KR102623366B1 - Composition for preventing, improving or treating benign prostatic hyperplasia comprising deer antler fermentation products as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for preventing, improving or treating benign prostatic hyperplasia containing fermented deer antler as an active ingredient.
The composition of the present invention contains deer antler fermentation as an active ingredient, thereby suppressing the enlargement of prostate tissue in benign prostatic hyperplasia as well as suppressing the expression of AR, PSA, and 5-alpha reductase, thereby improving benign prostatic hyperplasia. , it can be effectively used for treatment or prevention. In addition, since the present invention is a composition derived from natural products, it can be used safely without side effects, and can be usefully used in medicine or food.

Description

Composition for preventing, improving or treating benign prostatic hyperplasia comprising deer antler fermentation products as an active ingredient}

The present invention relates to a composition for preventing, improving, or treating benign prostatic hyperplasia (BPH), comprising fermented deer antler as an active ingredient.

The prostate is a walnut-shaped organ that surrounds the beginning of the urethra in men and produces mucous substance located below the bladder. The sole function of the prostate is to produce semen, which protects and stimulates sperm for reproduction. The prostate has the characteristic of growing and enlarging with age, and in most mammals, the urethra, a passage that discharges urine from the bladder, passes through the prostate. Due to these anatomical characteristics of the prostate, men often have problems related to urine discharge as they age, and various diseases, including prostate-related diseases, may occur.

In the past, prostatic hypertrophy was defined as a condition in which the prostate enlarges and blocks the passage through which urine comes out from the lower part of the bladder, causing urethral obstruction and reducing the flow of urine. Histologically, it was defined as the proliferation of prostatic stroma or epithelial tissue cells of the prostate. , Recently, the pathophysiology of the disease has become too complex to be explained with such definitions or concepts, and currently, 'frequent urination (urinating more than 8 times a day, nocturia, strong and sudden urge to urinate)' in men over 50 years of age Symptoms of bladder storage disorders such as uncontrollable urgency to urinate, urge incontinence, nocturia, and pain during urination, delayed urination (a phenomenon in which urine comes out only after a pause when urinating), and stop urine flow (a phenomenon in which urine flow is interrupted). ), lower urinary tract symptoms (LUTS), a general term for symptoms of bladder discharge disorders such as abdominal pressure urination (a phenomenon requiring strain during urination), urinary gland weakness, feeling of urination, and urinary retention, are defined as benign prostatic hyperplasia. there is.

One of the major causes of such enlarged prostate is caused by an imbalance in the contraction and relaxation of prostatic smooth muscle or urethral smooth muscle, and the biggest triggers are increasing age and the presence of male hormones. In addition, race, environment, diet, and genetic factors can also affect the disease. Symptoms generally begin in the 30s or 40s, usually appear after the age of 60, and with age, more than 50% of microscopic hypertrophic prostates develop into palpable hypertrophic prostates (Prostate 1996 (suppl. 6) 67-73). ).

Diagnosis of benign prostatic hyperplasia is made by palpation of the prostate within the rectum, and the degree of bladder outlet obstruction and bladder irritation is referred to through detailed examination through urodynamics or ultrasound, and an appropriate treatment method is selected based on this.

Currently, alpha-adrenergic blockers and 5α-reductase inhibitors are widely used as treatments for benign prostatic hyperplasia.

Representative alpha-adrenergic nerve blockers include alfuzosin, terazosin, doxazosin, prazosin, tamsulosin, and silodosin, which It reduces the symptoms of obstruction by reducing the tension of smooth muscles in the prostate tissue. However, alpha-adrenergic blockers have the disadvantage of resolving urinary problems by directly dilating the urethra, so they return to their original state when the medication is discontinued. Additionally, it is not an appropriate treatment for patients with benign prostatic hyperplasia and erectile dysfunction due to the side effects of retrograde ejaculation and causing orthostatic hypotension. Additionally, cautious administration should be considered in patients with severe hepatic and renal dysfunction.

In addition, 5α-reductase inhibitors such as finasteride block dihydrotestosterone (DHT), which thickens the prostate, thereby reducing differentiation of prostate tissue and reducing its size, thereby increasing urinary flow and urination in the long term. Alleviates symptoms of disability. However, it requires long-term use of at least 6 months to be effective, and has the disadvantage of causing side effects such as erectile dysfunction, decreased libido, ejaculation disorders, and gynecomastia.

Accordingly, research is being actively conducted to develop compositions effective in preventing or treating benign prostatic hyperplasia using natural products that have no side effects on the human body.

Meanwhile, antler ( Cornu cervi pantotrichum , antler) refers to the cartilage tissue that is regenerated every year from young non-keratinized antlers with dense hair from plum blossoms, marrocks, and related animals belonging to the deer family. The non-keratinized ones are antlers, and the non-keratinized ones are antlers, and the keratinized ones are It is called deer antler, and in Oriental countries such as Korea, China, and Japan, deer antler has been widely used as the best blood-replenishing tonic for a long time, and its properties and efficacy are listed in literature such as Materia Medica and Donguibogam.

Deer antler has been reported to have pharmacological effects on the immune system, hematopoietic system, sugar metabolism, and cardiovascular system. Its pharmacologically active ingredients include ganglioside, pantocrin (70% ethanol extract), amino acid, and chondroitin. ), glucosamine, hyaluronic acid, calcium phosphate, calcium carbonate, collagen, phospholipids, etc. are known. In addition, it has been scientifically reported that it has immune-boosting, blood pressure-lowering, hematopoietic function, hypercholesterolemia, and anti-stress effects.

Herbal medicines with tonic efficacy, such as deer antler, do not have a strong effect as they act only on specific organs. The effect is weaker than that of single-ingredient medicines, but is characterized by a complex effect throughout the body. Deer antler has side effects such as diarrhea, so in oriental medicine, it is used in combination with herbal medicine with an astringent effect, but in this case, the expected effect is reduced. During the extraction process using solvents such as ethanol, many of the active ingredients of deer antler are lost. A way to improve these problems is through fermentation, which increases the efficiency of deer antler and allows new pharmacological effects to be expected.

The purpose of fermentation is generally to cultivate useful microorganisms to produce ingredients that are helpful to the human body and to remove ingredients that cause toxicity or side effects. The process of fermentation has various advantages such as imparting new physiological activity, increasing useful intestinal microorganisms, increasing absorption rate, and reducing or eliminating residual pesticides, so it has recently been used as a biological conversion method using fermentation. However, to date, no data has been disclosed or taught regarding the treatment of benign prostatic hyperplasia using fermented deer antler fermented using lactic acid bacteria or Bacillus subtilis.

Accordingly, the present inventors made efforts to develop a substance for treating benign prostatic hyperplasia derived from natural products that is safe for the human body. As a result, they confirmed that fermented deer antler was significantly effective in improving, treating, or preventing benign prostatic hyperplasia, and completed the present invention.

KR 10-2037385 B

The purpose of the present invention is to provide a composition for preventing, improving or treating benign prostatic hyperplasia containing fermented deer antler as an active ingredient.

In order to achieve the above object, the present invention provides a food composition for preventing or improving benign prostatic hyperplasia containing fermented deer antler as an active ingredient.

According to one embodiment of the present invention, the fermented antler antler product may be obtained by fermenting antler extract with lactic acid bacteria or Bacillus subtilis .

According to one embodiment of the present invention, the deer antler extract may be an extract of deer antler with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

According to one embodiment of the present invention, the lactic acid bacteria are selected from the lactic acid bacteria of the genus Lactobacillus, Bifidobacterium, Leuconostoc, Pediococcus, Weissella, Staphylococcus, Erococcus and Enterococcus. It may be any one or more selected lactic acid bacteria.

According to one embodiment of the present invention, the lactic acid bacteria may be any one or more selected from Leuconostoc mesenteroides, Lactobacillus plantarum, and Lactobacillus paraplantarum.

According to one embodiment of the present invention, the deer antler fermentation product can inhibit the production of one or more types selected from prostate-specific antigen (PSA), androgen receptor (AR), and 5-alpha reductase-2.

In addition, the present invention provides a pharmaceutical composition for preventing or treating benign prostatic hyperplasia containing fermented deer antler as an active ingredient.

