CN103641871A - Macrocyclic polyphenol compound, and preparation and application thereof in pharmacy - Google Patents
Macrocyclic polyphenol compound, and preparation and application thereof in pharmacy Download PDFInfo
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- CN103641871A CN103641871A CN201310610785.XA CN201310610785A CN103641871A CN 103641871 A CN103641871 A CN 103641871A CN 201310610785 A CN201310610785 A CN 201310610785A CN 103641871 A CN103641871 A CN 103641871A
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- compound
- ethanol
- water
- methyl alcohol
- large ring
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- 229960002930 sirolimus Drugs 0.000 description 1
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical class [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 description 1
- 235000010234 sodium benzoate Nutrition 0.000 description 1
- 231100000240 steatosis hepatitis Toxicity 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 229960005187 telmisartan Drugs 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 230000007923 virulence factor Effects 0.000 description 1
- 239000000304 virulence factor Substances 0.000 description 1
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- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The invention provides a new use of a macrocyclic polyphenol compound separated from a penthorum chinense pursh extract in pharmacy. The macrocyclic polyphenol compound comprises a compound 1, a compound 2, a compound 3 and a compound 4, wherein the structural formulas of the compounds are as shown in the specification; the compounds have the following pharmacological activities that (1) the alpha-amylase activity is significantly inhibited, and the compounds have certain hypoglycemic effects, and (2) TGF (transforming growth factor)-beta1-induced hepatic stellate cell proliferation is significantly inhibited, and the compounds have anti-liver-fibrosis effects by pharmacology and efficacy experiments. The invention provides a novel medicament for preventing and treating diabetes mellitus, and an anti-liver fibrosis medicament. The medicament is abundant in raw material source, small in side effect, and beneficial to environmental protection, and has a relatively great clinical application value.
Description
Technical field
The present invention relates to Chinese medicine, be specifically related to the purposes of Chinese medicine, relate in particular to large ring polyphenolic substance and preparation thereof and prevent and treat the purposes in diabetes and hepatic fibrosis medicines in preparation.
Background technology
Diabetes be a kind of by Different types of etiopathogenises, caused take the metabolic disturbance diseases that chronic hyperglycemia is feature.Along with human life style's variation, the sickness rate of diabetes progressively raises, and expects 2025 and reaches 300,000,000 people, is seriously endangering human health and social development.Abnormal carbohydrate metabolism is the main pathological factor of diabetes, and wherein participate in glycometabolic relevant enzyme, carbohydrate metabolism is had to important impact, therefore regulate the activity of carbohydrate metabolism enzyme to significant (the Traditional Chinese Medicine Anti diabetes study progress based on carbohydrate metabolism enzyme regulation of the treatment of diabetes, CHINA JOURNAL OF CHINESE MATERIA MEDICA, 2012,37 (23): 3519-3525).Ofhypoglycemic medicine has Chinese medicine and Western medicine conventionally.Wherein conventional orally-taken blood sugar reducing medicine be take non-insulin class medicine as main, is divided into non-insulin class Drugs Promoting Insulin Secretion class, biguanides, alpha-glucosidase inhibitor class, insulin sensitizer etc.Modern pharmacological research proves, a lot of Chinese medicine has blood sugar reducing function.Chinese medicine is widely studied and applied because of its high-efficiency low-toxicity effect, and plurality of Chinese extract and composition are proved to be the conditioning agent of carbohydrate metabolism enzyme, compares with Western medicine, and Diabetes Mellitus Treated With Traditional Chinese Medicine has the advantages such as side effect is little and cheap, has good clinical prospect.
