CN103641871B - Big ring polyphenolic substance and its preparation and the application in pharmacy - Google Patents
Big ring polyphenolic substance and its preparation and the application in pharmacy Download PDFInfo
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Abstract
The invention provides the new application from the isolated big ring polyphenolic substance of Herba Lysimachiae Clethroids extract in pharmacy.Described big ring polyphenolic substance includes：Compound 1, compound 2, compound 3 and compound 4, structural formula is as follows：
Description
Technical field
The present invention relates to Chinese medicine, and in particular to the purposes of Chinese medicine, more particularly to big ring polyphenolic substance and its prepare with
Prepare the purposes in preventing and treating diabetes and hepatic fibrosis medicines.
Background technology
Diabetes are a kind of metabolic disturbance diseases as caused by Different types of etiopathogenises characterized by chronic hyperglycemia.With the mankind
The change of life style, the incidence of disease of diabetes are stepped up, it is contemplated that were reached 300,000,000 people by 2025, severely compromised the mankind
Health and social development.Abnormal carbohydrate metabolism is the major pathogenetic factor of diabetes, and wherein participates in the relevant enzyme of glycometabolism to sugar
Metabolism has important influence, therefore it is significant to adjust treatment of the activity of glycometabolism enzyme to diabetes（Based on glycometabolism
The Chinese medicine anti-diabetic progress of enzyme regulation, CHINA JOURNAL OF CHINESE MATERIA MEDICA, 2012,37 (23)：3519-3525）.Hypoglycemic medicine
Generally there are Chinese medicine and Western medicine.Wherein conventional oral hypoglycemic drug is divided into non-insulin class rush based on non-insulin class medicine
Insulin secretion agent class, biguanides, alpha-glucosidase inhibitor class, insulin sensitizer etc..Modern pharmacological research proves, a lot
Chinese medicine has blood sugar reducing function.Chinese medicine is widely studied and applied because its high-efficiency low-toxicity acts on, plurality of Chinese extract and composition
The conditioning agent of glycometabolism enzyme is proved to be, compared with Western medicine, Diabetes Mellitus Treated With Traditional Chinese Medicine has Small side effects and cheap etc. excellent
Point, there are preferable clinical landscapes.
Liver fibrosis (hepatic fibrosis, HF) is a variety of virulence factors, such as virus, chronic hepatitis, Chronic Alcohol
Poisoning, liver hemostasis, fatty liver etc. cause in hepar damnification and liver caused by inflammation the extensive hyperplasia of fibrous connective tissue and heavy
Product.Famous American hepatopathy scholar Friedman SL once pointed out that who can block or delay the generation of liver fibrosis, and who will just be cured
Most of chronic liver diseases.The activation of HSCs (hepatic stellate cell, HSC) is simultaneously converted into flesh into fiber finer
Born of the same parents (myofibroblast, MFB) are the key links that liver fibrosis occurs, and the HSC after activation synthesizes substantial amounts of extracellular matrix
(extracellular matrix, ECM), excessive ECM is constantly deposited in liver, causes the destruction of liver normal configuration and liver
The disorder of function simultaneously ultimately results in liver fibrosis.TGF (TGF-β 1) is to belong to one group of newly discovered regulation cell
Growth and the TGF-β superfamily of differentiation, research show that TGF-β 1 plays an important role in the occurrence and development of liver fibrosis（Thunder
The correlative study 2013,17 (2) that pa mycin develops with rat liver fibrosis：259-261；Liver fibrosis progress, 2012,8
(10)：199-201）.Substantial amounts of clinical practice and experimental study show, Chinese medicine from many aspects, multipath, too many levels there is difference
The effect of anti hepatic fibrosis of degree, and curative effect affirmative, few side effects.