The composition of the present invention contains deer antler fermentation as an active ingredient, thereby suppressing the enlargement of prostate tissue in benign prostatic hyperplasia as well as suppressing the expression of AR, PSA, and 5-alpha reductase, thereby improving benign prostatic hyperplasia. , it can be effectively used for treatment or prevention. In addition, since the present invention is a composition derived from natural products, it can be used safely without side effects, and can be usefully used in medicine or food.

Hereinafter, the present invention will be described in detail.

The present invention relates to a composition for preventing, improving or treating benign prostatic hyperplasia containing fermented deer antler as an active ingredient.

“Prostate hypertrophy” as used herein refers to a disease in which the prostate grows abnormally and puts pressure on the urethra, making it difficult to urinate. It is a general term for symptoms and diseases that are mainly accompanied by lower urinary tract symptoms due to enlargement of the prostate. Prostate thickening, urethral obstruction, dysuria, polyuria, enuresis, urinary incontinence, dysuria, delayed urination, interrupted urine, abdominal pressure urination, urinary duct weakness, and residual urination. , urinary retention, or a combination of these symptoms.

According to a specific embodiment of the present invention, a composition containing the fermented deer antler product of the present invention can suppress the enlargement of prostate tissue in benign prostatic hypertrophy. In addition, according to another specific embodiment of the present invention, the composition containing the fermented antler antler of the present invention acts on Prostate Specific Antigen (PSA), Androgen Receptor (AR), and 5-alpha-reduction in benign prostatic hyperplasia. It can inhibit the expression of enzyme-2 (5α-reductase22). Therefore, the fermented deer antler product of the present invention can be usefully used as an active ingredient in a composition for preventing, improving, or treating benign prostatic hyperplasia.

In one embodiment, the present invention provides a food composition for preventing or improving benign prostatic hyperplasia comprising fermented deer antler as an active ingredient.

In this specification, “fermented deer antler” may be a deer antler extract fermented with lactic acid bacteria or Bacillus subtilis . Preferably, the antler is mixed with one or more lactic acid bacteria selected from Lactobacillus, Bifidobacterium, Leuconostoc, Pediococcus, Weissella, Staphylococcus, Erococcus, and Enterococcus. , or it may be fermented with Bacillus subtilis. More preferably, the deer antler is grown with one or more lactic acid bacteria or Bacillus subtilis selected from among Leuconostoc mesenteroides , Lactobacillus plantarum , and Lactobacillus paraplantarum . It may have been fermented. Most preferably, the deer antler is prepared from Leuconostoc mesenteroides SST-9962 [Accession number: KCCM12712P], Lactobacillus paraplantarum SST-9961 [Accession number: KCCM12711P] and Bacillus subtilis. It may be fermented with any one strain selected from Bacillus subtilis SST-9960 [Accession number: KCCM 11402P].

The “fermented deer antler product” can be fermented using a common fermentation method known in the art without limitation.

Meanwhile, the antler extract includes not only the crude extract obtained by treating the antler, which is the raw material for extraction, with an extraction solvent, but also processed products of the crude extract. For example, deer antler extract can be prepared in powder form by additional processes such as reduced pressure distillation, freeze-drying, or spray drying. The extract also includes fractions obtained by additional fractionation of the crude extract.

The antler extract is an extract of water, alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, and the extraction method does not need to be particularly limited.

As a preferred embodiment of the present invention, hot water extraction is preferred when water is used as a solvent. For example, extraction can be performed in water at 80 to 130°C, preferably 90 to 110°C, for 0.5 to 24 hours, preferably 1 to 6 hours. And, even if hot water is not used as a solvent, that is, deer antler components can be extracted from a diluted solution obtained by diluting deer antler powder in cold water or room temperature water. When extracting deer antler components using cold water or water at room temperature, the extraction may be done separately from fermentation, or the components of the antler may be eluted from the antler during the fermentation process by inoculating it with lactic acid bacteria or Bacillus subtilis.

As a preferred embodiment of the present invention, an extract using an aqueous alcohol solution having 1 to 4 carbon atoms can be used. For example, it can be extracted with an aqueous alcohol solution such as ethanol, methanol, or isopropanol, preferably an aqueous alcohol solution of 20 to 80 wt%, and more preferably an aqueous alcohol solution of 50 to 70 wt%. When using an alcohol extract, before inoculating lactic acid bacteria or Bacillus subtilis, the alcohol contained in the alcohol extract can be vaporized to lower the alcohol content, or the alcohol extract can be concentrated and dried and then dissolved in water.

In addition, in the present invention, deer antler extract in a broad sense includes a diluted solution obtained by diluting deer antler powder in water. In the test example of the present invention, compared to the fermented antler antler product in which the antler antler hot water extract or the aqueous antler antler solution extract was fermented with lactic acid bacteria or Bacillus subtilis, the antler antler fermentation product in which the antler water dilution was fermented with lactic acid bacteria or Bacillus subtilis prevented and improved prostate hypertrophy. Alternatively, although the therapeutic effect was somewhat low, it was confirmed that the effect was significantly increased compared to unfermented deer antler extract.

Before inoculating the antler extract with lactic acid bacteria or Bacillus subtilis, lactic acid nutrient sources such as protein sources, carbohydrate sources, vitamins, or minerals may be additionally mixed to enhance the growth of the strain. As the strain nutrient source, commercially available media can be used, or only the necessary nutrients can be added individually.

Additionally, the antler extract, or a mixture of antler extract and strain nutrient source, can be sterilized by heat before inoculation with lactic acid bacteria or Bacillus subtilis.

The solid content of the deer antler extract does not need to be specifically limited, but is 1 to 15% by weight if there is no other concentration process after extraction. It can be used directly, concentrated, or diluted after concentration or drying.

In the present invention, the lactic acid bacteria are one or more lactic acid bacteria selected from the genus Lactobacillus, Bifidobacterium, Leuconostoc, Pediococcus, Weissella, Staphylococcus, Erococcus, and Enterococcus. It may be lactic acid bacteria. Preferably, the lactic acid bacteria may be at least one selected from Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus paraplantarum, Lactobacillus sakei, and Pediococcus pentosaceus, and more preferably the above The lactic acid bacteria may be any one or more selected from among Leuconostoc mesenteroides, Lactobacillus plantarum, and Lactobacillus paraplantarum, and more preferably, the lactic acid bacteria are selected from among Leuconostoc mesenteroides and Lactobacillus paraplantarum. There may be more than one selected.

The strain may be added as the strain itself or as a culture medium. The culture medium may include the culture medium itself obtained by culturing the strain in a liquid medium, a concentrate obtained by concentrating the culture medium, etc.

In a preferred embodiment of the present invention, the fermentation strain, especially the lactic acid bacteria or Bacillus subtilis strain, is 1×10 ⁶ to 1×10 ⁹ cfu/mL, preferably 1×10 ⁶ to 1×10 ⁸ cfu/mL, More preferably, the culture medium contained at a concentration of 1 × 10 ⁷ to 1 × 10 ⁸ cfu/mL is 0.3 to 3.0 parts by weight, preferably 1.0 to 2.5 parts by weight, more preferably 1.5 parts by weight, based on 100 parts by weight of the deer antler extract. It may be added in an amount of from 2.5 parts by weight.

The "fermented antler antler" for significantly producing the desired effect in the present invention is (1) placing antler extract in a fermentation tray and adding 0.3 to 3.0 weight of lactic acid bacteria culture medium or Bacillus subtilis culture medium per 100 parts by weight of deer antler extract. After addition, (2) at 20 to 40°C for 18 to 96 hours, preferably 18 to 96 hours, more preferably 18 to 72 hours, more preferably 18 to 48 hours, more preferably 18 to 36 hours. It may be manufactured by fermenting for 20 to 30 hours, more preferably 20 to 30 hours.

In particular, (3) after the fermentation, low-temperature aging at 2 to 10°C for 5 to 24 hours, and then (4) air drying at 25 to 35°C, preferably 28 to 33°C for 4 to 8 hours. Additional performance is more desirable in terms of effectiveness. According to one embodiment of the present invention, it was specifically confirmed that the effect of improving benign prostatic hyperplasia was further enhanced when the fermented antler antler was aged at low temperature.

