In Silico Evaluation of Anti-Inflammatory Flavonoids from Marrubium vulgare L.: DFT Reactivity, Molecular Docking with COX Isozymes, Molecular Dynamic Simulation and Toxicity Prediction

Abstract Chronic inflammation is a major contributor to the pathophysiology of numerous diseases, including cardiovascular disorders, neurodegeneration, and cancer. Although non-steroidal anti-inflammatory drugs (NSAIDs) such as Diclofenac are widely used, their long-term administration is associated with significant adverse effects. This has led to a growing demand for safer, plant-based alternatives with comparable efficacy. Marrubium vulgare L. (white horehound), a medicinal plant native to the Mediterranean region, is traditionally recognized for its therapeutic benefits and is particularly rich in flavonoids with potential anti-inflammatory properties.This study employed comprehensive and integrated computational approaches combining Density Functional Theory (DFT) calculations, Molecular Docking, Molecular Dynamics simulations, and Toxicity prediction to evaluate fourteen flavonoids based compounds that were selected from recent phytochemical analysis of Marrubium vulgare L. against the COX-1 and COX-2 isoforms, key enzymes in the inflammatory cascade. The results revealed four compounds, Luteolin, Cynaroside (Luteolin-7-O-glucoside), Quercetin, and Rhoifolin (Apigenin-7-O-neohesperidoside), with strong binding affinities and favorable interaction profiles. Among them, Rhoifolin demonstrated the highest binding affinities (− 10.6 kcal/mol for COX-1 and − 10.1 kcal/mol for COX-2), significantly outperforming diclofenac, while also exhibiting excellent predicted biocompatibility and low toxicity risk. Molecular Dynamics simulations further confirmed the stability of its interactions within the active sites.These findings highlight the therapeutic potential of Marrubium vulgare L., flavonoids, particularly Rhoifolin, as effective and safer alternatives to conventional NSAIDs.