KR20220126621A - Composition for preventing, ameliorating or treating andropause syndrome comprising deer antler fermentation products as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating or treating andropause syndrome comprising a deer antler fermented material as an active ingredient. The composition of the present invention, by comprising the deer antler fermented material as the active ingredient, increases the secretion of the male hormone testosterone, and thus can prevent, alleviate or treat the andropause syndrome. In addition, since the present invention is a composition derived from natural products, the composition can be safely used without side effects, and can be usefully used for medicines or foods and the like.

Description

Composition for preventing, ameliorating or treating andropause syndrome comprising deer antler fermentation products as an active ingredient

The present invention relates to a composition for preventing, improving, or treating male menopausal syndrome, comprising a fermented antler ferment as an active ingredient.

Usually, the term menopause is limited to women, but in men, the concept of androgenic syndrome is emerging due to endocrine changes due to the gradual decrease in male hormone production with aging.

In this regard, according to Werner in 1939, men after the age of 50 had similar sensitivity to menopausal women, depression, memory impairment, concentration loss, fatigue, insomnia, hot flashes, decreased muscle mass and bone density, periodic sweating and decreased sexual vitality. Complaining of the same complex symptoms has been reported. After that, it is defined as male menopause as a contrasting concept to female menopause, and these names were designated as late-onset hypogonadism (late-onset) at the 4th ISSAM Congress in 2004 jointly by the International Society for Menopause and the International Society for Menopause. hypogonadism, LOH). Currently, LOH is the most widely used, but it has been called by various names such as andropause, androgen deficiency for aging male (ADAM), partial androgen deficiency for aging male (PADAM), male climateric, etc.

For the treatment of these androgenic syndromes, the possibility of using male hormone replacement therapy has been claimed.

According to one study, it was reported that a significant improvement effect was obtained when menopausal patients were treated with testosterone undecanoate 40 × 2 mg/day. However, the risk that the administration of male hormones may adversely affect liver, lipid status, cardiovascular and prostate diseases, etc. has also been reported.

Recently, in order to overcome the problems caused by the treatment of these drugs, the development of andropause syndrome improving agents containing natural herbal medicines as a main component is being attempted.

On the other hand, antler ( Cornu cervi pantotrichum , antler ) refers to the cartilage tissue that is regenerated every year with young antlers that are dense and not ossified with the hairs of plum, maroc, and closely related animals belonging to the deer family. It is called antler, and in Eastern countries such as China and Japan, including Korea, antler has been widely used as the best blood tonic for a long time.

Deer antler has been reported to have pharmacological effects on the immune system, hematopoietic system, glucose metabolism, and cardiovascular system. ), glucosamine, hyaluronic acid, calcium phosphate, calcium carbonate, collagen, phospholipids, and the like are known. In addition, scientifically, it has been reported that there are immune enhancement, blood pressure lowering, hematopoietic function, hypercholesterolemia, and anti-stress effect.

Herbal drugs with tonic effects such as deer antlers do not exert a strong effect by being limited to a specific organ, and the effect is weaker than that of single-component drugs, but it is characterized by complex effects throughout the body. Deer antlers have side effects such as diarrhea, so in oriental medicine, they are used in combination with herbal medicines that have astringent action. In this case, the expected effect is reduced. Many active ingredients and active ingredients of antler are lost during the extraction process using a solvent such as ethanol. Fermentation is a method that can improve these problems, thereby increasing the efficiency of deer antlers and expecting new pharmacological effects.

Fermentation is generally aimed at culturing useful microorganisms to produce components that benefit the human body and to remove components that cause toxicity or side effects. When it goes through the process of fermentation, it has various advantages such as imparting new physiological activity, increasing useful intestinal microorganisms, increasing absorption, and reducing or removing residual pesticides, so it is recently used as a biological conversion method using fermentation. However, to date, there has been no disclosure or teaching of data on the improvement or therapeutic efficacy of male menopausal syndrome through fermented deer antler fermented using lactic acid bacteria or Bacillus subtilis.

Accordingly, the present inventors have endeavored to develop an agent or therapeutic agent for male menopausal syndrome derived from natural products that are safe for the human body, and as a result, confirmed that the fermented antler has a remarkable testosterone enhancing effect, and completed the present invention.

US 10-2122408 B

It is an object of the present invention to provide a composition for preventing, improving or treating male menopausal syndrome comprising a 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 male menopausal syndrome comprising fermented antler as an active ingredient.

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

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

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

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

According to an embodiment of the present invention, the menopausal syndrome is hypogonadism, prostatic hypertrophy, lower urinary tract symptoms, urogenital function deterioration, joint or muscle pain, osteoporosis, dizziness, heat sensation, flushing, hyperhidrosis, decreased physical strength, athletic ability Decreased muscle strength, decreased body hair, hypotestosterone, decreased bone density, increased arteriosclerosis and increased carotid artery thickness, increased insulin resistance, decreased libido, erectile dysfunction, premature ejaculation, decreased stamina, decreased sperm count and decreased sperm motility, nervousness, emotions It may be one or more selected from anxiety, depression, hot flashes, sleep disturbance, decreased vitality, lethargy, fatigue, decreased work ability, decreased concentration, decreased cognitive ability, decreased spatial perception, and decreased memory.

According to an embodiment of the present invention, the composition can increase the blood testosterone content.

According to an embodiment of the present invention, the fermented antler may enhance one or more selected from the group consisting of male sexual desire, penis erection, ejaculation control ability, and sexual life satisfaction.

According to an embodiment of the present invention, the fermented deer antler may increase sperm count and sperm motility.

In addition, the present invention provides a pharmaceutical composition for preventing or treating menopausal syndrome comprising a fermented antler as an active ingredient.

The composition of the present invention contains the fermented antler as an active ingredient, thereby enhancing the secretion of testosterone, which is a male hormone, thereby preventing, improving or treating androgenic syndrome. In addition, since the present invention is a composition derived from a natural product, it can be safely used without side effects, and thus can be usefully used in medicines or foods.

Hereinafter, the present invention will be described in detail.

The present invention provides a food composition for preventing or improving male male menopausal syndrome comprising fermented antler as an active ingredient.

