INTRODUCTION:

Lung cancer remains the most common cause of cancer-related deaths worldwide, accounting for around 85% of all lung cancer cases^1-5. The problem of late diagnosis and the limited effectiveness of conventional treatments are the main factors driving the need for new anti-tumour drugs^6-8. Natural substances derived from medicinal plants are a new way to discover drugs, especially when examined with in silico tools^9-14.

Bermuda grass or Cynodon dactylon is a pantropical herb containing bioactive phyto-chemicals such as flavonoids (e.g., luteolin, quercetin), phytosterols (e.g., β-sitosterol, stigmasterol), terpenoids, and phenolic compounds^15,16. There is growing evidence that these can be used to treat lung and other cancers. They also have anti-inflammatory, antioxidant, and anticancer properties ^17-19. Recent computational studies show that C. dactylon phytosterols, such as stigmasterol acetate and β-sitosterol, have stable conformations in molecular dynamics simulations and efficient interactions with cancer-associated proteins such as MAPK3 and PARP1, with a docking energy of -10.1 to -10.9kcal/mol^16,20,21. In addition, symbiotic endophytic metabolites of C. dactylon have been shown to have strong binding affinity to anti-apoptotic proteins such as Bcl-2²². Flavonoids such as kaempferol, luteolin, and quercetin have shown binding affinity to EGFR, including the drug-resistant mutations L858R and T790M, as well as to ALK, Bcl-2, AKT1, and MAPK3, targets of critical importance in the treatment of non-small cell lung cancer ^23-27. Pharmacological and ADMET profiling, which demonstrate pharmacokinetic viability and minimal toxicity, further confirm the findings^28,29.

In addition, the same in silico tool was used for the evaluation of related botanical plant phytochemicals (Moringa oleifera, Terminalia arjuna, and Withania somnifera), which are used as reference points for the docking method C. dactylon^30-34. These include virtual screening platforms for multi-target cancer therapies, molecular docking, molecular dynamics simulation, and profiling by GC-MS^17,35-37.

MATERIAL AND METHODS:

Selection of Phytoconstituents:

The plant-phytochemical components of C. dactylon have been selected following a careful literature review of published ethnopharmacological studies and PubChem databases. The selected compounds belong to different classes of structural compounds, namely flavonoids (e.g., vitexin, orientin, luteolin, apigenin), phenolic acids (e.g., ferulic acid, p-coumaric acid, hydroquinone), and phytosterols (beta-sitosterols).

Ligand Preparation:

The three-dimensional structures of the selected phytoconstituents were obtained in SDF format from PubChem. After the import of these structures into PyRx, a universal force field (UFF) was applied to minimise energy consumption. For docking, the truncated structures have been recorded in PDBQT format.

Protein Preparation:

The target protein structure with PDB ID 6JXT was obtained from the RCSB protein data bank. Structure has been created by removing the cocrystalline ligands, water molecules, and other heteroatoms using the Discovery Studio Visualizer. After purification, the structure was imported into PyRx, where the receptor was converted to PDBQT format for docking, and polar hydrogens were added to the receptor.

Molecular Docking Using PyRx:

The molecular docking was performed with AutoDock Vina integrated in PyRx. The grid box surrounding the receptor site has been determined using known binding residues. Using the default setting, docking was performed, and binding affinity was quantified as docking energy in kcal per mole. The conformation of each ligand with the lowest binding energy was selected for further investigation.

Visualization and Interaction Analysis:

The optimal binding positions of each ligand have been displayed and explored using the Discovery Studio Visualizer. The key interactions used to interpret binding specificity and potential anticancer relevance were hydrophobic contacts, π-π stacking, and hydrogen bonding.

Data Interpretation:

For the assessment of the relative binding affinity of compounds, docking scores have been ranked. High binding affinity compounds have been defined as compounds with a score less than -8.0kcal/mol. Based on the body of research on the anticancer properties of C. dactylon, the results were reviewed, with particular attention to trends within the phytochemical class^38-41.

RESULTS:

Orientin and Vitexin, the glycosylated flavonoids, exhibited the strongest binding affinities to the 6JXT receptor with docking scores of -9.2, followed closely by Beta sitosterol (-8.9) and Luteolin (-8.7), indicating high-strength interactions (Table 1).

Table 1: Molecular docking scores of phytoconstituents with the PDB ID: 6JXT. The table lists the phytoconstituents, their respective chemical classes, and the binding affinities (Dock scores in kcal/mol). Lower (more negative) docking scores indicate stronger binding affinity.

S. No.	Receptor	Phytoconstituents	Class	Dock Score
1.	6JXT	Orientin	Flavonoid glycoside	-9.2
2.	6JXT	Vitexin	Flavone glycoside	-9.2
3.	6JXT	Beta sitosterol	Phytosterols	-8.9
4.	6JXT	Luteolin	Flavonoid	-8.7
5.	6JXT	Apigenin	Flavonoid	-8.4
6.	6JXT	Ferulic acid	Phenolic compound	-6.0
7.	6JXT	p-Coumaric acid	Phenolic compound	-5.9
8.	6JXT	Hydroquinone	Phenolic compound	-4.5
9.	6JXT	Levoepicatechin	Flavonoid	-8.0

In contrast to phenolic compounds like ferulic acid, p-coumaric acid, and hydroquinone, which showed relatively weaker bindings with docking scores ranging from -6.0 to -4.5, flavonoids such as apigenin and levoepicatechin also showed strong affinities (-8.4 and -8.0, respectively). This suggests that flavonoids are probably more effective at targeting this receptor than phenolic compounds.

