Integrated experimental and computational evaluation of novel benzopyran-2-one congeners as reactive oxygen species scavengers in inflammation.

Reactive oxygen species (ROS) are key drivers of oxidative stress and inflammation. They damage cells and trigger pathways that further amplify the inflammatory response. To examine the dual antioxidant and anti-inflammatory effects of a series of benzopyran-2-one derivatives (2-10) by a combination of experimental and computational methods, a new series of benzopyran-2-one derivatives were designed, synthesized, and characterized. Carrageenan induced paw edema, formaldehyde induced arthritis, turpentine induced pleurisy, and biochemical tests like lipid peroxidation inhibition, SOD activity and DPPH radical scavenging were used as screening assays. Of all the congeners, compound 6b repeatedly demonstrated the best anti-inflammatory and antioxidant properties, better than diclofenac sodium and quercetin with minimal ulcerogenic or toxicity. Further, Molecular docking showed strong binding of 6b to inflammation and ROS related targets. Molecular dynamics simulations further confirmed its stable binding, with MM/PBSA free energy suggesting a more favorable binding energy than quercetin. Moreover, Steered Molecular Dynamics (SMD) pull-force analysis revealed that 6b had a greater rupture force to dissociate indicating stronger intermolecular forces and increased mechanical stability in the binding pocket. SwissADME/StopTox predictions suggested a favorable pharmacokinetic and predicted safety profile within the evaluated parameters. Overall, these combined computational and experimental results identify compound 6b as a promising dual antioxidant and anti-inflammatory lead candidate in preclinical models, which can serve as a promising scaffold in designing next generation therapeutics against ROS mediated inflammatory disease.