Chemical Constituents And Biological Activities From The Roots Of Cynanchum Sibiricum

Objectives:（1）To conduct a systematic study on the chemical composition of the n-butanol extracted fraction of 95%ethanol extract of the roots of the Xinjiang local medicinal plant Cynanchum sibiricum Willd.to obtain abundant data and lay the foundation for the elucidation of its medicinal substances.（2）To evaluate the biological activity of the compounds isolated from the n-butanol part of the root of Cynanchum sibiricum Willd.to discover its connotation as a purgative and detoxifying herb and to provide clues for the development of modern anti-infective drugs.Methods:（1）Separation and purification by forward and reverse silica gel chromatography,gel column chromatogra phy,thin layer preparation and recrystallization,and modern spectroscopic techniques such as IR,UV,TOF-MS,HR-MS,1D NMR(¹H NMR,¹³C NMR,DEPT,NOED),2D NMR（^IH-^IH COSY,HMBC,NOESY）and CD.In combination with comparison with standard compounds and comparison with relevant literature data to determine the structures of compounds,the Scifinder database was used to retrieve structural information of compounds.（2）Using Pubchem database and other relevant data research platforms,the active ingredients and anti-infla mmatory target proteins of Cynanchum sibiricum were analyzed,and a network structure from ingredients to targets was constructed and correlated.The binding ability of some of the active compounds to the major anti-inflammatory targets was assessed by using the Auto Dock molecular docking tool.（3）In vitro cytolytic assay was used to determine the anti-complement activity of the monomeric compounds isolated from the n-butanol part of the roots of Cynanchum sibiricum.Results:（1）Twenty-two monomeric compounds were isolated and purified from the n-butanol extracted fraction of95%ethanolic extract of the roots of Cynanchum sibiricum.fifteen structures were identified,namely:uracil（1）,tachoside（2）,1-O-methyl-guaiacyl-glycerol（3）,1-（4-Hydroxyphenyl）-1,2,3-propanetriol（4）,methylβ-glaucobioside（5）,β-D-digitoxopyranosyl-（1→4）-β-D-glucopyranosly-（1→4）-β-D-glucopyrano sly-（1→4）-α-D-digitoxopyranoside（6）,Me-adoligose E（7）,isotachioside（8）,deacetylmetaplexigenin3-O-β-D-digitoxopyranosyl-（1→4）-β-D-cymaropyranoside（9）,sarcostin 3-O-β-D-oleandropyranoside（10）,deacetylmetaplexigenin 3-O-β-D-oleandropyranoside（11）,deacetylmetaplexigenin 3-O-β-D-oleandropyra noside（12）,deacetylmetaplexigenin 3-O-β-D-cymaropyranoside（13）,Daucosterol（14）and 2-3 dihydroxy-2-methyl butyric acid（15）.（2）The anti-complement activity of the monomeric compounds isolated from the n-butanol extracted part of the ethanolic extract of the roots of Cynanchum sibiricum was determined in vitro using the classical route,cytolytic assay.Among them,compounds 9,10,11,12 and 13 showed weak anti-complement activity with CH₅₀ values of 1.613,1.817,1.576,1.797 and 1.649 m M,respectively.（3）From the chemical structure of Cynanchum sibiricum isolated by our group,27 active chemical components were screened and cross-referenced with the human anti-inflammatory-related gene set,and a total of 82 cross-repeat targets were obtained.The complex interconnections between VEGFA,PTGS2,PPARG,EGFR were observed.The molecular docking results showed that compound 13（Cynotophyllo side B）could bind well to VEGFA,PTGS2 and PPARG,the main anti-inflammatory targets of the plant.Conclusions:（1）Among the 15 monomeric compounds isolated and identified from the n-butanol extracted part of 95%ethanolic extract of the roots of Cynanchum sibiricum,there were four new structural compounds（compounds 7,9,10,12）;among the compound structural types,five were C₂₁ steroidal glycosides（compounds 9-13）;three were deoxy-oligosaccharide compounds（compounds 5-7）.Compounds 7,9,10,12 were first isolated from plants of this family,compounds 2,3,4,5,8 and 15 were first isolated from plants of this genus,and all compounds were isolated for the first time from this plant;This provides a reference for the chemical taxonomy of this genus.（2）The anti-inflammatory target genes of Cynanchum sibiricum are mainly focused on The main anti-inflammatory target genes of the Cynanchum sibiricum are the Pathways in cancer,Lipid and atherosclerosis,Chemical carcinogenesis-receptor activation,Calcium signaling pathway and JAK-STAT signaling pathway.ion signaling pathway,JAK-STAT signaling pathway and many other signaling pathways.（3）The molecular docking results showed that compound 13（Cynotophylloside B）could bind well with VEGFA,PTGS2 and PPARG.The above results suggest that the C₂₁ steroidal glycosides isolated from this plant have relatively good anti-inflammatory activity,which may be the main material basis for the efficacy of Cynotophylloside B as a purgative and detoxifying agent.