As what diabetes-related complication was studied further gos deep into, the hepatic injury of diabetes correlation is also increasingly learned
The attention of persons.Have to exist between many data confirmation diabetes and liver fibrosis at present and associate, but diabetes correlation liver fiber
Change pathogenesis not yet to illustrate.Because the research at present to diabetes correlation liver fibrosis is few, therefore intervene for its specificity
Medicine report it is very few, include Telmisartan and Simvastatin etc. currently for the medicine of diabetes correlation liver fibrosis.
Compared with Western medicine, Chinese medicine has the characteristics that Mutiple Targets, multipath, efficient, less toxic, for the treatment of diabetes correlation liver fibrosis
With good clinical landscapes.
Chinese medicine penthorum chinense pursh (Penthorum chinense Pursh）Also known as root vegetables, Chinese penthorum herb, mountain delicacy pearl etc. are pulled, according to《My god
Precious book on Chinese herbal medicine》、《Herbal for Relief of Famines》Deng record, penthorum chinense pursh has the effect such as dredging collateral and promoting blood circulation, dissolving stasis dehumidifying, promoting blood circulation to remove blood stasis, document report
Penthorum chinense pursh has preventing and treating hepatitis, the pharmacological activity of Liver protection.A series of the present inventor's therefrom isolated big ring Polyphenols chemical combination
Thing.Big ring polyphenol compound has the pharmacological action of anti-hepatitis（Two antiviral compounds from the
Plant Stylogne cauliflora as inhibitors of HCV NS3protease, Bioorganic＆
Medicinal Chemistry Letters,2003,13(17)：2925-2928）, but report both domestic and external is taken a broad view of, it is showed no
Such compound is hypoglycemic and the pharmacology of anti-hepatic fibrosis is reported.The present inventor shows to extract from penthorum chinense pursh by research to be separated
Obtained big ring polyphenolic substance has anti-diabetic and fibrosis effect, therefore, can further research and develop anti-diabetic
And the new drug of liver fibrosis.
The content of the invention
The technical problems to be solved by the invention in purpose extracted in research and design from penthorum chinense pursh it is isolated
The preparation of big ring polyphenolic substance and pharmaceutical applications.
The invention provides a kind of big ring polyphenolic substance, has following structural formula I：
Wherein, R1For H or
R2For。
Big ring polyphenolic substance of the present invention is prepared from the extraction of Chinese medicine penthorum chinense pursh.
Big ring polyphenolic substance of the present invention includes following compounds：
Compound 1:Yellow powder, C35H28O17,ESI-MS m/z:721[M+H]+, compound 1 is Pinocembrin-7-
O-[4”,6”-hexahydroxydiphenoyl]-β-D-glucose.（Pinocembrin -7-O- [4 ", 6 "-hexahydroxy biphenyl diformazans
Acyl group]-β-D-Glucose glycosides）
Compound 2：C42H34O21, yellow crystal formation powder.ESI-MS(m/z873.42[M-H]-), compound 2 is
Thonningianins A be Pinocembrine dihydrochalcone-7-O- [3 "-O-galloyl-4 ", 6 "-
hexahydroxydiphenoyl]-β-glucose.（Pinocembrin dihydrochalcone -7-O- [3 "-O- galloyls -4 ", 6 " -
Hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides）
Compound 3:White powder, C42H32O21,ESI-MS m/z:871[M-H]-, compound 3 is Pinocembrin-7-
O- [3 "-O-galloyl-4 ", 6 "-hexahydroxydiphenoyl]-β-glucose.（[3 "-O- are not eaten pinocembrin -7-O-
Sub- acyl group -4 ", 6 "-hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides）
Compound 4:White powder, C35H30O17,ESI-MS m/z:721[M-H]-, compound 4 is Thonningianins
B is Pinocembrin dihydrochalcone-7-O- [4 ", 6 "-hexahydroxydiphenoyl]-β-D-glucose.
