The Research On Saponins And Anti-allergy Of Belvedere Fruit

Belvedere fruit is the dry fruits ofKochia Scoparia (L.) Schrad., which has beentraditionally used as a herbal plant. It has been effective medicine because its abilitieson removing dampness heat, wind-dispelling and itching-arresting. It has been in use,oral and topical, for over 1,000 years in China, safe and harmless. According topharmacology, belvedere fruit has an antibacterial ability to dermatomyces on somedifferent level. Water extractions of belvedere fruit can water extract inhibitedmonocyte-macrophage function and the delayed-type hypersensitivity (DTH) [7].70% ethanol extractions of belvedere fruit can inhibit the allergic reactions bothimmediate and DTH. Some triterpenoid saponin and sapogenin had been purificatedfrom the belvedere fruit. By now, there have been very few reports on both itschemical constituents and biological activities, and the structure-activity relationshipis still uncovered. The information about the structure-activity relationship is veryimportant on the searching of lead compounds from nature (leadcompounds), thestructural transformation or modification and creation of a new drug. Thus, this thesisis mainly aim on the problems on the belvedere fruit study. The active site was locatedby active tracing method. Biological active the monomers of saponins compoundswere colleted by the purification of active site. GE had been used to determine itsstructures in order to detect the antiallgergic agents activities of every compound. Theinformation on the structure-activity of total saponins from Fructus Kochiae wereacquired by the analysis of the chemical structures and the antiallgergic agentsactivities. The medicinal value and the pharmacology of total saponins from FructusKochiae was understood by the pharmacological experiments. The active situs ofFructus Kochiae and the Structure-activity relationship of its' components wasinvestigated in order to provide the references to the further study of effectivecomponents and the developments of new anti-allergic drugs. In this paper, the theoretical references were provided to the reasonable and effective development tobelvedere fruit. The experimental results are summarized as follows:1. the selections to the active site of saponins from Fructus KochiaeThe active situs of silicates gel column chromatography of total saponins fromFructus Kochiae was located by filtered using experimental model of the miceallergic cutaneous pruritus. The Structure-activity relationship was investigated bycomparation of anti-allergic effects among the collections of GC of total saponins, theactivity situs of CH₃Cl—MeOH eluante was (2BV) changed from 9:1 to 6:4. Theanti-allergic effects of 3-O-β-D-Xylopyranoy 1 (1→3)-β-D-glucuronopyranosylOleanolic acid was more effective than xylopyranoy. The anti-allergic effect wasn'tdetected in the 3-O-β-D-Xylopyranoyl(1→3)-[6-O-Methyl-glucurono- pyranosyl]Oleanolic acid. Conclusion: the anti-allergic effective components were mainly exitedon the eluante of CH₃Cl—MeOH 9:1(2.3.BV)8:2(1-3BV) and 7:3(1-3BV). Theanti-allergic effects would be strengthened when 3-O-β-D-Xylopyranoyl(1→3)-β-D-glucuronopyranosyl Oleanolic acid connected on xylopyranoy 3C. The carboxyofβ-D-glucuronopyranosyl Oleanolic acid was an important functional group whosemodification would cause the changes of compounds' anti-allergic effects.2. The location and identification of the active ingredientSilica gel column chromatography of 200-300 head (6×90cm) was used toselect the active compound from saponin in scoparia fruits. Conditions of gradientelution, depending on the results of the screening to the active situs, were 9:1→6:4chloroform-methanol (active site) gradient elution. Using the TLC of different polarsolvents, the active situs' components of Fructus Kochiae were purified in order tocollect the monomeric compounds as many as possible.Results: from the 4th, 5th and6th position (chloroform-methanol 8:2 resolution 1BV,2BV,3BV) for compounds;From 3,4 site (chloroform-methanol 9:1 in the first 3BV,8:2 1BV) for compoundsâ…¡.From parts 2,3 (chloroform-methanol 9:1 resolution 2BV,3BV) complexesâ…¢.Through physical and chemical reactions, IR,1 H-NMR and TLC. 13C-NMR spectraidentification means that three compounds Compoundâ… oleanolic acid 3-O-beta-D -glucopyranosyl (1→3)-β- D-glucuronide (3-D-Xylopyranoyl-β(1→3)-β-D-glucuronopy ranosyl Oleanolic acidâ… ) Compoundâ…¡as oleanolic acid 3-O-beta-D-glucopyranosyl (1→3)-β- D-glucuronide methyl glycosides (-D- Xylopyranoyl O-(1→3) -[6-O-Methyl-g lucuronopyranosyl] Oleanolic acidâ…¡)â…¢of oleanolic acid(oleanolic acid,â…¢). the research on the anti-allergic effect of total saponins fromFructus KochiaeThe research on the anti-allergic effects and the dose-effect relationship ofsaponin in Fructus Kochiae. Method: The models of allergic skin itch in mice causedby 4-AP and the experiments on feet swelling of mice caused by histamine. Theinhibitory effects of total saponins in Fructus Kochiae on allergic skin itch in micecaused by 4-AP and the feet swelling of mice caused by histamine were shown whenthe dose rose up to 32mg/kg. The anti-allergic effects rose up when the dosage ofgastric perfusion grew up. When the dosage came up to 150mg/kg, the anti-allergicreaction was almost 100%. The immediate allergic reaction can be inhibited by totalsaponins in Fructus Kochiae. The mechanism of actionmay may related by thestabilization hypertrophy cell membrane, reducing the reflection of histamine and theproinflammatory action of anaphylactic mediatorThe anti-allergic effect had andose-effect relationship. It was shown that the dosage of effectual value was 32mg/kgand the dosage of final affection was 150mg/kg.Using the isolated compounds for materials, 4-AP-induced allergic pruritusmodel were used to study its anti-allergy effect size, compare their structuralcharacteristics, in order to explore structure-activity relationship. The results showedNobilisidenol oleanolic acid has anti-allergy effect. But instead of saponin oleanolicacid 3-O-beta-D-glucopyranosyl (1→3) -β-D-glucuronide compared Anti-allergiccapability much weaker oleanolic acid 3-O-beta-D-glucopyranosyl (1→3)-β-D-glucuronide methyl RP has shown resistance allergic. Oleanolic acid Note 3 carbonatoms linked oligosaccharidesβ- D-glucuronide the carboxyl functional group isimportant. Methyl after its anti-allergy disappeared. This may be modified functionalgroups suggest anti-allergic effect of a larger change. The references were provided to develop a new type of anti-allergic drugs.3. the research on the purification technology of saponins with macroreticular resinIt is investigated that the technological parameters of purifying of total saponin fromFructus Kochiae Scopariae with macroreticular resin in order to provide theinstructions to the industrial production of total saponin. Using the yield, elution rateand purity for indicators, it is detected the abstraction and elution parameters ofpurifying total saponin from Fructus Kochiae Scopariae with macroreticular resin.The 70% Fructus Kochiae Scopariae ethanolic fluidextract 260mL in volume wasflowed through macroreticular resin D-101-â… , (1.5×15mm, 2.238g sample.mL^-1). Therate of solution flowing through the column is 1.2mL·min^-1. The impurity was washedout with 3BV dd-H₂O. Then the total saponin was eluted with 3BV 50% ethanolicelution. After the purifying with macroreticular resin, it is tested that the elution rate is90.56%, the yield is 42.89%, and the purified efficiency is 263.59%. It is thatcertificated that this method can purify the total saponin from Fructus KochiaeScopariae efficiently.