CN108129528B - Compound, cassia seed extract containing compound and application of compound and cassia seed extract - Google Patents

Abstract

The embodiment of the invention provides a compound, wherein the chemical structural formula of the compound is as follows:

the embodiment of the invention also provides a cassia seed extract containing the compound; the embodiment of the invention also provides a pharmaceutical composition, which comprises the compound or the cassia seed extract and a pharmaceutically acceptable carrier or excipient. The compound provided by the invention or the cassia seed extract or the pharmaceutical composition containing the compound can inhibit the transport effect of a transporter; based on this, it is expected that the compound provided by the present invention or the extract of cassia seed or the pharmaceutical composition containing it can be used for the preparation of a medicament for inhibiting the action of a transporter; further, the medicament is used for preventing and/or treating at least one of the following diseases: hyperuricemia, gout, viral infection, hypertension, and renal mercury poisoning.

Description

Compound, cassia seed extract containing compound and application of compound and cassia seed extract

Technical Field

The invention relates to the technical field of traditional Chinese medicine extracts, in particular to a compound, a cassia seed extract containing the compound and application of the compound.

Background

The semen Cassiae is dry mature seed of Cassia obtusifolia L or Cassia tora L of Cassia of Leguminosae, is slightly cold in nature, sweet in taste, bitter, salty, and can enter liver, kidney and large intestine channels, and has effects of clearing liver-fire, improving eyesight, and loosening bowel to relieve constipation. Can be used for treating conjunctival congestion, photophobia, lacrimation, headache, vertigo, dim eyesight, and constipation. Semen Cassiae is planted in Shen nong Ben Cao Jing, listed as one of 120 kinds of top-quality drugs, and is called as "treating eye diseases, replenishing vital essence and relieving weight after long-term use". The cassia seed is also called cassia seed, sheep bone, sheep kidney bean, etc. The cassia seed is widely applied as one of 69 medicinal and edible traditional Chinese medicines published by the national ministry of health, is mainly distributed in Anhui, Guangxi, Sichuan, Zhejiang, Guangdong and the like, and contains various chemical components, such as anthraquinone, flavonoid, naphthopyrone, volatile oil, fatty acid, amino acid, polysaccharide and various inorganic elements. And has various pharmacological activities such as lowering blood pressure, reducing blood lipid, promoting urination, improving eyesight, resisting bacteria, resisting oxidation, resisting cancer, resisting aging, etc.

Transporters (also called transporters) are widely present in living bodies, and mainly include organic anion transporters (also called organic anion transporters, OAT) and organic cation transporters (also called organic cation transporters, OCT). OAT belongs to a member of solute transporter family (SLC)22A, plays an important role in the metabolism and clearance of various endogenous and exogenous anions and metabolites thereof in the liver and kidney, and mediates the transport of a large number of small molecule substrates. Among them, OAT1(SLC22a6) and OAT3(SLC22a8) are mainly expressed in renal proximal tubule basement membrane and play an important role in the clearance of exogenous substances, environmental toxins and endogenous metabolites. Many drugs such as diuretics, antihypertensive drugs, antibiotics, antiviral and anticancer drugs, etc. are substrates of OAT1 and OAT 3. In addition, substrates for their transport also include many endogenous substances (such as metabolic intermediates, by-products and hormones) and environmental toxins and poisons (such as mycotoxins and pesticides). Thus, OAT1 and OAT3 have important pharmacokinetic effects that affect the dose, potency, and toxicity of some drugs.

OCT is also one of the important members of SLC22A, where OCT1 is primarily distributed in the liver, mediating the first step in biotransformation of organic cations from the blood into the liver. OCT2 is mainly distributed in the kidney and is involved in renal clearance of organic cations. Clinically, the transport of more than 120 drugs is related to OCT, and OCT also mediates the transport of endogenous substances. The interaction of OCT1 and OCT2 with clinical medications has attracted a great deal of attention.

In conclusion, there has been increasing interest in OAT/OCT in pharmaceutical research in recent years, which acts on many drugs and endogenous compounds of different sizes and structures as a multi-specific transporter. Understanding the effect of natural products on OAT/OCT will help to predict and prevent adverse drug interactions and toxic side effects of drugs, and can guide people to fully utilize beneficial drug interactions to achieve a safer and more effective treatment.

Disclosure of Invention

The inventors have conducted intensive studies on cassia seeds to prepare a cassia seed extract and further extract and isolate a brand new compound therefrom; the compound is unexpectedly found to have transporter inhibiting activity and can be used for preparing a medicament for inhibiting the transporter function; and have accomplished the present invention based on this.