Hepatic fibrosis (hepatic fibrosis, HF) is multiple virulence factor, as virus, chronic hepatitis, chronic alcoholism, liver hemostasis, fatty liver etc. cause the extensive hyperplasia of fibrillar connective tissue and deposition in the liver that hepar damnification and inflammation caused.Famous American hepatopathy scholar Friedman SL once pointed out, who can block or delay the generation of hepatic fibrosis, and who just will cure most of chronic hepatopathys.Hepatic stellate cell (hepatic stellate cell, HSC) activation is also converted into myofibroblast (myofibroblast, MFB) be the key link that hepatic fibrosis occurs, synthetic a large amount of extracellular matrix (the extracellular matrix of HSC after activation, ECM), too much ECM constantly deposits in liver, causes that liver normal configuration is destroyed and the disorder of liver function also finally causes hepatic fibrosis.Transforming growth factor (TGF-β 1) is the TGF-beta superfamily that belongs to one group of adjusting Growth of Cells of recently finding and differentiation, studies show that the TGF-β 1 (correlative study 2013,17 (2) that rapamycin and rat liver fibrosis develop: 259-261 that plays an important role in the developing of hepatic fibrosis; Hepatic fibrosis progress, 2012,8 (10): 199-201).A large amount of clinical practice and experimental studies show, Chinese medicine is from many aspects, multipath, too many levels have anti hepatic fibrosis in various degree, and curative effect certainly, few side effects.
Along with the further of diabetes related complication research goed deep into, the relevant liver injury of diabetes is also more and more subject to scholars' attention.Have at present many data to confirm to exist between diabetes and hepatic fibrosis associated, but diabetes are correlated with, hepatic fibrosis pathogenesis is not yet illustrated.Due to few to the research of the relevant hepatic fibrosis of diabetes at present, therefore the medicine of intervening for its specificity report is very few, the medicine for the relevant hepatic fibrosis of diabetes comprises telmisartan and Simvastatin etc. at present.Compare with Western medicine, Chinese medicine has the features such as many target spots, multipath, efficient, low toxicity, for the treatment of the relevant hepatic fibrosis of diabetes, has good clinical prospect.
Chinese medicine Penthorum chinense (Penthorum chinense Pursh) has another name called Herba Lysimachiae Clethroids, Herba Penthori chinensis, mountain delicacy pearl etc.; according to records such as < < Tian Bao book on Chinese herbal medicine > >, < < Herbal for Relief of Famines > >; Penthorum chinense has the dredging collateral effects such as stasis of blood dehumidifying, promoting blood circulation to remove blood stasis of invigorating blood circulation, dispel, and bibliographical information Penthorum chinense has the pharmacologically active of the hepatitis of preventing and treating, liver protection.The inventor therefrom separation obtains a series of large ring polyphenolic compounds.Large ring polyphenolic compound has pharmacological action (the Two antiviral compounds from the plant Stylogne cauliflora as inhibitors of HCV NS3protease of anti-the third liver, Bioorganic & Medicinal Chemistry Letters, 2003,13 (17): 2925-2928), but take a broad view of report both domestic and external, be showed no this compounds pharmacology hypoglycemic and anti-hepatic fibrosis and report.The inventor shows to extract the separated large ring polyphenolic substance obtaining from Penthorum chinense through research and has anti-diabetic and fibrosis effect, therefore, can further research and develop the new drug of anti-diabetic and hepatic fibrosis.
Summary of the invention
The object that technical problem to be solved by this invention is is that research and design extracts separated preparation and the pharmaceutical applications that obtains encircling greatly polyphenolic substance from Penthorum chinense.
The invention provides a kind of large ring polyphenolic substance, there is following structural formula I:
Large ring polyphenolic substance of the present invention is to extract and prepare from Chinese medicine Penthorum chinense.
