Preparation Of Active Compounds From The Flowers Of Callicarpa Rubella With The Inhibitory Effect On NLRP3 Inflammasome Activation

The thesis consists of two chapters.The first chapter introduces the isolation and identification of the chemical composi-tion of the flowers of Callicarpa rubella Lindl.in detail,a plant of Callicarpa in the Verbenaceae family.By silica gel chromatography,Sephadex LH-20 gel chromatography,MCI reverse chromatography,HPLC and other purification methods,combined with NMR,HR-ESI-MS,IR,UV,OR and other technical means,48 compounds were isolated from the ethyl acetate phase and the n-butanol phase of the flowers of C.rubella,including 22 diterpenoids（9 new compounds）.All the new compounds are pimarane（isopiramane）diterpenoids,of which compound 3,6 and 8 are norditerpenoids,compound 4 and 10,5 and 11 are two sets of epimers,while compound 17 and 18 are enantiomers.At the same time,based on the obtained compounds,its inhibition effect on NLRP3 inflammasome activation was studied.Firstly,compound 12 with the highest inhibition rate was screened by LDH release inhibition activity experiment,and its IC₅₀value was 3.99μM.Afterwards,the mechanism of compound 12（the most abundant compound）blocking the activation of NLRP3 inflammasome was further explored.Through PI staining,it was determined that 12 could suppress pyroptosis;the results of ELISA experiments proved that 12 significantly inhibited the release of IL-1β;cytotoxicity assay showed that the inhibitory effect of 12 on NLRP3 inflammasome activation in J774 A.1 cells was not related to cytotoxicity.As an isopiramane diterpenoid,12 provides a new skeleton for targeted therapy of NLRP3 inflammasome related diseases.Chapter 2 discusses a review of piramane（isopiramane）and labdane diterpenoids from Callicarpa,Vitex and Clerodendrum in the Verbenaceae family in recent ten years.This section also summarizes the pharmacological activities of Callicarpa in hemostasis and cardiovascular diseases,and on this basis,expands the research status of related pharmacological activities to the plants of other families and genera.