Ravi Lokesh, Jain Pratishtha, K Ajith Kumar, M Jeevan Kumar, Panda Mukti, S Harshitha, Kareem Mohammed Abdul, Sai K Sivani Yelchuri, Fathima Ayman, Jensha V, Rai Adarsh Anurag, Faizal Hida Amal, S R Shrivats, Ajith Ankitha, Yesudas Derick, S Vaarruni, S Varshini, Shetty Raksha, Jha Aditi, Janaki Gundlapalli Saradha, George Varsha Hannah, Nithaniyal Stalin, Kesavan Mookkandi Palsamy, A Sajith Ahamed, A Sankara Narayanan
Department of Biomedical Sciences, School of Health Sciences, The Apollo University, Chittoor, Andhra Pradesh, 517127, India.
Centre for Digital Health and Precision Medicine, The Apollo University, Chittoor, Andhra Pradesh, 517217, India.
J Comput Aided Mol Des. 2025 Jul 14;39(1):49. doi: 10.1007/s10822-025-00627-y.
Understanding the mechanism of action of anticancer agent plays a key role in effective clinical application of natural products. This study aims to identify an anti-cancer phytochemical with multi-target inhibition potential against non-small cell lung cancer. This study employs virtual screening of 8352 phytochemicals by molecular docking (AutoDock Vina & SeeSAR) to identify potential inhibitor of KRas, SMYD3, ALDH1 and GFPT2 proteins. Molecular Dynamics Simulation (Desmond) simulation for extensive 500 ns duration was performed to validate the inhibition potential. Followed by cell biology studies i.e., MTT assay, Flowcytometry and qRT-PCR analysis to confirm the mechanism of action. Molecular Docking and Molecular Dynamics Simulation studies predicted Heptadecanol as potential inhibitor of three drug targets, i.e., KRas, SMYD3 and GFPT2. In-vitro cytotoxicity assay confirmed the anti-cancer cytotoxicity of Heptadecanol with a significant IC value of 3.12 µg/ml selectively target cancer cells (A549), without substantial toxicity to non-cancerous cells (L929) with IC of > 100 µg/ml. Flowcytometry analysis with Annexin-V and Propedium Iodide staining further confirmed the apoptotic potential of Heptadecanol against A549 cells. qRT-PCR analysis demonstrated a robust increase in GFPT2 (25.6 × fold) and SMYD3 (16.98 × fold) gene expression, that conclusively confirmed the multi-target inhibition potential of Heptadecanol. Results of the study concludes that Heptadecanol is a significant inhibitor of GFPT2 and SMYD3 protein, there by exhibiting selective anti-cancer activity against the investigated non-small cell lung cancer cells. Further in-vivo studies are in demand to quantify the anti-cancer efficacy in a living system.
了解抗癌药物的作用机制对于天然产物在临床中的有效应用起着关键作用。本研究旨在鉴定一种对非小细胞肺癌具有多靶点抑制潜力的抗癌植物化学物质。本研究通过分子对接（AutoDock Vina和SeeSAR）对8352种植物化学物质进行虚拟筛选，以鉴定KRas、SMYD3、ALDH1和GFPT2蛋白的潜在抑制剂。进行了长达500纳秒的分子动力学模拟（Desmond）以验证抑制潜力。随后进行细胞生物学研究，即MTT法、流式细胞术和qRT-PCR分析，以确认作用机制。分子对接和分子动力学模拟研究预测十七烷醇是三种药物靶点即KRas、SMYD3和GFPT2的潜在抑制剂。体外细胞毒性试验证实了十七烷醇的抗癌细胞毒性，其显著的IC值为3.12微克/毫升，可选择性靶向癌细胞（A549），而对非癌细胞（L929）无实质性毒性，IC值>100微克/毫升。用膜联蛋白-V和碘化丙啶染色的流式细胞术分析进一步证实了十七烷醇对A549细胞的凋亡潜力。qRT-PCR分析表明GFPT2（25.6倍）和SMYD3（16.98倍）基因表达显著增加，这最终证实了十七烷醇的多靶点抑制潜力。研究结果得出结论，十七烷醇是GFPT2和SMYD3蛋白的重要抑制剂，从而对所研究的非小细胞肺癌细胞表现出选择性抗癌活性。需要进一步的体内研究来量化其在活体系统中的抗癌疗效。