Large ring polyphenolic substance of the present invention comprises following compounds:
Compound 1: yellow powder, C
35h
28o
17, 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 diformyl]-β-D-Glucose glycosides)
Compound 2:C
42h
34o
21, yellow crystal formation powder.ESI-MS (m/z873.42[M-H]
-), compound 2 for Thonningianins A be Pinocembrine dihydrochalcone-7-O-[3 "-O-galloyl-4 ", 6 "-hexahydroxydiphenoyl]-β-glucose.(pinocembrin dihydrochalcone-7-O-[3 "-O-galloyl-4 ", 6 " and-hexahydroxy-biphenyl diformyl]-β-D-Glucose glycosides)
Compound 3: white powder, C
42h
32o
21, ESI-MS m/z:871[M-H]
-, compound 3 is Pinocembrin-7-O-[3 " and-O-galloyl-4 ", 6 " and-hexahydroxydiphenoyl]-β-glucose.(pinocembrin-7-O-[3 "-O-galloyl-4 ", 6 " and-hexahydroxy-biphenyl diformyl]-β-D-Glucose glycosides)
Compound 4: white powder, C
35h
30o
17, ESI-MS m/z:721[M-H]
-, compound 4 for Thonningianins B be Pinocembrin dihydrochalcone-7-O-[4 ", 6 "-hexahydroxydiphenoyl]-β-D-glucose.(pinocembrin dihydrochalcone-7-O-[4 ", 6 "-hexahydroxy-biphenyl diformyl]-β-D-Glucose glycosides)
Another object of the present invention has been to provide the preparation method of described large ring polyphenolic substance, and the method comprises the following steps:
(1) Herba Lysimachiae Clethroids medicinal material extract and initial gross separation
Herba Lysimachiae Clethroids is pulverized, and 70% alcohol heat reflux extracts three times, and each extraction time is 1 hour, adds 70% ethanol W/L of 3 times of weight of raw material at every turn, and united extraction liquid, is evaporated to without alcohol taste and obtains crude extract medicinal extract (relative density is 1.12~1.18); Then after adding the water W/L suspendible of 1 times of weight of medicinal extract, pass through macroporous resin column, water, 10% ethanol, 20% ethanol, 30% ethanol, 50% ethanol, 70% ethanol and 95% ethanol elution successively, elutriant is 10~12 times of column volume, obtain respectively water, 10% ethanol, 20% ethanol, 30% ethanol, 50% ethanol, 70% ethanol and 95% ethanol eluate, be concentrated into respectively the medicinal extract (relative density 1.12~1.18) that obtains each wash-out position without alcohol taste;
(2) encircle greatly the separated preparation of polyphenolic substance
Get above-mentioned wherein 70% ethanol eluate, with methyl alcohol: water=4:1 dissolves, through MCI-CHP20P gel column chromatography, with methyl alcohol: water (1:9~1:0) gradient elution, obtains 10 stream part (Fr
1-Fr
10) (detection of TLC thin layer plate), get stream part Fr
8after dissolve with methanol, through reverse phase silica gel post (elution requirement: methyl alcohol: water=1:9~1:0) segmentation, obtain 6 streams of I-VI part (detection of TLC thin layer plate), wherein stream part III is through methyl alcohol: after water=1:4 mixing solutions dissolves, through reverse phase silica gel post (methyl alcohol: water=1:4~1:0 gradient elution) purifying, obtain compound 1 again, stream part IV is through methyl alcohol: after water=1:4 mixing solutions dissolves, again through gel column (methyl alcohol: water=1:4~1:0 gradient elution) repeatedly purifying (2-3 time) compound 2 and compound 4, stream part V is through methyl alcohol: after water=1:4 mixing solutions dissolves, through gel column (methyl alcohol: water=1:4~1:0 gradient elution), purifying (2-3 time) must compound 3 repeatedly again.
The inventor further adopts HPLC method, investigated the content in Herba Lysimachiae Clethroids medicinal material that above-mentioned preparation encircles greatly polyphenolic substance 1-4, recorded the content of compound 1-4 in Herba Lysimachiae Clethroids and be respectively: 4.27-4.63 ‰, 0.97-1.23 ‰, 2.56-2.78 ‰, 0.68-0.91 ‰.
Another object of the present invention has been to provide the application of described large ring polyphenolic substance in pharmacy.
The invention provides the application of described large ring polyphenolic substance in preparing preventing/treating Rezulin.
The invention provides the application of described large ring polyphenolic substance in preparing preventing/treating hepatic fibrosis medicines.
Large ring polyphenolic compound of the present invention, through in vitro tests, has the effect of the hepatic stellate cell proliferation of significant inhibition α-amylase and inhibition TGF-β 1 induction, can be used for preparing the medicine of anti-diabetic and hepatic fibrosis.
