Studies On The Anticancer Active Constituents Of Panax Ginseng C.A. MEYER. And The Degrade Grease Active Constituents Of Helichrysi Flos. Syn

Panax ginseng C.A.Meyer is an perennial herbal plant belonging to the Aralaceae family, which mainly distributes in the Northeast China,and has conspicuous pharmacologic action as a valuable traditional medicine.Panax Ginseng was used to enhance the health and treat the various diseases.According to the references,the dammarane-type panaxsaponins in Panax ginseng which were considered for the biological agents for lightening fatigue, delaying insenecence,anti-cancer,accommodating central nervous system,regulating immunity,improving insufficiency of blood in heart or brain.In order to find new active agent from this plant,the chemical study of the extract of panoxatriol saponins of the leaves of Panax ginseng was carried out.By various methods of isolation（silica gel column chromatography,ODS gel column chromatography ）,14 compounds were isolated.By means of spectroscopic analysis and physiochemical properties, they were identified as 20（R）-Protopanaxatriol（1）,20（R）-Dammar-3β,6α,12β,20,25-pentol （2）,Dammar-20（22）,24-diene-3β,6α,12β-triol（3）,20（S）-Dammar-25（26）-ene-3β,6α,12β.20, 24（R）-pentol（4）,3β,6α,12β-trihydroxy-22,23,24,25,26,27-hexanordammaran-20-one（5）, ginsenoside Rh₄（6）,20（R）-ginsenoside Rh₁（7）,20（S）-ginsenoside Rh₁（8）, 3β,6α,12β-trihydroxy-22,23,24,25,26,27-hexanordammaran-20-one,6-O-β-D-glucopyranosid e（9）,20（R）-Dammar-3β,6α,12β,20,25-pentol,6-O-β-D-glucopyranoside（10）,20（S）-Dammar -3β,6α,12β-triol-20,25-epoxy,6-O-β-D-glucopyranoside（11）,20（R）-dammar-3β,6α, 12β,20,25-pentol,6-O-（α-L- rhamnopyranosyl-（2→1）-O-β-D-glucopyranoside（12）, 20（R）-ginsenoside Rg₂（13）,20（S）-Dammar-24-ene,3β,6α,20-trihydroxy-12,23-epoxy,6,20-di-O -β-D-glucopyranoside（14）,respectively.Among them,compound 5 and 14 were new compounds.Meanwhile,the study of the quality determination methods on ginsenoside Rg₃（R,S） and Rh₂（R,S） which were transformed from the extract of panoxadiol saponins in the leaves of Panax ginseng by HPLC was carried out.Rg₃（R）,Rg₃（S）,Rh₂（R）,Rh₂（S） can reach the standard of chromatograms separation under the chromatograms conditions which mobile phase was CH₃CN-H₂O-H₃PO₄（44:56:0.028）,the detection wavelength was 203nm,the flow rate was 1.0ml/min,room temperature.The results of studied on the precision of constancy, reproduction quality and constancy of sample showed this method was accurate,sensitive, stable which could be used for the quality control the contents of ginsenoside Rg₃（R,S）, Rh₂（R,S）,and the establishment of this method was the substructure for the new drug study about ginsenosides Rg₃,Rh₂ and the evaluation of pharmacodynamics, Now adiposis is an increasingly serious public epidemic disease,and threaten human being’s health and normal life.During the prophase course of our active screening studies on some medicinal flowers,we found that the methanolic extract of the flowers of Helichrysum arenarium has the conspicuous activity to anti-obesity,the further study on the chemical constituents and the activity of reducing grease of Helichrysum arenarium were carried out.The inflorescence of Helichrysum arenarium（L.） MOENCH（Helichrysi flos syn. Stoechados flos） is a native perennial herbal plant of the Asteraceae family.And mainly distributes in the northern,southern and central region of Europe.In China,it mainly grows in north of Xin-jiang Province.In Europe,the therapeutic application of Helichrysum arenarium（L.） MOENCH has been used as a folk medicine due to its choleretic,diuretic,and detoxify function for long time. The sap of its flos could be used to treat the disease of digestive system,such as stimulating the gastric secretion,adjusting the bilifaction and gallbladder disorders and so on.In addition, the flos also showed the anti-inflammatoty activity,and it was used to treat cystitis,arthritis, rheumatism.Literatures reported the active ingredients of Helichrysi flos were mainly phenolics such as flavonoids,phenolic acids.The thesis contained the studies on the chemical constituents and the activity of reducing grease.The study on the chemical constituents:For the purpose of finding the chemical constituents with the bioactivity in Helichrysi flos,the further chemical study on its MeOH extract was carried out.By various methods of isolation（silica