Research On Anti-colon Cancer Active Constituents Of Hairy Grape Stem

Hairy grape stem is the root and bark of Ampelopsis brevipedunculata（Maxim）Tranev.This genus has anti-tumor, anti-viral, anti-inflammatory and other significant pharmacologicalactivity. As an effective anti-tumor herbs, hairy grape stem has been used widely in folk, butits active ingredients are rarely reported. The studies on the chemical constituents andpharmacological activities, can provide certain theory basis for making full use of the plantresources and researching and developing in depth.Firstly, the extraction of hairy grape stem power was undertaken three times with75%ethanol. After suspension procedure by adding water, different polar solvents were used forthe five fractions of petroleum ether, chloroform, ethyl acetate and n-butyl alcohol andaqueous phase were gotten. Meanwhile, the effect of anti-colon cancer was carried out for thefive fraction extracts of wild vines. The obtained results were that the ethyl acetate andn-butanol phase extraction have significantly stronger anti-cancer activity than the other phaseextraction.The ethyl acetate extract of hairy grape stem ethanol extract was subjected to separateand purify its anti-colon cancer active constituents by large polyamide column, sephadex andODS preparation column, combined with the screening experiment for active constituents.Two compounds were gotten from the ethyl acetate extract. Using mass spectroscopy,1H and13C nuclear magnetic resonance spectroscopy and comparison with literatures, the twomonomeric compounds were identified to be:3,4-dihydroxybenzoic acid and Scirpusin AThe butanol fractions of hairy grape stem ethanol extract was subjected to separate andpurify its anti-colon cancer active constituents by large macroporous resin column, silica gelcolumn, sephadex and ODS preparation column, combined with the screening experiment foractive constituents. Two compounds were gotten from the butanol fractions. Using massspectroscopy,1H and13C nuclear magnetic resonance spectroscopy and comparison withliteratures, one is determined to be ampelopsin B and the other one was possible to beampelopsin B trimer.The anti-colon cancer activity of the four compounds were evaluated through MTTexperiment, when the four compounds contact with the colorectal cancer cells of human body for24hours, the IC50were134.0μ g/mL、27.1μ g/mL、26.1μ g/mL and81.3μ g/mLrespectively；When the four compounds contact with the colorectal cancer cells of humanbody for48hours, the IC50were204.2μ g/mL、14.5μ g/mL、18.6μ g/mL and66.1μ g/mLrespectively. Finally scirpusin A and ampelopsin B were determined to be which of the mostanti-colon cancer active substances.