The pharmaceutical composition that medicine of the present invention is comprised of large ring polyphenolic substance and pharmaceutical excipient as activeconstituents.
The pharmaceutical composition that medicine of the present invention is comprised of the compound 1 as activeconstituents and pharmaceutical excipient; Or
The pharmaceutical composition that medicine of the present invention is comprised of the compound 2 as activeconstituents and pharmaceutical excipient; Or
The pharmaceutical composition that medicine of the present invention is comprised of the compound 3 as activeconstituents and pharmaceutical excipient.
The pharmaceutical composition that medicine of the present invention is comprised of the compound 4 as activeconstituents and pharmaceutical excipient.
Pharmaceutical composition of the present invention is tablet, capsule, granule, powder or pill etc.
The invention provides new control diabetes medicament and anti-hepatic fibrosis medicines, the raw material sources of medicine are abundant, and side effect is little, is conducive to environmental protection, has larger clinical value.
Accompanying drawing explanation
The HPLC contrast figure of Fig. 1 Herba Lysimachiae Clethroids medicinal material, compound 1-4
Ordinate zou is response intensity, unit (AU), and X-coordinate is the time, unit (minute)
A: Herba Lysimachiae Clethroids medicinal material; B: compound 2; C: compound 3; D: compound 1; E: compound 4
Embodiment
With indefiniteness embodiment, the present invention is further described below.
Raw materials used commercially available the obtaining of embodiment.
The preparation of the large ring polyphenolic substance of embodiment 1
(1) Herba Lysimachiae Clethroids medicinal material extract and initial gross separation
Herba Lysimachiae Clethroids 5kg pulverizes, and 70% alcohol heat reflux extracts three times, and each extraction time is 1 hour, each liquid feeding 15L, united extraction liquid, concentrating under reduced pressure (55 ℃) is to obtaining crude extract medicinal extract (relative density is 1.12~1.18) without alcohol taste, claim that its weight is 625g, yield is 12.5%.Then after water (650ml) suspendible, pass through macroporous resin column (D-101), water, 10% ethanol, 20% ethanol, 30% ethanol, 50% ethanol, 70% ethanol and 95% ethanol elution successively, elutriant is 10~12 times of column volume, obtain respectively water, 10% ethanol, 20% ethanol, 30% ethanol, 50% ethanol, 70% ethanol and 95% ethanol eluate, concentrating under reduced pressure (55 ℃), to obtain the medicinal extract (relative density is 1.12~1.18) at each wash-out position without alcohol taste, claims its weight to be respectively 112g, 108g, 75g, 55g, 66g, 60g and 55g respectively.
(2) encircle greatly the separated preparation of polyphenolic substance
Get the concentrated medicinal extract 60g of 70% ethanol eluate that embodiment 1 obtains, through MCI-CHP20P gel column chromatography, with methyl alcohol: water (1:9~1:0) gradient elution, through TLC(silica gel thin-layer plate) detect, obtain 10 stream part (Fr
1-Fr
10).Stream part Fr
8through reverse phase silica gel post (ODS-C18) segmentation, through TLC(silica gel thin-layer plate) detect, obtain 6 streams part of I-VI, wherein a stream part III obtains compound 1 (100.1mg) through reverse phase silica gel post (ODS-C18) purifying again, stream part IV obtains compound 2 (201.6mg) and compound 4 (62.8mg) after 2 purifying of gel column (LH-20), and stream part V purifying after 2 purifying of gel column (LH-20) obtains compound 3(50.5mg).
