Integrative Network Pharmacology, Molecular Docking, and in Vitro Validation of Senna Auriculata Phytoconstituents As Potential Multi-Target Candidates For Alzheimer’s Disease

Abstract

Background: Alzheimer's disease is a progressive neurodegenerative disorder characterized by cholinergic deficit, amyloid-beta and tau pathology, and oxidative stress, for which currently approved therapies offer only modest symptomatic relief without altering disease progression. Medicinal plants rich in polyphenolic constituents represent a promising source of multi-target anti-Alzheimer's agents. Objective: To evaluate the anti-Alzheimer's disease potential of the whole-plant ethanolic extract of Senna auriculata through an integrated in-silico and in-vitro approach. Methods: Reported phytoconstituents of S. auriculata were retrieved from the IMPPAT database and their canonical structures standardized from PubChem. ADME and drug-likeness properties were predicted using SwissADME, and molecular targets were predicted with SwissTargetPrediction. Alzheimer's disease-associated targets from GeneCards and OMIM were intersected with the predicted targets by Venn analysis, and the common targets were subjected to protein–protein interaction network analysis in STRING, visualization in Cytoscape, hub identification with CytoHubba, and Gene Ontology and KEGG pathway enrichment analyses. Molecular docking of prioritized constituents with hub proteins was performed using CB-Dock2 and AutoDock Vina, followed by normal mode analysis of the lead complexes with iMODS. In-vitro, SA-EE was evaluated for DPPH and ABTS radical scavenging activity, acetylcholinesterase and butyrylcholinesterase inhibition by Ellman's method, and cytoprotection against hydrogen peroxide-induced oxidative stress in SH-SY5Y neuroblastoma cells by MTT assay. Results: Network pharmacology identified AKT1 and SRC as principal hub targets, and the flavonoid glycoside rutin emerged as the lead constituent, forming energetically favorable complexes with both hubs whose stability was supported by normal mode analysis. In-vitro, SA-EE exhibited concentration-dependent DPPH and ABTS radical scavenging activity, inhibited acetylcholinesterase by up to 91.00% at 100 µg/mL (R² = 0.9259) and butyrylcholinesterase by up to 81.44% at 600 µg/mL (R² = 0.9616), and protected SH-SY5Y cells against H₂O₂-induced oxidative stress. Conclusion: SA-EE possesses appreciable dual cholinesterase-inhibitory, antioxidant, and cytoprotective activities that are consistent with its network pharmacology profile, positioning S. auriculata as a promising multi-target natural source warranting further isolation of active constituents and in-vivo evaluation in Alzheimer's disease research.