（Pinocembrin dihydrochalcone -7-O- [4 ", 6 "-hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides）
。
It is a further object of the present invention to provide the preparation method of the big ring polyphenolic substance, this method includes following step
Suddenly：
(1) root vegetables medicinal material extract and initial gross separation are pulled
Root vegetables crushing is pulled, 70% alcohol heat reflux extracts three times, and each extraction time is 1 hour, adds 3 times of weights of raw material every time
70% ethanol W/L of amount, merge extract solution, be concentrated under reduced pressure into no alcohol taste and obtain crude extract medicinal extract（Relative density is 1.12~1.18）；
Then plus 1 times of weight of medicinal extract water W/L be suspended after by macroporous resin column, successively with water, 10% ethanol, 20% ethanol, 30% second
Alcohol, 50% ethanol, 70% ethanol and 95% ethanol elution, eluent are 10~12 times of column volume, respectively obtain water, 10% ethanol,
20% ethanol, 30% ethanol, 50% ethanol, 70% ethanol and 95% ethanol eluate, no alcohol taste is concentrated into respectively and obtains each elution position
Medicinal extract（Relative density 1.12~1.18）；
(2) prepared by big ring polyphenolic substance separation
Above-mentioned wherein 70% ethanol eluate is taken, uses methanol：Water=4:1 dissolving, through MCI-CHP20P gel column chromatographies, with first
Alcohol：Water（1:9~1:0）Gradient elution, obtain 10 fraction (Fr1-Fr10)（TLC lamellaes detect）；Take fraction Fr8Methanol is molten
Xie Hou, inverted silicagel column（Elution requirement：Methanol：Water=1:9~1:0）Segmentation, obtains I-VI 6 fraction（TLC lamellaes are examined
Survey）, wherein fraction III is through methanol：Water=1:After the dissolving of 4 mixed solutions, then inverted silicagel column（Methanol：Water=1:4~1:0 gradient
Elution）Compound 1 is purified to obtain, fraction IV is through methanol：Water=1:After the dissolving of 4 mixed solutions, then through gel column（Methanol：Water=1:4~
1:0 gradient elution）Purify repeatedly（2-3 times）Compound 2 and compound 4 are obtained, fraction V is through methanol：Water=1:4 mixed solutions dissolve
Afterwards, then through gel column（Methanol：Water=1:4~1:0 gradient elution）Purify repeatedly（2-3 times）Obtain compound 3.
The present inventor further uses HPLC methods, investigated it is above-mentioned prepare big ring polyphenolic substance 1-4 pulling root vegetables
Content in medicinal material, measuring contents of the compound 1-4 in root vegetables is pulled is respectively：4.27-4.63‰、0.97-1.23‰、
2.56-2.78‰、0.68-0.91‰。
It is yet another object of the invention to provide application of the big ring polyphenolic substance in pharmacy.
The invention provides application of the big ring polyphenolic substance in preventing/treating Rezulin is prepared.
The invention provides application of the big ring polyphenolic substance in preventing/treating hepatic fibrosis medicines are prepared.
Big ring polyphenol compound of the present invention, by vitro test, there are significant suppression alpha-amylase and suppression
The effect for the hepatic stellate cell proliferation that TGF-β 1 induces, available for the medicine for preparing anti-diabetic and liver fibrosis.
The medicine group that medicine of the present invention is made up of the big ring polyphenolic substance as active component and pharmaceutic adjuvant
Compound.
The pharmaceutical composition that medicine of the present invention is made up of the compound 1 as active component and pharmaceutic adjuvant；Or
The pharmaceutical composition that medicine of the present invention is made up of the compound 2 as active component and pharmaceutic adjuvant；Or
The pharmaceutical composition that medicine of the present invention is made up of the compound 3 as active component and pharmaceutic adjuvant.
The pharmaceutical composition that medicine of the present invention is made up of the compound 4 as active component and pharmaceutic adjuvant.
The pharmaceutical composition of the present invention is tablet, capsule, granule, powder or pill etc..
The invention provides new preventing and treating diabetes medicament and anti-hepatic fibrosis medicines, the raw material sources of medicine enrich, secondary
Act on small, be advantageous to environmental protection, there is larger clinical value.
