Jingchuan Zhang

Abstract 5168: KO-947, a potent ERK inhibitor with robust preclinical single agent activity in MAPK pathway dysregulated tumors

The RAS/RAF/MEK pathway is a major driver of malignant progression, particularly in cancers arising from mutations in RAS, BRAF and NF1. Although both BRAF and MEK inhibitors have been approved for treatment of melanoma, their clinical activity is commonly limited by acquired resistance due to reactivation of the pathway downstream of their targets. Since the ERK1/2 kinases are the final node in the MAPK signaling pathway, they are not subject to the feedback reactivation mechanisms that can undermine RAF or MEK blockade, offering the possibility of clinical benefit in settings where earlier drugs are ineffective. Here we describe the characterization of KO-947, a potent and selective inhibitor of ERK1/2 kinases, in biochemical, cellular and in vivo antitumor activity assays. KO-947 was profiled in vitro in biochemical activity assays, competition binding assays, and a probe-based competition binding assay in cell lysates, and subsequently screened in a large panel of human tumor cell lines focused on MAPK-dysregulated tumor types. In vivo studies were carried out in well-characterized xenograft models of BRAF-, KRAS- and NRAS-mutant disease and subsequently extended to a large panel of early-passage patient-derived xenograft (PDX) models representative of a wide range of tumor types and molecular profiles. Biochemical assays reveal that KO-947 is a 10nM inhibitor of ERK with at least 50-fold selectivity against a panel of 450 kinases. KO-947 blocks ERK signaling and proliferation of tumor cells exhibiting dysregulation of MAPK pathway signaling, including mutations in BRAF, NRAS or KRAS, at low nanomolar concentrations. KO-947 is differentiated from other published ERK inhibitors by an extended residence time and high potency in cell engagement that translate into prolonged pathway inhibition in vitro and in vivo. In cell-line derived xenograft studies, the drug profoundly suppresses ERK signaling for up to five days after a single dose and induces regressions in RAS- and RAF-mutant melanoma, NSCLC and pancreatic cancer models on administration schedules ranging from daily to weekly. Intermittent dosing enables comparable antitumor activity at reduced dose-intensity. PDX screening confirms and extends these findings to include RAS and BRAF mutant colorectal, gastric and cervical carcinoma models, and robust activity is also seen on both weekly and Q2D schedules in tumor models lacking BRAF/RAS mutations but with other dysregulation of the MAPK pathway. Thus, the favorable ADME properties of KO-947 enable the achievement of optimal antitumor activity with intermittent dosing, which may provide an opportunity to maximize the therapeutic window with flexible administration routes and schedules. These results demonstrate the potential clinical utility of KO-947 in MAPK pathway dysregulated tumors. Citation Format: Francis Burrows, Linda Kessler, Jeffrey Chen, Xin Gao, Rasmus Hansen, Shuangwei Li, Carol Thach, Levan Darjania, Yvonne Yao, Yi Wang, Ata Zarieh, Ke Yu, Tao Wu, Jingchuan Zhang, Dana Hu-Lowe, Liansheng Li, Pingda Ren, Yi Liu. KO-947, a potent ERK inhibitor with robust preclinical single agent activity in MAPK pathway dysregulated tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5168. doi:10.1158/1538-7445.AM2017-5168

An Internally Controlled Quantitative Target Occupancy Assay for Covalent Inhibitors

Assessing target occupancy is critical for establishing proof-of-mechanism for novel inhibitors and to determine whether robust target inhibition can be achieved at tolerated doses. This is challenging in the clinic using conventional methods due to the need for untreated controls. We describe a new mass spectrometry approach to quantitatively assess target occupancy for covalent inhibitors that does not require untreated controls, and apply the method to the KRASG12C inhibitor ARS-1620.

Abstract 5077: A novel small molecule menin-MLL inhibitor for potential treatment of MLL-rearranged leukemias

Patients with MLL-rearranged leukemia typically have a poor prognosis. With chemotherapy and stem cell transplantation as current standard of care, the 5-year survival rate is estimated to be only about 35%. As the leukemogenic activity of MLL fusion proteins has been shown to be dependent on their direct interaction with menin, development of small molecules that block the menin-MLL interaction is a promising therapeutic strategy for the treatment of this disease. Although small molecule menin-MLL inhibitors have been reported, previously published compounds exhibited modest cellular potency and/or poor pharmacokinetic properties. We now describe a novel, potent, and selective small molecule menin-MLL inhibitor that effectively treats MLL leukemias in in vivo models and demonstrate its potential clinical utility. The compound selectively inhibits the growth of a panel of MLL-rearranged cell lines relative to non MLL-rearranged cell lines, displays favorable pharmacokinetic properties, and induces tumor regression in an MV4;11 subcutaneous xenograft mouse model, with durable tumor suppression even after dosing is discontinued. In both the MV4;11 and MOLM13 disseminated leukemia models, the compound confers a prolonged survival benefit compared to an untreated group when dosed orally once daily at doses that are well-tolerated in the animals. We have used this potent and selective menin inhibitor to investigate the functional cellular response of MLL-rearranged cell lines to menin inhibition, assessing cell growth, cell cycle and gene expression responses over time, and compared that response to other epigenetic inhibitors and demonstrate significantly increased apoptosis which correlates with tumor regression measured both in early and long term readouts in xenograft and systemic leukemia models. We describe a menin-MLL inhibitor with optimized drug-like properties that demonstrates potential clinical utility in preclinical models of MLL leukemias. The compound is currently under further preclinical evaluation. Citation Format: Tao Wu, Linda Kessler, Shuangwei Li, Trupta Purohit, Shisheng Li, Hongzhi Miao, Brian Linhares, Rasmus Hansen, Jeff Kucharski, Yi Wang, Ke Yu, Katarzyna Kempinska, Tess Ely, Szymon Klossowski, Ata Zarieh, Ulf Peters, Jun Feng, Yvonne Yao, Yuan Liu, Bo Wen, Francis Burrows, Duxin Sun, Jingchuan Zhang, Levan Darjania, Dana Hu-Lowe, Patrick Zarrinkar, Liansheng Li, Tomasz Cierpicki, Jolanta Grembecka, Pingda Ren, Yi Liu. A novel small molecule menin-MLL inhibitor for potential treatment of MLL-rearranged leukemias [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5077. doi:10.1158/1538-7445.AM2017-5077