In addition, the "fermented deer antler product" of the present invention prepared by the above method has a nutritional component content compared to the fermented deer antler product prepared by other methods, or the fermented deer antler product fermented and produced by strains other than lactic acid bacteria or Bacillus subtilis. It was specifically confirmed that not only was this further enhanced, but the effect of improving benign prostatic hyperplasia was further enhanced.

In addition, among the strains for fermentation, lactic acid bacteria have an excellent growth rate in deer antler extract, so they have the advantage of having a shorter fermentation time compared to other strains, including Bacillus subtilis strains.

Meanwhile, the fermented deer antler may be a fermented product containing microbial cells, or may be a fermented product from which the bacteria have been removed. The fermented product from which the bacterial cells have been removed may be sterilized to contain dead bacterial cells, or may be a filtrate or centrifuged supernatant in which the bacterial cells have been removed through filtration or centrifugation. The prevention, improvement or treatment effect of benign prostatic hyperplasia of the present invention is achieved by bioconverting the components contained in the deer antler extract into ingredients that are easy to use in the body as microorganisms proliferate, or by creating a novel product that has the effect of preventing, improving or treating benign prostatic hyperplasia. It is expected that this is because the active ingredient is produced, and the effect is significantly higher than the effect that can be obtained by adding the simple strain itself. The fermented deer antler product can be used as a liquid fermented product itself, or it can be dried and powdered.

In this specification, “containing as an active ingredient” means containing an amount sufficient to achieve the efficacy or activity of preventing, improving, or treating benign prostatic hyperplasia in the fermented antler antler product of the present invention.

The term “prevention” as used herein means anything that suppresses or delays the enlargement of the prostate.

The term "improvement" herein means at least reducing the degree of symptoms, for example, parameters related to the condition being treated by administration of a composition comprising a fermented deer antler product of the present invention.

The term “treatment” herein, unless otherwise stated, means reversing, alleviating, inhibiting the progression of, or preventing the symptoms of benign prostatic hyperplasia.

The composition of the present invention does not exhibit cytotoxicity by containing the fermented deer antler product as an active ingredient and has a significant effect in preventing or improving benign prostatic hyperplasia.

In a specific embodiment of the present invention, the fermented deer antler of the present invention not only inhibits the enlargement of prostate tissue in benign prostatic hyperplasia, but also has the effect of suppressing the expression of AR, PSA, and 5-alpha reductase, and thus has the effect of suppressing the expression of AR, PSA, and 5-alpha reductase. It was specifically confirmed that all of them could show significant improvement or treatment effect.

The food composition for preventing or improving benign prostatic hyperplasia of the present invention includes all forms such as nutritional supplements, health functional foods, food additives, and feed, including humans or livestock. Animals are used as food.

The food composition may be a formulation of the fermented deer antler product of the present invention in the form of capsules, tablets, powders, granules, liquid, pills, flakes, pastes, syrups, gels, jelly or bars. Unlike general drugs, the food composition has the advantage of having no side effects that may occur when taken for a long period of time.

Additionally, the food composition can be manufactured in various forms according to conventional methods known in the art. General foods include, but are not limited to, beverages (including alcoholic beverages), fruits and their processed foods (e.g. canned fruit, bottled foods, jam, mamalade, etc.), fish, meat and their processed foods (e.g. ham, sausages, etc.) corned beef, etc.), bread and noodles (e.g. udon, buckwheat noodles, ramen, spagate, macaroni, etc.), fruit juice, various drinks, cookies, taffy, dairy products (e.g. butter, cheese, etc.), edible vegetable oil, margarine , vegetable proteins, retort foods, frozen foods, and various seasonings (e.g., soybean paste, soy sauce, sauce, etc.) can be produced by adding the fermented deer antler. In addition, nutritional supplements are not limited to this, but can be prepared by adding the fermented deer antler to capsules, tablets, pills, etc. In addition, the health functional food is not limited to this, but for example, the fermented antler antler itself can be manufactured in the form of tea, juice, and drinks and liquefied for drinking (health drinks), granulated, encapsulated, and powdered. It can be digested and consumed. In addition, in order to use the fermented antler antler product in the form of a food additive, it can be prepared as a powder, or extracted and used in the form of a concentrate.

When the composition for preventing or improving benign prostatic hyperplasia of the present invention is used as a health drink composition, the health drink composition may contain various flavoring agents or natural carbohydrates as additional ingredients like ordinary drinks. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; polysaccharides such as dextrins and cyclodextrins; It may be a sugar alcohol such as xylitol, sorbitol, or erythritol. Sweeteners include natural sweeteners such as thaumatin and stevia extract; Synthetic sweeteners such as saccharin and 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 mL of the composition of the present invention.

The fermented deer antler product of the present invention may be contained as an active ingredient in the food composition, and the amount may be an amount effective to achieve a preventive, improvement, or treatment effect on benign prostatic hyperplasia. Specifically, the antler fermented product is used in an amount of 0.01 to 10% by weight, preferably 0.05 to 10% by weight, more preferably 0.1 to 5% by weight, more preferably 0.3 to 2% by weight, based on the total weight of the food composition. More preferably, it may be contained in 0.6 to 1.5% by weight, and even more preferably in 0.8 to 1.2% by weight. In addition, when the food composition is a health drink composition, the antler fermented product is added to the health drink composition in an amount of 0.1 to 100 mg/ml, preferably 0.5 to 100 mg/ml, more preferably 1 to 50 mg/ml, More preferably, the concentration may be 3 to 20 mg/ml, more preferably 6 to 15 mg/ml, and even more preferably 8 to 12 mg/ml. At this time, if the content of the antler fermentation product is less than the lower limit, the cell survival rate is excellent, but the effect of improving benign prostatic hyperplasia may not be achieved to the desired extent. Conversely, if the upper limit is exceeded, the effect of improving benign prostatic hyperplasia may not increase as the concentration increases or may be toxic. Meanwhile, as a result of in vitro experiments, when the concentration of the fermented deer antler product of the present invention was within the above range, a significant effect was observed in improving benign prostatic hyperplasia, but no side effects such as cytotoxicity were observed.

Also, according to a preferred embodiment, the daily dosage of the antler fermented product may be 1 to 100 mg/kg, preferably 5 to 50 mg/kg, and more preferably 10 to 30 mg/kg, based on dry weight. . At this time, if the daily dose of the fermented deer antler is less than the lower limit, the effect of improving benign prostatic hyperplasia may not be achieved to the desired extent. Conversely, if the upper limit is exceeded, the effect of improving benign prostatic hyperplasia may not increase as the concentration increases or may be toxic.

The food composition of the present invention can be prepared by mixing deer antler fermentation product with other active ingredients known to have preventive, ameliorating or therapeutic effects on benign prostatic hyperplasia.

In addition to the above, the food composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, It may contain glycerin, alcohol, or carbonating agent. In addition, the health food of the present invention may contain pulp for the production of natural fruit juice, fruit juice beverage, or vegetable beverage. These ingredients can be used independently or in combination. The ratio of these additives is not very important, but is generally selected in the range of 0.001 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

In another embodiment, the present invention provides a pharmaceutical composition for preventing or treating benign prostatic hyperplasia comprising fermented deer antler as an active ingredient. The pharmaceutical composition of the present invention can be prepared using pharmaceutically suitable and physiologically acceptable auxiliaries in addition to the antler fermentation product, and the auxiliaries include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, Lubricants or flavoring agents can be used. In the present invention, the terms “fermented deer antler” and “prostate hypertrophy” are as described above.

Additionally, the pharmaceutical composition for preventing or treating benign prostatic hyperplasia of the present invention may be formulated to include one or more pharmaceutically acceptable carriers.

Pharmaceutically acceptable carriers may further include, for example, carriers for oral administration or carriers for parenteral administration.

Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc.

Carriers for parenteral administration may also include water, suitable oils, saline solutions, aqueous glucose and glycols, and the like. Additionally, stabilizers and preservatives may be additionally included. Suitable stabilizers include antioxidants such as sodium bisulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. As for other pharmaceutically acceptable carriers, those described in the following literature may be referred to (Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).

The pharmaceutical composition of the present invention can be administered to mammals, including humans, by any method. For example, it can be administered orally or parenterally, and in the case of parenteral administration, it can be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc., and oral administration is preferred.

