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Updated: Jan 7, 2026

Covalent Fragment Screening Using the Quantitative Irreversible Tethering Assay
Published on: February 28, 2025
The Current Toolbox for Covalent Inhibitors: From Hit Identification to Drug Discovery
Mengke You1,2, Hong Liu1,2,3, Chunpu Li1,2,3
1State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
Abstract:
Covalent modification of therapeutic targets has emerged as a powerful platform for creating clinical drugs and chemical probes. Covalent drugs have evolved from serendipitous discoveries to rationally designed therapeutics, driven by advances in electrophile-first screening technologies. This perspective takes stock of alternative technologies currently available in laboratories and industry that collectively enable targeted covalent inhibitor development across historically "undruggable" targets. We highlight five such technologies: activity-based protein profiling (ABPP), provides functional proteomic mapping to identify ligandable residues; covalent tethering, exploits dynamic chemistry to capture transient pockets; covalent DNA-encoded libraries, leverages trillion-member libraries for multiresidue targeting; phage/mRNA display, which facilitates evolution of covalent macrocyclic peptides; and sulfur-(VI) fluoride exchange (SuFEx), engages residues beyond cysteine. Integration of these approaches with chemoproteomics and artificial intelligence accelerates the discovery of covalent inhibitors with enhanced selectivity and reduced off-target risks. This technological convergence establishes a new paradigm for precision covalent therapeutics, offering innovative solutions to overcome drug resistance and target challenging protein interfaces.
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