CN108129528A - A kind of compound includes its cassia seed extract and application thereof - Google Patents

Abstract

An embodiment of the present invention provides a kind of compound, wherein, the chemical structural formula of the compound is as follows：The embodiment of the present invention additionally provides the cassia seed extract for containing the compound；The embodiment of the present invention additionally provides a kind of pharmaceutical composition, and it includes aforementioned compound or aforementioned cassia seed extract and pharmaceutically acceptable carrier or excipient.Compound provided by the present invention or cassia seed extract containing it or pharmaceutical composition can inhibit the transhipment of transporter to act on；Based on this, it is contemplated that compound provided by the present invention or cassia seed extract containing it or pharmaceutical composition can prepare the drug for transporter to be inhibited to act on；Further, the drug is used to prevent and/or treat to include at least one of following disease：Hyperuricemia, gout, virus infection, hypertension and kidney mercury poisoning.

Description

A kind of compound includes its cassia seed extract and application thereof

Technical field

The present invention relates to Chinese medical extract technical fields, are extracted more particularly to a kind of compound, comprising its cassia seed Object and application thereof.

Background technology

Cassia seed is species Cassia platymiscium Cassia (Cassia obtusifolia L) or little Cassia tora (Cassia tora L dry mature seed), cold nature is sweet in flavor, bitter, salty, Return liver, kidney, and large intestine channel has and clears liver and improve vision, the work(to relax bowel Effect.For the puckery pain of hot eyes, the more tears of photophobia, dizziness of having a headache, mesh is secretly unknown, the diseases such as constipation.Cassia seed begin plant in《Legendary god of farming's book on Chinese herbal medicine Through》, one of 120 kinds of top grade medicines are classified as, referred to as " all eye illnesses can be controlled, long term usage strengthening the essence light is made light of one's life by commiting suicide ".The alias of cassia seed has grass to determine It is bright, sheep is bright, swordweed, goes back pupil, floating mung bean, climbing groundsel, wild green soya bean, Nightjasmine life etc..Cassia seed is announced as health ministry One of the Chinese medicine of 69 kinds of integration of drinking and medicinal herbs be widely used, be distributed mainly on the ground such as Anhui, Guangxi, Sichuan, Zhejiang, Guangdong, certainly Chemical composition there are many containing in pine torch, mainly there is Anthraquinones, flavonoids, aphthopyrans ketone, volatile oil, fatty acid, ammonia Base acids, polysaccharide and a variety of inorganic elements.And there is a variety of pharmacological activity such as blood pressure lowering, and reducing blood lipid, diuresis, improving eyesight, antibacterial, It is anti-oxidant, anticancer, anti-aging etc..

Transporter (also referred to as transport protein) is widely present in life entity, mainly include machine anion transport body ( Referred to as organic anion transporter, OAT) and Organic Cation Transporter Gene (also referred to as organic cation transporter, OCT). OAT belongs to the member of Solute Transport protein family (SLC) 22A, to various endogenous, exogenous anion and its metabolin in liver Dirty and kidney metabolism and removing play an important role, and mediate the transhipment of a large amount of small molecule substrates.Wherein, OAT1 (SLC22A6) it is mainly expressed in renal proximal tubules basilar memebrane with OAT3 (SLC22A8), is removing exogenous material, environment poison Important role is play in element and endogenous metabolism object.Many drugs such as diuretics, antihypertensive, antibiotic, antiviral And anticancer drug etc. is all the substrate of OAT1 and OAT3.In addition, the substrate of its transhipment also includes many endogenous materials (such as generation Thank intermediate, by-product and hormone) and environmental toxin and poisonous substance (such as mycotoxin and pesticide).Therefore, OAT1 and OAT3 In pharmacokinetics important role, dosage, drug effect and the toxicity of some drugs are affected.

OCT also belongs to one of important member of SLC22A, and wherein OCT1 is distributed mainly on liver, mediates organic cation Enter the first step of liver progress bioconversion from blood.OCT2 is distributed mainly on kidney, and the kidney for participating in organic cation is removed. Clinically the transhipment of about 120 kinds or more of drug is related to OCT, while OCT also mediates the transhipment of endogenous material. The interaction of OCT1 and OCT2 and clinical application has attracted a large amount of concern.