The pharmaceutical composition of the present invention can be formulated into a preparation for oral administration or parenteral administration according to the administration route described above. When formulated, one or more buffers (e.g. saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g. EDTA or glutathione), fillers, bulking agents, binders, adjuvants (e.g. aluminum hydroxide) side), suspending agents, thickening agents, wetting agents, disintegrants or surfactants, diluents or excipients.

Preparations for oral administration include tablets, pills, powders, granules, solutions, gels, syrups, slurries, suspensions or capsules, and such solid preparations include at least one excipient, for example, in the pharmaceutical composition of the present invention. , starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol, maltitol, cellulose It can be prepared by mixing methyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose, or gelatin. For example, tablets or dragees can be obtained by combining the active ingredient with solid excipients, grinding them, adding suitable auxiliaries and processing them into a granule mixture.

In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, or syrups. In addition to the commonly used simple diluents such as water or liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, or preservatives may be included. .

In addition, in some cases, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as a disintegrant, and anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, and preservatives may be additionally included. .

When administered parenterally, the pharmaceutical composition of the present invention can be formulated with a suitable parenteral carrier in the form of injections, transdermal administration, and nasal inhalation according to methods known in the art. The above injections must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. For injections, examples of suitable carriers include, but are not limited to, solvents or dispersion media including water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.), mixtures thereof, and/or vegetable oils. You can. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine, or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol, and 5% dextrose. etc. can be used. In order to protect the injection from microbial contamination, it may additionally contain various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. Additionally, in most cases, the injection may additionally contain an isotonic agent such as sugar or sodium chloride.

In the case of transdermal administration, forms such as ointments, creams, lotions, gels, external solutions, paste preparations, linear preparations, and aerol preparations are included. In the above, “transdermal administration” means administering a pharmaceutical composition topically to the skin so that an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin.

For inhalation administration, the composition according to the invention can be administered from a pressurized pack or nebulizer using a suitable propellant, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. It can be conveniently delivered in the form of an aerosol spray. For pressurized aerosols, the dosage unit can be determined by providing a valve that delivers a metered amount. For example, gelatin capsules and cartridges for use in inhalers or insufflators can be formulated to contain powder mixtures of suitable powder bases such as lactose or starch. Formulations for parenteral administration are described in Remington's Pharmaceutical Science, 15th Edition, 1975. Mack Publishing Company, Easton, Pennsylvania 18042, Chapter 87: Blaug, Seymour, a text generally known in all pharmaceutical chemistry.

The pharmaceutical composition of the present invention can provide desirable effects for preventing, improving, or treating benign prostatic hyperplasia when it contains an effective amount of deer antler fermentation product. In this specification, the term 'effective amount' refers to an amount that shows a greater response than the negative control, and preferably refers to an amount sufficient to improve or treat benign prostatic hyperplasia. The deer antler fermentation product is used in an amount of 0.01 to 10% by weight, preferably 0.05 to 10% by weight, more preferably 0.1 to 5% by weight, more preferably 0.3 to 2% by weight, even more preferably, based on the total content of the pharmaceutical composition. It may be included at 0.6 to 1.5% by weight, more preferably at 0.8 to 1.2% by weight. Alternatively, the deer antler fermentation product is used in an amount of 0.1 to 100 mg/ml, preferably 0.5 to 100 mg/ml, more preferably 1 to 50 mg/ml, more preferably 3 to 20 mg/ml, relative to the pharmaceutical composition. , more preferably 6 to 15 mg/ml, more preferably 8 to 12 mg/ml. At this time, if the content of the antler fermentation product is less than the lower limit, the cell survival rate is excellent, but the effect of improving benign prostatic hyperplasia may not be achieved to the desired extent. Conversely, if the upper limit is exceeded, the effect of improving benign prostatic hyperplasia may not increase as the concentration increases or may be toxic. Meanwhile, as a result of in vitro experiments, when the concentration of the fermented deer antler product of the present invention was within the above range, a significant effect was observed in improving benign prostatic hyperplasia, but no side effects such as cytotoxicity were observed. The effective amount of fermented deer antler contained in the pharmaceutical composition of the present invention will vary depending on the form in which the composition is commercialized.

The total effective amount of the pharmaceutical composition of the present invention can be administered to a patient as a single dose, or may be administered by a fractionated treatment protocol in which multiple doses are administered over a long period of time. . The pharmaceutical composition of the present invention may vary the content of the active ingredient depending on the severity of the disease.

The appropriate dosage of the pharmaceutical composition varies depending on factors such as formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity, and is usually A skilled physician can easily determine and prescribe an effective dosage for desired treatment or prevention. According to a preferred embodiment, the daily dosage of the fermented deer antler product may be 1 to 100 mg/kg, preferably 5 to 50 mg/kg, and more preferably 10 to 30 mg/kg, based on dry weight. At this time, if the daily dose of the fermented deer antler is less than the lower limit, the effect of improving benign prostatic hyperplasia may not be achieved to the desired extent. Conversely, if the upper limit is exceeded, the effect of improving benign prostatic hyperplasia may not increase as the concentration increases or may be toxic.

The pharmaceutical composition for preventing and treating benign prostatic hyperplasia of the present invention can also be provided in the form of an external preparation containing fermented deer antler as an active ingredient.

When the pharmaceutical composition for preventing and treating benign prostatic hyperplasia of the present invention is used as an external skin agent, it may additionally contain fatty substances, organic solvents, solubilizers, thickening and gelling agents, softeners, antioxidants, suspending agents, stabilizers, and foaming agents. agent), fragrance, surfactant, water, ionic emulsifier, non-ionic emulsifier, filler, metal ion sequestrant, chelating agent, preservative, vitamin, blocking agent, wetting agent, essential oil, dye, pigment, hydrophilic activator, lipophilic It may contain adjuvants commonly used in the field of dermatology, such as active agents or lipid vesicles or any other ingredients commonly used in topical skin preparations. Additionally, the ingredients may be introduced in amounts commonly used in the field of dermatology.

When the pharmaceutical composition for preventing and treating benign prostatic hyperplasia of the present invention is provided as an external skin preparation, it is not limited thereto, but may be in the form of an ointment, patch, gel, cream, or spray.

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments. However, these examples are for illustrating the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited thereto.

<Example>

Preparation Example 1: Preparation of deer antler hot water extract

Domestic antler was selected, washed, and evenly ground with a grinder to prepare antler powder with an average particle size of 300 ㎛, which was stored in the refrigerator and used.

The antler powder and 10 times the weight of distilled water were placed in a reflux extractor and extracted twice at 100°C for 3 hours, then allowed to cool and concentrated under reduced pressure to obtain antler hot water extract. The sweetness of the antler hot water extract was 1.6 brix, the reducing sugar content was 0.24%, and the protein content was 0.31%.

Preparation Example 2: Preparation of deer antler ethanol aqueous solution extract

The deer antler powder of Preparation Example 1 and 4 times the weight of the 50% by weight ethanol aqueous solution were added, extracted at room temperature for 3 days, and then concentrated under reduced pressure to obtain a deer antler ethanol aqueous solution extract. The sweetness of the antler ethanol aqueous solution extract was 1.7 Brix.

Preparation Example 3: Preparation of antler water dilution

An antler water dilution was prepared by mixing the antler powder of Preparation Example 1 with water to 2.0% by weight.

Test Example 1: Fermentation characteristics of strains in antler extract - change in viable cell count

In order to confirm the fermentation characteristics of lactic acid bacteria and Bacillus subtilis in deer antler extract, we attempted to determine changes in the number of viable bacteria at 12 and 24 hours after inoculation.

First, a lactic acid bacteria culture solution was prepared by inoculating the lactic acid bacteria shown in Table 1 below into MRS broth medium and activating them by culturing them statically at 37°C for 24 hours.

Lactobacillus strain name Deposit number Leuconostoc mesenteroides SST-9962 KCCM12712P Lactobacillus paraplantarum SST-9961 KCCM12711P

In addition, Bacillus subtilis culture medium was prepared by inoculating the Bacillus subtilis SST-9960 [Accession number: KCCM 11420P] strain into LB liquid medium and culturing it at 30°C for 2 days to activate it.

