Unravelling Anti-parkinson's Potential of Combined Extracts of Juglans regia L. and Cardiospermum halicacabum L. through an in silico Approach.

INTRODUCTION: Dopaminergic neuronal loss, mitochondrial dysfunction, oxidative stress, and neurotransmitter imbalance are the hallmarks of Parkinson's Disease (PD), a progressive neurodegenerative disorder. Despite their established antioxidant and neuroprotective qualities, the mechanistic significance of Juglans regia L. and Cardiospermum halicacabum L. in Parkinson's disease remains unclear, especially in the context of extract-based treatment approaches.

METHODS: Drug-likeness and ADMET properties were evaluated with SwissADME and ADMETlab 3.0, respectively. SwissTargetPrediction was used to predict potential targets of selected phytoconstituents, whereas gene sets associated with Parkinson's disease were mined from Dis- GeNET and GeneCards. Venn analysis was performed to identify overlapping targets for drugs, followed by Protein-Protein Interaction (PPI) network construction using STRING and topological analysis in Cytoscape. Gene Ontology (GO) and KEGG pathway enrichment analysis were performed using DAVID 2021, then a Bioactive Compound-Target-Pathway (BA-TAR-PATH) network was constructed. Molecular docking based on AutoDock Vina was used to predict the binding affinity between the protein and ligand for the top 10 hits; then, molecular dynamics simulation using GROMACS was applied to assess the stability of the protein-ligand complexes.

RESULT: Phytochemical characterization by HPTLC, FTIR, HPLC, and ESI-MS supported the presence of flavonoids, phenolic compounds, tannins, and lipid-associated constituents in the combined extract. Target prediction yielded 355 related compounds and 30 PD-associated genes, identifying five overlapping targets (ACHE, DRD2, MAOB, COMT, and SLC6A3). Molecular docking revealed favourable binding interactions, particularly quercetin-ACHE (-11.4 kcal/mol), while molecular dynamics simulations supported the stability of selected protein-ligand complexes.

DISCUSSION: Network analysis highlighted MAOB, COMT, and SLC6A3 as key hub genes. GO and KEGG analyses revealed significant enrichment of dopaminergic synapse, neuroactive ligand- receptor signalling, and Parkinson disease pathways. Molecular docking demonstrated stable binding interactions, notably quercetin-ACHE (-11.4 kcal/mol), supporting the therapeutic relevance of flavonoids.

CONCLUSION: Based on the above findings, we propose that both J. regia kernel and C. halicacabum seed extracts may contain phytoconstituents that interact with various PD-related targets and pathways linked to dopaminergic neurotransmission and oxidative stress. This extractanchored network pharmacology approach presents a mechanism-related framework to facilitate follow-up experiments targeting specific bioactive constituents and molecular targets. At the same time, the results come exclusively from computational analyses and should be treated as hypothesis-generating. In vitro, in vivo, and pharmacokinetic studies need to be performed to validate the biological activity and therapeutic relevance experimentally.