To sum up, study of pharmacy field is more and more to the concern of OAT/OCT in recent years, as a kind of turning for polyspecific Body is transported, OAT/OCT may act on many sizes drug different with structure and endogenous compound.Natural products is understood to OAT/ The effect of OCT, it will help prediction and the toxic side effect of the interaction that is harmful to of prophylactic agent and drug can instruct people abundant Reach more safely and effectively therapeutic purposes using advantageous drug interaction.

Invention content

Inventor conducts in-depth research cassia seed, is prepared for cassia seed extract and further therefrom extraction point Completely new compound is separated out；And it is surprised to find that the compound has transporter inhibitory activity, it can be used for preparing inhibition turn Transport the drug of body effect；And the present invention is completed based on this.

First aspect present invention provides a kind of compound, wherein shown in the chemical structural formula of the compound such as following formula (I)：

In a kind of specific embodiment of first aspect present invention, which extracts from cassia seed and obtains.

Second aspect of the present invention provides the cassia seed extract of the compound provided comprising first aspect present invention.

Third aspect present invention provides the preparation method of aforementioned cassia seed extract, including：

1) cassia seed is extracted with ethanol water；Coarse extract is obtained after concentration；

2) ethyl acetate and extracting n-butyl alcohol are used after gained coarse extract being leached Yu Shuizhong, obtains ethyl acetate extraction portion respectively Sub-dip cream, n-butanol portion medicinal extract and water layer medicinal extract；

3) each section medicinal extract obtained in step 2) is merged, it is dry after up to cassia seed extract.

In a kind of specific embodiment of third aspect present invention, the preparation method of aforementioned cassia seed extract, including：

1) by cassia seed ethanol water circumfluence distillation；Coarse extract is obtained after being concentrated into no alcohol；Wherein, ethyl alcohol is water-soluble Liquid preferably uses percentage by volume as 50-100%, more preferably using 70-95%, most preferably using 95% ethanol water； The dosage of ethanol water is preferably 2-15 times of cassia seed and measures, and more preferably 4-10 times is measured, most preferably 6 times amounts.During extraction Between preferably 1-4 hours, more preferably 2 hours；Extraction time is preferably 1-6 times, more preferably 2-4 times, most preferably 3 times；

2) gained coarse extract leaches in water to (water is measured for 3-10 times of coarse extract, and more preferably 4-6 times is measured, optimal It is selected as 4 times of amounts.) afterwards with ethyl acetate and extracting n-butyl alcohol, it goes to obtain ethyl acetate extraction part medicinal extract, n-butanol after solvent respectively Extract part medicinal extract and water layer medicinal extract；Wherein, it is preferably extracted 1-6 times respectively with ethyl acetate and n-butanol, more preferably 2-4 It is secondary, most preferably 3 times；Go solvent can by be evaporated under reduced pressure etc. mode commonly used in the art realize, the present invention herein without It limits.

3) each section medicinal extract obtained in step 2) is merged, it is dry after up to cassia seed extract.Drying may be used Mode commonly used in the art, the present invention is herein without limiting.

Fourth aspect present invention provides a kind of pharmaceutical composition, and it includes aforementioned compounds or aforementioned cassia seed to carry Take object and pharmaceutically acceptable carrier or excipient.

In a kind of specific embodiment of fourth aspect present invention, also include second with inhibition transporter effect and control Treat agent；In one embodiment, second therapeutic agent have inhibit organic anion transporter effect, such as inhibit OAT1 with/ Or OAT3 effects；More specifically, the second therapeutic agent is selected from, but not limited to, quinine, probenecid etc..

Fifth aspect present invention provides the compound of first aspect, the cassia seed extract of second aspect or fourth aspect Pharmaceutical composition prepare for inhibit transporter act on drug in purposes.

In a kind of specific embodiment of fifth aspect present invention, the transporter is organic anion transporter.

In another specific embodiment of fifth aspect present invention, the organic anion transporter for OAT1 and/ Or OAT3.

In another specific embodiment of fifth aspect present invention, the drug is used to prevent and/or treat following At least one of disease：Hyperuricemia, gout, virus infection, hypertension and kidney mercury poisoning.

As used herein, term " treatment " has its general sense, and particularly refers to having suffered from herein It is of the present invention using the present invention with the mammalian subject (being preferably people) of transhipment body function (or effect) relevant disease Drug is handled, to generate the effects that treatment, healing, alleviation, mitigation to the disease.Similarly, as used herein, Term " prevention " has its general sense, and particularly refers to that may suffer from of the present invention and transhipment body function herein (or effect) relevant disease moves the lactation of the present invention for having risk with the transhipment relevant disease of body function Object individual drug using the present invention is handled, to disease generation is prevented, prevent, prevent, separate the effects that.