Wherein, compound 1: yellow powder, C
35h
28o
17, ESI-MS m/z:721[M+H]
+, nuclear magnetic data and document (Journal of Natural Product, 1998,61:523-524) report is basically identical, so deterministic 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, C
42h
34o
21, 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, therefore deterministic compound 2 for Thonningianins A be Pinocembrine dihydrochalcone-7-O-[3 "-O-galloyl-4 ", 6 "-hexahydroxydiphenoyl]-β-glucose.(pinocembrin dihydrochalcone-7-O-[3 "-O-galloyl-4 ", 6 " and-hexahydroxy-biphenyl diformyl]-β-D-Glucose glycosides)
Compound 3: white powder, C
42h
32o
21, 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, therefore deterministic compound 3 is Pinocembrin-7-O-[3 "-O-galloyl-4 ", 6 " and-hexahydroxydiphenoyl]-β-glucose.(pinocembrin-7-O-[3 "-O-galloyl-4 ", 6 " and-hexahydroxy-biphenyl diformyl]-β-D-Glucose glycosides)
Compound 4: white powder, C
35h
30o
17, ESI-MS m/z:721[M-H]
-nuclear magnetic data and document (Journal of Natural Product, 2000,63:676-679) report is basically identical, therefore deterministic compound 4 for Thonningianins B be Pinocembrin dihydrochalcone-7-O-[4 ", 6 "-hexahydroxydiphenoyl]-β-D-glucose.(pinocembrin dihydrochalcone-7-O-[4 ", 6 "-hexahydroxy-biphenyl diformyl]-β-D-Glucose glycosides)
The large ring of embodiment 2 polyphenolic substance 1-4 in Herba Lysimachiae Clethroids medicinal material assay
Its method and operation steps are as follows
(1) HPLC condition
Chromatographic column: Diamonsil C18column (4.6mm * 250mm, 5 μ m, Dikma);
Moving phase: adopt gradient elution, A is acetonitrile mutually, and B is 0.2% aqueous formic acid mutually;
Flow velocity: 1.0ml/min, working time, 70min, detected wavelength 280nm,
Column temperature is 25 ℃; Sample size 10 μ l;
Table 1 gradient elution program
(2) assay of compound 1-4
Get compound 1-4 reference substance (embodiment 1 system), accurately weighed, with 60% methyl alcohol, be mixed with the compound 1-4 solution that contains 350 μ g, 100 μ g, 200 μ g, 50 μ g in every 2mL, in contrast product solution respectively.
Get Herba Lysimachiae Clethroids crude extract medicinal extract appropriate (being equivalent to 10g Herba Lysimachiae Clethroids medicinal material) prepared by step in embodiment 1 (1), accurately weighed, 60% dissolve with methanol is settled to 100ml, as need testing solution.Get respectively each 10 μ l injection liquid chromatographies of reference substance solution and need testing solution, record color atlas, see Fig. 1.49.65 ± 0.60,59.80 ± 0.80,55.55 ± 0.80,54.37 ± 0.70min the retention time of compound 1-4 is respectively:; By external standard method, with calculated by peak area, obtaining the content of compound 1-4 in medicinal material is respectively: 4.27-4.63 ‰, 0.97-1.23 ‰, 2.56-2.78 ‰, 0.68-0.91 ‰.
The external hypoglycemic activity experiment of embodiment 3 compound 1-4
1 materials and methods
1.1 trial drug
Embodiment 1 prepares gained monomeric compound 1-4
Experiment material
Acarbose (Beyer Co., Ltd, 50mg/ sheet, commercially available), glucose detection test kit (glucose oxidase-enzymatic peroxidation, Shanghai Rongsheng Bioisystech Co., Ltd,) α-amylase (Wako Pure Chemicals Ind., Osaka, Japan), microplate reader (EIx800, Biotek), 96 porocyte culture plates (Corning/Costar)
1.2 experimental technique
1.3.1 developer: first by 10.8 grams of Sodium Benzoates, 5 grams of potassiumiodides are dissolved in 400ml water, are then settled to 500ml.
1.3.2 1-4 is used respectively to DMSO(methyl-sulphoxide) be made into the solution of 10mg/ml, then water is diluted to respectively 0.556mg/ml, 0.185mg/ml, 0.062mg/ml, a 0.020mg/ml and 0.007mg/ml5 different concns; It is 0.16% starch solution that starch is mixed with to concentration with distilled water.Acarbose is mixed with to 2.5mg/ml, 0.83mg/ml, 0.28mg/ml, a 0.09mg/ml and 0.03mg/ml5 different concns with distilled water simultaneously; With the PIPES damping fluid of 25mM, α-amylase is mixed with to 27.8mg/ml solution (PH=6.9).