Brief description of the drawings
Fig. 1 pulls root vegetables medicinal material, compound 1-4 HPLC compares figures
Ordinate is response intensity, unit（AU）, abscissa is the time, unit（Minute）
A:Pull root vegetables medicinal material；B:Compound 2；C:Compound 3；D：Compound 1；E:Compound 4
Embodiment
With nonlimiting examples, the present invention is further described below.
Embodiment is raw materials used to be commercially available.
The preparation of the big ring polyphenolic substance of embodiment 1
（1）Pull root vegetables medicinal material extract and initial gross separation
Root vegetables 5kg crushing is pulled, 70% alcohol heat reflux extracts three times, and each extraction time is 1 hour, each liquid feeding 15L,
Merge extract solution, be concentrated under reduced pressure（55℃）Extremely crude extract medicinal extract is obtained without alcohol taste（Relative density is 1.12~1.18）, claim its weight
For 625g, yield 12.5%.Then water（650ml）Pass through macroporous resin column after suspension（D-101）, successively with water, 10% ethanol,
20% ethanol, 30% ethanol, 50% ethanol, 70% ethanol and 95% ethanol elution, eluent are 10~12 times of column volume, respectively
To water, 10% ethanol, 20% ethanol, 30% ethanol, 50% ethanol, 70% ethanol and 95% ethanol eluate, it is concentrated under reduced pressure respectively（55
℃）To the medicinal extract that each elution position is obtained without alcohol taste（Relative density is 1.12~1.18）, claim its weight be respectively 112g, 108g,
75g, 55g, 66g, 60g and 55g.
（2）It is prepared by big ring polyphenolic substance separation
The medicinal extract 60g for the 70% ethanol eluate concentration that Example 1 obtains, through MCI-CHP20P gel column chromatographies, with first
Alcohol：Water（1:9~1:0）Gradient elution, through TLC（Silica gel thin-layer plate）Detection, obtains 10 fraction (Fr1-Fr10).Fraction Fr8Through
Reverse phase silica gel post（ODS-C18）Segmentation, through TLC（Silica gel thin-layer plate）Detection, I-VI 6 fraction is obtained, wherein fraction III is again through anti-
Phase silicagel column（ODS-C18）Compound 1 (100.1mg) is purified to obtain, fraction IV is through gel column（LH-20）2 times after purification chemical combination
Thing 2 (201.6mg) and compound 4 (62.8mg), fraction V is through gel column（LH-20）Compound 3 is purified to obtain after purification 2 times
（50.5mg）.
Wherein, compound 1:Yellow powder, C35H28O17,ESI-MS m/z:721[M+H]+, nuclear magnetic data and document
(Journal of Natural Product, 1998,61：523-524) report is basically identical, it is thus determined that compound 1 is
Pinocembrin-7-O-[4”,6”-hexahydroxydiphenoyl]-β-D-glucose.（Pinocembrin -7-O- [4 ", 6 " -
Hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides）
Compound 2, C42H34O21, yellow crystal formation powder.ESI-MS(m/z873.42[M-H]-), nuclear magnetic data and document
（Journal of Agricultural and Food Chemistry,2012,60:11097-11103) report is consistent, because
This determination compound 2 is that Thonningianins A are Pinocembrine dihydrochalcone-7-O- [3 "-O-
galloyl-4”,6”-hexahydroxydiphenoyl]-β-glucose.（[3 "-O- do not have pinocembrin dihydrochalcone -7-O-
Infanticide acyl group -4 ", 6 "-hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides）
Compound 3：White powder, C42H32O21,ESI-MS m/z:871[M-H]-, nuclear magnetic data and document (Journal
of Asian Natural Products Research,2006,8(8):757-761) report is consistent, it is thus determined that chemical combination
