Structural rationalization of IPMK inhibitor potency

Inositol polyphosphate multikinase (IPMK) is a ubiquitously expressed kinase that has been linked to several cancers including glioblastoma. The recent development of a large panel of ATP-competitive IPMK inhibitors has re-invigorated enthusiasm for targeting IPMK pharmacologically. However, the structural basis for how these inhibitors achieve such high potency is unknown. Here, we report 14 novel co-crystal structures (1.7Å - 2.0Å resolution) of the human IPMK kinase domain complexed with these 1 ^st generation and 2 ^nd generation IPMK inhibitors. We also apply a ³³ P-radiolabeled binding assay that permits high-confidence estimations of the IC ₅₀ values for these compounds, and isothermal titration calorimetry to establish the K _D for compound binding to IPMK. The new structures reveal an unoccupied pocket in the ATP-binding site of human IPMK, that is engaged by the most potent 2nd generation inhibitors examined herein. Two ordered water molecules also participate in hydrogen-bonding networks associated with the most potent IPMK inhibitors. In addition to providing the molecular basis for observed increases in potency and selectivity of these inhibitors, the data presented here provide a toolbelt of 14 novel inhibitor-bound structures of human IPMK that can serve as a reference for all future IPMK structure-based inhibitor development efforts.