Exploring the Therapeutic Landscape: Synthesis, Characterization, and Anticancer Activity of Novel Pyrazine-Piperidine Amide Pharmacophores in Human Lung Carcinoma Treatment

The current study aimed to synthesize and identify the biological activities of pyrazine-piperidine amide pharmacophore derivatives against non-small lung carcinoma (Calu-6) cells.

The combinatorial formulation was prepared by an active mixture of different chemical substituents, and five (6A-E) different molecules were synthesized. The chemical structures were confirmed by Fourier transform infrared (FT-IR) spectroscopy and proton nuclear magnetic resonance (H1) spectroscopy.

These compounds were also screened for cytotoxicity against the Calu-6 cell line. Compounds 6B and 6D displayed potent cytotoxicity, with IC50 Values of 45.21 µM and 89.64 µM, respectively. Cellular uptake and apoptotic studies using compound microscopy and flow cytometry revealed that cell damage gradually increased, leading to cell death. Compound 6B at 25 µM and 50 µM had 75.3% and 65.3% viability, 8.61% and 9.85% apoptotic effects, 12.05% and 21.4% late apoptosis, and 4.02% and 3.4% necrosis, respectively.

Compound 6B was found to significantly enhance cell cycle arrest at the G2/M phase. Additionally, real-time RT-PCR and western blot analyses further confirmed the enhanced expression of apoptotic markers, such as caspase-3 and 8, as well as the antiproliferative gene p53.

These findings indicate that compound 6B has a promising anticancer effect on lung cancer.