As used herein, it is " pharmaceutically acceptable " represent when with usual dosage using when do not have tangible toxicity Effect, so as to ratify or be approved for animal, more specifically to people by government or with its comparable international organization, Or it is registered in pharmacopeia.

" pharmaceutically acceptable carrier or excipient " available in pharmaceutical composition of the present invention can be pharmaceutical preparation neck Any conventional carrier in domain, the selection of specific support is by the administering mode or disease type depending on being used to treat particular patient And state.For specific administration pattern said synthetic processes preparation method completely in the knowledge of drug field technical staff In the range of.For example, the solvent of pharmaceutical field routine, diluent, dispersant, suspending can be included as pharmaceutically acceptable carrier Agent, isotonic agent, thickener, emulsifier, adhesive, lubricant, stabilizer, hydrating agents, emulsification accelerator, is delayed surfactant Electuary, absorbent, colorant, ion-exchanger, releasing agent, smears, corrigent and antioxidant etc..It when necessary, can be with Flavouring agent, preservative and sweetener etc. are added in pharmaceutical composition.

As used herein, term " pharmaceutical composition " has its general sense.In addition, " pharmaceutical composition " of the present invention Can also exist or provide in the form of health products, functional food, food, food additives etc..It is special that pharmaceutical field can be used It is the routine techniques in formulation art, the medicine group of the present invention is obtained by extraction separation and purification means common in pharmaceutical production The active ingredient of the raw material of object is closed, is optionally mixed with one or more of pharmaceutically acceptable carriers, then needed for formation Dosage form, to prepare the pharmaceutical composition of the present invention.Pharmaceutical composition according to the present invention, for can be adapted for oral medication, Parenteral or local administration, the pharmaceutical preparation of topical administration.The present invention pharmaceutical composition can be made tablet, pulvis, The diversified forms such as granule, capsule, oral liquid.The drug of above-mentioned various dosage forms can be according to the conventional method system of pharmaceutical field It is standby.Specifically, pharmaceutical composition according to the present invention, the pharmaceutical dosage form includes but not limited to：Tablet, capsule, Granula, pulvis, parenteral solution, injectable powder, transdermal patch, ointment, gelling agent, suppository, oral administration solution, oral administration mixed suspension, Emulsion for injection, Orally taken emulsion etc., sustained-release tablet, Dospan.The drug of above-mentioned various dosage forms can be according to pharmaceutical field Conventional method prepare.

Dosage forms for oral administration may include such as tablet, pill, hard or soft capsule, solution, suspension, breast Agent, syrup, powder, pulvis, granula subtilis, granule, pilule, elixir etc., however it is not limited to this.Other than active constituent, this A little preparations also may include diluent (such as lactose, dextrose, sucrose, mannitol, D-sorbite, cellulose and glycine), Lubricant (such as silica, talcum, stearic acid or its magnesium salts, calcium salt and polyethylene glycol).Tablet also may include adhesive, example Such as aluminium-magnesium silicate, gelatinized corn starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine.When necessary, It also may include medical additive, such as disintegrant (such as starch, agar, alginic acid or its sodium salt), absorbent, colorant, perfume (or spice) Taste agent, sweetener etc..Tablet can be prepared according to common mixing, the method for being granulated or being coated.

Dosage form for parenterally application may include such as injection, medicinal drops, ointment, lotion, gelling agent, breast Cream, spray, suspension, emulsion, suppository, patch etc., however it is not limited to this.

According to the pharmaceutical composition of the disclosure is orally available or non-bowel such as per rectum, through part, through skin, through vein It is interior, through it is intramuscular, intraperitoneally or through subcutaneous administration.

As used herein, term " about " its typically refer to the error range of this field permission, such as ± 10%, such as ± 5%, such as ± 2%.

Research has shown that formula (I) compound represented provided by the present invention or the cassia seed extract or medicine containing it Compositions can inhibit the transhipment of transporter to act on；Based on this, it is contemplated that compound provided by the present invention contains it Cassia seed extract or pharmaceutical composition can prepare for inhibit transporter act on drug；Further, the medicine Object is used to prevent and/or treat to include at least one of following disease：Hyperuricemia, gout, virus infection, hypertension and kidney Dirty mercury poisoning.

