Delphinium roylei Munz是一种印度特有的药用植物，其抗癌活性尚未进行过先前的研究，且其生物活性化合物与易受攻击的乳腺癌药物靶标的具体相互作用仍大部分未知。因此，在本研究中，我们旨在评估D. roylei不同提取物对乳腺癌的抗癌活性，并通过网络药理学结合分子对接和体外验证来解析其分子机制。根据它们的极性指数，实验植物采用不同有机溶剂进行提取。采用高分辨率液相色谱-质谱（HR-LC/MS）技术鉴定了植物化合物，并通过SwissADME程序评估了它们的物化性质。接下来，通过公共数据库检索与所得生物活性物或与乳腺癌相关的靶点相关的靶点，并使用Venn图选择重叠的靶点。通过STRING程序和Cytoscape软件可视化、构建和分析重叠靶点与生物活性物之间的网络。最后，我们使用AutoDock Vina进行分子对接试验（MDT），以探索关键靶点和化合物。HR-LC/MS检测到了数百种植物化合物，经过虚拟筛选后，少数符合Lipinski规则，并被归类为药物样化合物（DLCs）。对于九种定量植物化合物，获得了总共464个潜在靶基因，并利用Gene Cards、OMIM和DisGeNET平台检索到了与乳腺癌相关的12063个疾病靶点。通过京都基因与基因组百科全书（KEGG）通路富集，共展现了20个信号通路，经过分子对接研究，从这20个信号通路中选择出一个中枢信号通路（PI3K-Akt信号通路），一个关键靶点（Akt1）和一个关键化合物（8-羟基香豆素）。分子对接研究发现，在这九种植物化合物中，8-羟基香豆素与Akt1乳腺癌靶点具有最好的结合能（-9.2 kcal/mol）。8-羟基香豆素通过SwissADME遵循了所有ADME性质预测，并且通过100纳秒（ns）的分子动力学模拟发现8-羟基香豆素与Akt1的复合物是稳定的。此外，D. roylei提取物还通过体外研究显示了显著的抗氧化和抗癌活性。我们的发现首次表明D. roylei提取物可用于BC的治疗。Copyright © 2023 Mir, Bhat, Kumar, Dhiman, Alkhanani, Almilaibary, Dar, Ganie and Mir.

Delphinium roylei Munz is an indigenous medicinal plant to India where its activity against cancer has not been previously investigated, and its specific interactions of bioactive compounds with vulnerable breast cancer drug targets remain largely unknown. Therefore, in the current study, we aimed to evaluate the anti-breast cancer activity of different extracts of D. roylei against breast cancer and deciphering the molecular mechanism by Network Pharmacology combined with Molecular Docking and in vitro verification. The experimental plant was extracted with various organic solvents according to their polarity index. Phytocompounds were identified by High resolution-liquid chromatography-mass spectrometry (HR-LC/MS) technique, and SwissADME programme evaluated their physicochemical properties. Next, target(s) associated with the obtained bioactives or breast cancer-related targets were retrieved by public databases, and the Venn diagram selected the overlapping targets. The networks between overlapping targets and bioactive were visualized, constructed, and analyzed by STRING programme and Cytoscape software. Finally, we implemented a molecular docking test (MDT) using AutoDock Vina to explore key target(s) and compound(s). HR-LC/MS detected hundreds of phytocompounds, and few were accepted by Lipinski's rules after virtual screening and therefore classified as drug-like compounds (DLCs). A total of 464 potential target genes were attained for the nine quantitative phytocompounds and using Gene Cards, OMIM and DisGeNET platforms, 12063 disease targets linked to breast cancer were retrieved. With Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway enrichment, a total of 20 signalling pathways were manifested, and a hub signalling pathway (PI3K-Akt signalling pathway), a key target (Akt1), and a key compound (8-Hydroxycoumarin) were selected among the 20 signalling pathways via molecular docking studies. The molecular docking investigation revealed that among the nine phytoconstituents, 8-hydroxycoumarin showed the best binding energy (-9.2 kcal/mol) with the Akt1 breast cancer target. 8-hydroxycoumarin followed all the ADME property prediction using SwissADME, and 100 nanoseconds (ns) MD simulations of 8-hydroxycoumarin complexes with Akt1 were found to be stable. Furthermore, D. roylei extracts also showed significant antioxidant and anticancer activity through in vitro studies. Our findings indicated for the first time that D. roylei extracts could be used in the treatment of BC.Copyright © 2023 Mir, Bhat, Kumar, Dhiman, Alkhanani, Almilaibary, Dar, Ganie and Mir.