Metabolic profiling and multitarget in silico docking reveal the anti inflammatory and anti arthritic potential of Ricinus communis L formulations

Abstract

While Ricinus communis L. is traditionally used for its anti-inflammatory properties, the molecular interactions of its topical formulations remain to be systematically characterized. This study investigates the phytochemical composition of R. communis methanolic oil and cream formulations and evaluates their structural stabilization effects on proteins alongside in silico binding predictions against pro-inflammatory targets. Metabolites were putatively identified (MSI Level 2) via LC–MS/MS profiling. Anti-inflammatory potential was assessed in vitro using egg albumin and bovine serum albumin (BSA) denaturation assays to measure protein stabilization under thermal stress. To investigate potential molecular interaction pathways, 24 metabolites were docked against COX-2 (PDB: 5IKR), IL-6 (PDB: 1ALU), and TNF-α (PDB: 2AZ5). Structural stability of the predicted complexes was evaluated using 200 ns molecular dynamics (MD) simulations. In vitro assays demonstrated that R. communis formulations significantly inhibited thermal-induced protein denaturation compared to the untreated control, with the cream formulation achieving 97.80% ± 0.60% inhibition at 6400 µg/mL. While these results indicate structural stabilization in cell-free systems, they do not establish therapeutic superiority over clinical NSAIDs or account for complex systemic pathways. Chemical profiling annotated 24 putatively identified metabolites, including indole alkaloids, terpenoids, and cyclic phosphatidic acids (CPAs). Computational modeling identified potential binding modes; for example, Hirsutine and Hirsuteine exhibited favorable docking scores within the COX-2 binding pocket (ΔG = − 9.3 and − 9.0 kcal/mol, respectively), stabilized by hydrogen bonds with residues ARG120, TYR355, and SER530. MD simulations confirmed that these complexes maintained structural equilibrium over 200 ns. Similar structural complementarity was observed for Tsangane L 3-glucoside and CPAs with the IL-6 receptor interface, and alkylresorcinols with the TNF-α binding pocket. These findings provide a structural and biochemical baseline, confirming that R. communis metabolites effectively stabilize protein structures in in vitro cell-free models and exhibit favorable docking configurations with key inflammatory proteins. These results serve as structural hypotheses; further experimental validation using kinetic enzymatic assays and cellular models is required to determine the actual biological activity and pharmacological potential of these formulations.