Composition for preventing, ameliorating or treating benign prostatic hyperplasia containing hovenia dulcis extract as active ingredient

Abstract

The present invention relates to a composition for preventing, ameliorating or treating benign prostatic hyperplasia, containing a Hovenia dulcis extract as an active ingredient. Specifically, the Hovenia dulcis extract of the present invention was confirmed to inhibit the proliferation of hyperproliferative prostatic cells induced by dihydrotestosterone (DHT), which is a male hormone, and reduce prostate weight in an animal model with induced benign prostatic hyperplasia, and thus can be effectively used as a composition for preventing, ameliorating or treating benign prostatic hyperplasia.

Description

Composition for preventing, improving or treating benign prostatic hyperplasia containing extract of the walnut tree as an active ingredient

[0001] The present invention relates to a composition for preventing, improving or treating benign prostatic hyperplasia containing an extract of the walnut tree as an active ingredient.

[0002]

[0003] This invention was carried out with the support of the Basic Project of the Korea Institute of Oriental Medicine (Project No.: KSN2412013, KSN2412011) of the Ministry of Science and ICT.

[0004] Benign prostatic hyperplasia (BPH) is a general term for lower urinary tract symptoms caused by an enlarged prostate and impaired bladder function, resulting in decreased urine flow due to urethral obstruction. Symptoms include decreased urinary output, as well as various bladder irritation symptoms such as frequent urination, residual urine, and urgency (the inability to hold urine). Treatment options include medication, non-invasive procedures, invasive surgery, and alternative therapies. Testosterone plays a significant role in the growth, development, and pathological state of the prostate through androgen receptors in the prostate. BPH is influenced by androgens, and dihydrotestosterone (DHT) is the most important androgen in the prostate, playing a crucial role in BPH. Recently, herbal medicine has emerged as a promising alternative to the side effects and shortcomings of conventional synthetic drug treatments for BPH and male pattern baldness.

[0005] Hovenia dulcis is a deciduous broad-leaved tree in the buckthorn family, also called hokkae tree or heorikke tree. It is also used as a jiguja, baekseokmok, mokmil, and hyeonpori. It grows to a height of 10 to 20 m and a diameter of 40 to 80 cm. It mainly grows in areas such as Seoraksan Mountain, Odaesan Mountain, Jiri Mountain, and Hallasan Mountain, and grows well in the temperate southern regions rather than the central and northern regions. The fruit stalk and stem have a sweet and fragrant taste, so they have been used for food, especially for fruit wine, and have been used medicinally to eliminate alcoholic toxins. According to the Compendium of Materia Medica, the raw juice of the hovenia fruit is said to relieve alcoholic toxins and stop nausea, and the seeds have traditionally been used for alcoholism, blood purification, urinary incontinence, and vomiting, and the fruit stem is known to be effective in strengthening the stomach and protecting nutrients.

[0006] Meanwhile, as prior art related to benign prostatic hyperplasia, Korean Patent No. 2583293 discloses a 'composition for treating or preventing benign prostatic hyperplasia or alopecia comprising a Salvia miltiorrhiza Bunge extract as an active ingredient', and Korean Patent No. 2556622 discloses a 'composition for preventing or treating benign prostatic hyperplasia comprising extracts of ginseng, licorice, and peony root'. However, no prior art has been disclosed regarding a composition for preventing, improving, or treating benign prostatic hyperplasia containing an extract of the present invention's mollusk tree as an active ingredient.

[0007] The present invention was derived from the above-mentioned needs, and relates to a composition for preventing, improving, or treating benign prostatic hyperplasia containing an extract of the walnut tree as an active ingredient. Specifically, the present invention was completed by confirming that the extract of the walnut tree inhibits the proliferation of prostate cells hyperproliferated by the male hormone DHT (dihydrotestosterone) and reduces the weight of the prostate in an animal model in which benign prostatic hyperplasia was induced.

[0008] In order to solve the above problem, the present invention provides a pharmaceutical composition for preventing or treating benign prostatic hyperplasia containing an extract of the walnut tree as an active ingredient.

[0009] In addition, the present invention provides a health functional food composition for preventing or improving benign prostatic hyperplasia containing an extract of the walnut tree as an active ingredient.

[0010] In addition, the present invention provides a veterinary composition for preventing or treating benign prostatic hyperplasia containing an extract of the walnut tree as an active ingredient.

