Exploration of Potential Anti‐inflammatory cum Anti‐Rheumatoid‐arthritis Phyto‐molecule through Integrated Green Approach: Network Pharmacology, Molecular Docking, Molecular dynamics, In‐vitro and Ex‐vivo study

Abstract Rheumatoid arthritis (RA) and associated inflammatory complications are the most prevalent illnesses and can turn into fatal conditions if left untreated. Allopathic medicine is not satisfactory for curing RA. Scientific literature reports reveal that several phyto‐compounds viz . flavonoids, saponins, and terpenoids, can heal joints and organs from auto‐inflammatory rheumatoid arthritis and pain. Gene ontology, gene network analysis, molecular clustering, and literature review were used to optimise RA‐specific highly expressed genes. In‐silico molecular docking was performed to short‐out potential phytomolecules (Neohesperidin dihydrochalcone (NHDC)) from 1000 datasets‐library against RA and validate using MD simulation running at 100 ns. In‐vitro anti‐inflammatory assays of NHDC inhibited egg‐albumin denaturation, IC 50 of 47.739±0.51 μg/ml. The ex‐vivo MTT assay with NHDC rendered 67.209 % inhibition at 100 μM against fd‐FLS‐cells. NHDC downregulated pro‐inflammatory cytokine IL‐17 A production by 61.11 % and 50 % at 300 and 200 μM, respectively. Thus, this Studies recommend that NHDC may be highlighted as a novel multi‐target PADI4 and JAK3 inhibitor with better efficacy and minimal toxicity in RA warranted to In‐Vivo and clinical investigation. The current findings have uncovered remarkable genes and signalling pathways linked to RA, which could enhance our existing comprehension of the molecular mechanisms that drive its development and progression.