Thing 3 is Pinocembrin-7-O- [3 "-O-galloyl-4 ", 6 "-hexahydroxydiphenoyl]-β-glucose.（It is tall
Song Su -7-O- [3 "-O- galloyls -4 ", 6 "-hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides）
Compound 4:White powder, C35H30O17,ESI-MS m/z:721[M-H]-, nuclear magnetic data and document (Journal
Of Natural Product, 2000,63：676-679) report is basically identical, it is thus determined that compound 4 is
Thonningianins B be Pinocembrin dihydrochalcone-7-O- [4 ", 6 "-
hexahydroxydiphenoyl]-β-D-glucose.（Pinocembrin dihydrochalcone -7-O- [4 ", 6 "-hexahydroxy biphenyl diformazans
Acyl group]-β-D-Glucose glycosides）
Assays of the big ring polyphenolic substance 1-4 of embodiment 2 in root vegetables medicinal material is pulled
Its method and operating procedure are as follows
（1）HPLC conditions
Chromatographic column：Diamonsil C18column(4.6mm×250mm,5μm,Dikma)；
Mobile phase：Using gradient elution, A phases are acetonitrile, and B phases are 0.2% aqueous formic acid；
Flow velocity：1.0ml/min, run time 70min, Detection wavelength 280nm,
Column temperature is 25 DEG C；The μ l of sample size 10；
The gradient elution program of table 1
（2）Compound 1-4 assay
Take compound 1-4 reference substances（Embodiment 1 is made）, it is accurately weighed, it is configured to contain 350 in every 2mL respectively with 60% methanol
μ g, 100 μ g, 200 μ g, 50 μ g compound 1-4 solution, as reference substance solution.
Step in Example 1（1）Prepare to pull root vegetables crude extract medicinal extract appropriate（Root vegetables medicinal material is pulled equivalent to 10g）, essence
Close weighed, the dissolving of 60% methanol is settled to 100ml, as need testing solution.Reference substance solution and need testing solution each 10 are taken respectively
μ l inject liquid chromatograph, record chromatogram, see Fig. 1.Compound 1-4 retention time is respectively：49.65±0.60、59.80
±0.80、55.55±0.80、54.37±0.70min；Compound 1-4 containing in medicinal material is obtained with calculated by peak area by external standard method
Amount is respectively：4.27-4.63‰、0.97-1.23‰、2.56-2.78‰、0.68-0.91‰.
The external hypoglycemic activity experiments of the compound 1-4 of embodiment 3
1 materials and methods
1.1 trial drug
Embodiment 1 prepares gained monomeric compound 1-4
Experiment material
Acarbose（Beyer Co., Ltd, 50mg/ pieces are commercially available）, glucose determination reagent box（Glucose oxidase-peroxidating
Enzyme process, Shanghai Rongsheng Bioisystech Co., Ltd,）Alpha-amylase（Wako Pure Chemicals Ind.,Osaka,
Japan）, ELIASA（EIx800, Biotek）, 96 porocyte culture plates（Corning/Costar）
1.2 experimental method
1.3.1 developer：First 10.8 grams of sodium benzoate, 5 grams of KI are dissolved in 400ml water, be then settled to
500ml。
1.3.2 1-4 is used into DMSO respectively（Dimethyl sulfoxide）10mg/ml solution is made into, then is diluted to respectively with water
0.556mg/ml, 0.185mg/ml, 0.062mg/ml, 0.020mg/ml and 0.007mg/ml5 various concentrations；Starch is steamed
Distilled water is configured to the starch solution that concentration is 0.16%.Acarbose is configured to 2.5mg/ml, 0.83mg/ with distilled water simultaneously
Ml, 0.28mg/ml, 0.09mg/ml and 0.03mg/ml5 various concentrations；Alpha-amylase is matched somebody with somebody with 25mM PIPES buffer solutions
27.8mg/ml solution is made（PH=6.9）.
1.3.3 it is divided into 6 groups with 96 porocyte culture plates, experiment, respectively blank group compares（Distilled water）, positive controls
（Acarbose）, 4 sample measure groups, every group sets 6 multiple holes.