[0011] In addition, the present invention provides a feed additive for preventing or improving benign prostatic hyperplasia containing an extract of the walnut tree as an effective ingredient.

[0012] In addition, the present invention provides a herbal medicine composition for preventing or treating benign prostatic hyperplasia containing an extract of the walnut tree as an active ingredient.

[0013] The present invention relates to a pharmaceutical composition for preventing or treating benign prostatic hyperplasia containing an extract of the walnut tree as an active ingredient. Specifically, the extract of the walnut tree of the present invention has an excellent effect of inhibiting the proliferation of prostate cells hyperproliferated by the male hormone DHT (dihydrotestosterone) and reducing the weight of the prostate in an animal model in which benign prostatic hyperplasia is induced.

[0014] Figure 1 shows the results of confirming cytotoxicity when prostate epithelial cells (RWPE-1) were treated with the ethanol extract of the branches of the present invention (A), the hot water extract of the branches of the present invention (B), the ethanol extract of the fruit of the present invention (C), or the hot water extract of the fruit of the present invention (D) at various concentrations. CON is an untreated control group.

[0015] Figure 2 shows the results of confirming the prostate cell proliferation inhibitory effect by treatment with the ethanol extract of the branch of the present invention (A), the hot water extract of the branch of the present invention (B), the ethanol extract of the fruit of the present invention (C), or the hot water extract of the fruit of the present invention (D) in prostate epithelial cells (RWPE-1) hyperproliferated by the androgen DHT (dihydrotestosterone). CON is the control group, DHT is the group in which hyperproliferation of prostate cells was induced by treatment with 10 nM dihydrotestosterone, and Fi is the positive control group treated with 30 μM finasteride, a DHT inhibitor. #, ### indicate that cell proliferation was statistically significantly increased in the DHT treatment group (DHT) compared to the control group (CON), # is p<0.05, and ### is p<0.001. *, **, *** indicate that cell proliferation was statistically significantly reduced in the group treated with the extract of the walnut tree (DHT + ethanol and hot water extracts of walnut branches or fruits) or the positive control group treated with finasteride (DHT + Fi) compared to the DHT treatment group (DHT). * is p<0.05, ** is p<0.01, and *** is p<0.001.

[0016] Figure 3 shows the results of confirming the effect of the ethanol extract of the branches of the walnut tree on prostate hypertrophy in an animal model induced with benign prostatic hyperplasia. (A) is a graph showing the daily body weight (g) of each experimental group, (B) is a photograph of the prostate of each experimental group, and (C) is the result of confirming the relative ratio of the prostate weight to the body weight of each experimental group. CON is the control group, TP is the group induced with benign prostatic hyperplasia by administering 10 mg / kg of testosterone propionate, TP+walnut tree branch is the group administered testosterone propionate (10 mg / kg) and the ethanol extract of the branches of the walnut tree of the present invention (50 mg / kg), and TP+Fi is the positive control group administered testosterone propionate (10 mg / kg) and finasteride (10 mg / kg), an inhibitor of benign prostatic hyperplasia. ### indicates that the ratio of prostate weight to body weight was statistically significantly increased in the testosterone propionate administration group (TP) compared to the control group (CON), and p<0.001. *** indicates that the ratio of prostate weight to body weight was statistically significantly decreased in the group administered the ethanol extract of the branches of the present invention (TP+branches of the branches of the present invention) or the group administered the finasteride (TP+Fi) compared to the testosterone propionate administration group (TP), and p<0.001.

[0017] In order to achieve the purpose of the present invention, the present invention provides a pharmaceutical composition for preventing or treating benign prostatic hyperplasia containing an extract of the walnut tree as an active ingredient.

[0018] The above extract of the walnut tree may be any one extract selected from the leaves, bark, branches and fruits of the walnut tree, and is preferably an extract of the branches or fruits of the walnut tree, but is not limited thereto.

[0019] The extraction solvent of the above-mentioned extract of the walnut tree is preferably selected from among water, C1~C4 lower alcohol or a mixture thereof, more preferably water or ethanol, but is not limited thereto.

[0020] The extraction of the above-mentioned walnut tree can be accomplished using any conventional method known in the art, including filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction. Concentration under reduced pressure is preferably accomplished using a vacuum concentrator or a vacuum rotary evaporator, but is not limited thereto. Furthermore, drying is preferably accomplished using reduced pressure drying, vacuum drying, boiling drying, spray drying, or freeze drying, but is not limited thereto.