Description of the drawings

In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, to embodiment or will show below There is attached drawing needed in technology description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this Some embodiments of invention, for those of ordinary skill in the art, without creative efforts, can be with Other attached drawings are obtained according to these attached drawings.

Fig. 1 is formula (I) compound represented¹H-NMR collection of illustrative plates；

Fig. 2 is formula (I) compound represented¹³C-NMR collection of illustrative plates；

Fig. 3 is the HSQC collection of illustrative plates of formula (I) compound represented；

Fig. 4 is the HMBC collection of illustrative plates of formula (I) compound represented；

Fig. 5 is formula (I) compound represented¹H-¹H COSY collection of illustrative plates；

Fig. 6 shows formula (I) compound represented in 100uM, and 6-CF is transported in HEK-OAT1 (OAT1) cell Influence；

Fig. 7 shows formula (I) compound represented in 100uM, and 6-CF is transported in HEK-OAT3 (OAT3) cell Influence；

Fig. 8 shows the concentration curve that formula (I) compound represented transports 6-CF in HEK-OAT1 (OAT1) cell；

Fig. 9 shows the concentration curve that formula (I) compound represented transports 6-CF in HEK-OAT3 (OAT3) cell.

Specific embodiment

Below in conjunction with the attached drawing in the embodiment of the present invention, the technical solution in the embodiment of the present invention is carried out clear, complete Site preparation describes, it is clear that described embodiment is only part of the embodiment of the present invention, instead of all the embodiments.It is based on Embodiment in the present invention, those of ordinary skill in the art are obtained every other without making creative work Embodiment shall fall within the protection scope of the present invention.

The preparation of 1 cassia seed extract of embodiment and the separation of formula (I) compound represented

Take the 20kg cassia seeds (place of production：Hebei), with 95% ethyl alcohol of 120L (by second alcohol and water by volume 95：5 prepare) heat Refluxing extraction, each 2 hours extraction times extract 3 times altogether.It is evaporated under reduced pressure at 50 DEG C, is concentrated into no alcohol, obtain coarse extract, Quality about 1.4kg.

Coarse extract is leached in 5L water, with equivalent ethyl acetate and extracting n-butyl alcohol 3 times, respectively ethyl acetate extraction Part medicinal extract 550g (being named as COB-E), n-butanol portion medicinal extract 150g (being named as COB-B), water layer medicinal extract 600g (life Entitled COB-W).

Ethyl acetate portion (COB-E) 550g, with 700g silica gel mixed samples, 1.5kg silica gel dress column carries out silica gel column chromatography, Column volume 4L.Eluant, eluent, which is made, with methylene chloride-methanol carries out gradient elution, gradient (dichloromethane and methanol volume ratio) point It Wei 1:0,20:1,10:1,5:1,2:1,1:1,0:1.Each gradient collects three column volumes.Each fraction of gained passes through TLC points Analysis detects, totally 5 fractions.

Wherein the 3rd fraction is named as COB-E3, and ODS-C is pressed in Flash₁₈Column chromatography (methanol-water gradient elution, 10: 90→100:0) 6 fractions, are obtained, are named as：COB-E3-F1~F6；Wherein COB-E3-F3 fractions (46mg) are through preparing liquid phase Chromatographic isolation obtains formula (I) compound represented (17mg).

Embodiment 2：The structure of formula (I) compound represented determines

By HRESIMS (high-resolution electrospray ionization mass spectrometry),¹H、¹³C NMR (hydrogen, nuclear magnetic resonance of carbon spectrum) and HSQC are (different Vouching quantum Correlated Spectroscopy), HMBC (heteronuclear multiple-bond Correlated Spectroscopy) and¹H-¹H COSY (hydrogen-hydrogen chemical shifts Correlated Spectroscopy) are composed, and determine formula (I) structure of compound represented, embodiments result are as follows：

