Thiophene-Pyridine Derivatives as Potential Anti-Proliferative and Anti-Inflammatory Apoptotic Agents: Synthesis, In Vitro Evaluation, Molecular Docking, and Dynamic Simulations

Inflammation is a key contributor to tumorigenesis and chemotherapy resistance. This underscores the need to develop monotherapeutic compounds with both anti-tumor and anti-inflammatory activities. To address this concern, a new series of thiophene derivatives linked to 1,2-dihydropyridine was prepared. These compounds were studied as promising dual-action compounds targeting apoptosis and inflammation. It was found that compound 5b showed notable cytotoxicity against human HepG2 liver cancer cells (IC50 = 20.4 μg/mL). It also had reduced cytotoxicity against normal human fibroblasts. Also, automated analysis showed that 5b induced apoptosis, confirmed by the overexpression of apoptosis-related proteins BAX and BID, and the downregulation of Bcl-2, as measured by flow cytometry. In anti-inflammatory studies, compound 5b dose-dependently inhibited protein denaturation by 94.3% and increased membrane stability by 78.0%. It also suppressed COX-2 gene expression. Molecular docking analyses revealed that compound 5b had a high binding affinity to BAX, Bcl-2, BID, and COX-2. Also, ADMET predictions and Molecular dynamics simulations over 100 ns showed improvements and confirmed the structural stability of the protein-ligand complexes, with Root Mean Square Deviation (RMSD) values below 0.35 nm and stable intermolecular hydrogen bonds. Unlike the other compounds, 5b had both anti-apoptotic and anti-inflammatory activities. Therefore, compound 5b is a promising candidate for treating inflammation-related hepatocellular carcinoma.