‘Characterisation and anti-angiogenic evaluation of Verbanone and Quercetin isolated from <i>Hemidesmus indicus’</i>

This study was designed to assess the anti-angiogenic and cytotoxic effects of Verbanone and Quercetin from <i>Hemidesmus indicus</i> using <i>in vitro</i> and <i>ex vivo</i> models. Bioactivity-guided processes led to the identification of Verbanone and Quercetin, proven through ultraviolet-visible (UV-Vis) spectroscopy, Fourier transform infra-red (FT-IR) spectroscopy, nuclear magnetic resonance (NMR) spectroscopy, and gas chromatography-mass spectrometry (GC-MS). The effect of these compounds on angiogenesis was measured by chorioallantoic membrane (CAM) assay and human umbilical vein endothelial cell (HUVEC) tube formation test. Cytotoxicity was evaluated in MCF-7 (hormone receptor-positive breast cancer) and MDA-MB-231 (triple-negative breast cancer) cells by performing an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Applying Annexin V-FITC/PI and quantitative real-time polymerase chain reaction (RT-qPCR), apoptosis and the levels of vascular endothelial growth factor (VEGF)/epidermal growth factor receptor (EGFR) mRNA were measured. The study found that Quercetin resulted in apoptosis for 54.8% of the MCF-7 cells and reduced VEGF expression by 0.07-fold. Verbanone, on the other hand, activated apoptosis for 38.6% of the cells and reduced VEGF by 38.6-fold. Verbanone and Quercetin were found to inhibit the growth of new blood vessels and induce apoptosis, making them suitable candidates for future breast cancer treatment.