1.3.3 use 96 porocyte culture plates, experiment is divided into 6 groups, is respectively blank group contrast (distilled water), positive controls (acarbose), and 4 sample determination groups, establish 6 multiple holes for every group.
1.3.4 respectively above-mentioned 4 compounds and acarbose solution are added to corresponding each hole of organizing into groups on plate hole, every hole 25ul sample solution, blank group adds equivalent distilled water, respectively at every group wherein three holes add 12.5ul α-amylase solution, another three holes add 12.5ul distilled water, 37 ℃ of temperature are incubated 10min, finally respectively at each hole, add 500ul colouring reagents termination reaction, under microplate reader 630nm wavelength, measure absorbancy (A), by following method, calculate inhibiting rate (%): inhibiting rate (%)=[1-(A
sample is not enzyme-added-A
sample is enzyme-added)/(A
blank is not enzyme-added-A
blank is enzyme-added)] * 100%, calculate corresponding half-inhibition concentration (IC
50).
2 experimental results
With the positive contrast medicine of acarbose, result shows that the compound 1-4 of above embodiment 1 preparation can suppress alpha-amylase activity, has significant hypoglycemic activity.The results are shown in Table 2
The restraining effect of 1~3 pair of α-amylase of table 2 compound
3 experiment brief summaries
Compound 1-4 has identical configuration: 4 of glucose with 6 hydroxyls and gallic acid Cheng Huan, the activity that can suppress α-amylase, there is hypoglycemic effect, the activity of each monomeric compound is all better than the activity of positive drug, visible this compounds is the good alpha-amylase inhibitor of a class, and infer hypoglycemic effect with on glucose, be connected with gallic acid number become positive correlation; Large ring polyphenol can directly be digested and assimilated by body as a part for medicinal plant, develops ofhypoglycemic medicine and also have the advantage that cost is low from natural product, is indicating that this compounds has a good application prospect.
Embodiment 4 compound 1-4 suppress hepatic stellate cell proliferation experiment
1 materials and methods
1.1 experiment material
TGF-β 1 (Cat
#240-B-002, purchased from R & D company); DMEM culture medium dry powder (Gibco, USA), foetal calf serum (FBS) (Gibco, USA), pancreatin (Trypsin) (Amresco, USA), Cell Counting Kit-8 test kit (green skies biotechnology research), people's hepatic stellate cells (HSC-T6 cell) (Shuguang Hospital of Shanghai hepatopathy institute), microwell plate spectrophotometer (PowerWave XS, BioTek)
1.2 trial drug
Embodiment 1 prepares gained monomeric compound 1-4
Experimental technique
1.3.1 cell cultures: with DMEM(Dulbecco ' s Modified Eagle ' the s Medium containing 10%FBS) nutrient solution is cultivated HSC-T6 cell, and every 3-4 days goes down to posterity 1 time
1.3.2 sample preparation: TGF-β 1 use 4mM HCl solution preparation is become to the stock solution of 1ng/ μ l, the used time is diluted to desired concn with PBS; Compound 1-4 is mixed with respectively respectively to the stock solution of 20mg/ml with DMSO, the used time adopts DMEM nutrient solution to be diluted to 5ug/ml, 10ug/ml, 20ug/ml, 40ug/ml, a 80ug/ml and 160ug/ml5 different concns.Positive drug colchicine is mixed with the stock solution of 1mg/ml with distilled water, the used time is diluted to 2.5ug/ml, 5.0ug/ml, 10ug/ml, a 20ug/ml and 40ug/ml5 different concns with DMEM nutrient solution.
