Isolation,Identification And Anti-tumor And Anti-inflammatory Activity Of Diterpenoids From Jatropha Curcas

The research and development of drug discovery is still a hot topic in the treatment of tumors and inflammation,and screening for lead compounds from natural products is currently a mainstream approach.Diterpenoids have been reported to show good activities against tumors and inflammation,and some of them have been approved for clinical use.According to our previous research and literature review,diterpenoids is abundant in Jatropha curcas.Therefore,it is a feasible way to obtain more diterpenoid lead compounds with anti-tumor and anti-inflammatory activities from J.curcas.The powdered air-dried roots of J.curcas were extracted with MeOH,and the crude extract was extracted with petroleum ether and CH₂Cl₂,respectively.CH₂Cl₂-soluble fraction was subjected to column chromatography.As a result,19new diterpenoids named dimericursones A?1*?,dimericursones B?2*?,dimerilathyrane A?3*?,lathyranephenol A?4*?,curcusone K?5*?,curcusone L?6*?,curcusone M?7*?,curcusone N?8*?,jatrophadiketone A?9*?,18-hydroxyjatrophadiketone?10*?,jatrocurcasenone F?11*?,jatrocurcasenone G?12*?,jatrocurcasenone H?13*?,jatrocurcasenone I?14*?,jatrocurcasenone J?15*?,jatropholone F?16*?,7?-hydroxyjatropholone A?17*?,7?-hydroxyjatropholone B?18*?,2-epi-macroripremyrsinone A?19*?together with 33 known diterpenoids were isolated.Their structures were identified by spectroscopic data.Compounds 5*and 25 showed cytotoxicity against PC-3 cell lines with inhibitions of cell viability>50%at 20?M;Compound 15*exerted anti-proliferation effect against HepG2 and MCF-7 cell lines,with IC₅₀0 values of 5.21?M and 2.46?M,respectively.Anti-inflammatory evaluation performed on LPS-induced RAW 264.7macrophages demonstrated that compounds 2*,7*,13*,and 14*significantly inhibited the production of NO with IC₅₀0 values of 5.65?M,11.81?M,11.28?M and7.71?M,respectively.Furthermore,7*,13*,and 14*inhibited the production of COX-2 and iNOS proteins at 5-20?M in a dose-dependent manner in western blotting experiments.In order to investigate the anti-inflammatory mechanism of 14*,we further analyzed its transcriptome profiles.The results of RNA-seq showed that 14*has a regulatory effect on the 587 differentially expressed genes?DEGs?.The 587 DEGs were mainly enriched in GO terms which were associated with inflammatory responses,such as response to stimulus,response to stress,and the production and regulation of cytokines.The KEGG enrichment analysis showed that cytokine-cytokine receptor interaction was the most significantly enriched pathway.KSEA analysis was performed on the gene sets of model group and the group treated with 14*.The KSEA-GO analysis and KSEA-KEGG analysis also showed that the anti-inflammatory effect of 14*was closely related to cytokines.It can be speculated that compound 14*may erxert its anti-inflammatory potency in LPS-induced inflammation models through inhibiting cytokines expression and regulating the cytokine-cytokine receptor interaction pathway,Subsequently,pathways such as PI3K/AKT,JAK/STAT,NF-?B,JNK may be adjusted.In conclusion,52 diterpenoids were isolated from the root bark of J.curcas.Among all the compounds obtained,3 diterpenoids were found to have anti-cancer potency,and 4 diterpenoids showed anti-inflammatory activity in vitro.Furthermore,potential mechanism of anti-inflammatory of 14*was discussed through transcriptome analysis.This study offers substances and theoretical basis for the development of anti-tumor and anti-inflammatory drugs with independent intellectual property rights.