CN101181287A - Application of puerarin in the preparation of medicament for restraining uric acid transportor URAT1 - Google Patents
Application of puerarin in the preparation of medicament for restraining uric acid transportor URAT1 Download PDFInfo
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- CN101181287A CN101181287A CNA2007101901363A CN200710190136A CN101181287A CN 101181287 A CN101181287 A CN 101181287A CN A2007101901363 A CNA2007101901363 A CN A2007101901363A CN 200710190136 A CN200710190136 A CN 200710190136A CN 101181287 A CN101181287 A CN 101181287A
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- puerarin
- uric acid
- urat1
- acid transporter
- restraining
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- RXUWDKBZZLIASQ-UHFFFAOYSA-N Puerarin Natural products OCC1OC(Oc2c(O)cc(O)c3C(=O)C(=COc23)c4ccc(O)cc4)C(O)C(O)C1O RXUWDKBZZLIASQ-UHFFFAOYSA-N 0.000 title claims abstract description 50
- HKEAFJYKMMKDOR-VPRICQMDSA-N puerarin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1C1=C(O)C=CC(C2=O)=C1OC=C2C1=CC=C(O)C=C1 HKEAFJYKMMKDOR-VPRICQMDSA-N 0.000 title claims abstract description 50
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 title claims abstract description 28
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- XFSBVAOIAHNAPC-XTHSEXKGSA-N 16-Ethyl-1alpha,6alpha,19beta-trimethoxy-4-(methoxymethyl)-aconitane-3alpha,8,10alpha,11,18alpha-pentol, 8-acetate 10-benzoate Chemical compound O([C@H]1[C@]2(O)C[C@H]3[C@@]45C6[C@@H]([C@@]([C@H]31)(OC(C)=O)[C@@H](O)[C@@H]2OC)[C@H](OC)[C@@H]4[C@]([C@@H](C[C@@H]5OC)O)(COC)CN6CC)C(=O)C1=CC=CC=C1 XFSBVAOIAHNAPC-XTHSEXKGSA-N 0.000 description 1
- XFSBVAOIAHNAPC-UHFFFAOYSA-N Aconitin Natural products CCN1CC(C(CC2OC)O)(COC)C3C(OC)C(C(C45)(OC(C)=O)C(O)C6OC)C1C32C4CC6(O)C5OC(=O)C1=CC=CC=C1 XFSBVAOIAHNAPC-UHFFFAOYSA-N 0.000 description 1
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- STDXGNLCJACLFY-UHFFFAOYSA-N aconitine Natural products CCN1CC2(COC)C(O)CC(O)C34C5CC6(O)C(OC)C(O)C(OC(=O)C)(C5C6OC(=O)c7ccccc7)C(C(OC)C23)C14 STDXGNLCJACLFY-UHFFFAOYSA-N 0.000 description 1
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- WDIHJSXYQDMJHN-UHFFFAOYSA-L barium chloride Chemical compound [Cl-].[Cl-].[Ba+2] WDIHJSXYQDMJHN-UHFFFAOYSA-L 0.000 description 1
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to a new usage of a flavonoid monomer compound puerarin, in particular to an application of the puerarin in the preparation of uric acid transporter URAT1 suppressive drugs. The results of the animal test prove that: the puerarin has exact restraining function on abnormally increased expression of uric acid transporter URAT1 in hyperuricemia model animals, while has no significant restraining function on normal expression of uric acid transporter URAT1 in normal animals, and the invention has good safety; the puerarin is matched with the relevant pharmaceutical excipients to be prepared into the puerarin oral liquors, capsules, tablets, granules and other restraining drugs of uric acid transporter URAT1 by using a conventional preparation method, and the invention can be used in the treatment of the relevant diseases with abnormally high expression of uric acid transporter URAT1.
Description
Technical field: the present invention relates to the new purposes of flavonoid monomeric compound puerarin, specifically relate to the application of puerarin in the preparation medicament for restraining uric acid transportor URAT 1.