1.3.3 HSC-T6 cell is inoculated in to 96 orifice plates with 3000/Kong Wei unit, grow to sub-individual layer, be divided into blank group, model group, experimental group, blank group gives 0.5%FBS/DMED and cultivates, model group gives 2.5ng/ml TGF-β 1, experimental group gives 2.5ng/ml TGF-β 1 and gives respectively the compound 1-4 of 5 different concns, hatches altogether 24h.Hatch after end, inhale and abandon nutrient solution, every hole adds 100 μ l CCK-8 solution, hatches 2h, under microplate spectrophotometer 450nm wavelength, measures absorbancy (A), calculates inhibiting rate (%): inhibiting rate (%)=(A by following method
experimental group-A
blank group)/(A
model group-A
blank group) * 100%, calculates corresponding half-inhibition concentration (IC
50).
2 experimental results
Result shows that compound 1-4 can suppress TGF-β 1 induction hepatic stellate cell proliferation, has significant anti-hepatic fibrosis active.The results are shown in Table 3
The restraining effect of table 3 compound 1-3 to TGF-β 1 induction hepatic stellate cell proliferation
3 experiment brief summaries
Compound 1-4 has identical configuration: 4 of glucose with 6 hydroxyls and gallic acid Cheng Huan, there is the activity that suppresses TGF-β 1 induction hepatic stellate cell proliferation, there is the effect of anti-hepatic fibrosis, and infer to suppress active with on glucose, be connected with gallic acid number become positive correlation.
Claims (10)
2. large ring polyphenolic substance according to claim 1, it is characterized in that, described large ring polyphenolic substance is compound 1: pinocembrin-7-O-[4 ", 6 "-hexahydroxy-biphenyl diformyl]-β-D-Glucose glycosides, compound 2: pinocembrin dihydrochalcone-7-O-[3 "-O-galloyl-4 ", 6 "-hexahydroxy-biphenyl diformyl]-β-D-Glucose glycosides, compound 3: pinocembrin-7-O-[3 "-O-galloyl-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 is as follows:
3. the preparation method of large ring polyphenolic substance as claimed in claim 1, is characterized in that, the method comprises the following steps:
(1) Herba Lysimachiae Clethroids medicinal material extract and initial gross separation
Herba Lysimachiae Clethroids is pulverized, and 70% alcohol heat reflux extracts three times, and each extraction time is 1 hour, adds 70% ethanol W/L of 3 times of weight of raw material at every turn, united extraction liquid, and being evaporated to and obtaining crude extract medicinal extract relative density without alcohol taste is 1.12~1.18; Then after adding the water W/L suspendible of 1 times of weight of medicinal extract, pass through macroporous resin column, water, 10% ethanol, 20% ethanol, 30% ethanol, 50% ethanol, 70% ethanol and 95% ethanol elution successively, elutriant is 10~12 times of column volume, obtain respectively water, 10% ethanol, 20% ethanol, 30% ethanol, 50% ethanol, 70% ethanol and 95% ethanol eluate, being concentrated into respectively the medicinal extract relative density that obtains each wash-out position without alcohol taste is 1.12~1.18;
(2) encircle greatly the separated preparation of polyphenolic substance
Get above-mentioned wherein 70% ethanol eluate, with methyl alcohol: water=4:1 dissolves, through MCI-CHP20P gel column chromatography, with methyl alcohol: water 1:9~1:0 gradient elution, obtains 10 stream part Fr
1-Fr
10, TLC thin layer plate detects, get stream part Fr
8after dissolve with methanol, through reverse phase silica gel post, elution requirement: methyl alcohol: water=1:9~1:0 segmentation, obtain 6 streams part of I-VI, TLC thin layer plate detects, wherein stream part III is through methyl alcohol: after water=1:4 mixing solutions dissolves, again through reverse phase silica gel post methyl alcohol: water=1:4~1:0 gradient elution, purifying obtains compound 1, stream part IV is through methyl alcohol: after water=1:4 mixing solutions dissolves, again through gel column, methyl alcohol: water=1:4~1:0 gradient elution, purifying is 2-3 time repeatedly, obtain compound 2 and compound 4, stream part V is through methyl alcohol: after water=1:4 mixing solutions dissolves, again through gel column, methyl alcohol: water=1:4~1:0 gradient elution, purifying is 2-3 time repeatedly, obtain compound 3.
4. the application of large ring polyphenolic substance in pharmacy as claimed in claim 1.