Next, 1.5% by weight of glucose was added to the deer antler extract of Preparation Examples 1 to 3, each diluted with 4 times the weight of distilled water, and the lactic acid bacteria culture medium (1 × 10 ⁸ CFU/mL) or Bacillus subtilis The culture medium (1×10 ⁸ CFU/mL) was inoculated at 0.5% by weight, fermented at 37°C for 24 hours, and the number of viable bacteria was measured. The results are shown in Table 2 below.

For live bacterial water, dilute 1 mL of sample with 9 mL of sterile saline solution, mix well, dilute step by step, then take 0.1 mL, store in as anaerobic conditions as possible using an anaerobic jar (BBL Gas Pak Anaerobic System), and culture at 37°C. After that, the number of colonies produced was measured, and the average number of colonies was multiplied by the dilution factor to calculate the number of colonies per culture medium.

division Viable bacteria count (CFU/mL) Before fermentation After fermentation
(Production Example 1) After fermentation
(Production Example 2) After fermentation
(Production Example 3) Leuconostoc mesenteroides
SST-9962 5.0×10 ⁵ 7.2×10 ⁷ 6.8×10 ⁷ 3.3×10 ⁷ Lactobacillus paraplantarum SST-9961 5.0×10 ⁵ 2.6×10 ⁷ 2.2×10 ⁷ 1.0×10 ⁷ Bacillus subtilis
SST-9960 5.0×10 ⁵ 0.8×10 ⁷ 0.6×10 ⁷ 0.4×10 ⁷

As shown in Table 2, there was little difference between the antler hot water extract of Preparation Example 1 and the ethanol aqueous solution extract of Preparation Example 2 as the fermentation substrate, but when the antler water dilution of Preparation Example 3 was used, after fermentation The number of viable bacteria was slightly less than half of the number of viable cells in Preparation Examples 1 and 2.

In addition, it was confirmed that the number of viable bacteria in the culture medium prepared using the Leuconostoc mesenteroides SST-9962 strain was much higher (about 3 times) than that in the culture medium prepared using the Lactobacillus paraplant arum SST-9961 strain.

Test Example 2: Comparison of growth rates in deer antler extracts of lactic acid bacteria and Bacillus subtilis strains

We attempted to compare the growth rates of the lactic acid bacteria and Bacillus subtilis strains of the present invention in antler extract.

For this purpose, 1.5% by weight of glucose was added to the deer antler extract of Preparation Example 1 diluted with 2, 4, and 10 times the weight of distilled water, and the lactic acid bacteria culture medium (1 × 10 ⁸ CFU/mL) was added to 0.5%. After inoculation by weight percentage and fermentation at 37°C for 24 hours, the growth rate of the strain was measured at 600 nm using a spectrophotometer (Optizen 2120UV, Mecasys co., Ltd., Korea), and is shown in Table 3 below.

division Growth rate (OD ₆₀₀ ) Before dilution 2x dilution 4-fold dilution 10-fold dilution SST-9962 0.597 1.112 1.624 0.784 SST-9961 0.612 1.256 1.823 0.767 SST-9960 0.609 0.615 0.224 0.110

Looking at Table 3 above, there was no significant difference in the growth rates of the Leuconostoc mesenteroides SST-9962 strain and the Lactobacillus paraplant arum SST-9961 strain in the 4-fold diluted antler extract, but the growth rates of the Bacillus antler extract in the 4-fold diluted antler extract were not significantly different. The growth rate of the subtilis strain was relatively low compared to the growth rate of the SST-9962 strain and SST-9961 strain.

In addition, for the SST-9962 and SST-9961 strains, the growth rate was highest in the antler extract diluted 4 times compared to the antler extract diluted 2 times or 10 times before dilution, whereas for Bacillus subtilis, the growth rate was highest in the antler extract diluted 2 times or 10 times. The highest growth rate was found in whole or two-fold diluted deer antler extract.

Therefore, in the following examples, a fermented antler antler product was prepared using the antler antler hot water extract of Preparation Example 1. In addition, when fermenting with the SST-9962 strain or SST-9961 strain, the deer antler extract was diluted 4-fold and used, and when fermenting with the Bacillus subtilis strain, the antler extract was diluted 2-fold.

Examples 1-2: Preparation of fermented deer antler product

The deer antler extract of Preparation Example 1 was diluted with 4 times the weight of distilled water, and the antler extract was sterilized at 121°C, 1.5 atm, and 15 minutes. In addition, lactic acid bacteria culture medium was prepared by inoculating the SST-9962 and SST-9961 lactic acid bacteria into MRS broth medium and activating them by culturing them statically at 37°C for 24 hours.

Afterwards, 1.5% by weight of glucose was added to the antler extract, and each lactic acid bacteria culture (1 × 10 ⁸ CFU/mL) prepared above was inoculated at 1.0% by weight and incubated at 37° C. for 24 hours to obtain a fermented product. .

Example 3: Preparation of fermented deer antler fermented with Bacillus subtilis

The deer antler extract of Preparation Example 1 was diluted with twice the weight of distilled water and sterilized at 121°C, 1.5 atm, for 15 minutes.

In addition, Bacillus subtilis culture medium was prepared by inoculating the Bacillus subtilis SST-9960 [Accession number: KCCM 11420P] strain into LB liquid medium and culturing it at 30°C for 2 days to activate it.

Afterwards, 1.5% by weight of glucose was added to the antler extract, and the Bacillus subtilis culture (1 × 10 ⁸ CFU/mL) was inoculated at 1.0% by weight and incubated at 30° C. for 3 days under aerobic conditions with a stirring speed of 100 rpm. The fermented product was obtained by culturing.

Comparative example: Deer antler hot water extract

The antler antler hot water extract of Preparation Example 1 was used as a comparative example in the following test.

The antler extracts and fermentation strains of Preparation Examples 1 to 3 and Comparative Examples are summarized and shown in Table 4 below.

division deer antler extract Fermentation strain name Example 1 Manufacturing example 1 Leuconostoc mesenteroides SST-9962 Example 2 Manufacturing example 1 Lactobacillus paraplantarum SST-9961 Example 3 Manufacturing example 1 Bacillus subtilis SST-9960 Comparative example Manufacturing example 1 -

Test Example 3: Cytotoxicity

The effect of the deer antler fermentation products prepared in Examples 1 to 3 on the survival rate of LNCaP cells, a prostate cancer cell line, i.e., cytotoxicity, was confirmed by the following method.

LNCaP cells (Korea Cell Line Bank 21740) were cultured in RPMI-1640 (Gibco) medium supplemented with 10% FBS (fetal bovine serum), 1% penicillin, and streptomycin at 37°C under 5% CO ₂ conditions. The cultured cells were distributed in a ⁹⁶ _well plate at a concentration of 1 , were treated at concentrations of 100, 200, and 500 ㎍/mL. After 24 hours of incubation, MTS assay was performed using CellTiter 96 AQueous Non-Radioactive Cell Proliferation Assay kit (Promega, USA). Specifically, after removing the cell culture medium, it was replaced with a medium containing MTS solution, left at 37°C for 1 hour, and the absorbance was measured at a wavelength of 490 nm using a microplate reader. The cell viability of each test group was negative. The relative ratios to the control group are shown in Table 5 below. As a negative control group, an untreated group treated with only 0.1% DMSO was used instead of the sample.

division Cell viability (%) Sample processing concentration (㎍/mL) 10 50 100 200 500 Negative control group (DMSO 0.1%) 100 Example 1 113.1 132.3 116.5 99.7 94.9 Example 2 108.6 124.5 110.2 101.2 95.4 Example 3 108.7 126.1 109.9 102.4 93.8

As shown in Table 5 above, it was confirmed that the fermented deer antler product according to the example of the present invention did not exhibit cytotoxicity up to a concentration of 500 μg/mL.

Test Example 4: In vitro ( in vitro ) Inhibitory effect on PSA, AR and 5α-reductase 2 expression

After treating prostate cancer cells with testosterone and deer antler fermentation, the protein expression of prostate-specific antigen (PSA), androgen receptor (AR), and prostate-specific enzyme (5α-reductase 2) was confirmed, and it was confirmed that deer antler fermentation was associated with benign prostatic hypertrophy. It was confirmed whether it was effective against the disease.