gel column,ODS gel column, HPLC）,68 compounds were isolated,and by the methods of physico-chemical property,and spectrographic analysis,67 compounds were identified as 7-Hydroxy-5-methoxyphthalide,7-O-β-D-glucopyranoside（1）,（2R,3R）-1-Phenyl-2,3-butane diol-2-O-β-D-glucopyranoside（2）,Everlastoside A（3）,5,7-Dihydroxyphthalide-7-O-β-D-glucopyranoside （4）,Scopolin（5）,Syringin（6）,Dihydrosyringin（7）,Undulatoside A（8）, （E）-4-Hydroxybenzalacetone-3-O-β-D-glucopyranoside（9）,Tortoside B（10）, （2R）-Helichrysin（11）,（2S）-Helichrysin（12）,4-Allyl-2-methoxyphenyl 6-O-β-D-apiofurano syl（1-6）-β-D-glucopyranoside（13）,（2S）-Naringenin-7-O-β-D-glucopyranoside（14）, Kaempferol-3-O-β-D-glucopyranoside（15）,Apigenin-7-O-β-D-glucopyranoside（16）, Luteolin-7-O-β-D-glucopyranoside（17）,Luteolin-3’-O-β-D-glucopyranoside（18）, Orcinol-β-D-glucopyranoside（19）,Adenosine（20）,Everlastoside B（21）,Lcariside F2（22）, p-Methoxyphenyleth-anol-O-β-D-apiofuranosyl-（1-6）-β-D-glucopyranoside（23）,3-Methyl butanol-1-O-β-D-apiofuranosyl-（1-6）-β-D-glucopyranoside（24）,Icariside D1（25）, Phenethanol-β-vicianoside（26）,（2R,3R）- 1-Phenl-2,3-bunediol-2-O-β-D-apiofuranosyl-（1-6）-β-Dglucopyranoside（27）,3-（Z）-Hexenol-O-β-D-apiofuranosyl-（1-6）-β-D-glucopyranoside （28）,（7R,8S）-Dihydrodehydrodiconiferyl alcohol-4-O-β-D-Glucopyranoside（29）, Quercetin-3-O-β-D-glucopyranoside（30）,Chalonaringenin-2’-O-β-D-glucopyranoside（31）, （2R,3R）-Dihydrokaempferol-7-O-β-D-glucoside（32）,Benzylalcohot-β-vicianoside（33）, Everlastoside C（34）,Helicioside A（35）,Everlastoside D（36）, 5,7-Dihydroxyphthalide-7-O-β-D-gentiobioside（37）,Benzoic acid-β-D-gentiobioside（38）, （2S.3S）-1-Phenl-2,3-bunediol-2-O-β-D-glucopyranosyl-（1-6）-β-D-glucopyranoside（39）, Luteolin-3-methoxyl-6-hydroxy-7-O-β-D-glucopyranoside（40）,Aureusidin-6-O-β-D-glucpyranoside （41）,Scutellarein-7-O-β-D glucopyranoside（42）,Kaempferol-3-O-（3-β-D-glucopyranosyl） -β-D-glucopyranoside（43）,Phenethanol-β-D-gentiobioside（44）,Maltol-3-O-β-D-apiofuranosyl-（1-6）-O-β-D-glucopyranoside（45）（2R）-Eriodictyol-5-O-β-D-gluco pyranoside（46）,3-O-p-Coumaroylshikimicacid-4-O-β-D-glucopyranoside（47）, 2R）-Naringenin-5-O-β-D-gentiobioside（48）,（2S）-Naringenin-5-O-β-D-gentiobioside（49）, Luteolin-6-hydroxy-7-O-β-D-glucopyranoside（50）,Apigenin-7-O-β-D-gentiobioside（51）, Quercetin-3-O-rutinoside（52）,Kaempferol-3-O-gentiobioside（53）,2,4,6-Trihydroxylaceto phenone-2,4-di-O-β-D-glucopyranoside（54）,（2R）-Naringenin-5,7-di-O-β-D-glucopyranoside （55）,（2S）-Naringenin-5,7-di-O-β-D-glucopyranoside（56）,（2S）-Naringenin-5,4’-di-O-β-D -glucopyranoside（57）,Apigenin-4’,7-di-O-β-D-glucopyranoside（58）,Kaempferol-3,4’-di-O-β-D-glucopyranoside（59）,Chalonaringenin-2’,4’-O-β-D-glucopyranoside（60）, Quercetin-3,3’-di-O-β-D-glucopyranoside（61）,Chalonaringenin-2’-O-β-D-gentiobioside（62）, ArenariumosideⅡ（63）,Apigenin-7-O-β-D-glucopyranosiduronic acid methyl ester（64）, ArenariumosideⅢ（65）,Everlastoside E（66）,ArenariumosideⅣ（67）.Among them,compound 2,3,4,21,23,24,27,28,34,36,37,39,47,48,49,57,62,63, 65,66,67 were 21 new compounds,and compound 1、5、6、7、8、9、10、13、14、18、 19、20、22、25、26、29、32、33、35、38、40、41、42、43、44、45、46、50、51、52、54、55、56、58、59、60、61、64 were 38 comopounds isolated from Helichrysum for the first time,and compound 11 were isolated from Helichrysum arenarium for the first time.The study on the activity of reducing grease:1 The result of study on MeOH extract, EtOAc layer,MeOH eluted and H₂O eluted of Helichrysi flos on the neutral fat of primary culture liver cells in mice showed that MeOH extract,EtOAc layer,MeOH eluted had the noticeable activity of degrading the content of neutral fat of primary culture liver cells of mice.2 The same study on the compound 1,11,12,14,15,16,31,45,47 were valuated respectively,and the results indicated that the compound 11,14,15,31,47 had the marked activity of reducing grease,and among them,compound 31,47 showed the most conspicuous activity.On the above analysis,it could be concluded that the flavone, flavonone,chalcone and acids were the main constituents with bioactivity in Helichrysi flos.3 The study of the effect of compound 15 and its aglycone on the visceral lipopexia in mice was carried out,and investigated the weight,viscero-fat and the contents of glycerinate （TG）,total cholesterol（TC）,free fatty acids and glucose in blood plasma of mice,and the results of the glucoside showed better activity on reducing grease than that of aglycone.