5. application according to claim 4, is characterized in that, described in be applied as the application of large ring polyphenolic substance in preparing preventing/treating Rezulin.
6. application according to claim 4, is characterized in that, described in be applied as the application of large ring polyphenolic substance in preparing preventing/treating hepatic fibrosis medicines.
7. according to the application described in any one in claim 4-6, it is characterized in that the pharmaceutical composition that described medicine is comprised of large ring polyphenolic substance and pharmaceutical excipient as activeconstituents.
8. according to the application described in any one in claim 4-6, it is characterized in that the pharmaceutical composition that medicine of the present invention is comprised of the compound 1 as activeconstituents and pharmaceutical excipient.
9. according to the application described in any one in claim 4-6, it is characterized in that the pharmaceutical composition that described medicine is comprised of the compound 2 as activeconstituents and pharmaceutical excipient.
10. according to the application described in any one in claim 4-6, it is characterized in that the pharmaceutical composition that described medicine is comprised of the compound 3 as activeconstituents and pharmaceutical excipient; Or the pharmaceutical composition that formed by the compound 4 as activeconstituents and pharmaceutical excipient of described medicine.
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CN105125601A (en) * | 2015-10-13 | 2015-12-09 | 四川古蔺肝苏药业有限公司 | Penthorum chinense pursh extract and preparing method and application thereof |
CN105193833A (en) * | 2015-10-13 | 2015-12-30 | 四川古蔺肝苏药业有限公司 | Application of penthorum chinense pursh monomer in preparation of liver protection medicine |
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 |
CN108478610A (en) * | 2018-06-30 | 2018-09-04 | 浙江中医药大学 | A kind of penthorum chinense pursh extract and its application |
CN109125337A (en) * | 2018-07-23 | 2019-01-04 | 澳门科技大学 | The application of penthorum chinense pursh and its compound in preparation treatment atherosclerosis and the drug for mitigating Alzheimer disease |
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 |
CN110229204A (en) * | 2019-05-29 | 2019-09-13 | 四川大学 | A kind of compound PGHG preparation method treated in Parkinson disease drug |
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 |
-
2013
- 2013-11-26 CN CN201310610785.XA patent/CN103641871B/en not_active Expired - Fee Related
Non-Patent Citations (4)
Title |
---|
IKUKO I. OHTANI ET AL.: "Thonningianins A and B, New Antioxidants from the African Medicinal Herb Thonningia sanguinea", 《JOURNAL OF NATURAL PRODUCTS》 * |
K.K.OMPRAKASH ET AL.: "INSILICO STUDIES OF ANTIGEN-E (HEPATITS-B) AGAINST PRINCIPAL COMPONENTS OF 9 MEDICINAL PLANTS", 《ASIAN JOURNAL OF PHARMACEUTICAL AND CLINICAL RESEARCH》 * |
QUN LU ET AL.: "Isolation and Identification of Compounds from Penthorum chinense Pursh with Antioxidant and Antihepatocarcinoma Properties", 《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》 * |
YU-LIN HUANG ET AL.: "Two Tannins from Phyllanthus tenellus", 《JOURNAL OF NATURAL PRODUCTS》 * |
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CN105125601A (en) * | 2015-10-13 | 2015-12-09 | 四川古蔺肝苏药业有限公司 | Penthorum chinense pursh extract and preparing method and application thereof |
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CN105125601B (en) * | 2015-10-13 | 2018-10-26 | 四川古蔺肝苏药业有限公司 | A kind of extract of penthorum chinense pursh and its preparation method and application |
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 |
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CN108478610A (en) * | 2018-06-30 | 2018-09-04 | 浙江中医药大学 | A kind of penthorum chinense pursh extract and its application |
CN109125337A (en) * | 2018-07-23 | 2019-01-04 | 澳门科技大学 | The application of penthorum chinense pursh and its compound in preparation treatment atherosclerosis and the drug for mitigating Alzheimer disease |
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 |
CN110229204A (en) * | 2019-05-29 | 2019-09-13 | 四川大学 | A kind of compound PGHG preparation method treated in Parkinson disease drug |
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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