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
- Publication number
- 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
- Authority
- CN
- China
- Prior art keywords
- puerarin
- uric acid
- urat1
- acid transporter
- restraining
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 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
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 title claims abstract description 23
- 229940116269 uric acid Drugs 0.000 title claims abstract description 23
- 239000003814 drug Substances 0.000 title claims abstract description 14
- 230000000452 restraining effect Effects 0.000 title claims abstract description 13
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 101000821903 Homo sapiens Solute carrier family 22 member 12 Proteins 0.000 title abstract description 26
- 102100021495 Solute carrier family 22 member 12 Human genes 0.000 title description 21
- 150000001875 compounds Chemical class 0.000 claims abstract description 24
- 229930003935 flavonoid Natural products 0.000 claims abstract description 3
- 150000002215 flavonoids Chemical class 0.000 claims abstract description 3
- 235000017173 flavonoids Nutrition 0.000 claims abstract description 3
- 241001465754 Metazoa Species 0.000 abstract description 8
- 201000010099 disease Diseases 0.000 abstract description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 4
- 239000000546 pharmaceutical excipient Substances 0.000 abstract description 4
- 239000002775 capsule Substances 0.000 abstract description 3
- 201000001431 Hyperuricemia Diseases 0.000 abstract description 2
- 239000008187 granular material Substances 0.000 abstract description 2
- 239000000178 monomer Substances 0.000 abstract description 2
- 239000003826 tablet Substances 0.000 abstract description 2
- 238000012360 testing method Methods 0.000 abstract description 2
- 229940079593 drug Drugs 0.000 abstract 2
- 229940124531 pharmaceutical excipient Drugs 0.000 abstract 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 13
- 102100030935 Solute carrier family 2, facilitated glucose transporter member 9 Human genes 0.000 description 11
- 238000000034 method Methods 0.000 description 11
- 210000001519 tissue Anatomy 0.000 description 11
- 108010078530 urate transporter Proteins 0.000 description 11
- 241000699670 Mus sp. Species 0.000 description 9
- 108090000623 proteins and genes Proteins 0.000 description 9
- 210000002784 stomach Anatomy 0.000 description 9
- 102000004169 proteins and genes Human genes 0.000 description 6
- 238000011049 filling Methods 0.000 description 5
- 108020004999 messenger RNA Proteins 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- RYYCJUAHISIHTL-UHFFFAOYSA-N 5-azaorotic acid Chemical compound OC(=O)C1=NC(=O)NC(=O)N1 RYYCJUAHISIHTL-UHFFFAOYSA-N 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 229950000193 oteracil Drugs 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 238000001262 western blot Methods 0.000 description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 3
- 230000009102 absorption Effects 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 210000003734 kidney Anatomy 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 229920000858 Cyclodextrin Polymers 0.000 description 2
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 2
- 206010021131 Hypouricaemia Diseases 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000002961 anti-hyperuricemic effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 2
- 230000009103 reabsorption Effects 0.000 description 2
- 210000005084 renal tissue Anatomy 0.000 description 2
- 238000010839 reverse transcription Methods 0.000 description 2
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical group O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 210000005239 tubule Anatomy 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 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
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 238000009010 Bradford assay Methods 0.000 description 1
- 229930182476 C-glycoside Natural products 0.000 description 1
- 108010078791 Carrier Proteins Proteins 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 108700024394 Exon Proteins 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 201000005569 Gout Diseases 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- 238000012449 Kunming mouse Methods 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- 229920002565 Polyethylene Glycol 400 Polymers 0.000 description 1
- 108091006745 SLC22A12 Proteins 0.000 description 1
- 101150078067 SLC22A12 gene Proteins 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000005903 acid hydrolysis reaction Methods 0.000 description 1
- 229940039750 aconitine Drugs 0.000 description 1
- 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
- 239000003463 adsorbent Substances 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 230000006793 arrhythmia Effects 0.000 description 1
- 206010003119 arrhythmia Diseases 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- WDIHJSXYQDMJHN-UHFFFAOYSA-L barium chloride Chemical compound [Cl-].[Cl-].[Ba+2] WDIHJSXYQDMJHN-UHFFFAOYSA-L 0.000 description 1
