Studies On The Chemical Constituents And Antitumor Activity Of Panax Notoginseng Fruits

Panax notoginseng fruits(PNF)is a mature berry drupe of Panax notoginseng.its shape is similar to those of Panax ginseng and Panax quinquefolius fruits with globose or reniform.There are generally have 2 seeds in each PNF.At present,the main use of PNF is as the source of seeds,however,the by-product pulp has not been well utilized,and caused a great waste of this resource.In the interest of further exploitation and utilization of this precious medicinal plant,developing new medicinal parts of Panax notoginseng,and finding new trace active components,the chemical constituents and antitumor activity of PNF are systematically studied.This study is to develop a method for systematically extraction and isolation of the chemical constituents in the PNF.Using macroporous resin column chromatography,MCI column chromatography,silica gel column chromatography,RP-C₁₈ column chromatography and semi-preparative HPLC,respectively.The structures of the isolates were determined by MS,IR,and NMR experiments.A total of 41 compounds were isolated and identified,including malonylnotoginsenoside Fe(1),malonylnotoginsenoside Ra₁(2),malonylgypenoside LXXV(3),malonylginsenoside Mc(4),malonylfloralginsenoside Rc₁(5),malonylginsenoside Rc(6),and malonylgypenoside IX(7),20(R)-ginsenoside Rg₃(8),ginsenoside F₂(9),notoginsenoside SFt₃(10),notoginsenoside SFt₄(11),ginsenoside Rg₅(12),ginsenoside Mc(13),notoginsenoside Fe(14),ginsenoside Rd₂(15),gypenoside IX(16),gypenoside XIII(17),ginsenoside Ra_1a(18),notoginsenoside ST-4(19),notoginsenoside Ft₁(20),notoginsenoside K(21),ginsenoside Rb₂(22),ginsenoside Rd(23),ginsenoside Rb₃(24),notoginsenoside Fc(25),notoginsenoside FP₂(26),ginsenoside Rc(27),notoginsenoside Fa(28),notoginsenoside D(29),notoginsenoside S(30),notoginsenoside Q(31),ginsenoside Ra₁(32),ginsenoside Rb₁(33),notoginsenoside R₁(34),ginsenoside Rg₁(35),and ginsenoside Re(36),Penia-28-O-glucohemsloside Ma₃(37),quercetin(38),quercetin-3-O-(2?-?-D-glucopyranosyl)-?-D-galactopyranoside(39),kaempferol-3-O-(2?-?-D-glucopyranosyl)-?-D-galactopyranoside(40),mannitol(41),respctively.Among these isolates,compounds 1-4,7 were new malonylginsenosides,compound 18 was a new natural product,compound 37 was an oleanolic saponin,and isolated from Araliaceae family for the first time,compound 41 was first isolated from genus Panax,compounds 5-6 were malonylginsenosides,and isolated from Panax notoginseng for the first time.In addition,23 compounds were first isolated from PNF,including compounds 9-17,19-22,26-32,38-40,respctively.The UPLC-Orbitrap-MS and UPLC-UV techniques were first used for qualitative and quantitative determination of saponins in PNF,the fragmentation patterns of different saponin types were analyzed and summarized,60 triterpenoid saponins and 2 flavonoid glycosides were qualitatively analyzed from the ethanol extraction of PNF,among them,39 compounds were unambiguously characterized by the aid of their authentic standards with the retention time and fragment ion peak information.23 saponins were deduced by the aid of fragment mass spectrometry and reference data,including 3 new double malonylginsenosides.As to quantitative analysis,UPLC-UV method was used to determine the content of 8 major saponins in PNF from different producing regions in Yunnan Province.The results showed that,this method was good accuracy,stability,and repeatability,and it can be used for quantitative analysis of saponins in PNF.The content of 8 major saponins in PNF were no significant difference among the 9 regions.The above studies provide the pharmacodynamical material basis for the further development and utilization of PNF.The antitumor activity of 21 saponins was studied by MTT method in vitro,7 saponins were screened for cytotoxic activity against MCF-7 breast cancer cells.Among them,ginsenoside Rd₂ was the best cytotoxic activity against MCF-7 cells with IC₅₀ was 7.197?M.All of 7 saponins were time-and concentration-dependent on the cytotoxic activity of MCF-7 breast cancer cells,moreover,the antitumor activity in vivo and antitumor mechanism of these saponins still need to be further studied.