Background technology: uric acid discharge to reduce or generates and increases, and can cause that uric acid concentration increases in the blood, directly or indirectly causes relevant disease.The drainage of uric acid is a complex process, and when uric acid flows into glomerule with blood circulation, the uric acid of free type will all filter; Heavily absorbed (Diamond HS, MeiselAD.Postsecretory reabsorption of urate in man.Arthritis And Rheumatism, 1975,18 (6): 805-809.) in about 99% urate of near-end renal tubules.Urate transporter (albumen) URAT1 that is positioned at proximal convoluted tubule participates in the heavy absorption process of uric acid, by with anionic exchange urate being transported into cell by tube chamber.URAT1 albumen mainly is distributed in the renal cortex proximal tubular epithelial cells to the chamber film, is an electroneutral uric acid transporter albumen.The URAT1 gene is equaled at first to clone out from people's kidney in 2002 by Enomoto, be positioned at chromosome 11q13, by the SLC22A12 gene code, form by 9 exons of 10 introns, have 12 membrane spaning domains, its cDNA total length 2642bp, coding region 1659bp, coding contains 555 amino acid whose protein.Constitutional kidney Hypouricemia (hypouricemia) patient's the further reference's of SLC22A12 genetic flaw case URAT1 (hURAT1) participates in heavily absorption (the Atsushi Enomoto of uric acid, Hiroaki Kimura, ArthitChairoungdua, Yasuhiro Shigeta, et al.Molecular identification of a renal urate-anionexchanger that regulates blood urate levels.Nature 2002,417 (6887): 447-452).
Puerarin structural formula, molecular formula and the U.S. chemical abstract number of including (CAS No.) be as follows:
Puerarin can obtain by separating in the natural product, for example literature review has been reported and has been utilized solvent method, acid-hydrolysis method, salting out method, complexing abstraction, macroporous adsorbent resin method, polyamide column chromatography absorption method, cyclodextrin bonded immobile phase technology etc. to obtain puerarin (plum forests by the Radix Puerariae separation and purification, Shi Kaiyun, Yang Yuanjuan, Li Suili. the separating and purifying technology of puerarin progress in the Radix Puerariae. Guangzhou chemistry 2007; 32 (1): 73-76); Also there is report to change structure/synthetic puerarin (Lee, David Y.W. of obtaining by chemistry; Zhang, Wu-Yan; Karnati, Vishnu Vardhan R.Total synthesis of puerarin, an isoflavone C-glycoside.Tetrahedron Letters (2003), 44 (36), 6857-6859.).
Puerarin has been seen the report that multiple physiology and pharmacologically active are arranged, comprise effect (Qin Kaiyu such as arrhythmia effect that the effect of expanding coronary vasodilator, antagonism aconitine and barium chloride bring out, influence, microcirculation improvement obstacle, anticoagulant, blood sugar lowering to cardiac function and myocardial metabolism, Wu Min, the pharmacological action of puerarin and analysis of adverse reactions. Chinese Pharmaceutical 2007; 16 (3): 60-61).But the compound monomer puerarin is used to prepare the inhibition medicine at urate transporter URAT1, does not then appear in the newspapers.
Summary of the invention: the purpose of this invention is to provide the new purposes of monomeric compound puerarin, specifically is the application of puerarin in the preparation medicament for restraining uric acid transportor URAT 1.
One of technical solution of the present invention is: provide the monomeric compound puerarin to be used to prepare the new purposes of medicament for restraining uric acid transportor URAT 1.This new purposes has comprised with puerarin of the present invention, be equipped with the known pharmaceutic adjuvant of those skilled in the art (excipient, cosolvent, controlled release agent etc.), make the known dosage form of those skilled in the art (comprising oral liquid, injection, capsule, tablet, granule, microcapsule etc.), be used to prepare medicament for restraining uric acid transportor URAT 1.
Advantage of the present invention is, the puerarin that urate transporter URAT1 is had definite inhibition regulating action is provided, and is used to prepare medicament for restraining uric acid transportor URAT 1, can be used for treatment and the unusual high expressed relevant disease of urate transporter URAT1.Compare with the invention at gout or antihyperuricemic disease, monomeric compound puerarin provided by the invention prepares the new purposes of medicament for restraining uric acid transportor URAT 1 and has 3 significantly progressive and advantages: 1, effective ingredient definite (monomeric compound); 2, action target spot clear and definite (the urate transporter URAT1 of Reabsorption in the urate excretion process); 3, regulating action clear and definite (suppressing to regulate).
The present invention utilizes the antihyperuricemic animal model scientifically to estimate on gene transcription level and protein expression level and has determined the inhibition regulating and controlling effect of monomeric compound puerarin to urate transporter URAT1.
Monomeric compound puerarin involved in the present invention has definite inhibitory action to the urate transporter URAT1 that the animal pattern abnormal expression increases; And intact animal's normal level urate transporter is expressed no remarkable inhibitory action, have good safety.
Essence for a better understanding of the present invention will illustrate its application in the preparation medicament for restraining uric acid transportor URAT 1 with the pharmacological evaluation and the result of monomeric compound puerarin below.
