Abstract 2800: Design, synthesis, selection, and evaluation of a potent multi-kinase targeting N4-phenylsubstituted-7H-pyrrolo[2,3-d]pyrimidin-4-amine derivative with significant anticancer activity in P53 mutant and wild type models of upper gastrointestinal cancers
Authors: Samant Amruta , Visal Tanvi , Pancholi Priya , Kansara Dhvanir , Dichwalkar Tanmay , Senones Robert , Kurup Sonali , Sehdev Vikas 
Published: August 2018

Introduction: Upper Gastrointestinal Cancers (UGCs) exhibit resistance to conventional chemotherapy due to variable P53 status and constitutive overactivity of EGFR, ERBB2/HER-2, Aurora kinases, and JAK2 oncogenes. UGC is a leading cause of cancer related deaths worldwide and development of multi-kinase targeting inhibitors could pave the way for improved chemotherapy and better therapeutic outcomes. In this study, we investigated a series of N4-phenylsubstituted-7H-pyrrolo[2,3-d]pyrimidin-4-amines and selected a promising novel investigational derivative (RS-41) that targets multiple kinases and exhibits significant anticancer activity in P53 mutant and wild type models of UGC. Methods: Target kinase inhibition and selectivity screening (T-KISS) and MTT-cell viability assays were performed to determine potency and selectivity of kinase inhibition and anticancer efficacy of primary derivatives, respectively. RS-41 was identified as the most promising agent following analysis via T-KISS and MTT-cell viability assay. Next, clonogenic cell survival assay, cell cycle analyses, and western blot analyses were done to evaluate the effect of RS-41 treatment on cell survival, cell cycle progression, and expression of apoptotic markers in P53 mutant (FLO-1) and P53 wild type (AGS) UGC cells, respectively. The initial T-KISS and cell-viability data aided in the design of second-generation derivatives of RS-41. Results: The initial T-KISS and cell viability data aided in identification of structural moieties that were modified to make the second generation of derivatives with improved multi-kinase inhibition. Evaluation with T-KISS showed that RS-41 derivative selectively inhibits AURKA (IC50-0.96±0.03µM), JAK2 (IC50-1.21±0.17µM), and EGFR (IC50-5.92±0.75µM) kinases, respectively. The cell viability and clonogenic cell survival data indicate that treatment with RS-41 mediates significant (P≤0.05) inhibition of FLO-1 and AGS UGC cell viability and survival. The cell cycle data and western blotting data also exhibited a marked increase (P≤0.05) in the percentage of sub-G1-phase (cell death) and expression of various apoptotic markers in FLO-1 and AGS cells following treatment with RS-41 for 24 and 72 hrs. Conclusions: Our T-KISS data indicate that multi-targeted inhibition of EGFR, AURKA and JAK2 can be achieved with the pyrrolo[2,3-d]pyrimidine scaffold. In addition, we demonstrate that inhibition of various oncogenic kinases with RS-41 is an effective therapeutic strategy for inducing apoptosis in both P53 mutant and wild type UGC cells.

Citation Format: Amruta Samant, Tanvi Visal, Priya Pancholi, Dhvanir Kansara, Tanmay Dichwalkar, Robert Senones, Sonali Kurup, Vikas Sehdev. Design, synthesis, selection, and evaluation of a potent multi-kinase targeting N4-phenylsubstituted-7H-pyrrolo[2,3-d]pyrimidin-4-amine derivative with significant anticancer activity in P53 mutant and wild type models of upper gastrointestinal cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2800.