Evaluation of a novel Pan-RAS/β-Catenin inhibitor, ADT-030, for treatment of pancreatic cancer.

person Gary A Piazza Auburn University, Auburn, AL info_outline Gary A Piazza, Reddy DS Bandi, Pumachandra N Ganji, Jeremy B Foote, Adam B Keeton, Tyler Ernest Mattox, Xi Chen, Cherlene Hardy, Kristy Berry, Yulia Maxuitenko, Upender Manne, Donald J Buchsbaum, Bassel F. El-Rayes, Michael R Boyd

Authors person Gary A Piazza Auburn University, Auburn, AL info_outline Gary A Piazza, Reddy DS Bandi, Pumachandra N Ganji, Jeremy B Foote, Adam B Keeton, Tyler Ernest Mattox, Xi Chen, Cherlene Hardy, Kristy Berry, Yulia Maxuitenko, Upender Manne, Donald J Buchsbaum, Bassel F. El-Rayes, Michael R Boyd Organizations Auburn University, Auburn, AL, University of Alabama at Birmingham, Birmingham, AL, CARIS Life Sciences, Mobile, AL, ADT Pharmaceuticals, LLC., Orange Beach, AL Abstract Disclosures Research Funding National Cancer Institute/U.S. National Institutes of Health Background: Pancreatic ductal adenocarcinoma (PDAC) remains a formidable challenge due to its late diagnosis and limited treatment options. KRAS mutations and activation of Wnt/β-catenin pathway components that drive aberrant signaling cascades resulting in uncontrolled cellular proliferation and metastasis occur in a majority of PDAC patients. The development of mutant specific KRAS inhibitors has emerged as a promising approach to counter the oncogenic impact of KRAS mutations, although have limited use for PDAC given the complex mutational landscape, while inhibitors of Wnt/β-catenin signaling remain elusive. Novel target-directed drugs that effectively tackle the complex mutational status in KRAS mutant PDAC are urgently needed. A novel pan-RAS/β-catenin inhibitor, ADT-030, was evaluated in mouse tumor models of PDAC harboring KRAS G12C or KRAS G12D mutations. Methods: PDAC cell lines 2838c3 (KRAS G12D ), MIA PaCa-2 (KRAS G12C ), and BxPC-3 (RAS WT ) were utilized to evaluate in vitro growth inhibitory potency and selectivity of ADT-030. Assays for colony formation, apoptosis, and cell cycle, along with western blotting for RAS signaling were also conducted. A murine model involving 2838c3-Luc KRAS G12D PDAC cells injected into the pancreas of C57BL/6J mice was used to evaluate in vivo antitumor activity of ADT-030. ADT-030 was administered orally once daily at dosages of 50, 100, and 150mg/kg body weight, 5x/week for 4 weeks. Tumor growth was monitored by bioluminescence imaging using an IVIS Xenogen system. Body weights were measured twice weekly. Pancreatic tumors were collected and weighed at the end of treatment. Results: ADT-030 produced potent inhibition of proliferation of human and murine PDAC cells harboring KRAS G12D and KRAS G12C mutations. Annexin V assay revealed the capacity of ADT-030 to trigger apoptosis, specifically in KRAS mutant cells with minimal impact on BxPC3 RAS WT cells. Cell cycle analysis using Propidium Iodide staining revealed the ability of ADT-030 to arrest cells in the G2/M phase of the cell cycle. Western blot for detecting the key RAS signaling molecule P-ERK1/2 showed decreased levels following ADT-030 treatment. Other experiments revealed that ADT-030 and related analogs simultaneously suppress both MAPK/AKT signaling and β-catenin transcriptional activity (doi: 10.1186/s13048-022-01050-9). In an orthotopic xenograft model in which 2838c3-Luc cells were implanted in the pancreas, oral administration of 50, 100, and 150mg/kg of ADT-030 was well tolerated and inhibited tumor growth by 64.2%, 72.8%, and 80%, respectively, after 4 weeks of treatment. Conclusions: These compelling results position ADT-030 as a novel dual pan-RAS/β-catenin inhibitor for combating the challenges posed by complex mutations in PDAC and other RAS driven cancers and support its clinical evaluation as a valuable therapeutic alternative to mutant-specific KRAS inhibitors.

Pre-clinical