IP6 Kinase Inhibitor Screening Assays.

Inositol pyrophosphates, such as 5-diphosphoinositol pentakisphosphate (IP7), play crucial roles in various biological processes. Pharmacologic inhibition of IP6K has potential therapeutic benefits for treating type II diabetes, venous thrombosis, chronic kidney disease, and psychiatric disorders. This chapter describes the identification of IP6K inhibitors through kinase-activity-based high-throughput assays and dose-response assay. Additionally, it details cell-based assays on the determination of the effect of IP6K inhibitors on IP7 production in HCT116 cells and the binding of IP6K inhibitors to IP6K inside cells through cellular thermal shift assays (CETSAs). These robust assays provide a foundation for further exploration of IP6K inhibitors in various disease states.