Studies On The Chemical Constituents And Anti-inflammatory Activities Of The Active Part Of The Two-color Coreopsis Flower Head Polyacetylene

Objects:To optimize the extraction and enrichment process of polyacetylene in Coreopsis tinctoria Nutt..Purified polyacetylenes from the capitulum of C.tinctoria and determined their structures.Researched the anti-inflammatory activities of polyacetylenes is in order to explore the material basis of drug effect of C.tinctoria,so as to find out the mechanism of C.tinctoria,in the treatment of coronary heart disease(CAD),and provide a theoretical basis for studying of quality standards of C.tinctoria.Methods:Mono-factorial experiments combined with orthogonal design was used to investigate the extraction method of C.tinctoria.Mono-factorial experiments was used to optimize the enrichment process of the polyacetylenes.Various kinds of atmospheric pressure column chromatography and Semi-PHPLC technology,such as silica gel?Sephadex LH-20 and RP-18 reverse silica gel,were used for the isolation and purification of the polyacetylenes from C.tinctoria.The structures of the compounds were identified by 1H-NMR,13C-NMR,1H-1HCOSY,HSQC,HMBC,MS and other relevant technologies.The anti-inflammatory activities of polyacetylenes from C.tinctoria was screened by computer molecular simulation technology.The NO production inhibition method of RAW264.7 macrophage model induced by LPS was used to screen the anti-inflammatory activities of seven polyacetylenes in vitro.Based on the study of structure and activity,the structure-activity relationship of polyacetylenes in C.tinctoria was generally discussed.Results:The best extraction process of C.tinctoria is showed as following:get appropriate amount of herbs in the round-bottom flask,add 16 times amount of 70%ethanol to heated and reflux for 1 hour.The optimal enrichment conditions were as follows:AB-8 macroporous resins were used for enriching the polyaeetylenes in C.tinctoria.The largest sample amount was 2 BV of extracts of C.tinctoria(0.224 g·ml-1).The eluting solvent was 90%methanol,the ratio of diameter to height was 1:10 and the percolate rate was 3BV/h.In this study,25 compounds were isolated and 16 compounds in those identified.They are(2R)-(3E,5Z,11E)-triene-7,9-diyne-1,2-diol?(2R)-(3Z,5E;11E)-triene-7,9-diyne-1,2-diol?(2S)-(3Z,11Z)-tridecadiene-5,7,9-triyne-1,2-diol?coreoside A?(2R)-(3E,5E,11E)-triene-7,9-diyne-1,2-diol?(3E,11Z)-tridecadiene-5,7,9-triyne-1,2-diol?(2S)-(3E,11E)-tridecadiene-5,7,9-triyne-1,2-diol?(2S)-(3Z,11E)-tridecadiene-5,7,9-triyne-1,2-diol?(Z)-7-phenyl-2-heptene-4,6-diyn-1-ol?(E)-7-phenyl-2-heptene-4,6-diyn-1-ol(3E,11Z)-1,2,6-trihydroxytrideca-3,11-dien-7-yn-5-one?(3E,11E)-1,2,6-trihydroxytrideca-3,11-dien-7-yn-5-one;luteolin?butein?(-)-?-Viniferin and ampelopsin E.14 polyacetylenes were conducted with virtual screening,eventually,Five potential 5-LOX inhibitors were harvested,they are coreoside D?coreoside B?coreoside A?coreoside E and coreoside C.Seven potential LTA4H inhibitors were harvested,they are(2R)-(3E,5E,11E)-triene-7,9-diyne-1,2-diol?coreoside B and coreoside A.Two potential COX-2 inhibitors were harvested,they are coreoside C and(2R)-(3Z,5E,11E)-triene-7,9-diyne-1,2-diol.Three potential 12-LOX inhibitors were harvested,they arecoreoside C?(2R)-(3E,5Z,11E)-triene-7,9-diyne-1,2-diol and(2R)-(3Z,5E,11E)-triene-7,9-diyne-1,2-diol.Conclusion:This study found that the main components of polyacetylene fraction were C14-polyacetylenes?C13-polyacetylenes and C7-polyacetylenes,and In addition,polyacetylenes had some anti-inflammatory activity,so the polyacetylene fraction was one of the anti-inflammatory active parts.It is also the material basis of drug effect of the treatment on CAD and other cardiovascular diseases.