Studies On The Chemical Constituents Of The BuOH Fraction Of Smilax Trinervula Miq. And Their Cytotoxic Activities

The genus Smilax(Smilacaceae)comprises about 370 species,which are mainly distributed in the tropical and temperate zones throughout the world,especially in East Asia and North America.Smilax trinervula Miq.(S.trinervula),a perennial plant widely distributed in South of China,such as Jiangxi,Guizhou and Anhui Province.It has the substantial wild resources in Jiangxi Province.It is sweet,bitter,flat,and was documented the efficacy of expelling wind and dredging the meridians,expelling phlegm and arresting coughing in “The compilation of Chinese herbal medicine”,“Jiangxi herbs”,“Hunan drug volunteers”,“Guizhou herbs”and“Fujian Chinese herbal medicine”.Among the people,S.trinervula was mainly used for clinical treatment of gouty arthritis,aching pain,bruises,tuberculosis and other diseases.There is almost no report and study about the plant due to the lack of concern by investigators,and its medicinal value is also not fully understood and used.Modern pharmacological studies have shown that Smilax herbs have anti-inflammatory effects,and the treatment of arthritis is significant.However,so far,there has been no detailed report on the pharmacodynamic material basis and mechanism of action in vivo of S.trinervula.The research project is part of the National Natural Science Foundation –“Study the clarity and structure-activity relationship of the total saponins’ active substances of the Smilax trinervula Miq.for anti-gouty arthritis”(No.81160507).The topic of this paper focuses on two aspects: 1 Separation and purifition of the chemical constituents in the BuOH fraction of Smilax trinervula Miq.The BuOH subfraction of 70% EtOH extract of S.trinervula were investigated on the chemical constituents systematically by various kinds of chromatographic methods.The structure of 80 compounds was determined by means of chemical evidence and spectroscopic analysis.30 compounds were analyzed,including 22 steroidal saponins,2 sterols,6 other classes.They are listed as follows:16β-O-(25-methyl-26-O-β-D-glucopyranosyl-pentanosyl)-(25R)pregn-5-en-3β,16β,20-trihydroxy3-O-α-L-rhamnopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-β-D-glucopyranosi de,(trinervuloside A)(HF-26)、Prosapogenin A(HF-5)、HF-67、HF-71、HF-72、HF-121、HF-123(trinervuloside C、D、E、F、G),16β-O-(26,27-ol-pentanosyl)-pregn-5-en-3β,16β,20-trihydroxy3-O-α-L-rhamnopyranosyl-(1→2)-[α-L-rhamnopyra nosyl-(1→4)]-β-D-glucopyranoside,(trinervulosideB)(HF-20)、Prosapogenin B(HF-3)、Protodioscin(HF-35)、Methylprotod ioscin(HF-34)、Dioscin(HF-4、HF-6、HF-9)、dioscoreside E(HF-21)、dioscoresideC(HF-23)、pseudoprotodioscin(HF-25)、(25S)26-O-β-D-glucopyranosyl-pentanosyl-3β,20α,26-trihydroxyfurostan-5,22-diene 3-O-α-L-rhamnopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-O-β-D-glucopyran oside(HF-27)、Anguiviosides XV(HF-28)、taccoside A(HF-30)、isonarthogenin 3-O-α-L-rhamnopyranosyl-(1→2)-O-[α-L-rhamnopyranosyl-(1→4)]-β-D-glucopyran oside(HF-8)、hypoglaucinG(HF-19)、pregna-5,16-dien-3β-ol-20-one 3-O-α-L-rham nopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-β-D-glucopyranoside(HF-24)、(HF-37)、Daucosterol(HF-2)、β-sitosterol(HF-1、HF-7)、(HF-45)、(HF-48)、(HF-49)、(HF-50)、(HF-55)、(HF-58)、Daucosterol(HF-2)、β-sitosterol(HF-1、HF-7).HF-26 、HF-67、HF-71、HF-72、HF-121、HF-123 and HF-20 are seven new compounds,In addition to Prosapogenin A(HF-5)、 Dioscin(HF-4,HF-6,HF-9)、Protodioscin(HF-35)、Methylprotod ioscin(HF-34)、Daucosterol(HF-2)、β-sitosterol(HF-1,HF-7),other compounds were characterized in S.trinervula for the first time.2 Their cytotoxic activities of the chemical constituents in the BuOH fraction of S.trinervula using MTTBy using the method of MTT,to Africa green monkey kidney cell line(Vero)as a positive control,using 4 kinds of cells(human neuroblastoma cell line(SH-SY5Y),human gastric carcinoma cell line SGC-7901(SGC-7901),human colon cancer cell line HCT-116(HCT-116),colon cancer cells(Lovo)),to study the cytotoxic activity of 21 compounds(HF-3 HF-5,HF-4,HF-8,HF-13,HF-12,HF-14,HF-16,HF-15,HF-19,HF-21,HF-20,HF-22,HF-24,HF-23,HF-25,HF-27,HF-26,HF-28,HF-29,HF-30)and to investigate their structure-activity relationship.In summary,this topic is investigated chemical constituents of the BuOH subfraction of 70% EtOH extract of Smilax trinervula Miq.on the system and their cytotoxic activities.It provided the theoretical basis for the further development、utilization and the quality control.