In the present specification, "fermented antler" may be fermented with lactic acid bacteria or Bacillus subtilis ( Bacillus subtilis ) of the antler extract. Preferably, the deer antler is used in any one or more lactic acid bacteria selected from the genus Lactobacillus, Bifidobacterium, Leuconostok, Pediococcus, Weissella, Staphylococcus, Erococcus and Enterococcus. , or may be fermented with Bacillus subtilis. More preferably, the deer antler is used as any one or more lactic acid bacteria or Bacillus subtilis selected from Leuconostoc mesenteroides , Lactobacillus plantarum and Lactobacillus paraplantarum . It may be fermented. Most preferably, antler leuconostoc mesenteroides SST-9962 [Accession No.: KCCM12712P], Lactobacillus paraplantarum SST-9961 [Accession No.: KCCM12711P] and Bacillus subtili It may be fermented with any one strain selected from Bacillus subtilis SST-9960 [Accession No.: KCCM 11402P].

The "fermented antler" may be used without limitation, fermented by a conventional fermentation method known in the art.

On the other hand, the antler extract includes not only the crude extract obtained by treating the extraction solvent on the antler, which is an extraction raw material, but also the processed product of the crude extract. For example, the deer antler extract may be prepared in a powder state by an additional process such as distillation under reduced pressure, freeze-drying or spray-drying. The extract also includes a fraction obtained by additionally fractionating the crude extract.

The antler extract is an extract of water, an 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, when water is used as a solvent, hot water extraction is preferred. For example, it can be extracted in water of 80 to 130 ℃, preferably 90 to 110 ℃ 0.5 to 24 hours, preferably 1 to 6 hours. And, even if hot water is not used as the solvent, that is, the antler component can be extracted from the diluted solution obtained by diluting and mixing the antler powder with cold water or water at room temperature. When extracting the antler component using cold water or room temperature water, it may be extracted separately from fermentation, or may be inoculated with lactic acid bacteria or Bacillus subtilis so that the antler component is eluted from the antler during the fermentation process.

As a preferred embodiment of the present invention, an extract with an aqueous alcohol solution having 1 to 4 carbon atoms may be used. For example, extraction may be performed with an aqueous alcohol solution such as ethanol, methanol, isopropanol, etc., preferably 20 to 80% by weight of an aqueous alcohol solution, more preferably 50 to 70% by weight of an aqueous alcohol solution. In the case of using an alcohol extract, before inoculating lactic acid bacteria or Bacillus subtilis, the alcohol contained in the alcohol extract is vaporized to lower the alcohol content or the alcohol extract is concentrated and dried, and then dissolved in water for use.

Also, in the present invention, the antler extract includes a diluent obtained by diluting antler powder in water in a broad sense. In the test example of the present invention, the effect of fermented antler fermented with lactic acid bacteria or Bacillus subtilis on male menopausal syndrome compared to fermented antler antler hot water extract or antler ethanol aqueous solution extract with lactic acid bacteria or Bacillus subtilis. is slightly lower, but it was confirmed that the effect was significantly increased compared to the unfermented deer antler extract.

Before inoculating the antler extract with lactic acid bacteria or Bacillus subtilis, a lactic acid bacteria nutrient source such as a protein source, a carbohydrate source, a vitamin or a mineral may be additionally mixed in order to promote the growth of the strain. The strain nutrient source may use a commercially available medium or may be individually added only the necessary nutrient source.

In addition, the antler extract, or the mixture of the antler extract and the strain nutrient source may be heat-sterilized before inoculation with lactic acid bacteria or Bacillus subtilis.

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

In the present invention, the lactic acid bacteria is any one or more selected from Lactobacillus, Bifidobacterium, Leuconostok, Pediococcus, Weissella, Staphylococcus, Erococcus and Enterococcus lactic acid bacteria. It may be lactic acid bacteria. Preferably, the lactic acid bacterium may be any one or more selected from Leukonostok mecenteroides, Lactobacillus plantarum, Lactobacillus paraplantarum, Lactobacillus saccharis and Pediococcus pentosaceus, more preferably the The lactic acid bacteria may be any one or more selected from Leukonostok mecenteroides, Lactobacillus plantarum, and Lactobacillus paraplantarum, and more preferably, the lactic acid bacteria is Leukonostok mecenteroides and Lactobacillus paraplantarum. It may be any one or more selected.

The strain may be added as a strain itself or as a culture solution. The culture solution may include the culture solution itself in which the strain is cultured in a liquid medium, and a concentrate obtained by concentrating the culture solution.

In a preferred embodiment of the present invention, the fermentation strain, particularly 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, 0.3 to 3.0 parts by weight, preferably 1.0 to 2.5 parts by weight, more preferably 1.5 parts by weight of the culture solution contained at a concentration of 1×10 ⁷ to 1×10 ⁸ cfu/mL based on 100 parts by weight of the antler extract. to 2.5 parts by weight may be added.

In the present invention, the "fermented antler" for the desired effect to appear meaningfully, (1) put the antler extract in a fermentation tray, and 0.3 to 3.0 weight of the lactic acid bacteria culture medium or Bacillus subtilis culture medium based on 100 parts by weight of the antler extract After adding (2) at 20 to 40°C for 18 to 96 hours, preferably 18 to 96 hours, more preferably 18 to 72 hours, still more preferably 18 to 48 hours, still more preferably 18 to 36 hours. Time, more preferably, it may be prepared by fermentation for 20 to 30 hours.

In particular, (3) after the fermentation at 2 to 10 ° C. for 5 to 24 hours after low-temperature aging, (4) 25 to 35 ° C., preferably 28 to 33 ° C. for 4 to 8 hours air drying step It is more preferable in terms of effect to carry out additionally. According to an embodiment of the present invention, it was specifically confirmed that the effect on male menopausal syndrome was further enhanced when the fermented antler fermented product was aged at a low temperature.

In addition, the “fermented deer antler” of the present invention prepared by the above method has a nutritional component content compared to the fermented deer antler produced by another method or the fermented deer deer antler produced and fermented by a strain other than lactic acid bacteria or Bacillus subtilis. It was specifically confirmed that the effect on male menopausal syndrome was further enhanced as well as further enhanced.

In addition, since the lactic acid bacteria among the strains for the fermentation have excellent growth in the deer antler extract, there is an advantage in that the fermentation time is short compared to other strains including the Bacillus subtilis strain.