In a first aspect, the present invention provides a compound, wherein the compound has the following chemical structure formula (i):

in one embodiment of the first aspect of the invention, the compound is extracted from cassia seed.

In a second aspect, the present invention provides an extract of cassia seed comprising the compound provided in the first aspect of the invention.

The third aspect of the present invention provides a method for preparing the aforementioned cassia seed extract, which comprises:

1) extracting semen Cassiae with ethanol water solution; concentrating to obtain crude extract;

2) dissolving the obtained crude extract in water, and extracting with ethyl acetate and n-butanol to obtain ethyl acetate extract, n-butanol extract and water layer extract;

3) mixing the extracts obtained in step 2), and drying to obtain semen Cassiae extract.

In a specific embodiment of the third aspect of the present invention, the method for preparing the cassia seed extract comprises:

1) extracting semen Cassiae with ethanol water solution under reflux; concentrating until no alcohol exists to obtain a crude extract; wherein, the ethanol water solution is preferably 50-100% by volume, more preferably 70-95% by volume, and most preferably 95% by volume; the amount of the ethanol aqueous solution is preferably 2 to 15 times, more preferably 4 to 10 times, and most preferably 6 times that of the cassia seed. The extraction time is preferably 1 to 4 hours, more preferably 2 hours; the extraction times are preferably 1-6 times, more preferably 2-4 times, and most preferably 3 times;

2) the obtained crude extract is dissolved in water (3-10 times of water, preferably 4-6 times of water, and most preferably 4 times of water. ) Extracting with ethyl acetate and n-butanol, and removing solvent to obtain ethyl acetate extract, n-butanol extract and water layer extract; wherein, the ethyl acetate and the n-butanol are preferably used for extraction for 1 to 6 times respectively, more preferably for 2 to 4 times, and most preferably for 3 times; the solvent removal can be carried out by a method commonly used in the art such as distillation under reduced pressure, and the present invention is not limited thereto.

3) Mixing the extracts obtained in step 2), and drying to obtain semen Cassiae extract. Drying may be carried out in a manner commonly used in the art, and the present invention is not limited thereto.

In a fourth aspect, the present invention provides a pharmaceutical composition, which comprises the aforementioned compound or the aforementioned cassia seed extract, and a pharmaceutically acceptable carrier or excipient.

In a particular embodiment of the fourth aspect of the invention, it further comprises a second therapeutic agent having transporter-inhibiting action; in one embodiment, the second therapeutic agent has an organic anion transporter inhibitory effect, e.g., an OAT1 and/or OAT3 inhibitory effect; more specifically, the second therapeutic agent is selected from, but not limited to, quinine, probenecid, and the like.

In a fifth aspect, the present invention provides the use of a compound of the first aspect, an extract of cassia seed of the second aspect, or a pharmaceutical composition of the fourth aspect, in the manufacture of a medicament for inhibiting the action of a transporter.

In a particular embodiment of the fifth aspect of the invention, the transporter is an organic anion transporter.

In another specific embodiment of the fifth aspect of the invention, the organic anion transporter is OAT1 and/or OAT 3.

In a further embodiment of the fifth aspect of the invention, the medicament is for the prevention and/or treatment of at least one of the following diseases: hyperuricemia, gout, viral infection, hypertension, and renal mercury poisoning.

The term "treatment" as used herein has its ordinary meaning and refers herein in particular to the treatment of a mammalian subject, preferably a human, already suffering from a disorder related to transporter function (or effect) as described herein with a medicament of the present invention in order to produce a therapeutic, curative, palliative, etc. effect on said disorder. Similarly, the term "prevention" as used herein has its ordinary meaning and refers herein in particular to the treatment of a mammalian subject, who may suffer from or is at risk of suffering from a disorder associated with transporter function (or effects) as described herein, with a medicament of the present invention in order to produce a preventing, arresting, abrogating, etc. effect on said disorder.

As used herein, "pharmaceutically acceptable" means having no substantial toxic effect when used in the usual dosage amounts, and thus being approved by the government or equivalent international organization or approved for use in animals, more particularly in humans, or registered in the pharmacopoeia.

