Study On Antitumor Active Components And Content Determination Of The Ampelopsis

Malignant neoplasm is one of the most hazardous and severst diseases.Since clinical treatment for malignant neoplasm at present largely depends onchemical drugs, which has a strong side effect and drug resistance, theexploration of new antitumor drug with high effacy and low toxicity fromChinese herbal and natural medicine becomes the focus of drug reseachers.Chinese medicine the Ampelopsis is dry toot, it is acrid and bitter in taste,slightly cold in property. It has a function of heat-clearing and detoxicating,carbuncle disappearing and stagnation eliminating, furuncle stringing andpromoting tissue regeneration. the Ampelopsis was used to treat scarbies,dorsal ulcer, scrofula, burns and ascalds. Modern pharmacologies indicatedthat the Ampelopsis played a effective role in antibiosis, immunoregulationand antitumor. Previous study of our laboratory has demonstrated that theAmpelopsis was effective in antitumor and herbal literatures indicated that theAmpelopsis was non-toxic. An acute toxicity teat was performed in healtymice using different components of the Ampelopsis revealed the strongsecurity. Therefore, the Ampelopsis is a concern-worthy and a latent low toxic Chinese medicine for malignant neoplasm tratment. Our study was developedunder the guidance of bioactiviy. Active component seperation method wasused to stuty the chemical components in the antitumor patrs of theAmpelopsis. Quality stantards improved research was launched, providing ascientific basis for antitumor new drug development and improving qualitystantards. Folowing works has been accomplished in this paper:1. MTT method was used to screen antitumor activity in vitro. Human hepaticcarcinoma cell line HepG2was used for screening different parts of petroleumether, ethyl acetate, n-butanol and water of the Ampelopsis ethanol extract.The result indicated that the highest active part for tumor inhibition was ethylacetate the Ampelopsis ethanol extract. Petroleum ether and n-butanol partswere also antitumor active.2. Silica column chromatography was used to separate different parts. Foursolvents （petroleum ether, diethyl ether,ethyl acetate and methanol） withdifferent polarity were in sequnce as eluent to separate active part crudely.Active screening result indicated that both diethyl ether and ethyl acetate partshad a better antitumor active, and the highest antitumor active part was diethylehter part.3. Silica column chromatography, dextrangel colmn chromatography andpreparative chromatography seperation techniques were used to separate9monomers from diethyl ether and ethyl acetate parts under the basis ofantitumor active screening experiment（1-9）.6components were identified by Modern spectrum methods of1H-NMR、13C-NMR、H-HCOSY、C-HHMBC、 MS. The6components were β-sitosterol （1）,OleanolicAcid（2）,Gallic Acid （3）,Resveratrol （4）, Cianidanol（5）, Epicatechin（6）.4. MTT method was used to proceed antitumor active screening ofCianidanol（5）, Epicatechin（6） and component7. The result indicated thatcomponent7had high antitumor active. IC50value in24h,48h and72h were46.98mg·L^-1、57.79mg·L^-1、24.87mg·L^-1 respectively. Combined withprevious study,the main antitumor active components were Gallic Acid（3）,Resveratrol （4）, Oleanolic Acid and component7.5. HPLC with switching wavelength silmulataneous determination of thecontent of Gallic Acid, Protocatechuic aldehyde，Cianidanol and Resveratrol.The method was highly sensitive, reproducible, available in quality control forthe Ampelopsis, providing scientific basisi for improving the Ampelopsisquality stantards.