Composition for preventing, alleviating or treating prostatic hypertrophy containing hemp extract as active component

Abstract

The present invention relates to a composition for preventing, alleviating or treating prostatic hyperplasia, containing a hemp extract as an active component. Specifically, in the present invention, it has been confirmed that the hemp extract inhibits the proliferation of prostate cells hyperproliferated by dihydrotestosterone (DHT), which is a male hormone, and reduces weight of the prostate in an animal model in which prostatic hyperplasia is induced, and thus can be effectively used as a composition for preventing, alleviating or treating prostatic hyperplasia.

Description

Composition for preventing, improving or treating benign prostatic hyperplasia containing hemp extract as an active ingredient

[0001] The present invention relates to a composition for preventing, improving or treating benign prostatic hyperplasia, containing a hemp extract 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.: KSN2411013) 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] Hemp is an annual plant of the genus Cannabis in the family Cannabaceae. Its scientific name is Cannabis sativa L. It has been cultivated as a fiber crop in Korea and China, and in the West, it has been used as a medicinal plant for its cannabinoids, which are terpenoid components. Depending on the part of hemp, it has been used in various ways in the past. The hemp leaves, cannabis or hemp leaves, are said to have the effect of killing roundworms, and washing your hair with the water in which hemp leaves were boiled is said to make your hair grow long and shiny. It has also been used for asthma, chronic coughs, roundworms, as an analgesic, anesthetic, and as a diuretic. The root of hemp is said to treat dystocia and placental insufficiency, and to relieve blood stasis and promote the discharge of stone forests. It is said to be taken by boiling it in water and drinking it. Hemp's bark, called mapi, is said to treat bruises and open wounds, and its flower, mahwa, is said to be used for paralysis and itching. Hemp's flower spikes, called mabun, were used for dystocia, constipation, gout, sedation, and insomnia. Hemp contains various ingredients, including cannabinoids, of which the most representative ingredients are tetrahydrocannabinol (THC) and cannabidiol (CBD). These ingredients play a role in regulating various physiological functions of the body. In particular, CBD has been proven to be safe as it does not have psychoactive drug properties and is free of drug abuse or dependence. Currently, there are many cases known that CBD can be used as a useful treatment for many diseases.

[0006] Meanwhile, as for prior art related to hemp, Korean Patent No. 2269816 discloses a 'composition for anticancer, reduction of anticancer side effects and inhibition of cancer metastasis containing a hemp extract as an active ingredient', and Korean Patent Publication No. 2024-0016369 discloses a 'pharmaceutical composition for pain treatment containing hemp seed oil as an active ingredient'. However, there is no disclosure regarding a composition for preventing, improving or treating benign prostatic hyperplasia containing the hemp extract of the present invention 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 a hemp extract as an active ingredient. Specifically, the present invention was completed by confirming that the hemp extract 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] To achieve the above purpose, the present invention provides a pharmaceutical composition for preventing or treating benign prostatic hyperplasia containing a hemp extract as an active ingredient.

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

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

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

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

[0013] The present invention relates to a pharmaceutical composition for preventing or treating benign prostatic hyperplasia containing a hemp extract as an active ingredient, and more specifically, the hemp extract 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 (A) hemp root ethanol extract, (B) hemp root hot water extract, (C) hemp stem ethanol extract, or (D) hemp stem hot water extract of the present invention at various concentrations.

[0015] Figure 2 shows the results of confirming the prostate cell proliferation inhibition effect by treatment with (A) hemp root ethanol extract, (B) hemp root water extract, (C) hemp stem ethanol extract, or (D) hemp stem water extract of the present invention in prostate cells hyperproliferated by androgen DHT (dihydrotestosterone). CON is the control group, DHT is the prostate cell hyperproliferation-induced group treated 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<0.0001. *, **, *** indicate that cell proliferation was statistically significantly reduced in the hemp extract treatment group (DHT+hemp extract) or the positive control finasteride treatment group (DHT+Fi) compared to the DHT treatment group (DHT). * indicates p<0.05, ** indicates p<0.01, and *** indicates p<0.001.

[0016] Figure 3 shows the results of confirming the effect of hemp root ethanol extract on prostate hypertrophy in an animal model induced with benign prostatic hyperplasia. (A) Daily body weight (g) of each experimental group, (B) photograph of the prostate of each experimental group, (C) prostate weight (mg) of each experimental group, and (D) the relative ratio of prostate weight (PW) to body weight (BW) 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+Hemp Root is the group administered 10 mg / kg of testosterone propionate and 300 mg / kg of the hemp root ethanol extract of the present invention, and TP+Fi is the positive control group administered 10 mg / kg of testosterone propionate and 10 mg / kg of finasteride, an inhibitor of benign prostatic hyperplasia. ### indicates that the weight of the prostate or the PW / BW ratio was statistically significantly increased in the testosterone propionate administration group (TP) compared to the control group (CON), and p<0.001. **, *** indicate that the weight of the prostate or the PW / BW ratio was statistically significantly decreased in the hemp root ethanol extract administration group (TP+hemp root) or the finasteride administration group (TP+Fi) compared to the testosterone propionate administration group (TP), and ** indicates p<0.01, and *** indicates 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 a hemp extract as an active ingredient.

[0018] The extraction solvent of the above hemp extract is preferably selected from water, C1~C4 lower alcohol or a mixture thereof, more preferably ethanol, but is not limited thereto.