[0021] The pharmaceutical composition according to the present invention may further comprise a pharmaceutically acceptable carrier, excipient or diluent.

[0022] Carriers, excipients and diluents that may be included in the pharmaceutical composition of the present invention include various compounds or mixtures including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

[0023] The pharmaceutical composition according to the present invention can be formulated and used in the form of oral formulations such as capsules, powders, granules, tablets, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods.

[0024] When formulated, they are usually prepared using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrating agents, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, and capsules, and these solid preparations are prepared by mixing the pharmaceutical composition with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups, and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspending agents can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases can include withepsol, macrogol, Tween 61, cocoa butter, laurin butter, and glycerogelatin.

[0025] The appropriate dosage of the pharmaceutical composition of the present invention can be prescribed in various ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity.

[0026] The pharmaceutical composition of the present invention can be administered orally or parenterally, and in the case of parenteral administration, it can be administered by topical application to the skin, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc.

[0027] In addition, the present invention provides a health functional food composition for preventing or improving benign prostatic hyperplasia containing an extract of the walnut tree as an active ingredient.

[0028] The above composition is preferably manufactured in any one formulation selected from powder, granules, pills, tablets, capsules, candies, syrups, and beverages, but is not limited thereto.

[0029] When the health functional food composition of the present invention is used as a food additive, the effective ingredient may be added as is or used together with other foods or food ingredients, and may be used appropriately according to a conventional method. The amount of the effective ingredient mixed may be appropriately determined depending on the purpose of use (improvement). Generally, when manufacturing a food or beverage, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be below the above range, and since there is no problem in terms of safety, the effective ingredient may also be used in an amount exceeding the above range.

[0030] There are no specific restrictions on the types of the above foods. Examples of foods to which the above active ingredients can be added include meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and include all health functional foods in the conventional sense.

[0031] When the composition of the present invention is used as a health beverage, it may contain various flavoring agents or natural carbohydrates as additional ingredients, as in conventional beverages. The natural carbohydrates mentioned above are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As a sweetener, a natural sweetener such as thaumarin or stevia extract, or a synthetic sweetener such as saccharin or aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 g of the composition of the present invention. The composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the composition of the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These components may be used independently or in mixtures. The proportion of these additives is not particularly important, but the composition of the present invention is typically selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight.

[0032] In addition, the present invention provides a veterinary composition for preventing or treating benign prostatic hyperplasia containing an extract of the walnut tree as an active ingredient.

[0033] The veterinary composition comprising the extract of the Korean walnut tree of the present invention may further comprise suitable excipients and diluents according to conventional methods. Excipients and diluents that may be included in the veterinary composition comprising the extract of the Korean walnut tree of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, ethanol, stearyl alcohol, liquid paraffin, sorbitan monostearate, polysorbate 60, methylparaben, propylparaben, and mineral oil.

[0034] The veterinary composition comprising the extract of the walnut tree according to the present invention may further include fillers, anticoagulants, lubricants, wetting agents, spices, emulsifiers, preservatives, etc. The veterinary composition according to the present invention may be formulated using a method well known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to an animal, and the formulation may be in the form of a powder, granule, tablet, capsule, suspension, emulsion, solution, syrup, aerosol, soft or hard gelatin capsule, suppository, sterile injection solution, sterile external preparation, etc.

[0035] The veterinary composition according to the present invention may vary depending on the age, sex, and weight of the animal, but may be administered once or several times a day, and the dosage may be increased or decreased depending on the route of administration, severity of the disease, sex, weight, age, etc. Therefore, the above dosage does not limit the scope of the present invention in any way.

[0036] In addition, the present invention provides a feed additive for preventing or improving benign prostatic hyperplasia containing an extract of the walnut tree as an effective ingredient.

[0037] The feed additive of the present invention corresponds to supplementary feed under the Feed Management Act. The term "feed" in the present invention may mean any natural or artificial diet, meal, etc., or ingredients of the meal, which are intended for or suitable for animals to eat, ingest, and digest. The type of the feed is not particularly limited, and feed commonly used in the relevant technical field may be used. Non-limiting examples of the feed include plant feeds such as grains, roots, fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal, or grain by-products; and animal feeds such as proteins, inorganic substances, oils and fats, mineral substances, oils and fats, single-cell proteins, zooplankton, or food. These may be used alone or in combination of two or more.