Docking Dimensions:

The receptor 6jxt.pdbqt was used for molecular docking to ensure complete conformational sampling at the exhaustiveness level of 8. In order to efficiently assess ligand binding, the docking grid was strategically positioned at coordinates (x = -6.871, y = 62.152, z = 4.532) with dimensions of 60.68 Å × 78.43 Å × 25.0 Å along the x, y, and z axes, respectively. This allowed the grid to conveniently cover the receptor's active site region (Table 2 and Table 3).

Docking results interpretation

Table 3: Docking scores of phytoconstituents from Cynodon dactylon (Bermuda grass) against PDB ID: 6JXT with interpretations in the cancer context. The table correlates binding affinities with known anticancer mechanisms, highlighting possible roles of flavonoids, glycosides, phytosterols, and phenolic acids in apoptosis, oxidative stress regulation, angiogenesis inhibition, and signaling pathway modulation.

Compound	Phytochemical Class	Dock Score	Interpretation in Cancer Context
Vitexin	Flavone glycoside	-9.2	Demonstrates the highest affinity for binding. Recognized for causing apoptosis and preventing angiogenesis in cancerous models. Accordingly, it is possible that vitexin targeting signaling nodes such as PI3K/Akt or VEGF pathways contributes to C. dactylon's anticancer activity.
Orientin	Flavonoid glycoside	-9.2	The highest binding tie. Research backs up its function in preventing tumor cells from proliferating and experiencing oxidative stress. Its inclusion in cancer treatments is supported by the fact that it is found in C. dactylon.
Beta-sitosterol	Phytosterol	-8.9	A powerful binder that has been shown to have an impact on the integrity of cell membranes and to activate caspase, which promotes apoptosis. Its cytotoxic function in C. dactylon is supported by its docking score.
Luteolin	Flavonoid	-8.7	Suppresses pathways linked to cancer (e.g., 3. MAPK and NF-κB. Its high docking affinity here is consistent with reports of antiproliferative effects from extracts of C. dactylon that are rich in luteolin.
Apigenin	Flavonoid	-8.4	Binds well and is known to induce cell cycle arrest in several cancer types. The chemopreventive potential of C. dactylon is supported.
Levoepicatechin	Flavonoid	-8.0	Strong to moderate affinity; linked to pro-apoptotic and antioxidant properties, which could help justify C. dactylon's application in cancers linked to oxidative stress.
Ferulic acid	Phenolic acid	-6.0	Weaker binding, but through regulating ROS, it is known to indirectly prevent carcinogenesis. Possibly contributes to C. dactylon's anticancer synergy.
p-Coumaric acid	Phenolic acid	-5.9	Although there is little direct cytotoxicity, antioxidant effects could make cancer cells more sensitive to other therapies. Helps to create the plant's overall protective profile.
Hydroquinone	Phenolic compound	-4.5	Weakest score for docking. Despite being found in C. dactylon, hydroquinone's anticancer potential is uncertain because of its possible toxicity.

DISCUSSION:

According to the molecular docking analysis, the flavonoid glycosides Orientin and Vitexin showed the highest binding affinities toward the cancer-related receptor 6JXT among the assessed phytoconstituents of C. dactylon. Their docking scores of -9.2 indicated that they had strong and stable interactions with the active site. Due to their well-established functions in inducing apoptosis and inhibiting angiogenesis, these substances have the potential to be lead compounds in cancer treatments.

Following closely behind were luteolin (-8.7) and beta-sitosterol (-8.9), both of which have been shown to have cytotoxic and pathway-inhibitory effects in a variety of cancer models. Additional evidence for the importance of flavonoids in targeting this receptor came from the notable binding of other flavonoids like levoepicatechin (-8.0) and apigenin (-8.4). However, phenolic acids with a lower direct binding potential to 6JXT, such as hydroquinone (-4.5), p-coumaric acid (-5.9), and ferulic acid (-6.0), showed relatively weaker interactions. They may still, however, support anticancer mechanisms synergistically thanks to their antioxidant qualities. Using a grid dimension of 60.68 Å 78.43 Å × 25.0 Å and a rigorous exhaustiveness level of 8, the docking was carried out with a focus on (-6.871, 62.152, 4.532) to ensure accurate interaction profiling and thorough conformational sampling.

CONCLUSION:

The most promising phytoconstituents in C. dactylon for targeting the 6JXT receptor, which may be connected to pathways related to cancer, are flavonoids, particularly glycosylated forms like Orientin and Vitexin, according to this study's findings. These discoveries support the plant's traditional anticancer use at the molecular level and call for additional in vitro and in vivo testing to create targeted phytotherapeutics.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

ACKNOWLEDGMENTS:

The work was done in Maharishi School of Pharmaceutical Sciences, Maharishi University of Information Technology, Lucknow, Uttar Pradesh, India.

FUNDING:

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

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Received on 20.08.2025 Revised on 16.09.2025

Accepted on 11.10.2025 Published on 06.11.2025

Available online from November 11, 2025

Asian J. Research Chem.2025; 18(6):365-370.

DOI: 10.52711/0974-4150.2025.00056

This work is licensed under a Creative Commons Attribution-Non Commercial-Share Alike 4.0 International License. Creative Commons License.

Phytoconstituents	Ligand interaction	2D structure
Orientin
Vitexin
Beta sitosterol
Leuteolin
Apigenin
Ferrulic acid
p-coumaric acid
Hydroquinone
Levo-epicatechin