Meanwhile, the fermented deer antler may be a fermented product containing microorganisms, or may be a fermented product from which the cells are removed. The fermented product from which the cells have been removed may be sterilized to contain dead cells, or it may be a filtrate or centrifugation supernatant from which the cells are removed through filtration or centrifugation. The effect of improving the androgenic syndrome of the present invention is because the components contained in the antler extract are bioconverted into components that are easy to use in vivo while the microorganisms proliferate, or a novel active ingredient having an effect on andropause syndrome is generated. It is expected, and it is significantly higher than the effect that can be obtained by adding a simple strain itself. The fermented antler may be used as a liquid fermented product by itself, or may be dried and powdered for use.

As used herein, "comprising as an active ingredient" means including an amount sufficient to achieve prevention, improvement, or therapeutic efficacy or activity of the fermented antler of the present invention for andromenopausal syndrome.

The composition of the present invention has the effect of preventing, improving, or treating androgenic syndrome by enhancing the secretion of testosterone, a male hormone.

As used herein, "male menopausal syndrome" may be any one selected from the group consisting of physical conditions, sexual conditions and mental conditions caused by testosterone decrease.

The physical conditions include hypogonadism, enlarged prostate, lower urinary tract symptoms, decreased genitourinary function, joint or muscle pain, osteoporosis, dizziness, fever, flushing, hyperhidrosis, decreased physical strength, decreased exercise capacity, decreased muscle strength, decreased body hair, testosterone It may be one or more conditions selected from hypothyroidism, decreased bone density, increased arteriosclerosis, increased carotid artery thickness, and increased insulin resistance, but is not limited thereto.

The sexual condition may be one or more conditions selected from the group consisting of decreased libido, erectile dysfunction, decreased erection, premature ejaculation, decreased stamina, decreased ejaculation control ability, decreased sexual life satisfaction, decreased sperm count, and decreased sperm motility, but is not limited thereto.

The mental condition is one or more conditions selected from nervous irritability, emotional instability, depression, hot flashes, sleep disturbance, decreased vitality, lethargy, fatigue, decreased work ability, decreased concentration, decreased cognitive ability, decreased spatial perception, and decreased memory may be, but is not limited thereto.

As used herein, the term “prevention” refers to anything that suppresses or delays androgenic symptoms by administration of the composition.

As used herein, “improvement” means at least reducing a parameter related to a condition to be treated by administration of the composition, for example, the severity of symptoms.

As used herein, “treatment” refers to reversing, alleviating, inhibiting, or preventing androgenic symptoms by administration of the composition.

The composition of the present invention does not exhibit cytotoxicity by including the fermented deer antler as an active ingredient, and has a remarkable effect in preventing, improving, or treating male menopausal syndrome.

In a specific embodiment of the present invention, since the fermented antler of the present invention exhibits an effect of inhibiting phosphodiesterase 5 (PDE5) and increasing the testosterone content in blood, it is possible to prevent, improve or treat male menopausal syndrome. It was specifically confirmed that there is. In addition, in another specific embodiment of the present invention, it was specifically confirmed that the fermented antler of the present invention enhances athletic ability, particularly endurance. Further, in another specific example of the present invention, it was specifically confirmed that the fermented antler of the present invention increased sperm count and sperm motility. In addition, in another specific embodiment of the present invention, it was specifically confirmed that the fermented antler of the present invention enhances at least one selected from the group consisting of male sexual desire, penis erection, ejaculation control ability, and sexual life satisfaction.

The food composition for preventing or improving male menopausal syndrome of the present invention includes all forms of nutritional supplements, health functional foods, food additives and feed, including humans or livestock Animals are intended for food.

The food composition may be formulated in the form of capsules, tablets, powders, granules, liquids, pills, flakes, pastes, syrups, gels, jellies or bars of the fermented antler of the present invention. The food composition has the advantage of not having side effects that may occur during long-term administration, unlike general drugs.

In addition, the food composition can be prepared in various forms according to conventional methods known in the art. Common foods include, but are not limited to, beverages (including alcoholic beverages), fruits and their processed foods (eg, canned fruit, bottled, jam, marmalade, etc.), fish, meat and their processed foods (eg, ham, sausages) Corn beef, etc.), breads and noodles (eg udon noodles, soba noodles, ramen, spagate, macaroni, etc.), fruit juice, various drinks, cookies, syrup, dairy products (eg butter, cheese, etc.), edible vegetable oils and fats, margarine , vegetable protein, retort food, frozen food, various seasonings (eg, soybean paste, soy sauce, sauce, etc.) can be prepared by adding the fermented deer antler. In addition, the nutritional supplement is not limited thereto, but may be prepared by adding the fermented antler to capsules, tablets, pills, and the like. In addition, the health functional food is not limited thereto, but for example, the fermented deer antler itself can be prepared in the form of tea, juice, and drink and liquefied for drinking (health drink), granulated, encapsulated and powder It can be eaten raw. In addition, in order to use the fermented antler fermented product in the form of a food additive, it may be prepared as a powder or extracted and then used in the form of a concentrate.

When the composition for preventing or improving male menopausal syndrome 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 a conventional drink. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; It may be a sugar alcohol such as xylitol, sorbitol, or erythritol. Sweeteners include natural sweeteners such as taumatine, stevia extract; A synthetic sweetener such as saccharin or aspartame may 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 of the present invention may be contained as an active ingredient of the food composition, and the amount may be an effective amount to achieve the effect of preventing or improving androgenic syndrome. Specifically, the fermented deer antler is 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 0.6 to 1.5% by weight, more preferably 0.8 to 1.2% by weight may be included. In addition, when the food composition is a health drink composition, the fermented antler ferment is 0.1 to 100 mg/ml, preferably 0.5 to 100 mg/ml, more preferably 1 to 50 mg/ml, in the health drink composition, More preferably, it may be included at a concentration of 3 to 20 mg/ml, more preferably 6 to 15 mg/ml, more preferably 8 to 12 mg/ml. In this case, if the content of the fermented antler antler is less than the lower limit, the cell viability is excellent, but the effect of preventing or improving male menopausal syndrome may not appear to a desired degree. Conversely, if the upper limit is exceeded, the effect of improving the menopausal syndrome may not increase as much as the concentration increases, or there may be toxicity. On the other hand, as a result of the in vitro experiment, when the concentration of the fermented antler of the present invention was within the above range, a significant effect on blood testosterone enhancing activity was exhibited, but side effects such as cytotoxicity did not appear.