[0019] The extraction of the above hemp 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.

[0020] The above hemp extract is characterized in that it is an extract of the root or stem of hemp, preferably a root extract, 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 a hemp extract 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, sausage, 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 a hemp extract as an active ingredient.

[0033] The veterinary composition comprising the hemp extract 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 hemp extract 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 hemp extract according to the present invention may further comprise fillers, anticoagulants, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, etc., and the veterinary composition according to the present invention may be formulated using methods 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 powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, suppositories, sterile injectable solutions, sterile topical preparations, 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 a hemp extract as an active 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 a hemp extract 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 extraction using a conventional method using an extraction solvent selected from water, a lower alcohol having a concentration of C1 to C4, or a mixture thereof, or may be an extract in powder form 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 hemp extract

[0043] (1) Preparation of hemp ethanol extract

[0044] Dried hemp roots and stems were each crushed to a size of approximately 100 mesh using a crusher, and 3 L of 50% (roots) or 30% (stems) ethanol was added to 1 kg of each 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 a hemp ethanol extract.

[0045]

[0046] (2) Preparation of hemp hot water extract

[0047] Based on the weight of the cut hemp roots or stems, 10 times the amount of distilled water was added, and hot water extraction was performed at 115℃ 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℃ before being used in all analyses.

[0048]

[0049] Example 2. Confirmation of cytotoxicity according to hemp extract treatment

[0050] To determine the cytotoxicity of ethanol and hot water extracts of hemp root or stem, cell viability was measured using RWPE-1 cells, a prostate epithelial cell line. RWPE-1 cells, a prostate epithelial cell line, 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 seeded in a 96-well plate at a density of 10 μg / ml and treated with the ethanol and hot water extracts of the hemp root or stem of the present invention at concentrations (3.125, 6.25, 12.5, 25, 50, 100, 200 μg / ml). 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, both ethanol and hot water extracts of hemp roots or stems maintained cell viability up to a concentration of 200 μg / ml, confirming that there was no cytotoxicity.

[0052]

[0053] Example 3. Confirmation of the effect of hemp extract on improving benign prostatic hyperplasia.

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

[0055] To confirm the efficacy of hemp root and stem extracts 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. 4 The cells were seeded in a 96-well plate at a density of 10 μM 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 hemp extract and 30 μM of finasteride as a positive control. After culturing the cells for 24 hours, they were treated with CCK-8 solution, and 2 hours later, cell viability was measured at a wavelength of 450 nm using an Epoch® microvolume spectrophotometer system.

[0056] As a result, as disclosed in Fig. 2, prostate cells were hyperproliferated by DHT treatment, but it was confirmed that in the experimental group treated with the hemp root or stem ethanol extract of the present invention, proliferation of prostate cells was significantly suppressed compared to the DHT treatment group.

[0057]

[0058] (2) Changes in prostate weight following administration of hemp root ethanol extract

[0059] For the animal model of benign prostatic hyperplasia (BPH), 4-week-old male C57BL / 6J mice (18–20 g) were purchased from Dooyeol Bio (Korea) and used for experiments. The animals were randomly divided into four groups (n=5): a normal group (CON), a testosterone propionate-administered group (TP), a testosterone propionate + hemp root ethanol extract-administered group (TP + hemp root), 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 5 weeks to induce prostatic hypertrophy. Finasteride, a positive control, was administered intraperitoneally at a concentration of 10 mg / kg once daily for 5 weeks, and hemp root extract was administered orally at a concentration of 300 mg / kg once daily for 5 weeks.

[0061] To confirm the inhibitory effect of hemp root ethanol extract on prostate hypertrophy in experimental animals with benign prostatic hyperplasia, the weight of the prostate tissues of the experimental animals was measured after a total of 5 weeks of experiment (Fig. 3A). At the end of the 5th week, the experimental animals were sacrificed, and the weight of the prostate tissues (Fig. 3B) of the experimental animals was measured (Fig. 3C). The calculation of the ratio of prostate weight to body weight (PW / BW) is as follows (Fig. 3D).

[0062] Prostate weight relative to body weight (PW / BW) = (prostate weight of each experimental group) / (body weight of each experimental group) × 1000

[0063] As a result of measuring the weight of the prostate tissue of the experimental animals, as disclosed in Figures 3C and 3D, it was confirmed that the prostate weight of the TP group, which induced prostatic hypertrophy, significantly increased compared to the control group (CON). In contrast, in the TP+Fi group administered with the positive control drug finasteride or the group administered with TP+hemp root ethanol extract, the prostate weight and the prostate weight-to-body weight ratio were confirmed to be significantly reduced compared to the TP group.

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

Claims

1. A pharmaceutical composition for preventing or treating benign prostatic hyperplasia containing hemp extract as an active ingredient.

2. A pharmaceutical composition for preventing or treating benign prostatic hyperplasia, characterized in that the extraction solvent of the hemp extract in paragraph 1 is water, a lower alcohol of C1 to C4, or a mixture thereof.

3. A pharmaceutical composition for preventing or treating benign prostatic hyperplasia, characterized in that the hemp extract in paragraph 1 is an extract of the root or stem of hemp.

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 hemp extract 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 hemp extract as an active ingredient.

9. Feed additive for preventing or improving benign prostatic hyperplasia containing hemp extract as an active ingredient.

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

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