Pharmacological Evaluation of Anticonvulsant and Anxiolytic Effects of Mitragyna speciosa Leaf Alkaloid-Enriched Fraction through Experimental and Computational Studies

Abstract

The present study aimed to evaluate the anticonvulsant and anxiolytic potential of the alkaloid-enriched leaf fraction of Mitragyna speciosa (AEMS) using experimental and computational approaches. Powdered leaves were extracted by Stas-Otto method to obtain the alkaloid-enriched fraction, followed by LC-MS and preliminary phytochemical analysis using standard methods. AEMS was administered at doses of 50 mg/kg and 100 mg/kg and its effects were assessed using in vivo models over 15 days. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s test. In vitro antioxidant assays and in silico molecular docking studies were conducted to evaluate interactions with the GABA_A -BZD receptor. LC-MS and phytochemical screening confirmed the presence of alkaloids, viz. mitragynine, 7-hydroxymitragynine and corynoxine B. AEMS demonstrated significant (*p < 0.05) anticonvulsant and anxiolytic activities, with the 50 mg/kg dose exhibiting greater efficacy. The antioxidant activity was reported to be comparatively lesser (IC₅₀ = 399.93 ± 1.35 µg/mL) than the ascorbic acid standard (IC₅₀ = 49.6 ± 6.55 µg/mL) by DPPH method. Docking studies revealed strong binding affinities for all the three compounds wherein, 7-hydroxymitragynine demonstrated profound binding affinity (-7.7 kcal/mol), comparable to diazepam (-8.8 kcal/mol), suggesting GABAergic involvement. These findings indicate that AEMS possesses significant anticonvulsant and anxiolytic activity and may be useful in managing convulsions and anxiety.