KR101337672B1 - Composition comprising beta-d-glucose pentakis(3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl)oxy)benzoate) as a effective component for prevention and treatment of kidney stone disease - Google Patents

Abstract

The present invention prevents and treats kidney stones comprising beta-D-glucose pentacis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) as an active ingredient. The composition for the present invention, the composition of the present invention can be used for the prevention and treatment of kidney stones by inhibiting the migration of kidney cells, inhibit the growth of calcium hydroxide crystals, inhibit the absorption of calcium hydroxide crystals in kidney cells .

Description

A composition for preventing and treating kidney stones comprising beta-D-glucose pentakis (3,4-dihydroxy-5) ((3,4,5-trihydroxybenzoyl) oxy) benzoate as an active ingredient { COMPOSITION COMPRISING BETA-D-GLUCOSE PENTAKIS (3,4-DIHYDROXY-5-((3,4,5-TRIHYDROXYBENZOYL) OXY) BENZOATE) AS A EFFECTIVE COMPONENT FOR PREVENTION AND TREATMENT OF KIDNEY STONE DISEASE}

The present invention prevents and treats kidney stones comprising beta-D-glucose pentacis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) as an active ingredient. It relates to a composition for.

Nephrolithiasis or urolithiasis is a common disorder defined as the occurrence of stones in the urinary system, such as kidney stones. The disorder represents a serious health problem. Kidney stones are stones in the urine or bladder that form crystals of minerals in the urine. Kidney stones can be divided into kidney stones, ureter stones, bladder stones, urethral stones, and prostate stones, depending on the location of the stones. Kidney stones are thought to be formed by the complex interaction of several causative factors. In urine, saturation and supersaturation of substances forming urolithiasis occur, and these substances nucleate and crystallization forms through crystallization as crystals grow and aggregate. That is, crystals are formed through a process of supersaturation → crystal nucleation → crystal aggregation. Kidney stones are composed of hydroxyl and calcium, phosphoric acid and calcium, or uric acid. In particular, calcium stones (calcium oxalate, calcium phosphate) are the most common constituents in the kidney stones (75-80%), uric acid stones 5-10%, infected stones 5-15%, and cystine stones 1-2%. Occupies a degree. Kidney stones are one of the most common kidney diseases in the United States, estimated to have about 1 million people, and the number of patients continues to increase in Korea. This is thought to be the result of industrialization out of the natural environment, and it is a disease that is likely to develop into a modern disease in the future according to the change of diet and lifestyle according to urbanization. Calcium hydroxide stone is a major component of kidney stones. The amount of calcium hydroxide excreted from urine has a significant effect on calcium hydroxide supersaturation and kidney stone formation.

Thus, the inventors have focused on the fact that by reducing the calcium oxalate level in normal kidney cells can prevent or treat kidney stones, and as a result of intensive studies, beta-D-glucose pentakis (3, 4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) inhibits migration of kidney cells, inhibits the growth of calcium hydroxide crystals, and calcium hydroxide in kidney cells The present invention was completed by confirming that the adsorption of stone crystals can be suppressed.

An object of the present invention is to inhibit beta-D-glucose pentakis (3,4-dihydroxy-), which inhibits the migration of kidney cells, inhibits the growth of calcium oxalate crystals, and inhibits calcium oxalate crystal adsorption in kidney cells. It is to provide a composition for preventing and treating kidney stones comprising 5-((3,4,5-trihydroxybenzoyl) oxy) benzoate as an active ingredient.

In order to achieve the above object, the present invention provides a beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) It provides a composition for the prevention and treatment of kidney stones comprising (beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate)) as an active ingredient:

[Formula 1]

The beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) is CAS No. 1401-55-4, Molecular Formula C ₇₆ H ₅₂ O ₄₆ , a compound having a molecular weight of 1701.1985 g / mol, and is commonly referred to as gallotannin, tannic acid, tannin, gallotannic acid, or the like.

According to one embodiment of the invention, the beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) is a kidney cell The movement can be suppressed. In particular, beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) untreated control or 1,2, Renal cell migration compared to 3,4,6-penta-o-galloyl-beta-di-glucose (1,2,3,4,6-penta-O-galloyl-β-D-glucose, PGG) Inhibitory effect is remarkable.

(2)

According to one embodiment of the invention, beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) is calcium hydroxide crystal (cacium oxalate monohydrate crystal) the effect of inhibiting the growth may be remarkable.