The "pharmaceutically acceptable carrier or excipient" useful in the pharmaceutical compositions of the invention may be any conventional carrier in the art of pharmaceutical formulation, and the selection of a particular carrier will depend on the mode of administration or the type and state of the disease used to treat a particular patient. The preparation of suitable pharmaceutical compositions for a particular mode of administration is well within the knowledge of those skilled in the pharmaceutical art. For example, solvents, diluents, dispersing agents, suspending agents, surfactants, isotonic agents, thickening agents, emulsifiers, binders, lubricants, stabilizers, hydrating agents, emulsification accelerators, buffers, absorbents, colorants, ion exchangers, release agents, coating agents, flavoring agents, antioxidants, and the like, which are conventional in the pharmaceutical field, may be included as the pharmaceutically acceptable carrier. If necessary, a flavor, a preservative, a sweetener and the like may be further added to the pharmaceutical composition.

As used herein, the term "pharmaceutical composition" has its ordinary meaning. In addition, the 'pharmaceutical composition' of the invention can also be present or provided in the form of health products, functional foods, food additives and the like. The pharmaceutical composition of the present invention can be prepared by obtaining the active ingredients of the raw materials of the pharmaceutical composition of the present invention by extraction, separation and purification means commonly used in pharmaceutical manufacturing, optionally mixing with one or more pharmaceutically acceptable carriers, and then forming a desired dosage form, using conventional techniques in the pharmaceutical field, particularly in the field of formulation. The pharmaceutical composition according to the present invention is a pharmaceutical formulation which may be suitable for oral, parenteral or topical, topical administration. The pharmaceutical composition can be prepared into various forms such as tablets, powder, granules, capsules, oral liquid and the like. The medicaments in various dosage forms can be prepared according to the conventional method in the pharmaceutical field. Specifically, according to the pharmaceutical compositions of the present invention, the pharmaceutical dosage forms include, but are not limited to: tablet, capsule, granule, powder, injection, powder for injection, transdermal patch, ointment, gel, suppository, oral solution, oral suspension, emulsion for injection, oral emulsion, etc., sustained release tablet, and controlled release tablet. The medicaments in various dosage forms can be prepared according to the conventional method in the pharmaceutical field.

Dosage forms for oral administration may include, for example, tablets, pills, hard or soft capsules, solutions, suspensions, emulsions, syrups, powders, fine granules, pellets, elixirs and the like, without limitation. In addition to the active ingredient, these preparations may contain diluents (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and glycine), lubricants (e.g., silica, talc, stearic acid or its magnesium salt, calcium salt, and polyethylene glycol). Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, and polyvinylpyrrolidone. If necessary, it may further contain pharmaceutically acceptable additives such as disintegrating agents (e.g., starch, agar, alginic acid or sodium salt thereof), absorbents, coloring agents, flavoring agents, sweetening agents, and the like. Tablets may be prepared according to conventional mixing, granulating or coating methods.

Dosage forms for parenteral administration may include, for example, injections, drops for medical use, ointments, lotions, gels, creams, sprays, suspensions, emulsions, suppositories, patches and the like, without being limited thereto.

The pharmaceutical compositions according to the present disclosure may be administered orally or parenterally, for example rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, or subcutaneously.

As used herein, the term "about" generally refers to a range of error permitted in the art, such as ± 10%, e.g., ± 5%, e.g., ± 2%.

The research shows that the compound shown in the formula (I) or the cassia seed extract or the pharmaceutical composition containing the compound can inhibit the transport effect of a transporter; based on this, it is expected that the compound provided by the present invention or the extract of cassia seed or the pharmaceutical composition containing it can be used for the preparation of a medicament for inhibiting the action of a transporter; further, the medicament is used for preventing and/or treating at least one of the following diseases: hyperuricemia, gout, viral infection, hypertension, and renal mercury poisoning.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.

FIG. 1 is a schematic representation of a compound of formula (I)¹An H-NMR spectrum;

FIG. 2 is a schematic representation of a compound of formula (I)¹³A C-NMR spectrum;

FIG. 3 is an HSQC spectrum of the compound of formula (I);

FIG. 4 is an HMBC map of a compound of formula (I);

FIG. 5 is a drawing showing a method for producing a compound represented by the formula (I)¹H-¹H COSY map;

FIG. 6 shows the effect of compounds of formula (I) on 6-CF transport in HEK-OAT1(OAT1) cells at 100 uM;

FIG. 7 shows the effect of compounds of formula (I) on 6-CF transport in HEK-OAT3(OAT3) cells at 100 uM;

FIG. 8 shows a concentration curve of a compound of formula (I) versus 6-CF transport in HEK-OAT1(OAT1) cells;

FIG. 9 shows a concentration profile of compounds of formula (I) versus 6-CF transport in HEK-OAT3(OAT3) cells.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1 preparation of extract of Cassia seed and isolation of Compound of formula (I)

Extracting 20kg semen Cassiae (Hebei, origin) with 120L 95% ethanol (prepared from ethanol and water at volume ratio of 95: 5) under reflux for 2 hr for 3 times. Distilling at 50 deg.C under reduced pressure, concentrating to remove alcohol to obtain crude extract with a mass of about 1.4 kg.