- 229910001626 barium chloride Inorganic materials 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 230000000536 complexating effect Effects 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 239000003218 coronary vasodilator agent Substances 0.000 description 1
- 239000006184 cosolvent Substances 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 229910003460 diamond Inorganic materials 0.000 description 1
- 239000010432 diamond Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000004217 heart function Effects 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 description 1
- -1 isoflavone C-glycoside Chemical class 0.000 description 1
- 235000008696 isoflavones Nutrition 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 230000004089 microcirculation Effects 0.000 description 1
- 230000002107 myocardial effect Effects 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- JLFNLZLINWHATN-UHFFFAOYSA-N pentaethylene glycol Chemical compound OCCOCCOCCOCCOCCO JLFNLZLINWHATN-UHFFFAOYSA-N 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 238000005185 salting out Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000007901 soft capsule Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006257 total synthesis reaction Methods 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 210000004926 tubular epithelial cell Anatomy 0.000 description 1
Landscapes
- 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 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN101181287A true CN101181287A (en) | 2008-05-21 |
Family
ID=39446798
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CNA2007101901363A Pending CN101181287A (en) | 2007-11-15 | 2007-11-15 | Application of puerarin in the preparation of medicament for restraining uric acid transportor URAT1 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN101181287A (en) |
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 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN101181345B (en) | Application of flos Sophora chromocor extract in the preparation of medicament for restrainting uric acid transporter URAT1 | |
AU2014360040B2 (en) | Desmodium styracifolium (Osb.) Merr. flavonoids capsule, method of preparing same, and application thereof | |
CN1919856A (en) | Caulis trachelospermi total lignans extractive, extraction method and medicine use of the extractive and active constituent thereof | |
CN101896477B (en) | The five crystal forms of the nicousamide, the preparation methods, the pharmaceutical compositions and the uses thereof | |
CN101181287A (en) | Application of puerarin in the preparation of medicament for restraining uric acid transportor URAT1 | |
JP2020532501A (en) | Use of acteosides in the preparation of drugs to prevent or treat glomerular podocyte injury-type kidney disease | |
CN101181261A (en) | Application of waxberry element in the preparation of medicament for restraining uric acid transportor URAT1 | |
KR20190010578A (en) | New Dapagliflozoline Crystal Forms and Methods of Making and Uses Thereof | |
CN104940216B (en) | The application of galloyl glucose glycoside derivative and the pharmaceutical composition for treating hyperuricemia | |
TWI610676B (en) | Method for producing nerve cell death inhibitor, anti-Alzheimer's disease agent, anti-brain hypofunction agent, medicine having anti-Alzheimer's disease effect or anti-brain function, and nerve cell death inhibitor | |
CN100571692C (en) | The application of curcumin in the preparation medicament for restraining uric acid transportor URAT 1 | |
CN101181262A (en) | Application of coumarin monomeric compound in the preparation of medicament for restraining uric acid transportor URAT1 | |
JP2018509478A (en) | Methods and materials for treating cancer | |
CN105560302B (en) | Application of geranium water extract in preparation of anti-angiogenesis drugs | |
CN103864776B (en) | A kind of Tegafur derivative containing 1,3,4-thiadiazoles heterocycle and amide group | |
US20200222485A1 (en) | Medicinal ambrosia plant extracts | |
CN103113359B (en) | Silybin bis-bias succinate and pharmaceutical salts thereof | |
CN106977560B (en) | Preparation of 2S-cardiospermin-5-benzoate and application thereof in preparation of drugs for treating rheumatoid arthritis | |
CN103159710B (en) | Antiviral decalin derivate | |
CN106977561B (en) | Preparation of Sutherlandin-5-p-hydroxybenzoate and application thereof in preparation of drugs for treating rheumatoid arthritis | |
CN105311044A (en) | Application of pharmaceutical composition to prepare medicines treating cutaneous vasculitis | |
CN116036099B (en) | Application of phellodendron ketone combined pembrolizumab in preparation of pancreatic cancer resisting medicine | |
CN103172622B (en) | The active isomer of silybin bis-bias succinate | |
CN106974922B (en) | Preparation of 2S-cardiospermin-5-cis-p-coumarate and application thereof in preparation of medicines for treating rheumatoid arthritis | |
CN114588187B (en) | Traditional Chinese medicine total flavone extract for treating cataract, and composition, preparation method and medical application thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C12 | Rejection of a patent application after its publication | ||
RJ01 | Rejection of invention patent application after publication |
Open date: 20080521 |