Abstract 3087: KZR-8834: A novel, small molecule inhibitor of Sec61-dependent protein secretion with anti-tumor activity

Abstract

Abstract The majority of secreted and transmembrane (TM) proteins, including growth factors and TM oncogenes, require translocation through Sec61 into the ER for further processing. Sec61, therefore, represents a unique drug discovery opportunity through blockade of typically “undruggable” targets and modulation of protein homeostasis. Multiple inhibitors of protein secretion have been described that directly target Sec61 and have anti-tumor activity but lack adequate pharmaceutical properties and/or tolerability to be considered clinical candidates. We aimed to discover novel, selective inhibitors of Sec61 with increased tolerability as a potential treatment for various malignancies. Sec61 inhibitors were identified using HEK293 cell lines stably expressing secreted or TM proteins of interest fused to a luciferase reporter. Selective anti-tumor activity was assessed via HEK293 reporter cell viability in comparison to tumor cell lines with known levels of sensitivity to published Sec61-modulating compounds. An initial screen resulted in KZR-4152 (4152), which blocked secretion of several target proteins but had no impact on tumor cell viability. Following a medicinal chemistry campaign, KZR-8834 (8834) was identified, which exhibited a 60-fold increase in potency against target proteins relative to 4152. On-target activity of 8834 was demonstrated via protein secretion assays in cells overexpressing a previously described Sec61 mutant with resistance to published compounds. In addition, 8834 did not affect the viability of HEK293 cells or endotoxin- or TCR-stimulated PBMCs. 8834 induced cell death in H929 multiple myeloma cells in vitro (IC50 = 98 nM) with rapid activation (≤8 hours) of caspase 3/7 via dual activation of caspases 8 and 9. When assessed in a panel of 346 human cancer cell lines across 25 different tumor types, head and neck squamous cell carcinoma, acute lymphocytic leukemia, non-Hodgkin’s lymphoma, melanoma, multiple myeloma, and prostate cancer were among the most sensitive tumor types. Weekly and twice-weekly administration of 8834 was effective in an H929 mouse xenograft model at doses that resulted in >90% tumor growth inhibition without significant body weight loss or clinical signs of toxicity. Anti-tumor activity was >50% at doses 1/3rd of the maximum tolerated dose, suggesting a broad therapeutic index. 8834 was also effective in the B16F0 melanoma model, showing tumor growth inhibition similar to that of anti-PD1 therapy. KZR-8834 is a novel, small molecule inhibitor of Sec61-dependent translocation of multiple Sec61 client proteins. 8834 exhibits a broad anti-tumor profile in vitro and therapeutic activity in multiple mouse tumor models, demonstrating a potential new therapy for the treatment of malignant diseases. Citation Format: Eric Lowe, Janet L. Anderl, Andrea R. Fan, Jing Jiang, Henry W. Johnson, Christopher J. Kirk, Dustin McMinn, Tony Muchamuel, Jack Taunton, Jennifer A. Whang. KZR-8834: A novel, small molecule inhibitor of Sec61-dependent protein secretion with anti-tumor activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3087.