Study On Chemical Constituents Of Securidaca Inappendiculata And Its Anti-Multidrug Resistance Tumor Activity

Research on the chemical constituents and pharmacological activity of Securidaca inappendiculata Hassk.have been reported,about 30 kinds of xanthones have been isolated,and activity study showed that the compounds have anti-inflammatory,anti-tumor activity and so on.In addition,in recent years,the research found that xanthones can inhibit the proliferation of MDR tumor cells,but the structure-activity relationship has not been reported,and the mechanism of inhibit MDR tumor cells proliferation is not clear.Moreover,It has not been reported that the accurate and quantitative determination of xanthones.In order to ensure the quality of medicinal materials,a method for the determination of xanthonrs need establish.In this paper,study on the chemical constituents of Securidaca inappendiculata Hassk.,then several xanthones were select to study the inhibitory proliferation effect of MDR tumor cells in vitro,analyze and summarize the structure-activity relationship;and study on the mechanism of xanthones inhibiting MDR tumor cells based on metabonomics.The research mainly includes the following parts:Chapter 1.literature researchThe relevant literature was summarized,such as the reasons that multidrug resistance occurred,the advance in studies on the research progress on chemical constituents and pharmacological effects of Securidaca inappendiculata Hassk,the active components of traditional Chinese medicine reverse of MDR tumorand the research progress in cell metabonomics.Chapter 2.Study on chemical constituents of Securidaca inappendiculata Hassk.The 95%ethanol extract of Securidaca inappe-ndiculata Hassk was isolated and purified by silica,gel column,Sephadex LH-20 columns,et,then the structures of obtained compo-unds were identified by physicoch-emical properties and spectral data.Twenty-sewen compou nds were isolated and identifiedas 1,7-dihydroxyxyxanthone,1,3,8-trihydroxy-2-methoxyxantho ne,1,7-dihydroxy-3,4-dimethox-yxanthone,1,3,8-trihydroxy-4-methoxyxanthone,7-hydroxy-1,2-di-methoxyxanthone,1,3,6-trihyd-roxy-2,7-dimethoxyxanthone,1,4,8-trihydroxyxanthone,1,7-di hyd-roxy-3-methoxyxanthone,1,6-dihydroxy-7-methox-yxanthone,1,8-dihydroxy-3,4-dimethoxy xanthone,8-hydroxy-1,3,4-trimethoxyxanthone,1,2,7-trimethoxyxanthone,1,3,7-trihydroxyxyxan thon,1,3,7-trihydroxy-4-methoxyxanthone,1,3,7-trihydroxy-2-methoxyxanthone,3,8-dihydroxy-1,4-di-methoxyxanthone,2,7-dihydroxy-1-methoxyxanthone,3,7-dihydroxy-1,2,8-trimethoxy,1,7-dihydroxy-4-methoxyxanthone,2-hyd-roxy-1,7-dimethoxyxanthone,4-hydroxy-3,5-dimethoxyb enzaldehyde,sesamin,2-methoxy-3,4-methylenedioxybenzophenone,lignoceric acid,3-0-(3-D-(6-hexadecanoyl)-glucopyranosyl spinas-terol,a-spinasterol,?-sitosterol.Chapter 3.HPLC simultaneous determination of four xanthones in S.inappendiculata Hassk.Simple and reliable method using HPLC was established for the simultaneous determination of 4 xanthones(7-hydroxy-1,2-dimethoxyxanthone,1,7-dihydroxy-4-methoxyxanthone,2-hydroxy-1,7-dimethoxyxanthone and 1,7-dihydroxyxanthone).The developed method is proved by methodology validation that it is suitable for the quantitative analysis of four xanthones in S.inappendiculata Hassk.Chapter 4.The activity of xanthones inhibit MDR tumor cell and structure-activity relationship research.Thirteen xanthones were isolated naturally from the stem of Securidaca inappendiculata Hassk,and structure-activity relationships(SARs)of these compounds were comparatively predicted for their cytotoxic activity against three human multidrug resistant(MDR)cell lines MCF-7/ADR,SMMC-7721/Taxol,and A549/Taxol cells by MTT assay.The results showed that the selected xanthones exhibited different potent cytotoxic activity against the growth of different human tumor cell lines,and most of the xanthones exhibited selective cytotoxicity against SMMC-7721/Taxol cells.Furthermore,some tested xanthones showed stronger cytotoxicity than Cisplati.Among all the tested compounds,3,8-dihydroxy-1,4-dimethoxyxanthone showed the strongest activity against all the three kinds of MDR tumors,while the 1,3,6-trihyd-roxy-2,7-dimethoxyxanthone showed the weakest activity.The SARs analysis revealed that the cytotoxic activities of diverse xanthones were affected mostly by the number and position of methoxyl and hydroxyl groups,especially hydroxyl substituted in C-3,C-7 and methoxy substituted in C-4 show good inhibitory activity against SMMC-7721/Taxol,hydroxyl substituted in C-7and methoxy substituted in C-3,C-4 show good inhibitory activity against A549/Taxol,methoxy substituted in C-2,C-3,C-4 show good inhibitory activity against MCF-7/ADR.Chapter 5.Study on the mechanism of 3,8-dihydroxy-1,4-dimethoxyxanthone inhibiting A549/Taxo-1 cells based on metabonomicsIn this study,the mechanism of 3,8-dihydroxy-1,4-dimethoxyxanthone inhibiting A549/Ta-xol cells was studied by Cell metabonomics.Through multivariate statistical analysis to the metabolites of the group treated with 3,8-dihydroxy-1,4-dimethoxyxanthone and blank group,and identified 8 potential biomarkers were Adenine,L-Fucose,All-trans-13,14-dihydroretinol,D-Xylitol,L-Tyrosine,Lauroyl-CoA,bilirubin diglucuronide,3-Phosphoadenylylselenate.Com-pared with control group,Adenine,L-Fucose,D-Xylitol,L-Tyrosine in the group treated wit-h 3,8-dihydroxy-1,4-dimethoxyxanthone decreased significantly;while All-trans-13,14-dihydro-retinol,Lauroyl-CoA,bilirubin diglucuronide,3-Phosphoadenylylselenate in the group treated with3,8-dihydroxy-1,4-dimethoxyxanthone increased significantly.The metabolic pathway ana-lysisshowed four metabolic pathways that Tyrosine metabolism,Pentose and glucuronate interconver-sions,Fructose and mannose metabolism,Fatty acid elongation in mitochondria are closely related to t-he inhibition of A549/Taxol proliferation by 3,8-dihydroxy-1,4_dimethoxyxanthone.