Also, according to a preferred embodiment, the daily dose of the fermented antler may be 1 to 100 mg/kg, preferably 5 to 50 mg/kg, more preferably 10 to 30 mg/kg, based on dry weight. . In this case, if the daily dose of the fermented deer antler is less than the lower limit, the effect of preventing or improving male menopausal syndrome may not appear to a desired degree. Conversely, when the upper limit is exceeded, the effect of improving the androgenic syndrome may not increase as much as the concentration increases, or there may be toxicity.

The food composition of the present invention may be prepared by mixing the antler fermented product with other active ingredients known to be effective in preventing or improving male menopausal syndrome.

In addition to the above, the food composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, colorants, pectic acid, pectic acid salts, alginic acid, alginic acid salts, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, It may contain glycerin, alcohol or a carbonation agent and the like. In addition, the health food of the present invention may contain fruit for the production of natural fruit juice, fruit juice beverage, or vegetable beverage. These components may be used independently or in combination. The proportion of these additives is not critical, 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 male menopausal syndrome comprising a fermented antler as an active ingredient. The pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant other than the fermented antler, and the adjuvant includes an excipient, a disintegrant, a sweetener, a binder, a coating agent, a swelling agent, a lubricant, A lubricant or flavoring agent may be used. In the present invention, the terms “fermented antler” and “male menopausal syndrome” are the same as described above.

In addition, the pharmaceutical composition for preventing or treating male menopausal syndrome of the present invention may be formulated including one or more pharmaceutically acceptable carriers.

The pharmaceutically acceptable carrier may further include, for example, a carrier for oral administration or a carrier for parenteral administration.

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

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

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

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

Formulations 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 , methyl cellulose, sodium carboxymethyl cellulose, and hydroxypropylmethyl-cellulose or gelatin may be mixed and prepared. 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 mixture of granules.

In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid formulations for oral use include suspensions, solutions, emulsions or syrups, and various excipients, such as wetting agents, sweetening agents, fragrances, or preservatives, in addition to water or liquid paraffin, which are commonly used simple diluents, may be included. .

In addition, in some cases, cross-linked polyvinylpyrrolidone, agar, alginic acid or sodium alginate may be added as a disintegrant, and an anti-aggregating agent, a lubricant, a wetting agent, a flavoring agent, an emulsifier and a preservative may be further included. .

When administered parenterally, the pharmaceutical composition of the present invention may be formulated according to methods known in the art in the form of injections, transdermal administrations and nasal inhalants together with suitable parenteral carriers. In the case of the injection, it must be sterilized and protected from contamination of microorganisms such as bacteria and fungi. For injection, examples of suitable carriers include, but are not limited to, water, ethanol, polyols (eg, glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof and/or a solvent or dispersion medium containing vegetable oil. can More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) with 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 further include various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In addition, in most cases, the injection may further contain an isotonic agent such as sugar or sodium chloride.

In the case of transdermal administration, forms such as ointment, cream, lotion, gel, external solution, pasta, liniment, and air are included. As used herein, “transdermal administration” means that an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin by topically administering the pharmaceutical composition to the skin.

In the case of administration by inhalation, using a propellant suitable for the composition according to the invention, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas, it can be dispensed from a pressurized pack or nebulizer. It can be conveniently delivered in the form of an aerosol spray. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. For example, gelatin capsules and cartridges used in inhalers or insufflators may be formulated to contain a powder mixture of a suitable powder base 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 recipe commonly known to all pharmaceutical chemistry.

When the pharmaceutical composition of the present invention contains the fermented antler in an effective amount, it is possible to provide a desirable effect for preventing, improving, or treating male menopausal syndrome. As used herein, the term 'effective amount' refers to an amount that exhibits a higher response than that of a negative control group, and preferably refers to an amount sufficient to improve or treat androgenic syndrome. The fermented antler ferment is 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, more preferably based on the total content of the pharmaceutical composition. 0.6 to 1.5% by weight, more preferably 0.8 to 1.2% by weight may be included. Alternatively, the fermented deer antler is 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 of the pharmaceutical composition , more preferably 6 to 15 mg/ml, more preferably 8 to 12 mg/ml may be included at a concentration. At this time, if the content of the fermented deer antler is less than the lower limit, the cell viability is excellent, but the improvement or therapeutic effect of male menopausal syndrome may not appear to a desired degree. Conversely, if the upper limit is exceeded, the improvement or therapeutic effect of andropause syndrome may not increase as much as the concentration increases, or there may be toxicity. On the other hand, as a result of the in vitro experiment, when the concentration of the fermented antler of the present invention was within the above range, there was no side effect such as cytotoxicity while a significant effect was shown for the improvement or treatment of andromenopausal syndrome. The effective amount of the 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 may be administered to a patient as a single dose, and may be administered by a fractionated treatment protocol in which multiple doses are administered for 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.

A suitable dosage of the pharmaceutical composition varies depending on factors such as formulation method, administration mode, age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate, and response sensitivity of the patient, usually A skilled practitioner can readily determine and prescribe a dosage effective for the desired treatment or prophylaxis. According to a preferred embodiment, the daily dose of the fermented deer antler may be 1 to 100 mg/kg, preferably 5 to 50 mg/kg, more preferably 10 to 30 mg/kg, based on dry weight. In this case, if the daily dose of the fermented deer antler is less than the lower limit, the improvement or therapeutic effect of male menopausal syndrome may not appear to a desired degree. Conversely, if the upper limit is exceeded, the improvement or therapeutic effect of andropause syndrome may not increase as much as the concentration increases, or there may be toxicity.

The pharmaceutical composition for preventing and treating male menopausal syndrome of the present invention may also be provided in the form of an external preparation comprising fermented antler as an active ingredient.

When the pharmaceutical composition for preventing and treating androgenic syndrome of the present invention is used as an external preparation for skin, additionally, a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, and a foaming agent ( foaming agent), fragrance, surfactant, water, ionic emulsifier, non-ionic emulsifier, filler, sequestering agent, chelating agent, preservative, vitamin, blocker, wetting agent, essential oil, dye, pigment, hydrophilic active agent, hydrophilic It may contain adjuvants commonly used in the field of dermatology, such as oily active agents or any other ingredients commonly used in external preparations for skin, such as lipid vesicles. In addition, the above ingredients may be introduced in an amount generally used in the field of dermatology.

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

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments. However, these Examples are intended to illustrate the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited thereby.

<Example>

Preparation Example 1: Preparation of hot water extract of deer antler

After selecting and washing with water, domestic antler powder was uniformly pulverized with a grinder to prepare an antler powder with an average particle size of 300 μm, which was used while refrigerated.

