Exploring 4-morpholinopyrrolo-[2,3-d]-pyrimidine derivatives as novel anti-cancer agents: Design, synthesis and biological studies

Abstract

Despite extensive research efforts, cancer continues to pose a significant global health challenge. Continuing the anti-cancer research, twenty-four 4-Morpholinopyrrolo-[2,3-d]-pyrimidine derivatives were synthesized and characterized via HRMS, 1H, 13C NMR, and FTIR. A single crystal for one compound, PGT4, was developed for further confirmation. All synthesized compounds were assessed for their anti-cancer activity in three cancer cell lines, SCC-25 (oral squamous cell carcinoma), U87-MG (glioblastoma) and MCF-7 (breast cancer) and one normal cell line HEK-293T (human embryonic kidney) using WST assay. Fourteen compounds demonstrated IC50 values ranging from 2 to 10 µM against the SCC-25 cell line. PTT3 and PGT7 were the most effective compounds against the SCC-25 cell line, showing an IC50 of 2 µM. Cell cycle analysis was also performed using propidium iodide staining, and apoptosis studies were carried out using caspase 3/7 activity assay and Annexin V-FITC isothiocyanate assay. PTT3 and PGT7 arrested the cell cycle at the G1 phase and enhanced the apoptosis by 9.1 and 13.8-folds in SCC-25 cells, respectively. Treatment with PTT3 and PGT7 also resulted in a dose-dependent decrease in the colony formation of SCC-25 cells. We carried out the molecular docking and dynamic studies to find the insights on binding mechanisms of PTT3 and PGT7 within the active region of the PI3Kγ target protein (PDB ID:3IBE, 2.80Å). Additionally, in-silico ADME properties were predicted for the most active molecules, PTT3 and PGT7, aiding in assessing their pharmacokinetic profiles.