HRESIMS (anion) quasi-molecular ions of compound shown in formula (I)：m/z 513.1383[M-H]^-Prompt its molecular formula For C₂₁H₂₂O₁₀.Comprehensive analysis¹H-NMR (Fig. 1),¹³C-NMR (Fig. 2) and hsqc spectrum (Fig. 3), thus it is speculated that 2 should be contained in compound Ketone ester carbonyl, 10 olefinic carbons and 9 sp³The carbon of hydridization (wherein 6 are one group, 2 methyl, 1 methoxyl group).It can in hydrogen spectrum See characteristic signal：δ 12.75 (1H, s), 7.84 (1H, s), 7.75 (1H, t, J=8.0Hz), 7.65 (1H, d, J=8.0, 1.0Hz), 7.35 (1H, dd, J=8.0,1.0Hz), 6.72 (1H, m), 5.71 (1H, d-like, J=15.5,1.7Hz), 5.00 (1H, d, J=7.4Hz).From feature hydrogen signal, it can be found that following coherent signal in HMBC composes (Fig. 4)：δ12.75 (1-OH) and 124.3 (C-2), 161.5 (C-1), 116.9 (C-10a) are related；δ 7.35 (2-H) and 118.5 (C-4), 116.9 (C-10a) it is related；δ 7.75 (3-H) and 132.5 (C-4a), 161.5 (C-1) are related；δ 7.65 (4-H) and 181.5 (C-5), 124.3 (C-2), 116.9 (C-10a) are related；δ 7.84 (6-H) and (C-9a) phase of 181.5 (C-5), 154.5 (C-8), 124.5 It closes；2.36 (11-H) and 141.8 (C-7), 125.2 (C-6), 154.5 (C-8) are related；δ3.88(9-OCH₃) and 153.4 (C-9) Correlation, more than signal can determine the anthraquinone skeleton structure of compound shown in formula (I).¹H-¹H COSY compose (Fig. 5) signal：δ5.00 (1'-H)/3.3(2'-H)；3.34(2'-H)/3.30(3'-H)；3.34(3'-H)/3.20(4'-H)；3.34(5'-H)/4.24 (6'-Ha), 4.16 (6'-Hb) and HMBC signals：δ5.00(1'-H)/76.2(C-3'),74.0(C-5')；4.24(6'- Ha)/70.2 (C-4'), 3.30 (3'-H)/74.0 (C-5')], it finally can determine glycosyl structure, and true according to itself and nuclear magnetic signal Fixed its is a β-glucopyranose sugar.HMBC signals 5.00 (1'-H)/154.5 (C-8) confirm that glucose is linked at the C-8 of aglycon On position.¹H-¹H COSY signals：δ1.69(10'-H)/6.72(9'-H)；6.72 (10'-H)/5.71 (8'-H) and HMBC letters Number：6.72(9'-H)/165.3；4.24 (6'-Ha)/165.3 (C-7') confirm in glucose C-6' and are connected to a α-fourth Acidic group segment.According to the larger coupling constant 15.5Hz of δ 5.71 (C-8'), the double bond configuration for identifying iophenoxic acid base should be anti- Formula (E).According to information above, shown in the structure such as following formula (I) for finally identifying the compound, hydrocarbon attribution data is shown in Table 1.

(hydrogen composes 500MHz, carbon spectrum 125MHz, DMSO-d to the hydrocarbon signal of compound shown in 1 formula of table (I)₆)

The screening experiment that embodiment 3 formula (I) compound represented acts on transporter function inhibitio

Reagent：DMEM is produced for Cellgro companies；Article No. is R10-013-CV, and specification is 500ml/ bottles；Fetal calf serum FBS is produced for Gibco companies, article No. 10099-141, and specification is 500ml/ bottles；Penicillin streptomycin Solution (Penicillin Streptomycin Solution) is produced for Suo Laibao companies, article No. P1400, and specification is 100ml/ bottles；PBS is rope Lai Bao companies produce, and article No. is：P1210-500 specifications 500ml；0.05%Trypsin-EDTA (trypsase-EDTA) is Gibco companies produce article No.：1772640 specification 500ml；DMSO is produced for Suo Laibao companies, cell culture grade, article No.： 50ml/ bottles of D8371 specifications；TritonX-100 produces 100ml/ bottles of article No. T8200 specifications for Suo Laibao companies；BCA kits are Kang Wei companies produce article No.：CW0014S；Probenecid (probenecid) produces article No. for SIGMA companies：P8761 specifications 100mg；6- carboxyfluoresceins (6-CF) produce article No. for Aladdin companies：C105327 specifications 100mg；Hygromycin B is Suo Lai Precious company produces article No.：H8080 specifications 100ml；Poly-D-lysine produces article No. for SIGMA companies：P1024 specifications 50mg.