The specific embodiment: following examples are only as the usefulness of further setting forth invention, can not be used for limiting the present invention.
Embodiment 1: the monomeric compound puerarin is to the inhibition regulating action of urate transporter URAT1
Laboratory animal: Kunming mouse, the 15-20 gram, male
The medicine preparation: puerarin is dispersed in the normal saline, is used for gastric infusion, and dosage is 20mg/kg
Experiment material: Trizol Reagment (Invigen), chloroform, isopropyl alcohol, dehydrated alcohol, DEPC, M-MLV reverse transcription, PCR test kit, RIPA lysate etc.
Experimental apparatus: High speed refrigerated centrifuge, high-speed homogenization machine, Bio-rad vertical electrophoresis groove, MBI-PCR instrument, horizontal strip electrophoresis groove etc.
Experimental model: mice hyperuricemia model.
Experimental technique:
1, the foundation of model: animal conformed after 1 week, irritated stomach and gave oxonic acid potassium salt 250mg/kg; Contrast gives equal-volume normal saline, 10 days modeling cycles.
2, administration: administration simultaneously during the modeling, after giving oxonic acid potassium salt modeling/normal saline contrast 1h, irritate stomach and give monomeric compound puerarin 20mg/kg; Contrast gives normal saline.
3, the preservation of mice execution and tissue: mice gives puerarin/normal saline in the filling stomach and puts to death after 1 hour, separates renal tissue on the ice platform, and nephridial tissue is stored in-80 ℃ behind liquid nitrogen freezing.
4, western blotting (Western-blotting): get the RIPA lysate homogenate extraction that the about 100mg of tissue adds 1ml, 12000g4 ℃ centrifugal 15 minutes, get the renal tissue total protein extracting solution, the Bradford method is surveyed protein concentration, and diluted protein sample to final concentration is 5ug/ul.Mix sample in the sample-loading buffer degeneration, PVDF changes film behind the SDS-PAGE gel electrophoresis, the confining liquid sealing adds mURAT1 antibody and (prepares according to ncbi database mURAT1 protein sequence after 1 hour, working concentration 1: 4000) 4 ℃ of overnight incubation, two anti-(1: 4000) room temperature shaking tables were hatched 1 hour, HRP-ECL is luminous, the darkroom exposure imaging.After the film scanning picture is carried out gray analysis.
5, RT-polymerase chain reaction (RT-PCR): extract the total RNA of mouse kidney, reverse transcription obtains the cDNA template, according to the mURAT1 gene design primer of ncbi database, carries out the amplification of mURAT1 genes of interest.Amplified production is the UV imaging behind agarose gel electrophoresis, and picture is carried out gray analysis.
Experimental result:
A. western blotting Western-Blotting
| Group n＝5 | Blank+normal saline | Blank+puerarin | Model+normal saline | Model+puerarin |
| Gray value | 55.3±16.1 | 47.5±14.8 | 96.3±21.3 ++ | 60.7±13.9 ** |
++＜0.01 with blank+normal saline group relatively
*P＜0.01 and model+normal saline group filling in more continuous 10 days stomach gives oxonic acid potassium salt 250mg/kg modeling and causes URAT1 albumen high level expression in the mice nephridial tissue, and gray value reaches 96.3, is significantly higher than blank 55.3.The monomeric compound puerarin that the filling stomach gives 20mg/kg can significantly suppress the proteic high level expression of URAT1 in the model mice nephridial tissue, and gray value is 60.7, near the blank level.Illustrate that the monomeric compound puerarin significantly suppresses the urate transporter URAT1 protein expression that animal pattern increases unusually.The filling stomach gives the monomeric compound puerarin URAT1 protein expression level in the blank mice nephridial tissue is not made significant difference, and illustrates that the monomeric compound puerarin does not have remarkable inhibitory action to the URAT1 albumen of mice nephridial tissue normal level, has safety preferably.
B. RT-polymerase chain reaction RT-PCR
| Group n＝5 | Blank+normal saline | Blank+puerarin | Model+normal saline | Model+puerarin |
| Gray value | 26.6±7.1 | 25.9±10.8 | 58.4±17.6 ++ | 28.8±12.4 ** |
++Compare with blank+normal saline group P＜0.01
*P＜0.01 and model+normal saline group filling in more continuous 10 days stomach gives oxonic acid potassium salt 250mg/kg modeling and causes URAT1 mRNA high level expression in the mice nephridial tissue, and gray value reaches 58.4, is significantly higher than blank 26.6.Irritate monomeric compound puerarin that stomach gives 20mg/kg and can significantly suppress the high level expression of URAT1 mRNA in the model mice nephridial tissue, gray value is 28.8, near the blank level.Illustrate that the urate transporter URAT1 mRNA that the monomeric compound puerarin can suppress to increase unusually expresses on transcriptional level.Irritate stomach and give that URAT1 mRNA expression is 25.9 in the blank mice nephridial tissue of monomeric compound puerarin, there is not significant change with respect to the blank group, illustrate that the monomeric compound puerarin does not have remarkable inhibitory action to the URAT1 mRNA of mice nephridial tissue normal level, has safety preferably.
