Abstract 6246: The MTA-cooperative PRMT5 inhibitor AM-9747 exhibits robust antitumor activity in combination with clinically relevant chemotherapies and targeted agents in MTAP null tumor models
Authors: Slemmons Katherine , Belmontes Brian , Liu Siyuan , Moriguchi Jodi , Policheni Antonia , Hughes Paul E. 
Published: April 2023

Homozygous deletion of chr9p21, containing genes CDKN2A and MTAP, occurs in about 15% of cancers. MTAP loss, a key enzyme in the methionine and adenine salvage pathways, leads to the accumulation of its substrate MTA. MTA competes with the methyl donor SAM for binding to type II arginine methyltransferase PRMT5, placing PRMT5 in a hypomorphic state and vulnerable to further PRMT5 inhibition. MTA-cooperative PRMT5 inhibitors are an emerging class of therapeutics targeting MTAP null tumors. AMG 193, currently in Phase 1 trials, and its representative analog AM-9747 have broad spectrum activity in MTAP null tumor models across hematologic and solid tumor indications. Mechanistically, AM-9747 induces DNA damage, shown by increased phosphorylated H2AX, resulting in cell cycle arrest and senescence. Exploration of clinically relevant therapeutic combinations with standard of care (SOC) chemotherapies or targeted agents that could potentiate this DNA damage or target orthogonal pathways is a rational therapeutic approach. Here, we evaluated AM-9747 in combination with SOC chemotherapies and assessed synergy using the Chou-Talalay Method to generate Combination Index (CI) scores. SOC agents representing a variety of mechanisms of action (paclitaxel, carboplatin, gemcitabine, pemetrexed, irinotecan, 5-FU) were evaluated and results ranged from strongly synergistic (CI&lt;0.3) to additive (CI=1) in a panel of non-small cell lung carcinoma (NSCLC) and pancreatic cancer cell lines. The SOC combinations augmented AM-9747 induced DNA damage resulting in increased cell cycle arrest and nuclear count assays confirmed the synergistic antiproliferative effects of the combinations. Combinations of AM-9747 and one strongly synergistic agent (carboplatin) and one moderately synergistic agent (paclitaxel) were evaluated in H292 NSCLC xenografts. Significant reductions in tumor burden compared to single agent treatments were observed with AM-9747 + paclitaxel (67% TGI) and AM-9747 + carboplatin (82% TGI). Additionally, roughly 2-3% of NSCLC tumors harbor both MTAP loss and a KRAS G12C mutation. To target both pathways we combined the FDA-approved KRAS G12C inhibitor sotorasib with AM-9747 and observed synergy (CI&lt;0.6) in MIAPACA2 cells. Nuclear counts confirmed a greater cell growth inhibition in the combination and immunoblots confirmed on target inhibition of both pathways. Combination treatment of AM-9747 and sotorasib in LU99 NSCLC xenografts resulted in significant tumor regression compared to each single agent with 10/10 mice tumor free. This combination is being tested in additional MTAP null KRAS G12C mutant models. Overall, our data suggests that combining MTA-cooperative inhibitor AM-9747 with SOC or clinically relevant targeted agents is a compelling therapeutic strategy for the treatment of MTAP null cancers.

Citation Format: Katherine Slemmons, Brian Belmontes, Siyuan Liu, Jodi Moriguchi, Antonia Policheni, Paul E. Hughes. The MTA-cooperative PRMT5 inhibitor AM-9747 exhibits robust antitumor activity in combination with clinically relevant chemotherapies and targeted agents in MTAP null tumor models. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6246.