Integrated Phytochemical, Pharmacological, and In Silico Evaluation of the Methanolic Extract ofLeaves: Antioxidant, Anti‐Inflammatory, and Analgesic Activities
Source: PubMed Central Open Access, NCBI / U.S. National Library of Medicine
is traditionally used for treating pain, inflammation, and oxidative disorders, yet comprehensive experimental and computational validation of its pharmacological potential remains limited. The aim of this study is to investigate the phytochemical profile and evaluate antioxidant, anti‐inflammatory, and analgesic activities of methanolic and n‐hexane fractions ofleaves using integrated in vitro, in vivo, and in silico approaches. Leaves were extracted with methanol and fractionated via a modified Kupchan method. Phytochemical screening, DPPH radical scavenging, HRBC membrane stabilization, carrageenan‐induced paw edema, acetic acid‐induced writhing, and tail immersion assays were performed. In silico analysis included PASS prediction, ADME/T profiling, and molecular docking of 40 previously documented GC–MS compounds against COX‐2 (PDB: 6COX, 1CX2, 5IKR, and 1CQE) and CYP2C9 (PDB: 1OG5 and 4L7B) protein targets. Methanolic extract demonstrated potent antioxidant activity (IC= 2.69g/mL) and 87.46% HRBC membrane stabilization. Significant anti‐inflammatory effects were observed in vivo through reduced paw edema. The n‐hexane fraction produced notable analgesic responses, inhibiting writhing by 21.84% and increasing reaction time in the tail immersion test by 59.44% (< 0.0001). In silico analyses identified multiple bioactive compounds—especially 2‐cyclohexylethyl isobutyl ester and lup‐20(29)‐en‐3‐yl acetate—with binding affinities surpassing diclofenac, aspirin, and ascorbic a
Abstract
is traditionally used for treating pain, inflammation, and oxidative disorders, yet comprehensive experimental and computational validation of its pharmacological potential remains limited. The aim of this study is to investigate the phytochemical profile and evaluate antioxidant, anti‐inflammatory, and analgesic activities of methanolic and n‐hexane fractions ofleaves using integrated in vitro, in vivo, and in silico approaches. Leaves were extracted with methanol and fractionated via a modified Kupchan method. Phytochemical screening, DPPH radical scavenging, HRBC membrane stabilization, carrageenan‐induced paw edema, acetic acid‐induced writhing, and tail immersion assays were performed. In silico analysis included PASS prediction, ADME/T profiling, and molecular docking of 40 previously documented GC–MS compounds against COX‐2 (PDB: 6COX, 1CX2, 5IKR, and 1CQE) and CYP2C9 (PDB: 1OG5 and 4L7B) protein targets. Methanolic extract demonstrated potent antioxidant activity (IC= 2.69g/mL) and 87.46% HRBC membrane stabilization. Significant anti‐inflammatory effects were observed in vivo through reduced paw edema. The n‐hexane fraction produced notable analgesic responses, inhibiting writhing by 21.84% and increasing reaction time in the tail immersion test by 59.44% (< 0.0001). In silico analyses identified multiple bioactive compounds—especially 2‐cyclohexylethyl isobutyl ester and lup‐20(29)‐en‐3‐yl acetate—with binding affinities surpassing diclofenac, aspirin, and ascorbic acid, along with favourable ADME/T profiles. The combined experimental and computational findings indicate thatcontains phytoconstituents with strong therapeutic potential. Its methanolic fraction, supported by in silico validation, represents a promising source for developing natural antioxidant, anti‐inflammatory, and analgesic lead molecules.