Isolation Of The Sulfur-containing Metabolites Generated From Intestinal Bacteria Biotransformation Of Parthenolide And Identification Of In Vivo And In Vitro Metabolites Of Parthenolide

Parthenolide,a natural germacrane-type sesquiterpene lactone,was first isolated from traditional herb feverfew?Tanacetum parthenium?in 1965,which is the component of many composites.Recent studies have demonstrated that parthenolide has various biological properties,such as anti-inflammatory,anticancer,antimalarial,antiviral,antioxidative,antimigraine,and hepatoprotective effects.A pharmacokinetic study on rats showed that the bioavailability of parthenolide was just 7.78%after oral administration.Given these,this study aims to clarify the metabolites and metabolism of parthenolide in rats and rat intestinal bacteria biotransformation,and provide basis and ideas for the development of parthenolide and new drug discovery.Objective:With the help of silica gel column chromatography and preparative high performance liquid chromatography?Pre-HPLC?,the microbial transformation sample was isolated and purified.The structures of metabolic products were identified by HR-MS and NMR technologies.The metabolites and metabolism of parthenolide in rats and intestinal bacteria were identified and deduced by UPLC-MS technology and subsequent analysis strategies.Methods:1.Firstly,parthenolide was cultured with rat intestinal bacteria extracts in anaerobic culture solution.The mixture was extracted with ethyl acetate.Then the extract was separated by silica gel column chromatography and Pre-HPLC.The structures of the metabolites were identified with HR-MS and NMR technologies.2.Rats were administered intragastrically with parthenolide,and the bile,urine,feces of rats and the intestinal bacteria incubation solution in vitro were collected and measured by UPLC-MS.The combination of three adduct ions deduce the metabolism of parthenolide with Metabolite Pilot software.Results:In the rat intestinal bacteria culture solution of parthenolide,3sulfur-containing metabolites were separated and identified respectively as?11S?-13-sulfydryl-11,13-dihydroparthenolide?PA-H₂S?,13,13'-thiobis??11S?-11,13-dihydroparthenolide??PA₂-S?and13,13'-dithiobis??11S?-11,13-dihydroparthenolide??PA₂-S₂?.After UPLC-MS analysis,33 metabolitesof parthenolide including 18 sulfur-containing metabolites were identified.Conclusions:In this study,the metabolism of parthenolide was clarified using the combination of intestinal bacteria biotransformation and UPLC-MS for data analysis and metabolites identification.We demonstrated sufficiently the characteristic biotransformation pathway of sesquiterpene lactones in intestinal bacteria transformation.