Integrating Phytochemical Profiling with Pharmacological Evaluation of Himalayan Mango (Mangifera sylvatica Roxb.) Leaves: GC-MS/MS Insights into Antioxidant, Anti-inflammatory, and Analgesic Potentials

Objectives: Mangifera sylvatica Roxb. (Himalayan mango) is a wild fruit species available in Bangladesh. Its therapeutic properties have been scientifically less explored; thus, the current research aimed to assess the phytochemical profiling, acute toxicity, antioxidant, analgesic, and anti-inflammatory properties of the ethanolic extract derived from Mangifera sylvatica leaves (EMS). Method: The acute toxicity was evaluated through the in vivo method on Swiss albino mice. The antioxidant capacity was assessed using the DPPH radical scavenging assay and reducing power assessment. In vitro anti-inflammatory effects were determined through a protein denaturation test. The analgesic activity of EMS was examined using acetic acid-induced writhing and formalin-induced paw-licking tests on Swiss albino mice, along with a histopathological test. GC-MS/MS analysis was employed to identify bioactive compounds in the extract. Additionally, in silico studies, including pass prediction, ADME/T analysis, and molecular docking of various secondary metabolites, were conducted using different online tools. Results: At high doses, the extract did not exhibit any notable toxic effects in the acute toxicity. There were no significant changes in the histopathological parameters of vital organs when treated with the extract. The results showed that EMS exhibited notable antioxidant activity in the DPPH assay, with an IC ₅₀ value of 348.72 µg/mL, and demonstrated a concentration-dependent increase in reducing power. The extract also displayed significant anti-inflammatory effects, with an IC ₅₀ value of 142.52 µg/mL, comparable to the standard drug diclofenac in the protein denaturation assay. At doses of 100, 200, and 400 mg/kg, EMS significantly reduced acetic acid-induced writhing and formalin-induced paw licking in mice in a dose-dependent manner (p<0.01). Molecular docking analysis revealed that the compounds in the extract had binding affinities ranging from -0.3 to -10.8 kcal/mol toward human target receptors, indicating potential pharmacological activity. ADME/T analysis also supports the drug-likeliness characteristics of identified phytocompounds. Conclusion: Overall, the EMS exhibited no significant toxic effects in vital organs, excellent pharmacological activities with antioxidant, anti-inflammatory, and analgesic properties, which demands further extensive research.