Extended Data Fig. 2: Discovery of potent and selective KIF18A inhibitors.
a, AM-7710 series analogs were assessed in kinesin-8 microtubule (MT)-ATPase motor assays and 24-h mitotic image assay in MDA-MB-157 cells. Scatter plots show MT-ATPase IC50 values for KIF18A versus KIF19A motors (n = 97 analogs), KIF18A versus KIF18B motors (n = 77 analogs), or cellular EC50 values for pH3 versus PCM foci endpoints (n = 150 analogs) (n ≥ 1 experiment (s) per analog). b and c, KIF18A compounds were assessed in a panel of kinesin MT-ATPase motor assays and 24-h mitotic image assay in MDA-MB-157 cells. b, Graphs show compound profiles for each motor presented as MT-ATPase luminescence signal relative to the percentage of DMSO control (POC), as assessed by ADP-Glo (n = 2 or 4 independent experiments). Ispinesib and GSK923295 were included as motor inhibitor controls. c, Upper table shows compound MT-ATPase IC50 values for KIF18A, KIF18B, KIF19A, Eg5, CENP-E, and KIFC1 motors. Middle table shows compound (1 µM) MT-ATPase POC values for a panel of motors (n = 8), as assessed by enzyme-linked inorganic phosphate assay (ELIPA), results presented as a percentage of DMSO control (POC) (n = 1 or 2 independent experiments in duplicate). Lower table shows the compound cellular EC50 values for mitotic image assay (n = 2 independent experiments). d, AM-0277 and AM-1882 were assessed in KIF18A ATPase motor assays (± MTs) or varied ATP or MT concentrations, as assessed by ADP-Glo. Graphs show compound profiles in KIF18A motor assays presented as ATPase luminescence signal relative to the percentage of DMSO control (POC) (± MTs n = 2 independent experiments in duplicate, varied ATP, or MT concentrations n = 1 experiment).
