Isolation And Purification Of Anti-tumor Active Compound SIPI-ZQ2 From Hovenia Dulcis Thunb And Its In Vitro And In Vivo Antitumor Activity Evaluation And Acute Toxicity

Nowadays,cancer is one of the main threats to human beings and their health,and the main approaches to the treatment of cancer have surgical,radiotherapy and chemotherapy methods.But surgical treatment can not cure middle-stage and terminal cancers,and known chemotherapeutic drugs have strong side effects.Plant-derived natural compounds are one of the important sources of drug development and it has become one of hotspots that isolates and screens the high-performance,low-toxicity,broad-spectrum natural compounds with anti-tumor activity.In this study,the 7 different kinds of Chinese herbs with anti-tumor activity reported previously were selected for anti-tumor activities using in vitro cell MTT colorimetric assay and the effective parts of Hovenia acerba Lindl were investigated,in which the main active compound was isolated and prurified.Later,the structure,antitumor activity and acute toxicity of the active compound were carried out using in vitro cell MTT colorimetric assay and H22 tumor bearing mice methods.First,water extracts and ethanol extracts of Glycyrrhiza uralensis Fisch,Coix lacryma-jobi L.var.mayuen Stapf,mulberry leaf,Platycodon grandiflorus(Jacq.)A.DC,Taraxacum mongolicum Hand.-Mazz.,Phyllanthus emblica Linn and Hovenia acerba Lindl were prepared and detected for their in vitro inhibitory activity on tumor cells by MTT assay.The results showed that each extract had certain antitumor activity,in which 60% ethanol extract of Hovenia acerba Lindl exhibited the highest activity.The different solvent extraction fractions were prepared and assayed on the 60% ethanol extract of Hovenia acerba Lindl.Results showed that the ethyl acetate fraction had the strongest anti-tumor activity.A main active compound named as SIPI-ZQ2 was isolated and purified from the ethyl acetate fraction of Hovenia acerba Lindl by silica gel column chromatography and preparative high performance liquid chromatography.Compared with the literature,the mass spectrometry,nuclear magnetic absorption and UV absorption of SIPI-ZQ2 were identical to those of dihydromyricetin.In vitro antitumor activity assay showed that the inhibitory effect of SIPI-ZQ2 on tumor cell proliferation was close to that of cisplatin and showed strong antitumor activity.SIPI-ZQ2 and L-arginine were complexed to obtain SIPI-ZQ2-arginine complex whose water solubility was much higher than that of SIPI-ZQ2,and the molar IC50 lower than that of SIPI-ZQ2 by MTT.Finally,the preliminary anti-tumor efficacy and acute toxicity of SIPI-ZQ2 were studied by H22 tumor-bearing mouse model.The SIPI-ZQ2 was administered by intraperitoneal injection,and the inhibitory activity of H22 tumor was the highest at the medium dose of 200 mg / kg,however the anti-tumor activity was decreased at the high dose of 300 mg / kg.In vivo acute toxicity test showed that SIPI-ZQ2 had certain acute toxicity.This study suggests that the main anti-tumor effective active component of Hovenia acerba Lindl may be dihydromyricetin,which lays the foundationfor the development of anti-tumor products of Hovenia acerba Lindl.