First, LNCaP cells were distributed in a 6-well plate at a concentration of 1X10 ⁵ cells/well and cultured at 37°C. After 24 hours, the cultured cells were treated with 1 μM of testosterone, and at the same time, the antler fermented products of Examples 1 to 3 were treated at concentrations of 25, 50, 100, and 200 μg/ml, and then cultured for 24 hours and then collected. As a negative control group (benign prostatic hyperplasia, BPH), fermented antler antler was not treated and only testosterone 1μM was treated. As a positive control group (Fina), instead of deer antler fermentation product, Finasteride (Sigma-Aldrich Inc. (MO, USA), a treatment for benign prostatic hyperplasia, was used to treat benign prostatic hyperplasia. )) 10 μM was treated with 1 μM testosterone. In addition, the untreated group was used as a normal control group (Control). After culturing, the collected cells were treated with RIPA buffer (50mM Tris-HCl, pH 8.0, with 150mM sodium chloride, 1% NP-40, 0.5% sodium deoxycholate, and 0.1% sodium dodecyl sulfate, with a protease inhibitor cocktail). The protein was separated.

The separated proteins were quantified using a protein assay kit (Bio-Rad protein assay kit, Bio-Rad, USA), and the quantified proteins were subjected to SDS-PAGE on a 10% polyacrylamide gel and transferred to a PVDF membrane. .

Afterwards, the PVDF membrane was reacted with 5% skim milk for about 1 hour to remove non-specific binding proteins on the surface, and the primary antibodies, anti-AR antibody, anti-PSA antibody, anti-5α-reductase 2 antibody, Then, the reaction was performed at 4°C for one day using an anti-β-actin antibody. Afterwards, the secondary antibody was treated at room temperature for 1 hour, and the protein expression levels of AR, PSA, and 5α-reductase 2 were confirmed using an ECL kit (Thermo scientific, USA). Tables 6 to 8 below quantify the protein expression levels of AR, PSA, and 5α-reductase 2 in LNCaP cells treated with samples according to the negative control group, positive control group, examples, and comparative examples, respectively, and then show the relative band density (%) of negative control group).

division AR (% of control) Sample processing concentration (㎍/mL) 50 100 200 Negative control group 100 Positive control (Finasteride, 10 μM) 4.1±0.5 Example 1 32.6±1.8 11.0±0.9 9.2±0.3 Example 2 41.3±1.2 16.2±0.7 14.3±2.1 Example 3 43.5±3.2 21.4±2.2 17.0±1.3 Comparative example 75.1±2.7 53.5±0.5 44.6±1.8

Looking at Table 6 above, it can be seen that the fermented deer antler products of Examples 1 to 3 of the present invention inhibit AR production in a concentration-dependent manner.

division PSA (% of control) Sample processing concentration (㎍/mL) 50 100 200 Negative control group 100 Positive control (Finasteride, 10 μM) 21.2±0.6 Example 1 55.3±1.4 33.1±2.1 28.5±0.7 Example 2 63.8±2.1 42.5±2.4 37.9±1.0 Example 3 63.5±3.2 48.4±2.2 42.0±1.3 Comparative example 92.1±1.6 82.1±0.7 77.5±1.1

Looking at Table 7 above, it can be seen that the fermented deer antler products of Examples 1 to 3 of the present invention inhibit PSA production in a concentration-dependent manner.

division 5α-reductase 2 (% of control) Sample processing concentration (㎍/mL) 50 100 200 Negative control group 100 Positive control (Finasteride, 10 μM) 44.5±1.1 Example 1 63.1±2.4 56.2±2.6 49.3±1.3 Example 2 72.6±2.8 59.8±3.5 57.6±2.5 Example 3 78.2±1.6 63.7±1.9 61.4±3.3 Comparative example 96.3±3.4 89.8±2.1 86.6±0.7

Looking at Table 8 above, it can be seen that the fermented deer antler products of Examples 1 to 3 of the present invention inhibit the production of 5α-reductase 2 in a concentration-dependent manner.

Examples 4-42: Preparation of fermented deer antler product

A fermented deer antler product was prepared in the same manner as in Examples 1 and 2, except that the lactic acid bacteria in Examples 1 and 2 were replaced with the lactic acid bacteria in Table 9 below. The lactic acid bacteria shown in Table 9 below were isolated from traditional foods such as yeast and kimchi and commercially available yogurt, and are strains owned by the applicant. The strain names below are abbreviations of the scientific name of each strain, or are assigned their own numbers.

division Strain name Example 4 Leuconostoc mesenteroides LM-2 Example 5 Leuconostoc mesenteroides LM-3 Example 6 Leuconostoc lactis LL Example 7 Lactobacillus plantarum LP-2 Example 8 Lactobacillus paraplantarum LP-3 Example 9 Lactobacillus plantarum LP-4 Example 10 Lactobacillus plantarum LP-5 Example 11 Lactobacillus sakei LS-1 Example 12 Lactobacillus sakei LS-2 Example 13 Lactobacillus sakei LS-3 Example 14 Lactobacillus sakei LS-4 Example 15 Lactobacillus sakei LS-5 Example 16 Lactobacillus arizonensis LA-1 Example 17 Lactobacillus arizonensis LA-2 Example 18 Lactobacillus johnsonii LJ Example 19 Lactobacillus pentosus LPE Example 20 Lactobacillus brevis L.B. Example 21 Lactobacillus hayakitensis LH-1 Example 22 Lactobacillus hayakitensis LH-2 Example 23 Lactococcus taiwanensis L.T. Example 24 Pediococcus pentosaceus PP-1 Example 25 Pediococcus pentosaceus PP-2 Example 26 Pediococcus pentosaceus PP-3 Example 27 Pediococcus pentosaceus PP-4 Example 28 Pediococcus pentosaceus PP-5 Example 29 Pediococcus pentosaceus PP-6 Example 30 Pediococcus pentosaceus PP-7 Example 31 Pediococcus pentosaceus PP-8 Example 32 Pediococcus pentosaceus PP-9 Example 33 Pediococcus pentosaceus PP-10 Example 34 Pediococcus pentosaceus PP-11 Example 35 Weissella confusa W.C. Example 36 Enterococcus raffinosus ER-1 Example 37 Enterococcus raffinosus ER-2 Example 38 Enterococcus faecalis EF Example 39 Staphylococcus warneri SW Example 40 Staphylococcus xylosus SX Example 41 Bifidobacterium bifidum BB Example 42 Aerococcus urinaeequi AU

Test Example 5: In vitro ( in vitro ) Inhibitory effect on the expression of AR, PSA, and 5α-reductase 2

In the same manner as Test Example 4, LNCaP cells were treated with deer antler fermentation product fermented with lactic acid bacteria other than the lactic acid bacteria of Examples 1 and 2 at a concentration of 200 μg/mL, and then protein expression levels of AR, PSA, and 5α-reductase 2 was confirmed. Afterwards, the protein expression levels of AR, PSA, and 5α-reductase 2 in LNCaP cells treated with samples according to the negative control group, positive control group, Examples, and Comparative Examples were quantified, respectively, and then calculated as relative band density (% of negative control group). The comparison is shown in Table 10 below.

division strain AR
(% of control group) PSA
(% of control group) 5α-
reductase 2
(% of control group) Negative control group - 100 100 100 Positive control group - 4.1±0.5 21.2±0.6 44.5±1.1 Comparative example - 44.6±1.8 77.5±1.1 86.6±0.7 Example 4 Leuconostoc mesenteroides LM-2 26.2±1.3 44.4±1.2 65.3±2.4 Example 7 Lactobacillus plantarum LP-2 30.4±0.9 52.7±0.6 62.7±3.1 Example 8 Lactobacillus paraplantarum LP-3 27.1±0.6 50.5±0.7 67.4±2.8 Example 12 Lactobacillus sakei LS-2 34.4±1.4 53.2±1.0 70.3±3.2 Example 13 Lactobacillus sakei LS-3 36.1±0.8 53.5±0.9 71.4±0.9 Example 21 Lactobacillus hayakitensis LH-1 38.3±1.4 59.5±1.7 68.2±4.7 Example 22 Lactobacillus hayakitensis LH-2 37.5±0.6 57.2±0.8 69.1±2.0 Example 23 Lactococcus taiwanensis L.T. 32.4±.07 61.2±1.6 71.3±3.6 Example 28 Pediococcus pentosaceus PP-5 31.6±0.4 53.6±1.5 70.9±4.2 Example 33 Pediococcus pentosaceus PP-10 39.4±1.1 54.1±0.3 72.6±2.1 Example 34 Weissella confusa W.C. 36.7±2.6 58.0±0.7 68.2±1.6 Example 40 Staphylococcus xylosus SX 40.6±1.7 56.8±1.3 75.2±2.2

Looking at Table 10, it was confirmed that the fermented deer antler product fermented with lactic acid bacteria other than the strains of Examples 1 and 2 of the present invention had lower production amounts of AR, PSA, and 5α-reductase 2 compared to the comparative example. This means that even in the case of deer antler fermentation products fermented with lactic acid bacteria other than the strains of Examples 1 and 2, the activity of suppressing the expression of AR, PSA, and 5α-reductase 2 was excellent.