Dissolving the crude extract in 5L water, and extracting with equal amount of ethyl acetate and n-butanol for 3 times to obtain 550g (named COB-E) of ethyl acetate extract part, 150g (named COB-B) of n-butanol extract part, and 600g (named COB-W) of water layer extract respectively.

An ethyl acetate fraction (COB-E)550g was stirred with 700g of silica gel and packed in a column of 1.5kg of silica gel, followed by column chromatography on silica gel in a volume of 4L. Performing gradient elution by using dichloromethane-methanol as an eluent, wherein the elution gradient (the volume ratio of dichloromethane to methanol) is 1:0, 20:1, 10:1, 5:1, 2:1, 1:1 and 0:1 respectively. Three column volumes were collected for each gradient. The fractions were checked by TLC analysis for a total of 5 fractions.

Wherein the fraction 3 is named COB-E3 and Flash medium-pressure ODS-C₁₈Column chromatography (methanol-water gradient elution, 10:90 → 100:0) yielded 6 fractions, designated: COB-E3-F1-F6; wherein the COB-E3-F3 fraction (46mg) was subjected to preparative liquid chromatography to give the compound represented by the formula (I) (17 mg).

Example 2: determination of the Structure of the Compound of formula (I)

By HRESIMS (high resolution electrospray ionization mass spectrometry),¹H、¹³C NMR (hydrogen, carbon nuclear magnetic resonance spectrum) and HSQC (heteronuclear single quantum correlation spectrum), HMBC (heteronuclear multiple bond correlation spectrum) and¹H-¹h COSY (hydrogen-hydrogen chemical shift correlation) spectrum, determining the structure of the compound shown in the formula (I), and specifically characterizing the result asThe following:

HRESIMS (negative ion) ion peak of the compound represented by formula (I): m/z 513.1383[ M-H]^-Indicating that its molecular formula is C₂₁H₂₂O₁₀. Comprehensive analysis¹H-NMR (FIG. 1),¹³C-NMR (FIG. 2) and HSQC spectra (FIG. 3) presume that the compound should contain 2 ketoester carbonyl groups, 10 olefinic carbons and 9 sp³Hybridized carbons (6 of which are in one group, 2 methyl groups, 1 methoxy group). Characteristic signals are visible in the hydrogen spectrum: 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 ═ 7.4 Hz). Starting from the characteristic hydrogen signal, the following relevant signals can be found in the HMBC spectrum (fig. 4): 12.75(1-OH) is associated with 124.3(C-2), 161.5(C-1), 116.9(C-10 a); 7.35(2-H) is related to 118.5(C-4), 116.9(C-10 a); 7.75(3-H) is related to 132.5(C-4a), 161.5 (C-1); 7.65(4-H) is related to 181.5(C-5), 124.3(C-2), 116.9(C-10 a); 7.84(6-H) is related to 181.5(C-5), 154.5(C-8), 124.5(C-9 a); 2.36(11-H) is related to 141.8(C-7), 125.2(C-6), 154.5 (C-8); 3.88 (9-OCH)₃) In relation to 153.4(C-9), the above signals allow the determination of the anthraquinone framework structure of the compounds of formula (I).¹H-¹H COSY spectrum (fig. 5) signal: 5.00(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: 5.00(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')]Finally, the sugar structure was determined and determined to be an β -glucopyranose based on the nuclear magnetic signal and the HMBC signal 5.00(1' -H)/154.5(C-8) confirmed that the glucose was linked to the aglycone at the C-8 position.¹H-¹H COSY signal 1.69(10' -H)/6.72(9' -H), 6.72(10' -H)/5.71(8' -H), and HMBC signal 6.72(9' -H)/165.3; 4.24(6' -Ha)/165.3(C-7'), confirming the attachment of a butyrate α -fragment at glucose C-6', identifying that the double bond configuration of α -butyrate should be trans (E) according to the larger coupling constant of 15.5Hz of 5.71(C-8'), the structure of the compound was finally identified as shown in the following formula (I) according to the above information, and the hydrocarbon data are shown in Table 1.