According to one embodiment of the invention, beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) is hydroxyl in kidney cells The effect of inhibiting adsorption of calcium oxalate monohydrate crystal may be remarkable.

Prevention and treatment of kidney stones comprising beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) of the present invention as an active ingredient Composition for administration can be administered. The composition for preventing and treating kidney stones can confirm the inhibitory effect of in vitro renal cell migration and the inhibition of calcium oxalate monohydrate crystal adsorption on kidney cells.

The composition for preventing and treating kidney stones of the present invention may include additives such as pharmaceutically acceptable diluents, binders, disintegrants, lubricants, pH adjusting agents, antioxidants, and dissolution aids within the scope of not impairing the effects of the present invention. .

The diluent may be sugar, starch, microcrystalline cellulose, lactose (lactose monohydrate), glucose, di-mannitol, alginate, alkaline earth metal salts, clay, polyethylene glycol, anhydrous calcium hydrogen phosphate, or a mixture thereof; The binder may be selected from the group consisting of starch, microcrystalline cellulose, highly disperse silica, mannitol, di-mannitol, sucrose, lactose hydrate, polyethylene glycol, polyvinylpyrrolidone (povidone), polyvinylpyrrolidone copolymer , Hydroxypropylcellulose, natural gum, synthetic gum, copovidone, gelatin, or a mixture thereof.

The disintegrant may be a starch or modified starch such as sodium starch glycolate, corn starch, potato starch or pregelatinized starch; Clays such as bentonite, montmorillonite, or veegum; Cellulose such as microcrystalline cellulose, hydroxypropylcellulose or carboxymethylcellulose; Alginates such as sodium alginate and alginic acid; Crosslinked celluloses such as croscarmellose sodium; Guar gum and xanthan gum; Crosslinked polymers such as crosslinked polyvinylpyrrolidone (crospovidone); Sodium bicarbonate, citric acid and the like, or a mixture thereof.

The lubricant may be selected from the group consisting of talc, stearic acid, magnesium stearate, calcium stearate, sodium lauryl sulfate, hydrogenated vegetable oil, sodium benzoate, sodium stearyl fumarate, glyceryl behenate, glyceryl monolate, glyceryl monostearate, Stearate, colloidal silicon dioxide, or a mixture thereof may be used.

The pH adjusting agent may be an acidifying agent such as acetic acid, adipic acid, ascorbic acid, sodium ascorbate, sodium ethoxide, malic acid, succinic acid, tartaric acid, fumaric acid, citric acid (citric acid) and precipitated calcium carbonate, ammonia water, meglumine, Basic agents such as sodium, magnesium oxide, magnesium carbonate, sodium citrate, and tribasic calcium phosphate can be used.

As the antioxidant, dibutylhydroxytoluene, butylated hydroxyanisole, tocopheryl acetate, tocopherol, propyl gallate, sodium hydrogen sulfite, sodium pyrophosphate and the like can be used. In the prior-release compartment of the present invention, Polyoxyethylene sorbitan fatty acid esters such as sodium lauryl sulfate and polysorbate, docusate sodium, poloxamer and the like can be used.

In addition, an enteric polymer, a water-insoluble polymer, a hydrophobic compound, and a hydrophilic polymer may be included to form a delayed-release preparation.

The enteric polymer is insoluble or stable under acidic conditions below pH 5 and is dissolved or decomposed under specific pH conditions of pH 5 or higher. For example, (Hydroxypropylmethylcellulose phthalate), hydroxymethylethylcellulose phthalate, cellulose acetate phthalate, cellulose acetate succinate, cellulose acetate maleate, cellulose benzoate phthalate, cellulose propionate phthalate, methylcellulose phthalate, carboxymethylethylcellulose and An enteric cellulose derivative such as ethylhydroxyethylcellulose phthalate and methylhydroxyethylcellulose; Styrene-acrylic acid copolymers such as styrene-acrylic acid copolymers, acrylic acid methyl-acrylic acid copolymers, acrylic acid methyl methacrylic acid copolymers (e.g., acrylic acid), butyl acrylate-styrene-acrylic acid copolymers, and methyl methacrylate- Said enteric acrylic acid-based copolymer; Poly (methacrylic acid methyl acrylate) copolymers (e.g., Eudragit L100-E), poly (methacrylic acid methyl methacrylate) copolymers such as Eudragit L, Eudragit S, An enteric polymethacrylate copolymer such as < RTI ID = 0.0 > Maleic anhydride copolymers, styrene-maleic anhydride copolymers, styrene-maleic anhydride copolymers, styrene-maleic anhydride copolymers, vinyl styrene-maleic anhydride copolymers, styrene-maleic anhydride copolymers, styrene-maleic anhydride copolymers, Maleic anhydride copolymers such as styrene-maleic anhydride copolymer, maleic anhydride copolymer, maleic anhydride copolymer, maleic anhydride copolymer, maleic anhydride copolymer, maleic anhydride copolymer, maleic anhydride copolymer, And enteric polyvinyl derivatives such as polyvinyl alcohol phthalate, polyvinyl acetal phthalate, polyvinyl butyrate phthalate and polyvinylacetacetal phthalate.

