sanguiin-h-6 and ellagitannin

The vascular endothelial growth factor (VEGF) plays a key role in angiogenesis, which is a process where new blood vessels develop from the endothelium of a pre-existing vasculature. VEGF exerts its activity by binding to its receptor tyrosine kinase, KDR/Flk-1, which is expressed on the surface of endothelial cells. A methanol extract and organic solvent (n-hexane, ethyl acetate, n-butanol, aqueous) fractions from Rubus coreanus were examined for their inhibitory effects on VEGF binding to the VEGF receptor. The methanol extract from the crude drug were found to significantly inhibit VEGF binding to the VEGF receptor (IC50 approximately = 27 microg/mL). Among the fractions examined, the aqueous fraction from the medicinal plant showed potent inhibitory effects against the binding of KDR/Flk-1-Fc to immobilized VEGF165 in a dose-dependent manner (IC50 approximately = 11 microg/mL). Sanguiin H-6 was isolated as an active principle from the aqueous fraction, and inhibited the binding of KDR/Flk-1-Fc to immobilized VEGF165 in a dose-dependent manner (IC50 approximately = 0.3 microg/mL). In addition, sanguiin H-6 efficiently blocked the VEGF-induced HUVEC proliferation in a dose-dependent manner (IC50 approximately = 7.4 microg/mL) but had no effect on the growth of HT1080 human fibrosarcoma cells. This suggests that sanguiin H-6 might be a potential anti-angiogenic agent.

Topics: Carbohydrate Sequence; Cell Proliferation; Cells, Cultured; Endothelial Cells; Fruit; Humans; Hydrolyzable Tannins; Molecular Sequence Data; Plant Extracts; Receptors, Vascular Endothelial Growth Factor; Umbilical Veins; Vascular Endothelial Growth Factor A

Analysis of extracts of Glen Ample raspberries (Rubus idaeus L.) by gradient, reverse phase HPLC with diode array and tandem mass spectrometry identified eleven anthocyanins, including cyanidin-3-sophoroside, cyanidin-3-(2(G)-glucosylrutinoside), cyanidin-3-glucoside, cyanidin-3-rutinoside, pelargonidin-3-sophoroside, pelargonidin-3-(2(G)-glucosylrutinoside), and pelargonidin-3-glucoside. Significant quantities of an ellagitannin, sanguiin H-6, with an M(r) of 1870 were detected along with lower levels of a second ellagitannin, lambertianin C, which has an M(r) of 2804. Other phenolic compounds that were detected included trace levels of ellagic acid and its sugar conjugates along with one kaempferol- and four quercetin-based flavonol conjugates. Fractionation by preparative HPLC revealed that sanguiin H-6 was a major contributor to the antioxidant capacity of raspberries together with vitamin C and the anthocyanins. Vasodilation activity was restricted to fractions containing lambertianin C and sanguiin H-6.

Topics: Animals; Anthocyanins; Antioxidants; Chemical Fractionation; Chromatography, High Pressure Liquid; Electron Spin Resonance Spectroscopy; Flavonoids; Fruit; Hydrolyzable Tannins; Male; Mass Spectrometry; Phenols; Plant Extracts; Rabbits; Rosaceae; Tannins; Vasodilator Agents

Many tannins were previously identified as candidate topoisomerase poisons. Here we report further studies on sanguiin H-6, a dimeric ellagitannin isolated from Sanguisorba officinalis as an inhibitor of DNA topoisomerases. Catalytic strand-passing activities of topoisomerases I and II were inhibited in vitro with IC50 values of 1 microM and 0.01 microM, respectively. This inhibition was not associated with stabilization of covalent enzyme-DNA complexes but rather by a mechanism preventing formation of such covalent intermediates, as measured by interference with drug-induced cleavage in vitro. The IC50 values for topoisomerase I-DNA complexes induced by camptothecin and with topoisomerase II-DNA complexes induced by VP-16 were 0.02 microM and 0.16 microM, respectively. Pre-incubation studies followed by drug-dilution revealed that the in vitro inhibitory effects of sanguiin H-6 were irreversible, and for topoisomerase I, the test compound prevented enzyme-DNA interaction as seen by shifts in mobility on agarose gels. By measuring interference with drug-induced protein-linked DNA breaks in isolated HeLa nuclei, inhibition of topoisomerases I and II on a natural chromatin template was demonstrated with IC50 values of 5 microM and > 10 microM, respectively. Sanguiin H-6 inhibited HeLa cell growth with an ED50 of 12 microM and also interfered in a dose-dependent fashion with intracellular topoisomerase activities but with lower potencies than those observed using subcellular assay systems. Based on these studies, sanguiin H-6 could be broadly classified as a type of poison which does not stimulate the formation of cleavable-complexes, with intracellular activity but without any marked selectivity.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Catalysis; Cell Division; Cell Nucleus; DNA; DNA Topoisomerases, Type I; HeLa Cells; Humans; Hydrolyzable Tannins; Molecular Structure; Plants; Tannins; Topoisomerase I Inhibitors; Tumor Cells, Cultured