Integrated study of HR-LC/MS and network pharmacology to identify breast cancer-related molecular targets of Fritillaria cirrhosa D. Don active constituents in combination with molecular dynamic simulation and experimental evaluation

Abstract

AbstractFritillaria cirrhosaD. Don is a well-known medicinal plant in Kashmir Himalya. Traditionally, it has been used to treat several diseases, most notably in the treatment of various cancers particularly lung cancer. However, there is a significant gap between scientific research and its application in conventional medicine. The aim of the current work is to provide first-hand evidences bothin-vitroandin silicoshowing thatF. cirrhosaextracts exerts anti-cancer effects against breast cancer. Bulbs ofF. cirrhosawas extracted with various solvents of increasing polarity. Compounds were identified by High resolution-liquid chromatography-mass spectrometry (HR-LC/MS) technique. Phytocompounds were studied for protein targets involved in pathogenesis of breast cancer using Binding 1DB (similarity index > 0.7). Later, the protein-protein interactions (PPI) network was studied using STRING programme and compound-protein interactions using Cytoscape. In addition, molecular docking was used to investigate intermolecular interactions between the compounds and the proteins software using Autodock tool. Molecular dynamics simulations studies were also used to explore the stability of the representative CDK2 + Peiminine complex. In addition, standardin-vitrobiochemical assays were used to evaluate thein-vitroantiproliferative activity of active extracts ofF. cirrhosaagainst several breast cancer cell lines. Bioactive components and potential targets in the treatment of breast cancer were validated through network pharmacology approach. HR-LC/MS detected the presence of several secondary metabolites. Afterward, molecular docking was used to verify the effective activity of the active ingredients against the prospective targets. Additionally, Peiminine showed the highest binding energy score against CDK2 (-12.99 kcal/mol). CDK2 + Peiminine was further explored for molecular dynamics simulations. During the MD simulation study at 100 nanoseconds (ns), a stable complex formation of CDK2 + Peiminine was observed. According to molecular docking results predicted, several key targets of breast cancer bind stably with the corresponding phytocompounds ofF. cirrhosa. Lastly,F. cirrhosaextracts exhibited momentous anticancer activity throughin vitrostudies. Overall, the most important constituents were Imperialine-3-β-glucoside and Peiminine from theF. cirrhosabulbs has effective anti-cancer efficacy by deactivating Akt1 on the PI3K-Akt signaling pathway. Therefore, these findings emphasized the momentous anti-breast cancer activity ofF. cirrhosaextracts. This may open a new window and provide a theoretical foundation for further development and utilization ofF. cirrhosamedicinal plant in the treatment of breast cancer.