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

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

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

Preparation Example 3: Preparation of antler water diluted solution

Water was mixed with water so that the antler powder of Preparation Example 1 was 2.0% by weight to prepare a water dilution solution for antler.

Test Example 1: Fermentation characteristics of strains in deer antler extract - Change in number of viable cells

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

First, each of the lactic acid bacteria of Table 1 below was inoculated into the MRS broth medium and activated by stationary culture at 37 ° C. for 24 hours to prepare a lactic acid bacteria culture solution.

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

In addition, Bacillus subtilis SST-9960 [Accession No.: KCCM 11420P] strain was inoculated in LB liquid medium and then cultured for 2 days at 30 ° C. and activated to prepare a Bacillus subtilis culture.

Next, 1.5% by weight of glucose was added to the antler extract obtained by diluting the antler extract of Preparation Examples 1 to 3 with distilled water at a weight of 4 times, respectively, and the lactic acid bacteria culture solution (1×10 ⁸ CFU/mL) or Bacillus subtilis After inoculating a culture solution (1×10 ⁸ CFU/mL) at 0.5 wt%, and fermenting at 37° C. for 24 hours, the number of viable cells was measured, and the results are shown in Table 2 below.

For live cell water, dilute 1 mL of sample in 9 mL of sterile physiological saline, mix well, dilute step by step, take 0.1 mL, use an anaerobic jar (BBL Gas Pak Anaerobic System), and store in anaerobic condition as much as possible and incubate at 37 °C. Then, the number of colonies generated was counted, and the average number of colonies was multiplied by a dilution factor to calculate colonies per culture.

division Number of viable cells (CFU/mL) before fermentation after fermentation
(Production Example 1) after fermentation
(Production Example 2) after fermentation
(Production Example 3) Leukonostok mecenteroides
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 use of the hot water extract of Preparation Example 1 and the ethanol aqueous extract of Preparation Example 2 as the fermentation substrate, but in the case of using the diluted antler water of Preparation Example 3 after fermentation, The number of viable cells was slightly less than half the number of viable cells in Preparation Examples 1 and 2.

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

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

To the lactic acid bacteria and Bacillus subtilis strains of the present invention, it was attempted to compare the strain growth in the deer antler extract.

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

division Viability (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, the growth degree of the Leuconostoc mesenteroides SST-9962 strain and the Lactobacillus paraplant arum SST-9961 strain with respect to the 4-fold diluted antler extract did not differ significantly, but Bacillus against the 4-fold diluted antler extract. The viability of the subtilis strain was relatively low compared to the viability of the SST-9962 strain and the SST-9961 strain.

In addition, in the case of SST-9962 strain and SST-9961 strain, 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 in the case of Bacillus subtilis, the growth rate was highest. The highest growth rate was found in the antler extract diluted before or twice.

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

Examples 1-2: Preparation of fermented deer antler

The antler extract obtained by diluting the deer antler extract of Preparation Example 1 with distilled water by 4 times the weight was sterilized at 121° C., 1.5 atmospheres, and 15 minutes. In addition, the SST-9962 and SST-9961 lactic acid bacteria were each inoculated into the MRS broth medium and activated by stationary culture at 37 ° C. for 24 hours to prepare a lactic acid bacteria culture solution.

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

Example 3: Preparation of fermented antler fermented with Bacillus subtilis

The antler extract obtained by diluting the deer antler extract of Preparation Example 1 with distilled water at twice the weight was sterilized at 121° C., 1.5 atmospheres, and 15 minutes.

In addition, Bacillus subtilis SST-9960 [Accession No.: KCCM 11420P] strain was inoculated in LB liquid medium and then cultured for 2 days at 30 ° C. and activated to prepare a Bacillus subtilis culture.

After that, 1.5 wt% of glucose was added to the deer antler extract, and 1.0 wt% of the Bacillus subtilis culture solution (1×10 ⁸ CFU/mL) was inoculated, followed by aerobic conditions with agitation speed of 100 rpm at 30°C for 3 days. cultured to obtain a fermented product.

Comparative Example: Deer antler hot water extract

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

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

division deer antler extract Fermented strain name Example 1 Preparation Example 1 Leuconostoc mesenteroides SST-9962 Example 2 Preparation Example 1 Lactobacillus paraplantarum SST-9961 Example 3 Preparation Example 1 Bacillus subtilis SST-9960 comparative example Preparation Example 1 -

Test Example 3: Cytotoxicity

The effect of the fermented antlers prepared in Examples 1 to 3 on the viability of Leydig cells (TM3, Korea Cell Line Bank), a testicular cell line of male mice, was confirmed by the following method.

TM3 cells were cultured in RPMI-1640 (Gibco) medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin and streptomycin at 37° C. and 5% CO ₂ conditions. The cultured cells were aliquoted in a 96-well plate at a concentration of 1X10 ⁵ cell/well and cultured again for 12 hours at 37° C., 5% CO ₂ in an incubator, and then the fermented deer antlers prepared in Examples 1 to 3 were respectively 10 and 50. , 100, 200, and 500 μg/mL concentrations were treated. After culturing for 24 hours, 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 then absorbance was measured at a wavelength of 490 nm using a microplate reader, and the cell viability of each test group was negative. Relative ratios to the control group are shown in Table 5 below. As a negative control group, an untreated group treated with only DMSO 0.1% was used instead of the sample.

division Cell viability (%) Sample treatment concentration (㎍/mL) 10 50 100 200 500 Negative control (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, it was confirmed that the fermented deer antler according to the embodiment of the present invention did not show cytotoxicity up to a concentration of 500 μg/mL.

Test Example 4: In vitro ( in vitro ) Testosterone Increase Effect

After the TM3 cells were treated with the fermented antler, the testosterone concentration was checked to determine whether the fermented antler was effective in increasing testosterone.

First, TM3 cells were aliquoted in a 6-well plate at a concentration of 2X10 ⁵ cells/well and incubated at 37°C. After 24 hours, the cultured cells were treated with the fermented deer antlers of Examples 1 to 3 at concentrations of 25, 50, 100, and 200 μg/ml, respectively, and cultured for 48 hours.

