Design and Synthesis of Benzofuran‐1,4‐dihydropyridine Hybrids as Novel Anti‐Inflammatory Agents: Theoretical Exploration of a Potential Dual COX‐2/TNF‐α Mechanism

ABSTRACT This research presents the design, synthesis, and thorough evaluation of a new series of benzofuran‐1,4‐dihydropyridine hybrids ( 4a–i ) as potential multi‐target anti‐inflammatory drugs. These compounds were created through an optimized multistep process and examined using a combination of in silico, in vitro, and in vivo methods. Molecular docking revealed a potential dual‐target profile, with the leading compounds showing strong predicted binding to both COX‐2 and TNF‐α. Notably, compound 4i demonstrated the highest affinity for COX‐2 (MolDock score = −181.002 kcal/mol), with good selectivity over COX‐1 (Δ = −21.35), and also showed one of the strongest affinities for TNF‐α (−174.847 kcal/mol). Molecular dynamics simulations confirmed the stability of the 4i ‐COX‐2 complex. This in silico effectiveness was supported by strong in vitro anti‐inflammatory activity (eg. 10.09 µg/mL for IC 50 of 4b in hypotonicity‐induced hemolysis). Notably, the compounds showed excellent in vivo activity in a carrageenan‐induced paw edema model, with 4i achieving 91.5% inhibition at 90 min. Histological analysis confirmed that the lead compounds were highly gastrointestinal (GI)‐sparing, consistent with their predicted COX‐2 selectivity. No liver damage was observed, and platelet count remained normal ( 4i ). Overall, this study identifies compound 4i as a potent, orally active, and safe antinflammatory candidate with potential dual mechanisms.