Kinome profiling enabled development of a LATS1/2 inhibitor with wound healing potential

System-level mapping of kinase inhibitors’ polypharmacology can uncover opportunities for drug repurposing and identify starting points for medicinal chemistry campaigns aimed at novel targets. Here, we describe a large-scale profiling of 257 kinase inhibitors that revealed domains of promiscuity, a broad region of uninhibited targets and discrete pockets of selectivity. Focusing on a subcluster enriched for AGC group kinases and using the AKT inhibitor AT-7867 (an antiproliferative agent) as a starting point, we employed scaffold hopping and property-based optimization to discover NCGC-6782, a LATS1/2 inhibitor with robust proproliferative activity. In biochemical and cell-based assays, NCGC-6782 inhibited LATS1/2 activity, reduced YAP phosphorylation, and promoted rapid YAP nuclear translocation and transactivation of YAP-target genes. Kinome-wide profilings demonstrated good in vitro selectivity and exquisite chemoproteomic and kinetic selectivity. In vivo, NCGC-6782 exhibited favorable pharmacokinetic (PK) profiles following intraperitoneal (IP), intravenous (IV), or oral (PO) dosing, leading to rapid inhibition of YAP-phosphorylation in murine liver samples. Transcriptomics and proteomics following NCGC-6782 revealed upregulation of YAP-dependent genes (4 h) followed by activation of cell cycle programs (24 h). Consistently, NCGC-6782 accelerated proliferative regeneration in both cell-based and in vivo wound healing models of full-thickness excisional skin injury, significantly shortening wound-closure time without evidence of systemic toxicity. Altogether, we describe an optimized, potent, and selective LATS1/2 kinase inhibitor with desirable drug-like properties and broad potential for regenerative medicine applications following acute injury of nonproliferative, postmitotic tissues.