1.3.4, above-mentioned 4 compounds and acarbose solution are added on plate hole to the corresponding each hole organized into groups respectively, per hole
25ul sample solutions, blank control group add equivalent distilled water, and it is molten to add 12.5ul alpha-amylases respectively at every group of wherein three holes
Liquid, the other three hole add 12.5ul distilled water, and 37 DEG C of temperature incubate 10min, and finally adding 500ul colour reagents respectively at each hole terminates
Reaction, absorbance is determined under ELIASA 630nm wavelength（A）, inhibiting rate is calculated as follows（%）：Inhibiting rate（%）=[1-
(ASample is not enzyme-added-ASample is enzyme-added)/(ABlank control is not enzyme-added-ABlank control is enzyme-added)] × 100%, calculate corresponding half-inhibition concentration（IC50）.
2 experimental results
Using acarbose as positive control drug, as a result show that compound 1-4 prepared by above example 1 can suppress alphalise starch
Enzymatic activity, there is significant hypoglycemic effect.It the results are shown in Table 2
The inhibitory action of 1~3 pair of alpha-amylase of compound of table 2
3 experiment brief summaries
Compound 1-4 has identical configuration：4 of glucose and 6 hydroxyls and gallic acid cyclization, can suppress α-shallow lake
The activity of powder enzyme, there is hypoglycemic, the active activity for being better than positive drug of each monomeric compound, it is seen that this kind ofization
Compound is a kind of good alpha-amylase inhibitor, and speculate hypoglycemic effect be connected with glucose gallic acid number into just
It is related；Big ring polyphenol can be digested and assimilated directly as a part of of medicinal plant by body, and hypoglycemic is developed from natural products
Medicine also has the advantages of cost is low, imply that this kind of compound has a good application prospect.
The compound 1-4 of embodiment 4 suppresses hepatic stellate cell proliferation experiment
1 materials and methods
1.1 experiment material
TGF-β1(Cat#240-B-002, purchased from R＆D companies)；DMEM culture medium dry powders（Gibco, USA）, hyclone
（FBS）（Gibco, USA）, pancreatin (Trypsin) (Amresco, USA), Cell Counting Kit-8 kits（The green skies
Biotechnology research）, human liver microsome proteins system（HSC-T6 cells）(Shuguang Hospital of Shanghai hepatopathy research institute）, microwell plate light splitting light
Degree meter（PowerWave XS, BioTek)
1.2 trial drug
Embodiment 1 prepares gained monomeric compound 1-4
Experimental method
1.3.1 cell culture：With the DMEM containing 10%FBS（Dulbecco’s Modified Eagle’s Medium）Training
Nutrient solution culture HSC-T6 cells, passed on 1 time per 3-4 days
1.3.2 sample preparation：TGF-β 1 is configured to 1ng/ μ l stock solution with 4mM HCl solutions, the used time is dilute with PBS
It is interpreted into required concentration；Compound 1-4 is configured to 20mg/ml stock solution respectively with DMSO respectively, the used time is trained using DMEM
Nutrient solution is diluted to 5ug/ml, 10ug/ml, 20ug/ml, 40ug/ml, 80ug/ml and 160ug/ml5 various concentrations.Positive drug
Colchicin is configured to 1mg/ml stock solution with distilled water, and the used time is diluted to 2.5ug/ml, 5.0ug/ with DMEM nutrient solutions
Ml, 10ug/ml, 20ug/ml and 40ug/ml5 various concentrations.
1.3.3 HSC-T6 cells are inoculated in 96 orifice plates in units of 3000/hole, it is long to subband structures, it is divided into blank pair
According to group, model group, experimental group, blank control group is given 0.5%FBS/DMED and cultivated, and model group gives 2.5ng/ml TGF-
β 1, experimental group give 2.5ng/ml TGF-βs 1 and give the compound 1-4 of 5 various concentrations respectively, are incubated 24h altogether.It is incubated knot
Nutrient solution is abandoned in Shu Hou, suction, and 100 μ l CCK-8 solution are added per hole, 2h is incubated, in microplate spectrophotometer 450nm ripples
Long lower measure absorbance (A), calculates inhibiting rate as follows（%）：Inhibiting rate（%）=（AExperimental group-ABlank group）/（AModel group-ABlank group）×
100%, calculate corresponding half-inhibition concentration（IC50）.