[0038] In addition, the present invention provides a herbal medicine composition for preventing or treating benign prostatic hyperplasia containing an extract of the walnut tree as an active ingredient.

[0039] The herbal medicine composition according to the present invention refers to a conventional dried medicinal herb used in the manufacture of herbal medicine. The herbal medicine composition of the present invention may be an extract obtained by conventional methods using an extraction solvent selected from water, lower alcohols having a concentration of C1 to C4, or mixtures thereof, or may be a powdered extract obtained by drying the extract. There are no particular restrictions on the formulation. For example, the herbal medicine composition may be manufactured in the form of a pill, tablet, capsule, or liquid.

[0040] Hereinafter, the present invention will be described in detail by examples. However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited to the following examples.

[0041]

[0042] Example 1. Preparation of extract of the walnut tree

[0043] (1) Preparation of ethanol extract of the walnut tree

[0044] Dried branches and fruits of the Korean walnut tree were crushed using a crusher, and 10 L of 70% ethanol was added to each 1 kg of the crushed sample, and extracted at 75°C. The extracted solution was filtered under reduced pressure using a 0.45 μm microfilter to remove solids, and the filtrate was concentrated under reduced pressure to obtain the Korean walnut tree ethanol extract. The extract was freeze-dried at -80°C and stored at -20°C for use in all analyses.

[0045]

[0046] (2) Preparation of hot water extract of the walnut tree

[0047] Based on the weight of the cut branches and fruits of the Korean walnut tree, 10 times the amount of distilled water was added, and hot water extraction was performed at 100℃ for 180 minutes. After primary filtration using a 0.45㎛ microfilter, secondary filtration was performed using a 0.22㎛ microfilter to remove precipitates. Subsequently, the extract was freeze-dried at -80℃ and stored at -20℃ for use in all analyses.

[0048]

[0049] Example 2. Confirmation of cytotoxicity according to treatment with extract of the walnut tree

[0050] To determine the cytotoxicity of ethanol and hot water extracts of branches and fruits of the walnut tree, cell viability was measured using RWPE-1 cells, a prostate epithelial cell line. RWPE-1 cells were cultured in K-SFM (Keratinocyte serum-free medium) containing 1% penicillin / streptomycin (P / S). 5 × 10 cells were seeded per well. 4The cells were dispensed into 96-well plates at a density of 10 μg / mL and treated with ethanol and hot water extracts of the branches and fruits of the present invention at concentrations (3.125, 6.25, 12.5, 25, 50, 100, and 200 μg / mL), respectively. The cells were then cultured for 24 hours and cell viability was measured using the CCK (Cell Counting Kit)-8 assay. Cell viability was measured at a wavelength of 450 nm using an Epoch microvolume spectrophotometer system (BioTek Instruments Inc., Winooski, VT, USA).

[0051] As a result, as disclosed in Fig. 1, it was confirmed that cell viability was maintained up to a concentration of 200 μg / ml of ethanol and hot water extracts of branches and fruits of the walnut tree, indicating no cytotoxicity.

[0052]

[0053] Example 3. Confirmation of the effect of extracts from the walnut tree on improving benign prostatic hyperplasia.

[0054] (1) Measurement of prostate cell proliferation inhibition ability

[0055] To confirm the efficacy of extracts of the branches and fruits of the Korean walnut tree on inhibiting prostate cell proliferation, a CCK (Cell Counting Kit)-8 experiment was performed using RWPE-1 cells, which are prostate epithelial cells. RWPE-1 cells were seeded at 5 × 10 per well. 4The cells were seeded in a 96-well plate at a density of 10 μg / ml and pretreated with 10 nM of dihydrotestosterone (DHT), a male hormone, and then treated with various concentrations (3.125, 6.25, 12.5, 25, 50, 100, 200 μg / ml) of extracts of the branches and fruits of the Chinese juniper tree, respectively. After that, the cells were treated with 30 μM of finasteride, a positive control. After culturing the cells for 24 hours, they were treated with CCK-8 solution, and 2 hours later, the cell viability was measured at a wavelength of 450 nm using an Epoch microvolume spectrophotometer system.

