Synthesis and potential anticancer activity of 3-(Arylsulfonamido)-N-(pyridin-2/3-yl)benzamide derivatives in pancreatic ductal adenocarcinoma through modulation of AKT/mTOR/STAT3 signalling
Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive malignancies globally. PDAC remains a leading cause of cancer-related mortality due to a high rate of metastasis, late diagnosis, and higher therapeutic resistance. Therefore, the limited effectiveness of existing therapeutic regimens highlights the need for continued development of new anticancer agents. Recently, pyridine-linked benzamide compounds have been explored and reported to have promising anticancer activity. Henceforth, we synthesized 3-(Arylsulfonamido)-N-(pyridin-2/3-yl)benzamide derivatives for evaluating their anticancer potential against PDAC cells. The structures of the compounds (5a-d) were evaluated by 1H NMR, 13C NMR, and MALDI-TOF mass spectrometry. During initial screening in SW1990 cells, the 5a derivative demonstrated marked and significant anticancer activity among all the synthesized derivatives. Hence, the derivative 5a was selected for evaluation of its anticancer potential against a panel of PDAC cell lines. The treatment of SW1990, Panc10.05, Capan-2, CFPAC-1, and MIA PaCa-2 cells with the 5a derivative showed significant inhibition in cell viability and clonogenicity. The treatment of 5a derivative induced G2/M arrest with marked accumulation of SW1990 and Panc10.05 cells in the sub-G1 phase during cell cycle analysis. Further, Annexin V/PI staining displayed that 5a treatment significantly induced apoptotic cell death in SW1990 and Panc10.05 cells. Mechanistic studies revealed a marked downregulation of cyclin D1 and CDK-6 protein expression, associated with the regulation of cell cycle arrest. Interestingly, the 5a derivative showed either a low or a non-significant effect on the growth, clonogenicity, and apoptosis of the hTERT-HPNE, normal pancreatic cells. Furthermore, molecular docking of the 5a derivative showed binding affinity with the AKT1 (PDB ID: 4EJN), mTOR (PDB ID: 4JT6), and STAT3 (PDB ID: 6NJS) proteins. The molecular analysis, conducted through a Western blot experiment in PDAC cells, revealed that the 5a derivative suppressed the phosphorylation of AKT, mTOR, and STAT3 in SW1990 and Capan-2 cells. Taken together, the results of molecular docking, molecular dynamics, and biological assays demonstrated that 5a exhibited promising anticancer activity by regulating the AKT/mTOR /STAT3 axis in PDAC with therapeutic potential.