Instrument:CO₂Incubator is produced for Thermo companies；Adjustable speed can timing eddy mixer (SI-T256)；Electrothermal Thermostat water bath is produced for Tianjin Ou Nuo instrument companies；Liquid-transfering gun is produced for Eppendorf companies；Nikon ECLIPSE Ti-U fall Put biomicroscope；The desk-top multiplace centrifuges of 1.6R are produced for Thermo companies；Water-making machine is U.S. Mi Libo Milli-Q Century ultrapure water systems；4 DEG C of Haier's refrigerators；- 20 DEG C of Haier's refrigerators.

Compound sample is prepared：With the sample quality m divided by the relative molecular mass M of the sample weighed, the sample is obtained Substance amount n, then add in 10 times of n volumes DMSO, obtain 10^-1The initial concentration of mol/L.- 20 DEG C are kept in dark place.

The inhibition for investigating compound to OAT1/3 transhipment body functions is tested using cellular uptake.Organic anion transporter (organ anion transport protein, OAT) OAT1 and OAT3 is mainly expressed in proximal renal tubular epithelial cells substrate Side form, function are transported into cell for mediation organic anion, so as to participate in drug in the secretion metabolism of kidney, reabsorption etc.. Both transporters are by FDA《Drug interaction investigative technique guideline》During being classified as innovation drug research and declaring The indispensable research project of pharmacokinetic experiment.

Experimental method：

1st, cell and its culture

HEK-OAT1 and HEK-OAT3 overexpressing cells are by this laboratory structure and secondary culture.This two plants of cells are 37 DEG C, 5%CO₂Cell incubator in, with 10% fetal calf serum/DMEM, 1% penicillin/streptomycin, 50 μ g/ml hygromycin Bs Culture solution culture.

HEK-OAT1 and HEK-OAT3 overexpressing cells refer to transfect the expression matter comprising OAT1, OAT3, gene respectively The HEK cells of grain may be used this field conventional technical means and prepare, are not described in detail herein.

2nd, cellular uptake experimental procedure

At 37 DEG C, 5%CO₂Under conditions of, it is used in cell incubator containing 10% fetal calf serum, 1% penicillin/chain Mycin and the DMEM culture solutions of 50 μ g/ml hygromycin Bs cultivate HEK-OAT1 and HEK-OAT3 overexpressing cells respectively.When cell is close It spends when being 80% or so, cell is pressed into 5*10⁴The density in/hole is laid on 96 holes handled in advance with 0.05mg/ml poly-D-lysines In tissue culture plate, 24 hours are cultivated to the adherent completely rear progress cellular uptake experiment of cell.With intake buffer solution (OAT1 and OAT3：135mM NaCl,5mM KCl,2.5mM CaCl₂,1.2mM MgCl₂,0.8mM MgSO₄, 28mM glucose, 13mM Hepes it) is cultivated 10 minutes in 37 DEG C of balances, and cellular uptake experiment is carried out under the conditions of 37 DEG C, the intake time is 5 minutes. In 96 orifice plates, experimental port prepares the drug (formula (I) of the invention of a concentration of 100 μM of 100 μ l per hole with intake buffer solution dilution Compound, abbreviation compound 1) and transporter substrate (OAT1 and OAT3：5 μM of 6- carboxyfluoresceins) mixture；Positive control Hole is 100 μ l transporters substrates and positive control drug (OAT1：30 μM of probenecid, OAT3：10 μM of probenecid) mixture, blank Control wells are 100 μ l transporter substrates.Experiment sets three multiple holes.At the end of intake experiment, terminated with the intake buffer solution of precooling Experiment per 150 μ l of hole, then washes cell 3 times with intake buffer solution.The cell pyrolysis liquid that 1%TritonX-100 is added in per hole is split After solving cell half an hour, in microplate reader reading fluorescent value, the absorption launch wavelength of OAT1 and OAT3 are respectively 485 and 528.Experiment As a result it is mapped with Graphpad softwares, ANOVA processing data, as a result as shown in Figure 6, Figure 7.By to inhibitor concentration and substrate Speed into cell is mapped, and obtains the IC that formula (I) compound represented inhibits various transporters₅₀, as a result such as Fig. 8, Fig. 9 institute Show.