Test Example 6: Growth rate of other lactic acid bacteria in deer antler extract

In order to confirm whether other lactic acid bacteria other than the lactic acid bacteria of Examples 1 and 2 also showed high growth rates in deer antler extract (4-fold dilution), the growth rates of lactic acid bacteria were measured in the same manner as Test Example 2, and are shown in Table 11 below. .

division Strain name Growth rate (O.D. 600nm) Example 4 Leuconostoc mesenteroides LM-2 1.623 Example 5 Leuconostoc mesenteroides LM-3 1.522 Example 6 Leuconostoc lactis LL 1.526 Example 7 Lactobacillus plantarum LP-2 1.700 Example 8 Lactobacillus paraplantarum LP-3 1.550 Example 9 Lactobacillus plantarum LP-4 1.646 Example 10 Lactobacillus plantarum LP-5 1.544 Example 11 Lactobacillus sakei LS-1 1.695 Example 12 Lactobacillus sakei LS-2 1.584 Example 13 Lactobacillus sakei LS-3 1.676 Example 14 Lactobacillus sakei LS-4 1.584 Example 15 Lactobacillus sakei LS-5 1.637 Example 16 Lactobacillus arizonensis LA-1 1.649 Example 17 Lactobacillus arizonensis LA-2 1.530 Example 18 Lactobacillus johnsonii LJ 1.851 Example 19 Lactobacillus pentosus LPE 1.570 Example 20 Lactobacillus brevis L.B. 1.564 Example 21 Lactobacillus hayakitensis LH-1 1.595 Example 22 Lactobacillus hayakitensis LH-2 1.563 Example 23 Lactococcus taiwanensis L.T. 1.643 Example 24 Pediococcus pentosaceus PP-1 1.635 Example 25 Pediococcus pentosaceus PP-2 1.631 Example 26 Pediococcus pentosaceus PP-3 1.618 Example 27 Pediococcus pentosaceus PP-4 1.602 Example 28 Pediococcus pentosaceus PP-5 1.560 Example 29 Pediococcus pentosaceus PP-6 1.560 Example 30 Pediococcus pentosaceus PP-7 1.524 Example 31 Pediococcus pentosaceus PP-8 1.516 Example 32 Pediococcus pentosaceus PP-9 1.548 Example 33 Pediococcus pentosaceus PP-10 1.515 Example 34 Pediococcus pentosaceus PP-11 1.537 Example 35 Weissella confusa W.C. 1.918 Example 36 Enterococcus raffinosus ER-1 1.792 Example 37 Enterococcus raffinosus ER-2 1.518 Example 38 Enterococcus faecalis EF 1.606 Example 39 Staphylococcus warneri SW 1.710 Example 40 Staphylococcus xylosus SX 1.654 Example 41 Bifidobacterium bifidum BB 1.574 Example 42 Aerococcus urinaeequi AU 1.546

Looking at Table 11 above, the growth rate (OD ₆₀₀ ) of lactic acid bacteria in the deer antler fermented product ranged from 1.515 to 1.918, confirming that the lactic acid bacteria had a very high growth rate in the deer antler extract.

Test Example 7: Fermentation characteristics of antler fermented product - pH change

The pH change according to the type of lactic acid bacteria and culture time is shown in Table 12 below. The pH was measured at room temperature using a pH meter (Seven Compact, Mettler Toledo, Switzerland) after taking 10 mL of sample.

division Strain name 0 hours 24 hours Example 2 Leuconostoc mesenteroides SST-9962 6.42 5.28 Example 6 Leuconostoc lactis LL 6.42 5.40 Example 7 Lactobacillus plantarum LP-2 6.42 5.22 Example 11 Lactobacillus sakei LS-1 6.41 5.38 Example 16 Lactobacillus arizonensis LA-1 6.42 5.47 Example 17 Lactobacillus johnsonii LJ 6.42 5.50 Example 18 Lactobacillus pentosus LPE 6.42 5.46 Example 20 Lactobacillus brevis L.B. 6.42 5.41 Example 21 Lactobacillus hayakitensis LH-1 6.42 5.51 Example 23 Lactococcus taiwanensis L.T. 6.41 5.42 Example 28 Pediococcus pentosaceus PP-5 6.42 5.45 Example 35 Weissella confusa W.C. 6.42 5.49 Example 36 Enterococcus raffinosus ER-1 6.42 5.44 Example 38 Enterococcus faecalis EF 6.42 5.47 Example 39 Staphylococcus warneri SW 6.42 5.64 Example 40 Staphylococcus xylosus SX 6.42 5.58 Example 41 Bifidobacterium bifidum BB 6.42 5.65

Looking at Table 12 above, comparing the pH change for each lactic acid bacteria, Leuconostoc mesenteroides SST-9962 and Lactobacillus plantarum LP-2 decreased by a relatively large amount compared to other strains.

On the other hand, when the lactic acid bacteria were the same, there was little difference in the pH of the culture medium using the antler antler hot water extract of Preparation Example 1, the antler ethanol extract of Preparation Example 2, and the antler water dilution of Preparation Example 3 as substrates, and in Preparation Example 3, the remaining Compared to that, the pH at 24 hours was somewhat higher.

Test Example 8: Confirmation of the prevention, treatment, and improvement effects of benign antler fermentation on benign prostatic hyperplasia using an animal model inducing benign prostatic hyperplasia

laboratory animals

The experimental animals used were 7-week-old female C57BL/6J normal mice weighing approximately 20 g. All mice were purchased from Daehan Biolink (Eumseong, North Chungcheong Province) and used in this experiment after an acclimatization period of one week. During the entire experiment, all mice were supplied with standardized solid food and water, and the room temperature was maintained at 23 ± 2 ℃, humidity was 50 ± 10%, and the light-dark cycle was maintained at 12 hours to 12 hours. It has been done.

Induction and drug treatment of prostatic hyperplasia

The experimental animals were divided into a total of 7 groups (n = 7), and 6 of the 7 groups received TP (Testosterone propionate) (5 mg/kg/day) dissolved in corn oil and injected subcutaneously for 2 weeks. Prostatic hypertrophy was induced, and the other group, which was a normal control group, was injected with the same amount of vehicle (distilled water).

After inducing prostatic hyperplasia for 2 weeks, a normal control group (NC) was injected with vehicle (distilled water) for an additional 4 weeks, a negative control group was injected with TP for an additional 4 weeks, and finasteride (1 mg/mg/kg), a treatment for benign prostatic hyperplasia, was administered along with TP for an additional 4 weeks. kg/day), a positive control group (Fina), experimental groups 1-3, which were orally administered the deer antler fermentation product of Examples 1 to 3 (20 mg/kg in dry weight) along with TP injection for an additional 4 weeks, and additional 4. The experiment was conducted under different conditions with a comparison group in which the deer antler extract of the comparative example was orally administered along with weekly TP injection. The test group classification and dosage settings of administered substances are shown in Table 13 below.

