Phosphatidylinositol 5-phosphate 4-kinases (PI5P4Ks) are important molecular players in a variety of diseases, such as cancer. Currently available PI5P4K inhibitors are reversible small molecules, which may lack selectivity and sufficient cellular on-target activity. In this study, we present a new class of covalent pan-PI5P4K inhibitors with potent biochemical and cellular activity. Our designs are based on THZ-P1-2, a covalent PI5P4K inhibitor previously developed in our lab. Here, we report further structure-guided optimization and structure-activity relationship (SAR) study of this scaffold, resulting in compound 30, which retained biochemical and cellular potency, while demonstrating a significantly improved selectivity profile. Furthermore, we confirm that the inhibitors show efficient binding affinity in the context of HEK 293T cells using isothermal CETSA methods. Taken together, compound 30 represents a highly selective pan-PI5P4K covalent lead molecule.

Manz, Theresa D; Sivakumaren, Sindhu C; Yasgar, Adam; Hall, Matthew; Davis, Mindy I; Seo, Hyuk-Soo; Card, Joseph D; Ficarro, Scott B; Shim, Hyeseok; Marto, Jarrod A; Dhe-Paganon, Sirano; Sasaki, Atsuo T; Boxer, Matthew B; Simeonov, Anton; Cantley, Lewis C; Shen, Min; Zhang, Tinghu; Ferguson, Fleur M; Gray, Nathanael S;