Targeting Tracking Study Of Anti-tumor Active Ingredients Of Sambora Serrata In Vitro

Trigonostemon lutescens Y.T.Chang et J.Y.Liang,a species of the Trigonostemon genus from Euphorbiaceae,is mainly distributed in the southern of Guangxi Province.It is a special medicinal plant of China which grows in the shrubbery of limestone mountains.Scientific researches show that there are lots of chemical constituents in Trigonostemon,such as terpenoids,alkaloids,phenylpropanoids and so on.Many of these components are active substances with antibacterial activities,antitumor activities,antiinflammatory activities and antiviral activities.All of them showed that Trigonostemon were important plant sources for the development of new drugs.In vitro cytotoxicity of extracts from Trigonostemon lutescens were studied in this dissertation.CCK-8 method was used for cytotoxicity screening.The testing tumor cell lines were human colon cancer cell lines HCT116,huanman cervical carcinoma cell lines HELA and human hepatoma cell lines HepG2.The results showed that petroleum ether extract and the ethyl acetate extract had significantly inhibited to the proliferation of tumor cells.According to the amount of each section,ethyl acetate extracts were selected as the main section to separate.The open silica gel,MCI,Sephadex LH-20 column chromatography,medium pressure ODS column chromatography,preparative thin layer chromatography and the semi-preparative HPLC were used to isolate and purify the chemical constituents from the EtOAc extract of T.lutescens.The structures of these constituents were elucidated by their physiochemical properties,NMR,and MS spectral datas,as well as comparison with literature datas.Twenty two compounds were isolated from the 95%ethanol extract of the twigs of T.lutescens,and their structures were identified as 5-(2"-hydroxy-3"-methylbut-3"-en-1"-yl)-8-hydroxyfurocoumarin(1),xanthotoxin(2),bergapten(3),isoimpinellin(4),imperatorin(5),alloisoimperatorin(6),isodemethyl-furopinarine(7),auraptenol(8),meranzin hydrate(9),murrayone(10),3,4-dihydroxyallyl-benzene-4-O-?-D-glucopyranoside(11),1-O-?-D-gslucopyranosyl-4-allyl-benzene(12),3-O-acetyloleano lic acid(13),methyl 3-acetoxy-12-enly-oleanen-28-oate(14),taraxerone(15),simiarenol(16),spiciflorin(17),stigamasterol(18),?-sitosterol(19),?-sitosterol palmitate(20),palmitic acid(21)? 3,3',4-tri-O-methylellagicacid(22)?Among them,compound 1 is a new compound,and compounds 1-7 with furanocoumarin nucleus were firstly isolated from genus Trigono-stemon;compounds 1-17 were separated from the genus Trigonostemon for the first time and compounds 18 and 22 were firstly found from T.lutescens.By the way of UV-guided targeting,we separated ten coumarins,and summarized the regular pattern of UV absorption.Based on the method of LC-MS and biosynthesis pathway,five microconstituents has been successfully inferred.The method based on HPLC-DAD-MS which has a positive significance for the targeted separation.The in vitro cytotoxic activity of twenty-two compounds on human cancer lines were evaluated by CCK-8.As a result,compound 22 showed moderate cytotoxicities to HCT116 cells with a dose-dependent manner with an half maximal inhibitory concentration(IC50)of 17.11?g/mL.Compounds 10 and 20 showed strong proliferation inhibition to HELA cells with IC50 values of 9.63 and 3.72 ?g/mL,respectively.