The water-insoluble polymer refers to a polymer that does not dissolve in a pharmaceutically acceptable water to control the release of the drug. For example, the water-insoluble polymer can be selected from the group consisting of polyvinyl acetate (e.g., Colicot SR30D), water insoluble polymethacrylate copolymer (e.g., poly (ethyl acrylate-methyl methacrylate) , Poly (ethyl acrylate-methyl methacrylate-trimethylaminoethyl methacrylate) copolymer (such as Eudragit RSPO), ethyl cellulose, cellulose ester, cellulose ether, cellulose acylate, cellulose diacylate, Cellulose triacylate, cellulose acetate, cellulose diacetate and cellulose triacetate.

The hydrophobic compound refers to a pharmaceutically acceptable water-insoluble substance that controls the release of the drug. For example, fatty acid and fatty acid esters such as glyceryl palmitostearate, glyceryl stearate, glyceryl behenate, cetyl palmitate, glyceryl monooleate, and stearic acid; Fatty acid alcohols such as cetostearyl alcohol, cetyl alcohol and stearyl alcohol; Waxes such as carnauba wax, beeswax, and microcrystalline wax; Inorganic materials such as talc, precipitated calcium carbonate, calcium dihydrogen phosphate, zinc oxide, titanium oxide, kaolin, bentonite, montmorillonite, and gum.

The hydrophilic polymer refers to a polymeric substance that dissolves in a pharmaceutically acceptable water to control the release of a drug. For example, there may be mentioned sugars such as dextrin, polydextrin, dextran, pectin and pectin derivatives, alginates, polygalacturonic acid, xylan, arabinozaylan, arabinogalactan, starch, hydroxypropyl starch, amylose and amylopectin ; Cellulose derivatives such as hypromellose, hydroxypropylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, methylcellulose and carboxymethylcellulose sodium; Gums such as guar gum, locust bean gum, tragacanth, carrageenan, acacia gum, gum arabic, gellan gum, and xanthan gum; Proteins such as gelatin, casein, and zein; Polyvinyl derivatives such as polyvinyl alcohol, polyvinyl pyrrolidone, and polyvinyl acetal diethylaminoacetate; Poly (ethyl acrylate-methyl methacrylate-methyl methacrylate) copolymer (e.g., Eudragit E100, Ebonic, Germany), poly (butyl methacrylate- (2-dimethylaminoethyl) Hydrophilic polymethacrylate copolymers such as triethylaminoethyl-methacrylate chloride) copolymer (e.g., Eudragit RL, RS, Evonik, Germany); Polyethylene derivatives such as polyethylene glycol, and polyethylene oxide; And carbomers.

In addition, the pharmaceutical composition of the present invention can be formulated by selecting pharmaceutically acceptable additives as various additives selected from coloring agents and perfumes.

The range of the additive in the present invention is not limited to the use of the additive, and the additive may be formulated to contain the usual range of capacity by selection.

The pharmaceutical compositions according to the invention can be used in the form of oral dosage forms, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations, suppositories or sterile injectable solutions according to conventional methods. have.