TABLE 1 Hydrocarbon signals (hydrogen spectrum 500MHz, carbon spectrum 125MHz, DMSO-d) of the compounds of formula (I)₆)

Example 3 screening test for inhibitory Effect of Compounds represented by formula (I) on transporter function

Reagent: DMEM is produced by Cellgro; the product number is R10-013-CV, and the specification is 500 ml/bottle; the fetal bovine serum FBS is produced by Gibco company, the product number is 10099-; penicilin streptomycicinolysis (Penicillin streptomycin solution) is produced by Solebao corporation, the product number is P1400, and the specification is 100 ml/bottle; PBS is produced by Solebao company, and the product number is: 500ml of P1210-500 specification; 0.05% Trypsin-EDTA (Trypsin-EDTA) is a product number produced by Gibco corporation: 1772640 specification 500 ml; DMSO, produced by solibao corporation, cell culture grade, cat #: d8371 specification 50 ml/bottle; TritonX-100 is 100 ml/bottle of the product number T8200 specification of Solebao company; the BCA kit is kang for company production lot number: CW 0014S; probenecid (Probenecid) is a product number produced by SIGMA company: p8761 specification 100 mg; 6-carbonyl fluorescein (6-CF) is manufactured by Aladdin company under the product number: specification of C105327 is 100 mg; hygromycin B is product number of Solebao company: h8080 specification 100 ml; polylysine is SIGMA company product number: p1024 format 50 mg.

Instrument CO₂The incubator is produced by Thermo company; a speed adjustable, timing swirl mixer (SI-T256); the electric heating type constant temperature water bath kettle is produced by Tianjin Euro instruments company; the liquid-transfering gun is produced by Eppendorf company; nikon ECLIPSE Ti-U inverted biological microscope; 1.6R desk-top multi-purpose centrifuge is manufactured by Thermo corporation; the water generator is an American Milli-QCenture ultrapure water system; heier ice at 4 DEG CA tank; a Haier refrigerator at-20 ℃.

Compound sample preparation: dividing the weighed mass M of the sample by the relative molecular mass M of the sample to obtain the amount n of the substance of the sample, and adding 10 times of n volume of DMSO to obtain 10^-1Initial concentration of mol/L. Stored at-20 ℃ in the dark.

The inhibition of OAT1/3 transporter function by compounds was examined using a cellular uptake assay. Organic anion transport proteins (OAT) OAT1 and OAT3 are mainly expressed in the basal-lateral membrane of proximal tubular epithelial cells and have the function of mediating the transport of organic anions into cells, thereby participating in the secretory metabolism, reabsorption and the like of drugs in the kidney. Both of the two transporters are listed as necessary research projects for the research and the application process of pharmacokinetics of the medicine by FDA (drug interaction research and guidance principles).

The experimental method comprises the following steps:

1. cells and culture thereof

HEK-OAT1 and HEK-OAT3 overexpressing cells were constructed and subcultured in this laboratory. These two cells were cultured at 37 ℃ in 5% CO₂The cell culture chamber of (1) was cultured in a culture medium of 10% fetal bovine serum/DMEM, 1% penicillin/streptomycin, and 50. mu.g/ml hygromycin B.

HEK-OAT1 and HEK-OAT3 overexpression cells refer to HEK cells transfected with expression plasmids containing OAT1, OAT3 and genes, respectively, and can be prepared by means of the conventional technology in the field, and are not described in detail herein.

