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. 2025 Jul-Aug;16(4):101088.
doi: 10.1016/j.jaim.2024.101088. Epub 2025 Jun 26.

Network pharmacology and in-silico studies for molecular mechanisms of analgesic, anti-inflammatory and anti-arthritic effects of Withania somnifera (L.) Dunal phytoconstituents

Mukul S Tambe  1 , Shreyas R Shinde  1 , Akshay M Baheti  1 , Shuchi Nagar  2 , Anil T Pawar  3
Affiliations

Network pharmacology and in-silico studies for molecular mechanisms of analgesic, anti-inflammatory and anti-arthritic effects of Withania somnifera (L.) Dunal phytoconstituents

Mukul S Tambe et al. J Ayurveda Integr Med. 2025 Jul-Aug.

Abstract

Background: Withania somnifera (L.) Dunal, commonly known as ashwagandha, is an Ayurvedic herb belonging to the family Solanaceae.

Objectives: This study aims to explore the analgesic, anti-inflammatory and anti-arthritic potential of phytoconstituents of Withania somnifera (L.) Dunal (W. somnifera) by network pharmacology and in-silico docking studies.

Methods: Five major phytoconstituents, namely ashwagandhanolide, quercetin, withaferin A, withanone and withanolide A, were selected for the network pharmacology study. All five phytoconstituents were further evaluated for their binding properties using molecular docking (MD) and simulation tools. The compounds that exhibited significant binding affinities were further studied for pharmacokinetic and toxicity (ADMET) predictions.

Results: The network pharmacology study showed that out of the five selected constituents, withaferin A, withanolide A and quercetin can interact with various inflammation and pain-related genes. In in-silico studies, all five constituents were found to have significant interactions with inflammatory and nociception proteins cyclooxygenases, lipoxygenase, myeloperoxidase and cathepsin B. Further, ADMET studies predicted that all five phytoconstituents could not cross the blood-brain barrier but have high gastrointestinal absorption and bioavailability. Quercetin was predicted to have mutagenic potential and the other three constituents (withaferin A, withanone and withanolide A) were predicted to have immunotoxicity. The MD simulation studies showed that the complexes lipoxygenase_ashwagandhanolide and cathepsin B_ashwagandhanolide exhibit lower RMSD, RMSF, and higher H-bonding, indicating greater stability of ashwagandhanolide with lipoxygenase and cathepsin B.

Conclusion: Ashwagandhanolide, quercetin, withaferin A, withanone, and withanolide A from W. somnifera may show the potential for analgesic, anti-inflammatory, and anti-arthritic activities. These findings provide a foundation for future in-vitro and in-vivo studies to confirm the therapeutic efficacy of these phytoconstituents from W. somnifera.

Keywords: Anti-inflammatory; Ashwagandha; In-silico; Molecular dynamics; Network pharmacology; Withania somnifera.

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Conflict of interest statement

Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1
Fig. 1
Phytoconstituents of W. somnifera: (A) ashwagandhanolide, (B) quercetin, (C) withaferin A, (D) withanone, and (E) withanolide A.
Fig. 2
Fig. 2
Network pharmacology of phytoconstituents of W. somnifera.
Fig. 3
Fig. 3
String plot of protein-protein interaction network of genes regulated by phytoconstituents of W. somnifera. Node count: 44; Edge count: 17; PPI enrichment P-value: 0.0865.
Fig. 4
Fig. 4
AGE-RAGE pathway obtained from the ShinyGo database indicating genes regulated by phytoconstituents of W. somnifera.
Fig. 5
Fig. 5
VEGF pathway obtained from the ShinyGo database indicating genes regulated by phytoconstituents of W. somnifera.
Fig. 6
Fig. 6
Interactions of ashwagandhanolide with A) 3D3L, B) 1MHL, C) 1GMY, D) 1CVU, E) 1A3Q and F) 1TNF.
Fig. 7
Fig. 7
ADME radars of phytoconstituents of W. somnifera. A) quercetin, B) withaferin A, C) withanone and D) withanolide A.
Fig. 8
Fig. 8
Toxicity predictions of phytoconstituents of W. somnifera. AhR: Aryl hydrocarbon receptor, AR: Androgen receptor, AR-LBD: Androgen receptor ligand binding domain, ER: Estrogen receptor alpha, ER-LBD: Estrogen receptor ligand binding domain, PPAR-Gamma: Peroxisome proliferator-activated receptor gamma, NrF2/ARE: Nuclear factor (erythroid-derived 2)-like 2/antioxidant responsive element, HSE: Heat shock factor response element, MMP: Mitochondrial membrane potential, ATAD5: ATPase family AAA Domain Containing 5.
Fig. 9
Fig. 9
RMSD of ashwagandhanolide with A) 3D3L, B) 1MHL, C) 1GMY and D) 1CVU.
Fig. 10
Fig. 10
RMSF of ashwagandhanolide with A) 3D3L, B) 1MHL, C) 1GMY and D) 1CVU.
Fig. 11
Fig. 11
H-bond formation of ashwagandhanolide with A) 3D3L, B) 1MHL and C) 1GMY.
Fig. 12
Fig. 12
Radius of gyration of ashwagandhanolide with A) 3D3L, B) 1MHL and C) 1GMY
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