The present invention also relates to the prevention of kidney stones comprising beta-D-glucose pentacis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) as an active ingredient Provided is a method for preventing or treating kidney stones by administering a therapeutic composition to a subject comprising a mammal. As used herein, the term "administration" refers to beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy of the present invention in any suitable manner. Means to introduce a composition for preventing and treating kidney stones comprising (benzoate) as an active ingredient, beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4) The route of administration of the composition for the prevention and treatment of kidney stones comprising, 5-trihydroxybenzoyl) oxy) benzoate) as an active ingredient can be administered through any general route as long as the target tissue can be reached. But are not limited to, oral administration, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, endothelial administration, intranasal administration, intrapulmonary administration, intrarectal administration, intraperitoneal administration, intraperitoneal administration and intrathecal administration.

The composition for preventing and treating kidney stones according to the present invention may be administered once or twice a day at regular time intervals.

The dosage of the composition for preventing and treating kidney stones of the present invention varies depending on the weight, age, sex, health condition, diet, time of administration, administration method, excretion rate and severity of disease of the patient.

It is to be understood that the dosage of the composition for preventing and treating kidney stones of the present invention should be determined according to the activity, sex, time of administration, route of administration, duration and frequency of treatment, etc. The dosage does not limit the scope of the invention in any way.

Prevention of kidney stones comprising beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) according to the present invention; The therapeutic composition can be used for preventing and treating kidney stones by inhibiting the migration of kidney cells, inhibiting the growth of calcium hydroxide crystals, and inhibiting calcium hydroxide crystal adsorption in kidney cells.

1 is a histogram showing cytotoxicity measurement results (survival rate by treatment concentration) in MDCK, a normal kidney epithelial cell line of 1,2,3,4,6-penta-o-galloyl-beta-di-glucose by MTT method. to be.
Figure 2 shows the migration of MDCK, a normal renal epithelial cell line of beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) Histogram showing inhibitory effect.
Figure 3 is a histogram showing the inhibitory effect of migration in MDCK, a normal kidney epithelial cell line of 1,2,3,4,6-penta-o-galloyl-beta-di-glucose.
4 shows the migration of MDCK, a normal kidney epithelial cell line of beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) Micrograph showing inhibitory effect.
5 is a histogram showing the effect of inhibiting calcium hydroxide growth by beta-D-glucose pentacis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) to be.
Figure 6 Calcium hydroxide in MDCK, a normal kidney epithelial cell line of beta-D-glucose pentacis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) It is a histogram showing the effect of inhibiting stone adsorption.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are only for the purpose of clarifying the contents of the present invention, and the present invention is not limited by the examples.

Example  1. Cytotoxicity

Cytotoxicity against normal renal epithelial cell line was measured by MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide) method. The seeded-Darby canine kidney (MDCK) cell line (obtained from John C. Lieske, Mayo clinic, USA) in a 96-well culture plate was seeded in 1 × 10 ⁴ cell counts / well and beta- after 24 hours. D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) [gallotanine (sc-202619) from Santa-cruz, molecular weight (MW 1701.1985) was treated by concentration (0, 6.25, 12.25 and 25 μΜ). After 24 hours of incubation, the culture medium (DMEM medium) was discarded and 50 μl of MTT solution (1 mg / ml in phosphate buffered saline) was added and incubated for 1 hour. After incubation, the plate was washed with PBS, and 100 μl of DMSO (dimethyl sulfoxide) was added thereto and left at room temperature for 20 minutes. Absorbance was measured at a wavelength of 570 nm using a microplate reader (Molecular Devices Co.).

Example  2. Liquid  Formulation manufacturing

Beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) [gallotanine (sc-202619), molecular weight of Santa-cruz (MW) 1701.1985] was prepared by dissolving 10 mg of the dried powder in 0.058 ml of 100% ethanol to 100 mM to prepare a liquid according to the concentration.

Example  3. Beta-D- Glucose  Pentakis (3,4- Dihydroxy -5-((3,4,5- Trihydroxybenzoyl ) Oxy ) Benzoate Experiment in normal kidney epithelial cells

A normal kidney epithelial cell line, MDCK (Maddin-Darby canine kidney) cell line (obtained from John C. Lieske, Mayo clinic, USA) was added to a 35-mm plate, and serum-containing medium (10% FBS And DMEM medium supplemented with 1% penicillin / streptomycin) for 24 hours, and then exchanged with a serum-free medium (DMEM medium) for 6 hours. The center of the plate to which the cells were attached was scratched using a 200 μl tip. Beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) was treated by concentration (0, 5 and 10 μM) After incubation for 24 hours, it was stained with Diff-Quick Solution (Sysmex Co., Japan). Cells migrated on an Axiovert S 100 light microscope (carl Zeiss, USA) were counted (see FIGS. 2 and 4).

