Studies On Chemical Constituents And Bioactivities Of The Pheretima Aspergillum(E.Perrier) And Inula Lineariifolia

BACKGROUND Pheretima aspergillum?E.Perrier?is an animal of the genus Pheretima in Megascolecidae.In traditional Chinese medicine,it is used as medicine for all its dryness,commonly known as"Guang earthworm",and together with"Hu earthworm",it is collectively referred to as"earthworm".There are many natural active substances in earthworm,such as fatty acids,steroids,amino acids,nucleosides and furan sulfonic acids.It has a wide range of pharmacological activities,with significant anticoagulant,antithrombotic,antiasthmatic and wound healing promoting effects.As a traditional Chinese medicine in China,earthworm has been used for thousands of years,but there are few systematic studies on its chemical components,most of which are speculated by using analytical instruments.For example,after the first discovery of furan sulfonic acid compounds,they did not separate to get the same type of structure,but used UPLC-MS analysis to speculate that there were such series of compounds in earthworm.In order to get more furan sulfonic acid compounds,the further study of earthworm was carried out.Inula lineariifolia is a perennial herb of Inula in Asteraceae.In traditional Chinese medicine,its dried flower heads are used as medicine,also known as"JinFeiCao".It is mainly distributed in the north and northeast of China.There are a lot of terpenoids in this genus,which have various structures and good biological activities in anti-inflammatory and anti-tumor.In the previous study,several sesquiterpene lactone dimers with novel structure have been isolated from Inula linearifolia.In order to find more active terpenoids with novel structure and further improve the natural active substances in Inula linearifolia.METHOD In this paper,LC-MS was used to trace,thin-layer chromatography,sulfuric acid vanillin as the guide,and thin-layer chromatography silica gel,Sephadex LH-20 gel,C18 reverse phase silica gel,preparation-HPLC and other separation methods,as well as NMR,HR-ESI-MS,ESI-MS,infrared,ultraviolet,circular dichroism spectrum,optical rotation and other identification technologies,the chemical constituents of two kinds of Chinese medicinal materials were isolated and identified.At the same time,the antiplatelet aggregation rate was determined by enzyme labeling method,the cytotoxicity was determined by CCK-8,and the NO content was determined by Griess method,so as to study the biological activity of some new compounds.RESULTS Thirteen compounds,including 10 furan sulfonic acids,1 steroid,1 fatty acid and 1 nucleoside,were isolated and identified from Pheretima aspergillum?E.Perrier?.The new compounds are all furan sulfonic acid compounds?Pha-2?10?,the structure type is 2-hexyl-5-ethyl-3-furan sulfonic acid?Pha-1?as the basic skeleton,and the group substitution position occurs on the skeleton.The bioactivity of some furan sulfonic acid compounds against platelet aggregation was studied.The results showed that Pha-3 and Pha-4 had some antiplatelet aggregation activity,and the inhibition of Pha-4 was concentration dependent.The lower the concentration,the weaker the antiplatelet aggregation.For Inula lineariifolia,11 monomer compounds have been isolated and identified.It includes 5 sesquiterpene lactones dimers,1 diterpene and5 sesquiterpene lactones.Three of them are rare new sesquiterpene lactone dimers?IL-1?3?,which are linked by C-13/C-1'and C-11/C-3'.The anti-inflammatory biological activity of IL-1?3 was researched.The results showed that IL-1?3 had a significant inhibitory effect on LPS induced NO production in RAW 264.7 macrophages(IC₅₀ values of 1.421,1.087 and1.243?M,respectively),and had no significant cytotoxic activity on RAW264.7 macrophages(IC₅₀>50).CONCLUSION In this paper,the chemical constituents and activities of the two herbs were studied,which not only laid a foundation for the discovery of their effective parts,but also provided the research direction for the design of antiplatelet aggregation drugs and anti-inflammatory drugs,which has certain scientific research value and significance.