International Journal of Drug Delivery Technology
Volume 16, Issue 8s, 2026

Synergistic Anti-Prostate Cancer Activity of Withaferin-A and Guggulsterone-Z From Ashwagandha and Guggul Through Inhibition of the NIK/NF-κB Signalling Pathway

1 Vinayak Bhushan, 1* Satyender Kumar, 2 Lalit Gahlawat, 2 Priyanka Sharma, 2 Birender Singh, 3 Rakesh K Sindhu, 4 Sonia Kashyap

1School of Pharmacy, Sharda University, Plot no 32, 34, Knowledge Park III, Greater Noida, Uttar Pradesh, 201310

2Gateway College of Pharmacy, Sonipat, Haryana, 131001

3Guru Jambheshwar University of Science and Technology, Hisar, Haryana, 125001

4MM College of Pharmacy, Maharshi Markandeshwar University, Mullana (Ambala), 133001

*Corresponding Author:
Dr. Satyender Kumar, Associate Professor, School of Pharmacy, Sharda University, Plot no 32, 34, Knowledge Park III, Greater Noida, Uttar Pradesh 201310. Email: sjinagal@gmail.com
Graphical abstract
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Highlights

• Withaferin-A (WA) and Guggulsterone-Z (GZ) target NIK/NF-κB signalling in prostate cancer

• Molecular docking reveals strong binding with NF-κB and NIK proteins

• Combination treatment shows synergistic cytotoxicity in LNCaP cells

• Flow cytometry confirms apoptosis induction by phytochemical combination

• Study validates traditional medicinal relevance of Ashwagandha and Guggul

Abstract

Traditional medicinal plants are valuable sources of bioactive compounds with potential therapeutic effects against chronic diseases including cancer. Ashwagandha (Withania somnifera) and Guggul (Commiphora mukul) are widely used in Ayurvedic medicine for treating inflammatory disorders and tumor-related conditions.

Aim of the study: The present study aimed to evaluate the synergistic anti-prostate cancer potential of the phytochemicals Withaferin-A and Guggulsterone-Z, the main bioactive compounds of Ashwagandha and Guggul, respectively, using integrated computational and experimental approaches targeting the NIK/NF-κB signalling pathway.

Materials and methods: Molecular docking studies were conducted using AutoDock Vina to evaluate binding interactions of Withaferin-A and Guggulsterone-Z with prostate cancer targets including NF-κB and NF-κB-inducing kinase (NIK). Pharmacokinetic and toxicity profiles were predicted using SwissADME and ProTox-II. In-vitro anticancer activity was evaluated in LNCaP prostate cancer cells using MTT assays. Apoptosis was assessed using Annexin V-FITC/PI flow cytometry, while gene expression changes were analyzed using conventional PCR.

Results: Docking results revealed strong binding affinities of Withaferin-A and Guggulsterone-Z toward NF-κB and NIK proteins, suggesting potential inhibition of inflammatory signalling pathways associated with tumor progression. In-vitro studies demonstrated dose-dependent inhibition of LNCaP cell proliferation. Combination treatment showed significantly higher cytotoxicity compared with individual treatments. Flow cytometry analysis confirmed enhanced apoptosis induction following combination therapy. Gene expression analysis further indicated modulation of apoptosis-related genes and androgen-associated signalling pathways.

Conclusion: The findings provide scientific validation for the traditional medicinal use of Ashwagandha and Guggul and suggest that their bioactive phytoconstituents may serve as promising multi-target therapeutic candidates for prostate cancer treatment.

Keywords: Prostate cancer, Ashwagandha, Guggul, Molecular docking, Withaferin-A, Guggulsterone-Z, NF-κB signalling

How to cite this article: Bhushan V, Kumar S, Gahlawat L, Sharma P, Singh B, Sindhu RK, Kashyap S. Synergistic Anti-Prostate Cancer Activity of Withaferin-A and Guggulsterone-Z From Ashwagandha and Guggul Through Inhibition of the NIK/NF-κB Signalling Pathway. Int J Drug Deliv Technol. 2026;16(8s): 1055-1065; DOI: 10.25258/ijddt.16.8s.118

Source of support: Nil.

Conflict of interest: None