Extraction And Separation Of Effective Components From Saussurea Lappa Root And Derivatization Of Dehydrocostus Lactone

The dried roots of Saussurea lappa C.B Clarke, as a traditional Chinese herbalmedicine, had been widely used for the treatment of various kinds of disorders such asindigestion, diarrhea, vomit, cough, colic. It was shown that its effective componentswere sesquiterpenes, such as dehydrocostus lactone and costunolide, which had the ef-fect of anti-ulcer, antibacterial and anti-cancer. According to the characteristics of effec-tive components of Saussurea lappa root, this paper carried out a systematic studyabout the effective components of extraction and separation from Saussurea lappa root.Then, the derivatization of dehydrocostus lactone and the anti-cancer activity of the de-rivants were studied. These research provided technical guidance for extracting and se-parating dehydrocostus lactone, costunolide and other effective components rapidly andeconomically, explored medicinal components further, and provided basic data for thenew drug research and development.The main achievements are presented as follows:The extractive methods, chemical component, pharmacological activity and deriv-ative methods of Saussurea lappa root had been reviewed and evaluated systematically.On the basis, the general study and technological implement scheme of this paper wereput forward and formed.Dehydrocostus lactone and costunolide were selected as index components, the ex-traction efficiency of solvent extraction, steam distillation and microwave assisted ex-traction were compared. Meanwhile, the separation ability of extraction and macropor-ous resin absorption for low polar compounds were made a comparison. Researchshowed that3.5g of dehydrocostus lactone and costunolide from100g of Saussurealappa root can be gained in the case that microwave assisted extraction at the micro-wave power of400w, ratio of solid to liquid of1:15, and extraction time of10min for3times; the purity of dehydrocostus lactone and costunolide that separated from themethanol extraction reached43.1%and46.8%when using95%ethanol for eluting theAB-8type macroporous resin.11kinds of compounds from the petroleum ether extract of Saussurea lappa rootwere gained by using column chromatography on silica gel, thin layer chromatography(TLC), recrystallization and other methods. In addition the7kinds of them were identi-fied by TLC、IR、NMR and other identification technology, which were alantolactone, isoalantolactone, dehydrocostus lactone, costunolide, stigmasterol,12-methoxydihydrocostunolide and pregnenolone.Petroleum ether extract of Saussurea lappa root was separated and purified byHigh-speed counter-current chromatography (HSCCC). Partition coefficient of dehy-drocostus lactone and costunolide was measured by HPLC. And combining the separa-tion of the results of analytical type HSCCC, the solvent selection of HSCCC was ef-fectively guided. Solvent system, revolution velocity, flow rate and some other influ-ence factors were investigated by an orthogonal experimental design. Research showedthat the best condition of HSCCC was solvent system of petroleum eth-er-methanol-water (5:7:3), revolution velocity of1000rpm and flow rate of3ml/min.In this condition, high purity dehydrocostus lactone and costunolide could be collectedwithin150min, whose purity was more than95%and whose quantities of preparationwere able to achieve more than140mg. When partition coefficient was measured, thetest values of HPLC cannot reflect real distribution of the solvent system in the HSCCC.On the other hand, thoroughly separating petroleum ether extract of Saussurea lapparoot, a new compound10-methoxy-artemisinic acid was gained.In order to improve the dehydrocostus lactone’s anticancer acitivity and bioavaila-bility, structural modification and derivatization of it was studied. According to theα-methylene-γ-lactone of dehydrocostus lactone and the theory of Michael additionreaction, different compounds were added to the C-13of dehydrocostus lactone, andthus series of derivants were prepared. This research finished7kinds of derivative reac-tion such as the reaction of piperidine, morpholine and so on. The derivants were sepa-rated by column chromatography on silica gel and their structure were identified by IR,NMR and other identification technology. The anticancer activity of dehydrocostus lac-tone, costunolide and the derivants was measured by MTT method. The experimentalresults showed the4-OH piperidine, morpholine, pyrrolidine, methanol and diethyla-mine derivants had obvious antitumor activity and their activities were better than de-hydrocostus lactone, so they can be treated as candidate drugs to continue to studyingfurther.