Multitarget anticancer profiling of Withania somnifera - Dual herbal combination with P. emblica, B. monnieri, and O. basilicum through integrated In silico docking and In vitro Jurkat cell line analysis.

BACKGROUND

Combinatorial Ayurvedic Rasayana extracts represent a rich source of bioactive compounds with anticancer potential. Withania somnifera (W) and its dual herbal Rasayana partners-Phyllanthus emblica (WP), Bacopa monnieri (WB), and Ocimum sanctum (WO)-are traditionally used to restore immune function, enhance resilience, and improve systemic health. Their multi-targeted phytoconstituents provide a promising approach for cancer therapy.

OBJECTIVE

This study aimed to identify bioactive compounds from W. somnifera and its dual Rasayana extracts, evaluate their in silico binding affinity with cancer-related targets, and assess their in vitro antiproliferative potential compared with cladribine.

MATERIALS AND

METHODS

Phytocompounds were identified by GCMS and pharmacological activities were predicted using PASS-BAS. Molecular docking (Schrodinger- Glide) against 23 cancer-relevant proteins curated from TCGA and COSMIC datasets. Drug-likeness and ADME were evaluated with QikProp 3.0 and SwissADME. Molecular Dynamics (MD) simulations (100 ns, Desmond) were conducted for selected protein-ligand complexes to validate binding stability through RMSD, RMSF, SSE, and protein-ligand interaction analysis. Jurkat E6-1 leukemia cells were treated with W, WP3, WB2, and WO3 extracts for 24 h, and antiproliferative activity assessed via MTT assay.

RESULTS

Of 62 screened phytocompounds, four key ligands -Campesterol (W), WP ligand, WB ligand, and Retinol (WO)-based on drug-likeness, pharmacokinetics, and compared with Cladribine. Docking revealed high affinities: WP3 with FAAH (-10), WB2 with PPARD (-9.08) and RBP4 (-9.5), WO3 with PPARA (-9.85) and RBP4 (-10.23), and W with CYP17A1 (-8.74). Enrichment analysis linked these targets to MAPK, JAK-STAT, and PPAR pathways. The MD results of both the complexes (5NUJ-445354 and 4D03-9592405) showed acceptable stability with dynamic adaptability, supporting the robustness of docking predictions. WP3 and WO3 had the highest antiproliferative activity (IC50 20.27 and 20.40 μg/mL) against Jurkat cells at 24hr and at 25 μg/mL.

CONCLUSION

This first integrative in silico-invitro analysis of dual herbal Rasayana extracts reveals multi-target anticancer activity and validates the stability of the key protein-ligand interactions suggesting synergistic therapeutic potential of W. somnifera with P. emblica as promising lead formulations for further drug development.