The function of OAT1 is produced when can be seen that formula (I) compound represented at 100 μM from Fig. 6, Fig. 8 relatively strong Inhibiting effect, inhibiting rate reached more than 50%, IC50 for 51 μM.The change shown in formula (I) is can be seen that from Fig. 7, Fig. 9 Closing object has the function of OAT3 at 100 μM very strong inhibiting effect, and IC50 is 46 μM.

Numerous in the prior art it has been reported that transporter OAT1 compares other organ ratios with expression of the OAT3 in kidney It is higher, they for the kidney discharge of uric acid play key effect (Capasso et al., 2005；So and Thorens, 2010).In fact, the drug probenecid for being used for treating hyperuricemia and gout at present is exactly the inhibitor of OAT1 and OAT3. The knock-out mice blood pressure lower than wild-type mice 15% of OAT3, inhibit OAT3 play the role of blood pressure lowering (Vallon et al., 2008).In addition, the inhibition of OAT3 can be with compound for preventing flu virus (Perwitasari et al., 2013).It therefore, can be with It is expected that these compounds (compound for having inhibiting effect to OAT1 and OAT3) for gout, hyperuricemia, hypertension and Common cold virus is infected with prevention and/or therapeutic effect.

Bibliography

1、Capasso,G.,Jaeger,P.,Robertson,W.G.,Unwin,R.J.,2005.Uric acid and the kidney:urate transport,stone disease and progressive renal failure.Current pharmaceutical design 11,4153-4159.

2、So,A.,Thorens,B.,2010.Uric acid transport and disease.The Journal of clinical investigation 120,1791-1799.

3、Vallon,V.,Eraly,S.A.,Wikoff,W.R.,Rieg,T.,Kaler,G.,Truong,D.M.,Ahn, S.Y.,Mahapatra,N.R.,Mahata,S.K.,Gangoiti,J.A.,Wu,W.,Barshop,B.A.,Siuzdak,G., Nigam,S.K.,2008.Organic anion transporter 3contributes to the regulation of blood pressure.Journal of the American Society of Nephrology 19,1732-1740.

4、Perwitasari,O.,Yan,X.Z.,Johnson,S.,White,C.,Brooks,P.,Tompkins, S.M.,Tripp,R.A.,2013.Targeting Organic Anion Transporter 3with Probenecid as a Novel Anti-Influenza A Virus Strategy.Antimicrobial agents and chemotherapy 57,475-483.

It has been carried out in detail to a kind of compound provided by the present invention, comprising its cassia seed extract and application thereof above It introduces.Specific embodiment used herein is expounded the principle of the present invention and embodiment, and above example is said The bright method and its central idea for being merely used to help understand the present invention.It should be pointed out that for those of ordinary skill in the art For, without departing from the principle of the present invention, can also to the present invention some improvement and modification can also be carried out, these improve and repair Decorations also fall into the protection of the claims in the present invention.

Claims (10)

1. shown in the chemical structural formula of a kind of compound, the wherein compound such as following formula (I)：

2. compound as described in claim 1, the wherein compound are extracted from cassia seed and are obtained.

3. include the cassia seed extract of compound described in claim 1-2.

4. the preparation method of cassia seed extract as claimed in claim 3, including：

1) cassia seed is extracted with ethanol water；Coarse extract is obtained after concentration；

2) ethyl acetate and extracting n-butyl alcohol are used after gained coarse extract being leached Yu Shuizhong, obtains ethyl acetate extraction part leaching respectively Cream, n-butanol portion medicinal extract and water layer medicinal extract；

3) each section medicinal extract obtained in step 2) is merged, it is dry after up to cassia seed extract.

5. a kind of pharmaceutical composition, it includes the cassia seed extractions described in the compound or claim 3 described in claim 1-2 Object and pharmaceutically acceptable carrier or excipient.

6. pharmaceutical composition as claimed in claim 5 is also included with the second therapeutic agent for inhibiting transporter effect.

7. described in the cassia seed extract described in compound, claim 3 or claim 5-6 described in claim 1-2 Purposes of the pharmaceutical composition in the drug for inhibiting transporter effect is prepared.

8. purposes as claimed in claim 7, wherein the transporter is organic anion transporter.

9. purposes as claimed in claim 8, wherein the organic anion transporter is OAT1 and/or OAT3.

10. the purposes as described in claim 7-9, wherein the drug is used to prevent and/or treat in following disease extremely Few one kind：Hyperuricemia, gout, virus infection, hypertension and kidney mercury poisoning.