanomolar range is of particular interest, with compounds <b>THIA-6</b>, <b>-7</b>, and <b>-10</b> proving to be the most potent inhibitors of the series, and compounds <b>THIA-1</b>, <b>-2</b>, and <b>-8</b> proving to be the most selective with respect to the other proteases. We explored the molecular bases of the observed activity profile of <b>THIA-1</b>-<b>10</b> through computational studies, which supported and complemented the experimental findings, paving the way for future structure optimization. The results highlight that inhibitory potency depends not only on electrophilicity but also on the ability to access the catalytic cysteine within the active site. The dual functionality of <b>THIA-1</b>-<b>10</b> as H₂S donors and selective cysteine protease inhibitors underscores its potential as a promising lead for therapeutic development.