100 μl of the culture supernatant was transferred to a microtiter plate containing goat anti-mouse immunoglobulin G (goat anti-mouse IgG), and testosterone EIA antibody was added thereto, followed by incubation for 1 hour. Thereafter, a conjugate was added, incubated for 1 hour, and washed with a washing solution. After washing, a pNpp substrate solution was added, and after reaction for 1 hour, the reaction was stopped with a stop solution, and absorbance was measured at 405 nm.

division Testosterone Concentration (% of Control) Sample treatment concentration (㎍/mL) 25 50 100 200 Control (DMSO 0.1%) 100 Example 1 101.4±4.7 182.7±2.8 221.4±3.9 253.3±5.7 Example 2 100.7±5.1 159.4±3.7 195.7±6.2 219.4±3.8 Example 3 101.3±3.6 118.9±5.6 133.5±4.8 158.3±5.4 comparative example 100.2±4.5 100.9±4.4 102.7±2.6 103.6±2.7

Referring to Table 6, it can be seen that the fermented antlers of Examples 1 to 3 of the present invention significantly increased the testosterone concentration. These results confirm that the fermented antler of the present invention promotes the secretion of testosterone in the testis.

Test Example 5: In vitro ( in vitro ) Measurement of phosphate diesterase 5 (PDE5; Phosphodiesterase5) inhibitory ability

Sildenafil (Viagra), developed as an oral erectile dysfunction treatment and used worldwide, inhibits the activity of Phosphodiesterase5 (PDE5), which is specifically distributed in the corpus cavernosum. Since PDE5 has cGMP degradation activity, when the activity of PDE5 is inhibited, the concentration of intracellular cGMP increases and the level of intracellular calcium decreases. This induces scaphoid smooth muscle relaxation, which causes cavernous volume expansion and induces penile erection. Accordingly, it was attempted to confirm the PDE5 inhibitory activity using the samples of the above Examples.

A standard curve was obtained using 5'-GMP according to the protocol of BIOMOL GREENTM reagent (Cyclic Nucleotide Phosphodiesterase Assay Kit, BML-AK800, Enzo Life Sciences), and PDE and samples of the above examples using 3,5-cGMP as a substrate 200 μg/mL was also added to the assay buffer. 5-nucleotidase enzyme treatment and incubation for 30 minutes at 30 ℃. The reaction was terminated by adding 100 μL BIOMOL® GREEN Reagent, and absorbance was measured at 620 nm after 20-30 minutes. The amount of 5-GMP produced was compared with the control, and the average of the PDE inhibition rate was calculated and shown in Table 7 below.

division PDE5 inhibition rate (%) Control (DMSO 0.1%) 0 Example 1 66.0±7.3 Example 2 52.4±6.2 Example 3 50.3±5.9 comparative example 12.7±4.8

Referring to Table 7, it can be seen that the fermented deer antler according to the embodiment of the present invention exhibits significantly higher PDE5 inhibitory activity than the comparative example.

Examples 4-42: Preparation of fermented deer antler

It was carried out in the same manner as in Examples 1 and 2, except that the lactic acid bacteria of Examples 1 and 2 were replaced with the lactic acid bacteria of Table 8 below to prepare a fermented deer antler. The lactic acid bacteria in Table 7 below are isolated from traditional foods such as yeast and kimchi and commercially available yogurt, and are strains owned by the applicant. The following strain names are an abbreviation of the scientific name of each strain, or an abbreviation of its own number.

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 LB Example 21 Lactobacillus hayakitensis LH-1 Example 22 Lactobacillus hayakitensis LH-2 Example 23 Lactococcus taiwanensis LT 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 WC 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 6: In vitro ( in vitro ) Effect of increasing testosterone concentration

In the same manner as in Test Example 4, TM3 cells were treated with fermented antler 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 the testosterone concentration was confirmed. Specifically, the testosterone concentrations in the TM3 cells treated with the samples according to the control group, Examples, and Comparative Examples were measured, respectively, and calculated as relative values and shown in Table 9 below.

division strain Testosterone Concentration (% of Control) control - 100 comparative example - 103.6±2.7 Example 4 Leuconostoc mesenteroides LM-2 162.5±3.4 Example 7 Lactobacillus plantarum LP-2 151.7±4.7 Example 8 Lactobacillus paraplantarum LP-3 163.6±5.9 Example 12 Lactobacillus sakei LS-2 142.3±1.8 Example 13 Lactobacillus sakei LS-3 160.9±4.6 Example 21 Lactobacillus hayakitensis LH-1 143.8±2.7 Example 22 Lactobacillus hayakitensis LH-2 161.9±6.4 Example 23 Lactococcus taiwanensis LT 151.7±3.8 Example 28 Pediococcus pentosaceus PP-5 150.6±4.9 Example 33 Pediococcus pentosaceus PP-10 149.8±5.3 Example 34 Weissella confusa WC 132.7±6.2 Example 40 Staphylococcus xylosus SX 131.6±5.7

Referring to Table 9, it was confirmed that the fermented antler fermented with lactic acid bacteria other than the strains of Examples 1 and 2 of the present invention had a higher testosterone concentration than the Comparative Example. This means that even in the case of the fermented antler fermented with lactic acid bacteria other than the strains of Examples 1 and 2, the activity of promoting testosterone secretion is excellent.

Test Example 7: In vitro ( in vitro ) Measurement of phosphate diesterase 5 (PDE5; Phosphodiesterase5) inhibitory ability

In the same manner as in Test Example 5, PDE and fermented antler fermented with lactic acid bacteria other than the lactic acid bacteria of Examples 1 and 2 using 3,5-cGMP as a substrate were treated at a concentration of 200 μg/mL, and then PDE5 inhibitory ability was confirmed. did. The amount of 5-GMP produced was compared with the control, and the average of the PDE inhibition rate was calculated and shown in Table 10 below.

division strain PDE5 inhibition rate (%) control - 0 comparative example - 12.7±4.8 Example 4 Leuconostoc mesenteroides LM-2 44.6±7.2 Example 7 Lactobacillus plantarum LP-2 41.3±5.7 Example 8 Lactobacillus paraplantarum LP-3 47.6±6.3 Example 12 Lactobacillus sakei LS-2 40.4±2.8 Example 13 Lactobacillus sakei LS-3 41.6±3.7 Example 21 Lactobacillus hayakitensis LH-1 46.3±6.6 Example 22 Lactobacillus hayakitensis LH-2 44.9±4.8 Example 23 Lactococcus taiwanensis LT 39.2±6.1 Example 28 Pediococcus pentosaceus PP-5 42.6±1.7 Example 33 Pediococcus pentosaceus PP-10 40.7±5.0 Example 34 Weissella confusa WC 43.8±10.4 Example 40 Staphylococcus xylosus SX 38.1±8.2

Looking at Table 10, it can be seen that the fermented deer antler according to the embodiment of the present invention exhibits significantly higher PDE5 inhibitory activity than the comparative example.

