Abstract LB202: Discovery and preclinical characterization of novel small molecule inhibitors of kinesin KIF18A motor protein with potent activity against chromosomally unstable cancers

Abstract

Abstract KIF18A is a mitotic kinesin that localizes to the plus-end tips of kinetochore microtubule (MT) spindle fibers during metaphase, where it regulates chromosome alignment, and promotes the viability of chromosomally unstable cancer cells. KIF18A is overexpressed in a subset of human cancers, and its elevated expression is associated with tumor aggressiveness. Chromosomal instability (CIN) is a hallmark of human cancers and is caused by persistent errors in chromosome segregation during mitosis. Aggressive types of human cancer such as high-grade serous ovarian cancer (HGSOC) and triple-negative breast cancer (TNBC) have elevated levels of CIN and frequently harbor alterations in TP53 tumor suppressor gene. These two CIN+ cancer subtypes share molecular similarities but have limited treatment options at present. The rationale of pharmacological inhibition of KIF18A motor activity is to selectively target a tumor-specific mitotic spindle vulnerability in CIN+ cancer cells while largely sparing normal diploid dividing somatic cells. Here, we describe the identification of a novel series of potent and selective small molecule inhibitors of KIF18A MT-ATPase motor activity exemplified by AM-1882, that disrupt the mitotic spindle and selectively kill chromosomally unstable cancer cells. Our KIF18A inhibitors phenocopy genetic ablation of KIF18A and trigger spindle assembly checkpoint activation, multipolarity, and apoptosis in sensitive CIN+ cancer cell lines. The sensitivity profile of AM-1882 is focal-in-nature with cell potency in the low double-digit nanomolar range across a panel of breast and ovarian cancer cell lines, including lines that harbor genetic alterations (e.g., TP53, CCNE1, RB1, BRCA1, whole genome doubling) frequently enriched in CIN+ cancers and in HGSOC and TNBC tumor subtypes. Furthermore, the sensitivity profile of AM-1882 is distinct from comparator test agents ispinesib (Eg5, pan cytotoxic) and palbociclib (CDK4/6, focal cytostatic). The combination of AM-1882 with PARP inhibitor olaparib is synergistic in BRCA1-deficient cancer cell lines, with evidence of increased double-strand DNA breaks (p-H2AX) and apoptosis (cl-PARP). Importantly, KIF18A inhibitors have minimal toxicity on normal dividing somatic cell types in vitro, including proliferating human bone marrow mononuclear cells, distinct from paclitaxel and small molecule inhibitors of essential mitotic kinases and kinesins. In vivo, we demonstrate that administration of KIF18A inhibitors AM-1882 and AM-5308 induce a robust pharmacodynamic response (pH3, mitotic marker) and frank tumor regressions in two TP53 mutant human HGSOC xenograft models (OVCAR-3, OVCAR-8) at well-tolerated doses. Collectively, our preclinical data provides the first example of a therapeutic strategy to selectively target CIN+ cancers through inhibition of KIF18A motor protein. Citation Format: Jan Sun, Brain Belmontes, Jodi Moriguchi, Grace Chung, Kui Chen, John D. McCarter, Upendra P. Dahal, Andrew S. Boghossian, Matthew G. Rees, Melissa M. Ronan, Jennifer A. Roth, Sheroy Minocherhomji, Matthew P. Bourbeau, Jennifer R. Allen, Angela Coxon, Paul E. Hughes, Nuria Tamayo, Marc N. Payton. Discovery and preclinical characterization of novel small molecule inhibitors of kinesin KIF18A motor protein with potent activity against chromosomally unstable cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr LB202.