Repurposing-inspired discovery of novel sagitta-shaped covalent TEAD inhibitors from NSAID diflunisal

The YAP/TAZ-TEAD complex, a core downstream co-regulatory module of the Hippo pathway, drives tumorigenesis and progression upon aberrant activation. Targeting its protein-protein interaction (PPI) is a highly promising anticancer strategy. TEAD palmitoylation maintains TEAD homeostasis and facilitates YAP/TAZ binding, and this pocket has been successfully exploited to develop covalent and non-covalent TEAD inhibitors. To discover novel chemotype TEAD inhibitors, we established a strategy integrating virtual screening, drug repurposing and structure-based optimization, and recently reported several FDA-approved drugs as potential TEAD inhibitors. Herein, we identify NSAID diflunisal (DF) as a potent TEAD inhibitor, elucidating its undefined anticancer mechanism. Structure-based optimization of DF further yielded novel biphenyl-structured covalent TEAD inhibitors, and representative derivative DF-CIV-2 features a unique sagitta-shaped binding conformation, potently inhibiting TEAD palmitoylation, transcriptional activity and YAP-TEAD complex formation, downregulating Hippo target genes, suppressing YAP-dependent cancer cell proliferation and 3D spheroid growth, and synergizing with MEK/PI3Kα/mTORC1 inhibitors. Collectively, we established a feasible TEAD inhibitor discovery pipeline, and identified DF and its sagitta-shaped derivative DF-CIV-2 as promising TEAD inhibitors, providing novel molecular tools and insights for YAP/TAZ-TEAD-targeted cancer therapy.