Advanced technologies for screening and identifying covalent inhibitors

Covalent inhibitors, forming reversible or irreversible covalent bonds with nucleophilic groups in target protein active sites, effectively inhibit protein function for therapeutic purposes. They can also be used for target validation, biomarker identification, and as chemical probes. Boasting high bioefficiency, low drug resistance, and minimal off-target effects, covalent inhibitors possess significant application potential within the small molecule drug market. Their discovery usually involves rational drug design. Understanding screening and identification tools for covalent inhibitors aids in selecting suitable and efficient development methods. This review encompasses screening platforms, target-ligand covalent binding, and selective analysis tools used in covalent inhibitor development. It focuses on discovery strategies such as computer-aided drug design, library screening, and classic techniques for binding determination and selectivity. Examples highlight the advantages and limitations of screening platforms, offering insights for covalent inhibitor discovery and advancing drug development.