2. Cell uptake assay procedure

At 37 ℃ 5% CO₂Under the conditions of (1), HEK-OAT1 and HEK-OAT3 overexpression cells were cultured in a cell culture incubator using DMEM medium containing 10% fetal bovine serum, 1% penicillin/streptomycin and 50. mu.g/ml hygromycin B, respectively. When the cell density is about 80%, the cells are arranged at 5 x 10⁴The density per well was plated in 96-well cell culture plates previously treated with 0.05mg/ml polylysine and cell uptake experiments were performed after 24 hours of culture until cell attachment was complete. With uptake buffer (OAT1 and OAT 3: 135mM NaCl,5mM KCl,2.5mM CaCl₂,1.2mM MgCl₂,0.8mM MgSO₄28mM glucose, 13mM hepes) was incubated at 37 ℃ for 10 minutes in equilibrium, and then cell uptake experiments were performed at 37 ℃ for 5 minutes. In a 96-well plate, each well of the assay was diluted with uptake buffer to prepare 100. mu.l of a mixture of drug (compound of formula (I) of the invention, abbreviated as Compound 1) and transporter substrate (OAT1 and OAT 3: 5. mu.M 6-carbonyl fluorescein) at a concentration of 100. mu.M; the positive control wells were a mixture of 100. mu.l transporter substrate and positive control drugs (OAT 1: 30. mu.M probenecid, OAT 3: 10. mu.M probenecid), and the blank control wells were 100. mu.l transporter substrate. Three duplicate wells were set for the experiment. At the end of the uptake experiment, the experiment was stopped with pre-cooled uptake buffer, 150 μ l per well, and the cells were washed 3 times with uptake buffer. After cell lysis solution of 1% TritonX-100 is added into each well for half an hour, the fluorescence value is read by a microplate reader, and the absorption emission wavelengths of OAT1 and OAT3 are 485 and 528 respectively. Results of the experiment were plotted using Graphpad software and data were processed using ANOVA, and the results are shown in fig. 6 and 7. The IC of the compound of formula (I) for inhibiting various transporters was obtained by plotting the concentration of the inhibitor and the rate of entry of the substrate into the cells₅₀The results are shown in fig. 8 and 9.

As can be seen from FIGS. 6 and 8, the compound of formula (I) has strong inhibitory effect on OAT1 function at 100 μ M, the inhibitory rate is more than 50%, and the IC50 is 51 μ M. As can be seen from FIGS. 7 and 9, the compound of formula (I) has a strong inhibitory effect on the function of OAT3 at 100. mu.M, and its IC50 is 46. mu.M.

It has been reported in numerous prior art that the expression of the transporters OAT1 and OAT3 is higher in the kidney compared to other organs, which play a critical role in renal excretion of uric acid (Capasso et al, 2005; So and Thorens, 2010). In fact, probenecid, a drug currently used to treat hyperuricemia and gout, is an inhibitor of OAT1 and OAT 3. Knockout mice of OAT3 have 15% lower blood pressure than wild-type mice, and inhibition of OAT3 has a hypotensive effect (Vallon et al, 2008). In addition, inhibition of OAT3 may prevent influenza virus (perwitassari et al, 2013). Therefore, these compounds (compounds having inhibitory effects on OAT1 and OAT3) are expected to have preventive and/or therapeutic effects on gout, hyperuricemia, hypertension, and cold virus infection.

Reference to the literature

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

2、So,A.,Thorens,B.,2010.Uric acid transport and disease.The Journalof 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 ofblood 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 asa Novel Anti-Influenza A Virus Strategy.Antimicrobial agents and chemotherapy57,475-483.

The compound, the cassia seed extract containing the compound and the application of the compound are described in detail above. The principles and embodiments of the present invention are explained herein using specific examples, which are presented only to assist in understanding the method and its central concept. It should be noted that it would be apparent to those skilled in the art that various changes and modifications can be made in the invention without departing from the principles of the invention, and such changes and modifications are intended to be covered by the appended claims.

Claims (8)

1. A compound, wherein the compound has the chemical structure shown in formula (i):

2. the compound of claim 1, wherein the compound is extracted from cassia seed.

3. A cassia seed extract comprising the compound of claims 1-2.

4. The method for preparing the extract of cassia seed as claimed in claim 3, which comprises:

1) extracting semen Cassiae with ethanol water solution; concentrating to obtain crude extract;

2) dissolving the obtained crude extract in water, and extracting with ethyl acetate and n-butanol to obtain ethyl acetate extract, n-butanol extract and water layer extract;

3) mixing the extracts obtained in step 2), and drying to obtain semen Cassiae extract.

5. A pharmaceutical composition comprising a compound of claims 1-2 or an extract of cassia seed of claim 3, and a pharmaceutically acceptable carrier or excipient.

6. The pharmaceutical composition of claim 5, further comprising a second therapeutic agent having transporter-inhibiting action.

7. Use of a compound of claims 1-2, a cassia seed extract of claim 3, or a pharmaceutical composition of claims 5-6 in the manufacture of a medicament for inhibiting the action of a transporter; the transporter is an organic anion transporter OAT1 and/or OAT 3.

8. The use according to claim 7, wherein the medicament is for the prevention and/or treatment of at least one of the following diseases: hyperuricemia, gout, viral infection, hypertension, and renal mercury poisoning.

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