Neuroprotective and Anti-Migraine Potential of Piper Betel: A Multi-Dimensional Study Involving In Vitro, In Silico Studies

AbstractMigraine is a complex neurovascular disorder associated with oxidative stress,neuroinflammation, trigeminovascular activation, and neuronal hyperexcitability. The presentstudy evaluated the potential neuroprotective and anti-migraine properties of ethanolic extractof Piper betle leaves (PBLEE) using integrated in vitro and in silico approaches.Preliminary phytochemical screening revealed the presence of alkaloids, flavonoids,phenols, tannins, glycosides, steroids, and saponins, indicating a diverse phytochemicalprofile with potential antioxidant and anti-inflammatory activities. GC–MS analysisidentified several bioactive constituents, including eugenol, hydroxychavicol, methyleugenol, β-caryophyllene, and phytol. ADME evaluation indicated favorablepharmacokinetic characteristics for selected constituents, including gastrointestinalabsorption and potential blood–brain barrier permeability, supporting their possible centralnervous system activity. The antioxidant potential of PBLEE was demonstrated by DPPHfree-radical scavenging activity, suggesting its ability to counteract oxidative stressimplicated in migraine pathogenesis. Molecular docking was subsequently performed againstthe selected target protein (PDB ID: 3N7P). Among the investigated compounds, eugenolacetate exhibited the strongest binding affinity (-6.4 kcal/mol), followed by eucalyptol (-6.2kcal/mol) and methyl eugenol (-5.8 kcal/mol). Bicyclogermacrene, germacrene, andlimonene showed moderate binding affinities, whereas myrcene, safrole, and linalooldemonstrated comparatively weaker interactions. The observed activities may be attributed tosynergistic antioxidant, anti-inflammatory, and neuroprotective effects of the plant'sphytoconstituents. Overall, the findings provide preliminary scientific evidence supportingthe anti-migraine potential of Piper beetle and identify promising bioactive compoundsfor further investigation. Future in vivo and clinical studies are necessary to validateefficacy, safety, mechanisms, and therapeutic applicability.