2 experimental results
As a result show that compound 1-4 can suppress TGF-β 1 and induce hepatic stellate cell proliferation, there is significant anti-hepatic fibrosis
Activity.It the results are shown in Table 3
The compound 1-3 of table 3 induces TGF-β 1 inhibitory action of hepatic stellate cell proliferation
3 experiment brief summaries
Compound 1-4 has identical configuration：4 of glucose and 6 hydroxyls and gallic acid cyclization, have and suppress
TGF-β 1 induces the activity of hepatic stellate cell proliferation, has the function that anti-hepatic fibrosis, and speculate on inhibitory activity and glucose
The number of gallic acid is connected with into positive correlation.
Claims (8)
1. the preparation method of big ring polyphenolic substance, the compound is compound 1：[4 ", 6 "-hexahydroxy joins pinocembrin -7-O-
Phenyl-diformyl base]-β-D-Glucose glycosides, compound 2：Pinocembrin dihydrochalcone -7-O- [3 "-O- galloyls -4 ", 6 " -
Hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides, compound 3：Pinocembrin -7-O- [3 "-O- galloyls -4 ", 6 "-six
Xenol diformyl]-β-D-Glucose glycosides, compound 4：Pinocembrin dihydrochalcone -7-O- [4 ", 6 "-hexahydroxy biphenyl
Diformyl]-β-D-Glucose glycosides, structural formula is as follows：
Characterized in that, the preparation method comprises the following steps：
(1) root vegetables medicinal material extract and initial gross separation are pulled
Root vegetables crushing is pulled, 70% alcohol heat reflux extracts three times, and each extraction time is 1 hour, adds 3 times of weight of raw material every time
70% ethanol W/L, merge extract solution, be concentrated under reduced pressure into no alcohol taste and obtain crude extract medicinal extract relative density as 1.12~1.18；Then
Add the water W/L of 1 times of weight of medicinal extract be suspended after by macroporous resin column, successively with water, 10% ethanol, 20% ethanol, 30% ethanol,
50% ethanol, 70% ethanol and 95% ethanol elution, eluent are 10~12 times of column volume, respectively obtain water, 10% ethanol,
20% ethanol, 30% ethanol, 50% ethanol, 70% ethanol and 95% ethanol eluate, no alcohol taste is concentrated into respectively and obtains each elution
The medicinal extract relative density at position is 1.12~1.18；
(2) prepared by big ring polyphenolic substance separation
Above-mentioned wherein 70% ethanol eluate is taken, uses methanol：Water=4:1 dissolving, through MCI-CHP 20P gel column chromatographies, with first
Alcohol：Water 1:9~1:0 gradient elution, obtain 10 fraction Fr1-Fr10, the detection of TLC lamellaes；Take fraction Fr8After methanol dissolving,
Inverted silicagel column, elution requirement：Methanol：Water=1:9~1:0 segmentation, I-VI 6 fraction being obtained, TLC lamellaes detect, wherein
Fraction III is through methanol：Water=1:After the dissolving of 4 mixed solutions, then inverted silicagel column methanol：Water=1:4~1:0 gradient elution is pure
Change to obtain compound 1, fraction IV is through methanol：Water=1:After the dissolving of 4 mixed solutions, then through gel column, methanol：Water=1:4~1:0 ladder
Degree elution, purifies 2-3 times, obtains compound 2 and compound 4, fraction V is through methanol repeatedly：Water=1:After the dissolving of 4 mixed solutions, then
Through gel column, methanol：Water=1:4~1:0 gradient elution, purify 2-3 times repeatedly, obtain compound 3.