The administration of the sample was repeated once a day for a total of 4 weeks, and on the 29th day, the mouse was sacrificed according to the purpose of the experiment. Afterwards, the prostate tissue from the lesion site was extracted for analysis, and the extracted prostate tissue was used in the following experiments. The experimental results below are expressed as mean ± standard error (mean ± S.E.M.), and the difference in mean values between groups was calculated by calculating ANOVA (analysis of variance) through each experimental result and then using Duncan's multiple range test. It was verified at the p < 0.05 level.

army number of animals
(number of animals) administered substance Dosage amount
(mg/kg/day) Number of administrations G1 Jeongsang-gun 7 Distilled water 5 1 time/day G2 Negative control group 7 TP 5 1 time/day G3 Positive control group 7 TP + finasteride One 1 time/day G4 Test group 1 7 TP + deer antler fermentation product of Example 1 20 1 time/day G5 Test group 2 7 TP + deer antler fermentation product of Example 2 20 1 time/day G6 Test group 3 7 TP + deer antler fermentation product of Example 3 20 1 time/day G7 comparison group 7 TP + deer antler extract of comparative example 20 1 time/day

8-1: Inhibitory effect of prostate tissue hypertrophy of deer antler fermentation

The volume and weight of the prostate tissue of the mice sacrificed after the experiment for a total of 8 weeks were measured. In addition, in order to correct for differences in tissue weight depending on body weight, the prostate index was calculated by dividing the prostate weight (mg) of each experimental group by the value for each 100 g of body weight, and is shown in Table 14 below.

division Prostate Index Jeongsang-gun 195.3±5.4 Negative control group 462.7±21.6 Positive control group 303.4±13.2 Test group 1 290.2±14.6 Test group 2 320.5±11.7 Test group 3 338.6±16.2 Comparative example 416.9±22.3

Looking at Table 14 above, it can be seen that the fermented deer antler product according to Examples 1 to 3 of the present invention significantly suppresses the hypertrophy of prostate tissue compared to the negative control group. In particular, in the case of test group 1 administered with the fermented deer antler product of Example 1, it was confirmed that the effect of suppressing prostate tissue hypertrophy was greater than that of the positive control group.

8-2: Inhibitory effect of deer antler fermentation on the expression of AR, PSA and 5α-reductase 2 in prostate tissue

The prostate tissue of the sacrificed mouse was washed once with physiological saline, 4 times phosphate-buffered saline pH 7.4 (Gibco, USA) was added, and homogenized for 1 minute using a Wise Stir homogenizer (Daihan Scientific, Korea). The prostate tissue homogenate was centrifuged at 4°C and 5,000 rpm for 5 minutes (Micro 17TR, Hanil Science, Korea) to separate the supernatant.

Thereafter, the supernatant was analyzed for the expression levels of AR, PSA, and 5-alpha reductase using AR ELISA kit (Mybiosource, USA), PSA ELISA kit (Mybiosource, USA), and 5-alpha reductase ELISA kit (Mybiosource, USA). was measured, and it is shown in Table 15.

division AR
(% of control group) PSA
(% of control group) 5α-
reductase 2
(% of control group) Jeongsang-gun 18.6±1.2 16.7±0.8 76.7±5.4 Negative control group 100 100 100 Positive control group 47.3±2.6 39.3±3.7 23.5±2.8 Test group 1 22.4±1.7 46.3±4.5 70.3±1.9 Test group 2 36.4±3.1 54.8±2.7 79.6±3.2 Test group 3 42.7±2.5 58.2±1.4 83.4±1.6 comparison group 79.6±4.5 88.2±4.6 96.3±2.2

Looking at Table 15 above, it can be seen that the fermented deer antler product according to the example of the present invention exhibits an excellent expression inhibition effect in the body for AR, PSA, and 5-alpha reductase.

Hereinafter, an example of manufacturing a food or medicine containing the sample of the above example according to the present invention as an active ingredient will be described, but the present invention is not intended to be limited, but merely explained in detail. Using the fermented antler antler product, which is highly effective in preventing and treating benign prostatic hyperplasia, foods and medicines were prepared according to conventional methods in Preparation Examples 4 and 5 according to the composition and ratio as follows.

<Manufacturing example>

Preparation Example 4. Preparation of food

Production Example 4-1. Manufacturing of health functional foods

100 mg of deer antler fermentation product of Example 1

Vitamin mixture dosage

Vitamin A acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 ㎍

Vitamin C 10 mg

Biotin 10 μg

Nicotinamide 1.7 mg

Folic acid 50 μg

Calcium pantothenate 0.5 mg

Mineral mixture appropriate amount

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Monobasic Potassium Phosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

The composition ratio of the above vitamin and mineral mixture is a mixture of ingredients relatively suitable for health functional foods in a preferred embodiment, but the mixing ratio may be modified arbitrarily, and the above ingredients are mixed according to a normal health functional food manufacturing method. Then, granules can be prepared and used to manufacture a health functional food composition according to a conventional method.

Production Example 4-2. Manufacturing of functional beverages

100 mg of deer antler fermentation product of Example 1

1,000 mg citric acid

100 g oligosaccharides

2 g plum concentrate

1 g taurine

Add purified water to make a total of 900 mL.

After mixing the above ingredients according to the normal health drink manufacturing method, stirring and heating at 85°C for about 1 hour, the resulting solution was filtered, placed in a sterilized 2 L container, sealed, sterilized, stored in the refrigerator, and then refrigerated. Used for manufacturing the functional beverage composition of the invention.

The composition ratio is a preferred embodiment of mixing ingredients that are relatively suitable for beverages of preference, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as demand class, country of demand, and intended use.

Production Example 4-3. candy

Candy was prepared in a conventional manner by mixing 60% by weight of sugar, 39.8% by weight of starch syrup, 0.1% by weight of flavoring, and 0.1% by weight of the fermented deer antler product of Example 1.

Preparation Example 5. Preparation of pharmaceuticals

Production Example 5-1. Sanje

200 mg of deer antler fermentation product of Example 1

100 mg lactose

Talc 10 mg

The above ingredients are mixed and filled into an airtight bubble to prepare a powder.

Production Example 5-2. refine

500 mg of deer antler fermentation product of Example 1

Corn starch 100 mg

100 mg lactose

Magnesium stearate 2 mg

After mixing the above ingredients, tablets are manufactured by compressing them according to a typical tablet manufacturing method.

Production Example 5-3. capsule

500 mg of deer antler fermentation product of Example 1

3 mg crystalline cellulose

Lactose 14.8 mg

Magnesium stearate 0.2 mg

Capsules are prepared by mixing the above ingredients and filling them into gelatin capsules according to a typical capsule manufacturing method.

Production Example 5-4. liquid

100 mg of deer antler fermentation product of Example 1

10 g isomerized sugar

5 g mannitol

Proper amount of purified water

According to the usual liquid preparation method, add and dissolve each ingredient in purified water, add an appropriate amount of lemon flavor, mix the above ingredients, add purified water, adjust the total to 100 by adding purified water, and then fill in a brown bottle and sterilize. to produce a liquid.

Although preferred embodiments of the present invention have been described above, the present invention is not limited thereto, and can be implemented with various modifications within the technical scope of the present invention, which also fall within the scope of the appended patent claims. It is natural.

Claims (7)

Contains deer antler fermentation product obtained by fermenting deer antler extract with lactic acid bacteria as an active ingredient,
The lactic acid bacteria are at least one selected from among Leuconostoc mesenteroides SST-9962 strain (Accession number: KCCM12712P) and Lactobacillus paraplantarum SST-9961 strain (Accession number: KCCM12711P) A food composition for preventing or improving benign prostatic hyperplasia. delete According to paragraph 1,
The antler extract is a food composition for preventing or improving benign prostatic hyperplasia, characterized in that it is an extract of antler antler with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. delete delete According to paragraph 1,
The fermented deer antler is a food composition for preventing or improving benign prostatic hyperplasia, characterized in that it inhibits the production of one or more types selected from prostate specific antigen (PSA), androgen receptor (AR), and 5-alpha reductase-2. . Contains deer antler fermentation product obtained by fermenting deer antler extract with lactic acid bacteria as an active ingredient,
The lactic acid bacteria are at least one selected from among Leuconostoc mesenteroides SST-9962 strain (Accession number: KCCM12712P) and Lactobacillus paraplantarum SST-9961 strain (Accession number: KCCM12711P) A pharmaceutical composition for preventing or treating benign prostatic hyperplasia.