As a comparative example, 1,2,3,4,6-penta-o-galloyl-beta-di-glucose (1,2,3,4,6-penta-O-galloyl-β-D of Chemical Formula 2) -glucose, PGG), was experimented in substantially the same manner as above (see Figure 3).

Comparative example 1,2,3,4,6-penta-o-galloyl-beta-di-glucose (PGG) had a cell migration inhibitory effect of 25% at 5 μM and 50% at 10 μM. , Beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate), which is an embodiment of the present invention, is 40% at 5 μM, 10 It showed that the cell migration inhibitory effect of 70% at μM, the cell migration inhibitory effect is remarkable.

Example  3. Beta-D- Glucose  Pentakis (3,4- Dihydroxy -5-((3,4,5- Trihydroxybenzoyl ) Oxy ) Benzoate )On by Calcium hydroxide stone  Growth inhibition experiment

Calcium oxalate monohydrate crystal was added by adding 8 mg of calcium oxalate monohydrate crystal (BDH Limited, Poole, England) to 90 mM NaCl and 10 mM Tris buffer (pH 7.2). slurries). The slurry was allowed to mix well at 1,100 rpm by centrifugation to maintain equilibrium overnight. The slurry was added to 2 ml of metastable calcium hydroxide solution, and the metastable calcium hydroxide solution was 0.5 mM Ca (calcium cation), 0.5 mM oxalate, 90 mM NaCl. , 10 mM Tris buffer (pH 7.2). Continuous recording spectrophotometer (Model Cary) was monitored at 214 nm for 400 seconds, and data was compiled using a Lambda 7 spectrophotometer (Perkin Elmer Corp., Norwalk CT, USA). The samples were alternately placed in the measuring instrument.

Referring to FIG. 5, beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) markedly increased calcium hydroxide stone growth. It was confirmed that inhibition.

Example  4. Beta-D- Glucose  Pentakis (3,4- Dihydroxy -5-((3,4,5- Trihydroxybenzoyl ) Oxy ) Benzoate Calcium in normal kidney epithelial cells Olsalate Monohydride  Adsorption experiment

After 24 hours treatment with gallotanine at various concentrations (0, 1 and 10 μM) in the normal kidney epithelial cell line MDCK (Maddin-Darby canine kidney) cell line (obtained from John C. Lieske of Mayo clinic, USA) After removing the DMEM medium was washed three times with PBS (phosphate buffered saline). [ ¹⁴ C] COM crystal suspension (calcium oxalate crystal suspension) was added to MDCK cells, and after 2 minutes, the suspension was removed, and then washed three times with PBS. MDCK cells are scraped and placed in 5 ml scintillation vials (Sigma-Aldrich). 3 ml of scintillation cocktail (scintillation cocktail, scintillation vial, Emulsifier Safe Scintillation mixture; PerkinEl-mer) was added to the scintillation vial and measured using a beta scintillation counter (Berkman Coulter, Fullerton, CA).

Referring to Figure 6, beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) is calcium in normal kidney epithelial cells It was confirmed that olsallate monohydride adsorption was significantly suppressed.

According to embodiments of the present invention, beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) transfers kidney cells It was confirmed that the present invention can inhibit the growth of calcium hydroxide crystals, inhibit the calcium hydroxide crystal adsorption in renal cells, and can be used for the prevention and treatment of kidney stones.

Claims (4)

Beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) (beta-D-glucose pentakis (3,4-dihydroxy- A composition for preventing and treating kidney stones comprising 5-((3,4,5-trihydroxybenzoyl) oxy) benzoate)) as an active ingredient. The method of claim 1, wherein the beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) inhibits the migration of kidney cells Composition for preventing and treating kidney stones, characterized in that. The method of claim 1, wherein the beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) is calcium hydroxide crystal (cacium) oxalate monohydrate crystal) composition for preventing and treating kidney stones, characterized by inhibiting the growth. The method of claim 1, wherein the beta-D-glucose pentakis (3,4-dihydroxy-5-((3,4,5-trihydroxybenzoyl) oxy) benzoate) is calcium hydroxide in kidney cells. Crystal (cacium oxalate monohydrate crystal) composition for preventing and treating kidney stones, characterized in that the adsorption.

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