Test Example 8: Growth of other lactic acid bacteria in deer antler extract

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

division strain name Viability (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 LB 1.564 Example 21 Lactobacillus hayakitensis LH-1 1.595 Example 22 Lactobacillus hayakitensis LH-2 1.563 Example 23 Lactococcus taiwanensis LT 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 WC 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, the lactic acid bacteria growth degree (OD ₆₀₀ ) of the fermented deer antler was in the range of 1.515 to 1.918, and it was confirmed that the lactic acid bacteria had a very high growth rate in the antler extract.

Test Example 9: Fermentation characteristics of fermented deer antler - pH change

The pH change according to the type of lactic acid bacteria and the 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 the sample.

division strain name 0 hours 24 hours Example 1 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 LB 6.42 5.41 Example 21 Lactobacillus hayakitensis LH-1 6.42 5.51 Example 23 Lactococcus taiwanensis LT 6.41 5.42 Example 28 Pediococcus pentosaceus PP-5 6.42 5.45 Example 35 Weissella confusa WC 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, when comparing the pH change for each lactic acid bacteria, Leuconostoc mesenteroides SST-9962 and Lactobacillus plantarum LP-2 were significantly decreased compared to other strains.

On the other hand, when the lactic acid bacteria are the same, there was almost no difference in the pH of the culture medium using the hot water extract of Preparation Example 1, the ethanol extract of Preparation Example 2, and the dilution of the antler water of Preparation Example 3 as substrates, and in Preparation Example 3, the remaining Compared to that, the pH at 24 hours was slightly higher.

Animal model manufacturing

After obtaining and acclimatizing 23-week-old male SD rats (Samtaco, Korea), when the weight of each individual was about 510 g (6 months old), 9 rats per test group were used, and 11-week-old males were used one week before the end of the test. and female SD rats (Samtaco, Korea) were obtained and acclimatized, respectively, and 9 rats were constituted at 12 weeks of age. All mice received standardized solid feed and water during the entire experiment period, and the temperature of the breeding room was 23 ± 2 ℃, humidity 50 ± 10%, and the photoperiod and light-dark cycle were maintained at 12 hours - 12 hours. became

The composition of the test group consisted of 12-week-old rats with sexual maturation completed and 24-week-old elderly rats as controls. A test group was prepared by oral administration. Table 13 below shows the test group classification and dosage setting of the administered substance. The following experimental results are expressed as mean ± standard error (S.E.M) values, and the average difference between groups is analyzed using the Duncan's multiple range test after obtaining ANOVA (analysis of variance) from each experimental result. Thus, it was verified at the level of p < 0.05.

army number of animals
(number of animals) dosing substance dose amount
(mg/kg/day) dosing frequency G1 12-week-old control group 9 - - 1 time/day G2 24-week-old control group 9 - - 1 time/day G3 test group 1 9 24 weeks old, fermented deer antler of Example 1 20 1 time/day G4 test group 2 9 24 weeks old, fermented deer antler of Example 2 20 1 time/day G5 test group 3 9 24 weeks old, fermented deer antler of Example 3 20 1 time/day G6 control group 9 24 weeks old, antler extract of comparative example 20 1 time/day

Test Example 10: Confirmation of the effect of increasing the blood testosterone content of the fermented antler

After completing the 30-day sample administration, the sacrificed experimental animals were anesthetized with CO ₂ , blood was collected from the abdominal aorta, and centrifuged at 3000 rpm for 10 minutes to separate the serum. Then, the blood testosterone content was measured by the competitive RIA method using Hitachi 7600-110/7170 (Tokyo, Japan), and is shown in Table 14 below.

division Blood Testosterone Concentration (ng/mL) 12-week-old control group 8.2±2.4 24-week-old control group 4.5±1.2 test group 1 8.9±2.1 test group 2 7.6±1.6 test group 3 6.3±2.0 control group 5.1±1.4

Referring to Table 14, it can be seen that test groups 1 to 3 orally administered with the fermented antler of the present invention have significantly increased blood testosterone concentration compared to the 24-week-old control group.

Test Example 11: Exercise ability improvement effect (improvement of endurance) of fermented deer antler

After starting the treadmill exercise for 5 minutes at an inclination of 10 degrees and a speed of 10 m/min for the experimental animals of each test group to which the samples were administered for a total of 30 days, the exercise intensity was increased by increasing the speed of 1 m/min every 1 minute Table 15 shows the duration of exercise from exhaustion to exhaustion at a maximum speed of 25 m/min. The time until exhaustion was defined as the first time that a lagging occurred without being able to follow three consecutive times, and one run was set to run continuously for more than 5 minutes without falling behind in speed. This test was measured three times one day after the end of administration.

division Time to exhaustion (sec) 12-week-old control group 412±38 24-week-old control group 167±16 test group 1 286±37 test group 2 259±25 test group 3 224±29 control group 175±31

Referring to Table 15, it can be seen that test groups 1 to 3 orally administered with the fermented deer antler of the present invention have significantly increased exercise capacity compared to the 24-week-old control group.

Test Example 12: Effect of improving sexual function of fermented deer antler

12-1: Effect of increasing the number of mounts of fermented antler

One male rat and three female rats of each test group, to which the samples were administered for a total of 30 days, were transferred to a tank, isolated with a transparent separator for 2 hours, adapted to meet, and then the separator was removed to meet each other. Thereafter, the number of mounts (number of mounts without or with penile insertion, number of attempts at copulation) of male rats to female rats was observed for 2 hours and shown in Table 16 below.

division number of mounts 12-week-old control group 6.2±0.3 24-week-old control group 1.9±0.2 test group 1 11.3±0.7 test group 2 9.2±0.6 test group 3 8.4±0.5 control group 3.8±0.3

Looking at Table 16, it can be seen that test groups 1 to 3 orally administered with the fermented antler of the present invention significantly increased the number of mounts compared to the control group.