2. application of the big ring polyphenolic substance in preventing/treating Rezulin is prepared, it is characterised in that the compound is change
Compound 1：Pinocembrin -7-O- [4 ", 6 "-hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides, compound 2：Pinocembrin dihydro is looked into
Ear ketone -7-O- [3 "-O- galloyls -4 ", 6 "-hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides, compound 3：Himalayan pine
Element -7-O- [3 "-O- galloyls -4 ", 6 "-hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides, compound 4：Pinocembrin
Dihydrochalcone -7-O- [4 ", 6 "-hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides, structural formula are as follows：
。
3. application of the big ring polyphenolic substance in preventing/treating hepatic fibrosis medicines are prepared, it is characterised in that the compound
For compound 1：Pinocembrin -7-O- [4 ", 6 "-hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides, compound 2：Pinocembrin two
Hydrogen chalcone -7-O- [3 "-O- galloyls -4 ", 6 "-hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides, compound 3：
Pinocembrin -7-O- [3 "-O- galloyls -4 ", 6 "-hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides, compound 4：It is tall
Plain dihydrochalcone -7-O- [4 ", 6 "-hexahydroxy biphenyl diformyl]-β-D-Glucose glycosides of pine, structural formula are as follows：
。
4. the application according to any one of Claims 2 or 3, it is characterised in that the medicine is by as active component
Big ring polyphenolic substance and pharmaceutic adjuvant composition pharmaceutical composition.
5. application according to claim 4, it is characterised in that described pharmaceutical composition is by the chemical combination as active component
The pharmaceutical composition of thing 1 and pharmaceutic adjuvant composition.
6. application according to claim 4, it is characterised in that described pharmaceutical composition is by the chemical combination as active component
The pharmaceutical composition of thing 2 and pharmaceutic adjuvant composition.
7. application according to claim 4, it is characterised in that described pharmaceutical composition is by the chemical combination as active component
The pharmaceutical composition of thing 3 and pharmaceutic adjuvant composition.
8. application according to claim 4, it is characterised in that described pharmaceutical composition is by the chemical combination as active component
The pharmaceutical composition of thing 4 and pharmaceutic adjuvant composition.
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| CN105193833B (en) * | 2015-10-13 | 2018-11-30 | 四川古蔺肝苏药业有限公司 | Penthorum chinense pursh monomer is preparing the purposes in Liver protection drug |
| CN108484694A (en) * | 2018-04-04 | 2018-09-04 | 新疆维吾尔自治区药物研究所 | Natural products isostrictiniin and its preparation method and application |
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| CN109125337B (en) * | 2018-07-23 | 2021-06-22 | 澳门科技大学 | Application of penthorum chinense pursh compound in preparation of medicine for treating atherosclerosis |
| CN109793073A (en) * | 2019-03-21 | 2019-05-24 | 苏州美稀生物科技有限公司 | The extracting method at Ilex paraguarensis drink Lipid-lowering activities position and hypoglycemic activity position |
| CN110229204B (en) * | 2019-05-29 | 2020-04-17 | 四川大学 | Preparation method of compound PGHG in Parkinson disease protection medicine |
| CN113143943A (en) * | 2021-06-15 | 2021-07-23 | 西南医科大学附属医院 | Application of Thonningianin A in preparation of medicines as L-type calcium ion channel activator |
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| INSILICO STUDIES OF ANTIGEN-E (HEPATITS-B) AGAINST PRINCIPAL COMPONENTS OF 9 MEDICINAL PLANTS;K.K.Omprakash et al.;《Asian Journal of Pharmaceutical and Clinical Research》;20111231;第4卷(第2期);55-58 * |
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| Thonningianins A and B, New Antioxidants from the African Medicinal Herb Thonningia sanguinea;Ikuko I. Ohtani et al.;《Journal of Natural Products》;20000504;第63卷(第5期);676-679 * |
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