Embodiment 2:
The monomeric compound puerarin according to the conventional formulation method, is added entry and an amount of solubilizing agent (PEG400) dissolving, packing, sterilization, being prepared into specification is the puerarin oral liquid of 20mg/ml;
According to the conventional formulation method, the soft capsule material is selected gelatin and sorbitol for use with the monomeric compound puerarin, and being prepared into specification is the puerarin capsule of 20mg/ grain;
The monomeric compound puerarin according to the conventional formulation method, is added the excipient cyclodextrin, and mix homogeneously is granulated, tabletting, and being prepared into specification is the puerarin tablet of 20mg/ sheet.
Claims (1)
1. the application of flavonoid monomeric compound puerarin in the preparation medicament for restraining uric acid transportor URAT 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101901363A CN101181287A (en) | 2007-11-15 | 2007-11-15 | Application of puerarin in the preparation of medicament for restraining uric acid transportor URAT1 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101901363A CN101181287A (en) | 2007-11-15 | 2007-11-15 | Application of puerarin in the preparation of medicament for restraining uric acid transportor URAT1 |
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| Publication Number | Publication Date |
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| CN101181287A true CN101181287A (en) | 2008-05-21 |
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| CNA2007101901363A Pending CN101181287A (en) | 2007-11-15 | 2007-11-15 | Application of puerarin in the preparation of medicament for restraining uric acid transportor URAT1 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102526263A (en) * | 2011-12-20 | 2012-07-04 | 瑞坝（北京）新能源科技有限公司 | Plant health care product for preventing and treating gout |
| CN106074540A (en) * | 2016-06-10 | 2016-11-09 | 青岛科瑞元生物科技有限公司 | A kind of pharmaceutical composition for hyperuricemia treatment and application thereof |
| WO2020031961A1 (en) * | 2018-08-10 | 2020-02-13 | サントリーホールディングス株式会社 | Composition for promoting uric acid excretion, composition for inhibiting urat1 and composition for lowering blood uric acid level |
| JP2021008445A (en) * | 2019-06-28 | 2021-01-28 | 国立大学法人 東京大学 | Urat1 inhibitor and urat1 inhibitory food and drink composition |
-
2007
- 2007-11-15 CN CNA2007101901363A patent/CN101181287A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102526263A (en) * | 2011-12-20 | 2012-07-04 | 瑞坝（北京）新能源科技有限公司 | Plant health care product for preventing and treating gout |
| CN106074540A (en) * | 2016-06-10 | 2016-11-09 | 青岛科瑞元生物科技有限公司 | A kind of pharmaceutical composition for hyperuricemia treatment and application thereof |
| WO2020031961A1 (en) * | 2018-08-10 | 2020-02-13 | サントリーホールディングス株式会社 | Composition for promoting uric acid excretion, composition for inhibiting urat1 and composition for lowering blood uric acid level |
| CN112566514A (en) * | 2018-08-10 | 2021-03-26 | 三得利控股株式会社 | Composition for promoting uric acid excretion, composition for inhibiting URAT1, and composition for lowering uric acid level in blood |
| JPWO2020031961A1 (en) * | 2018-08-10 | 2021-08-12 | サントリーホールディングス株式会社 | Composition for promoting uric acid excretion, composition for inhibiting URAT1 and composition for lowering blood uric acid level |
| JP7307073B2 (en) | 2018-08-10 | 2023-07-11 | サントリーホールディングス株式会社 | Composition for promoting uric acid excretion, composition for inhibiting URAT1 and composition for reducing blood uric acid level |
| JP2021008445A (en) * | 2019-06-28 | 2021-01-28 | 国立大学法人 東京大学 | Urat1 inhibitor and urat1 inhibitory food and drink composition |
| JP7376889B2 (en) | 2019-06-28 | 2023-11-09 | 国立大学法人 東京大学 | URAT1 inhibitor and food and drink composition for URAT1 inhibition |
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