[0056] As a result, although prostate cells were hyperproliferated by DHT treatment, it was confirmed that in the experimental group treated with the ethanol and hot water extracts of the branches and fruits of the walnut tree of the present invention, the proliferation of prostate cells was significantly suppressed compared to the DHT treatment group (Fig. 2).

[0057]

[0058] (2) Changes in prostate weight following administration of ethanol extract of the branches of the walnut tree

[0059] For the animal model of benign prostatic hyperplasia (BPH), 6-week-old male C57BL / 6J mice (18–20 g) were purchased from Dooyeol Bio (Korea) and used for experiments. The experimental animals were randomly divided into four groups (n=5): a normal group (CON), a testosterone propionate-administered group (TP), a testosterone propionate + ethanol extract of the branches of the genus Hovenia chinensis (TP + genus Hovenia chinensis), and a testosterone propionate + finasteride-administered group (TP + Fi).

[0060] Testosterone propionate was administered subcutaneously at a concentration of 10 mg / kg once daily for 8 weeks to induce prostatic hypertrophy. Finasteride, a positive control, was administered intraperitoneally at a concentration of 10 mg / kg once daily for 8 weeks, and the ethanol extract of the branches of the walnut tree was administered orally at a concentration of 50 mg / kg once daily for 8 weeks.

[0061] To confirm the inhibitory effect of the ethanol extract of the branches of the walnut tree in experimental animals with benign prostatic hyperplasia, the body weights of the experimental animals were measured for 8 weeks (Fig. 3A). At the end of the 8th week, the experimental animals were sacrificed to examine the prostate tissues (Fig. 3B), and the ratio of prostate weight to body weight compared to the control group (CON) was measured (Fig. 3C). The ratio of prostate weight to body weight was calculated using the following equation 1).

[0062] Equation 1) Prostate weight to body weight ratio = (prostate weight of each experimental group) / (body weight of each experimental group) × 1000

[0063] As a result of checking the ratio of prostate weight to body weight of experimental animals, as shown in Fig. 3C, the ratio of prostate weight to body weight of the TP group, which induced prostatic hypertrophy, was significantly increased compared to the control group (CON). In contrast, in the TP+Fi group or the TP+Hydrangea chinensis branch ethanol extract group administered with the positive control drug finasteride, the ratio of prostate weight to body weight was confirmed to be significantly decreased compared to the TP alone administered group.

[0064] From these results, it was confirmed that the extract of the walnut tree of the present invention has an effect of improving benign prostatic hyperplasia.

Claims

1. A pharmaceutical composition for the prevention or treatment of benign prostatic hyperplasia containing extract of the cinnamon tree as an active ingredient.

2. A pharmaceutical composition for preventing or treating benign prostatic hyperplasia, characterized in that the extract of the walnut tree in the first paragraph is an extract of a branch or fruit of the walnut tree.

3. A pharmaceutical composition for preventing or treating benign prostatic hyperplasia, characterized in that the extraction solvent of the extract of the walnut tree in the first paragraph is water, a lower alcohol of C1 to C4, or a mixture thereof.

4. A pharmaceutical composition for preventing or treating benign prostatic hyperplasia, characterized in that, in addition to the effective ingredient in paragraph 1, it further comprises a pharmaceutically acceptable carrier, excipient or diluent.

5. A pharmaceutical composition for preventing or treating benign prostatic hyperplasia, characterized in that the composition is prepared in any one formulation selected from among capsules, powders, granules, tablets, suspensions, emulsions, syrups, and aerosols in the first paragraph.

6. A health functional food composition for preventing or improving benign prostatic hyperplasia containing extract of the Chinese juniper tree as an active ingredient.

7. A health functional food composition for preventing or improving benign prostatic hyperplasia, characterized in that the composition is manufactured in any one formulation selected from powder, granules, pills, tablets, capsules, candies, syrups, and beverages in the 6th paragraph.

8. A veterinary composition for the prevention or treatment of benign prostatic hyperplasia containing an extract of the cinnamon tree as an active ingredient.

9. Feed additive for preventing or improving benign prostatic hyperplasia containing extract of the cinnamon tree as an active ingredient.

10. A herbal medicine composition for the prevention or treatment of benign prostatic hyperplasia containing extract of the Chinese juniper tree as an active ingredient.

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