12-2: Effect of increasing sperm count and sperm motility of fermented antler

After completion of the sample administration, the epididymis was extracted from the testis of the sacrificed experimental animal, and the epididymis was transferred to a Petri dish containing mineral oil (Sigma, USA) maintained at 37° C. to collect sperm from the tail. Sperm in mineral oil was liquefied for 20 minutes in a 37° C. CO ₂ incubator (Forma Co., USA). After that, liquefied sperm was collected, mixed with PBS, transferred to a slide glass, and analyzed for sperm concentration (number) per ml and sperm motility using SAIS (Semen Analysis Imagine System, Medical Supply Co. Ltd., Korea) program. The results are shown in Tables 17 and 18 below.

division Sperm count/mL (%) 12-week-old control group 76.7±1.2 24-week-old control group 29.6±6.4 test group 1 70.2±5.7 test group 2 64.3±4.9 test group 3 62.9±6.6 control group 35.4±3.8

Referring to Table 17, it can be seen that test groups 1 to 3, which were orally administered with the fermented antler of the present invention, significantly increased the number of sperm compared to the 24-week-old control group.

division sperm motility (%) 12-week-old control group 93.2±6.6 24-week-old control group 59.6±8.4 test group 1 85.2±7.5 test group 2 79.8±6.7 test group 3 77.5±8.3 control group 63.2±9.4

Referring to Table 18, the sperm motility of the 24-week-old control group was significantly reduced compared to the 12-week-old control group, and the sperm motility of Test Groups 1 to 3 orally administered with the fermented deer antler of the present invention compared to the 24-week-old control group. It can be confirmed that this significantly increased and improved to a value similar to that of 12-week-old young rats.

Test Example 13: Evaluation of sexual function improvement effect of fermented deer antler

Selection of subjects

In order to confirm the male sexual function improvement effect of the fermented antler of the present invention, a survey was conducted on male sexual function targeting men in their 30s and 50s who did not have special health problems such as diseases, and respondents who scored 16 or less out of 40 A total of 30 people were selected. (30-39 years old: 7 people, 40-49 years old: 12 people, 50-59 years old: 11 people)

The contents of the questionnaire are as shown in Table 19 below, and for each question, 'Very good' is given 5 points, 'Slightly good' is 4 points, 'Normal' is 3 points, 'Slightly bad' is 2 points, ' 'Very poor' was marked with 1 point, and 5 points were allocated to each question, and finally, the total of questions for each category was converted into a score of 10 for calculation.

classification content of the question converted score sexual desire A. The number of times you feel sexual desire for a stimulus that makes you feel sexual desire is the same as before. out of 10 B. After ejaculation, the time it takes to regain sexual desire is the same as before. erectile function A. I am confident in myself that I can get and maintain an erection. out of 10 B. It is possible to maintain an erection state to the end in more than half of the number of intercourse. C. There is a difference in the firmness of the penis during a complete erection compared to before. D. There is no difference in erection or erection power when waking up in the morning compared to before. ability to control ejaculation A. The ejaculation may be arbitrarily delayed. out of 10 B. I have never felt anxious or stressed about ejaculating sooner than I intended. sex life
satisfaction A. More than half of the number of times they felt sufficient satisfaction after recent intercourse. out of 10 B. I am satisfied with my current sex life pattern, such as the number of sexual intercourse and the quality of intercourse. C. I am confident that my partner is satisfied with my sex life.

Confirmation of sexual function improvement effect of fermented antler

The subjects selected above drank the fermented antler deer antler of Example 1 at a dry weight of 20 mg/kg/day, once a day, for a total of 30 days, followed by a questionnaire evaluation, and then after taking a break for 15 days, it was carried out The fermented deer antler of Example 2 was drank at the same dose and usage as in Example 1, and questionnaire evaluation was conducted. After that, after taking a 15-day break again, drinking the antler extract of Comparative Example in a manner of drinking the fermented antler of Example 3, and conducting a questionnaire evaluation, each score was converted and shown in Table 20 below.

classification sexual desire erectile function ability to control ejaculation Satisfaction with sex life control 3.40 3.20 2.43 3.33 Example 1 7.96 7.80 6.33 8.02 Example 2 7.40 7.20 5.94 7.60 Example 3 6.33 6.15 5.89 6.48 comparative example 4.13 4.23 3.50 4.76

Looking at Table 20, when the fermented deer antler according to the embodiment of the present invention was administered, it was confirmed that sexual functions such as libido, erectile function, ejaculation control ability and satisfaction with sex life were significantly improved compared to the case of administration of the control or comparative example. can

Hereinafter, a manufacturing example of a food or drug 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 limit the present invention, but merely to describe it in detail. Using the fermented deer antler, which is excellent in preventing, improving, or treating the male menopausal syndrome, foods and drugs were prepared according to a conventional method in Preparation Examples 4 and 5 according to the following composition ingredients and composition ratios.

<Production Example>

Preparation Example 4. Preparation of food

Preparation Example 4-1. Manufacturing of health functional food

100 mg of fermented deer antler of Example 1

appropriate amount of vitamin mixture

70 μg vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

0.5 mg of vitamin B6

0.2 μg of vitamin B12

Vitamin C 10 mg

Biotin 10 μg

Nicotinamide 1.7 mg

50 μg of folic acid

Calcium pantothenate 0.5 mg

Appropriate amount of inorganic mixture

ferrous sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

potassium phosphate monobasic 15 mg

Dibasic calcium phosphate 55 mg

Potassium citrate 90 mg

100 mg calcium carbonate

Magnesium chloride 24.8 mg

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

Preparation Example 4-2. Production of functional beverages

100 mg of fermented deer antler of Example 1

1,000 mg citric acid

100 g of oligosaccharides

2 g of plum concentrate

1 g taurine

Add purified water to total 900 mL

After mixing the above ingredients according to the usual health drink manufacturing method, after stirring and heating at 85°C for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2L container, sealed and sterilized, then refrigerated. It is used to prepare the functional beverage composition of the present invention.

Although the composition ratio is prepared by mixing ingredients suitable for relatively favorite beverages in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and national preferences such as demand class, demanding country, and use.

Preparation Example 4-3. candy

Sugar 60% by weight, starch syrup 39.8% by weight, and flavoring 0.1% by weight and 0.1% by weight of the fermented antler antler of Example 1 were mixed to prepare a candy in a conventional manner.

Preparation 5. Preparation of pharmaceuticals

Preparation Example 5-1. powder

200 mg of fermented deer antler of Example 1

Lactose 100 mg

talc 10 mg

The above ingredients are mixed and filled in an airtight bag to prepare a powder.

Preparation Example 5-2. refine

500 mg of fermented deer antler of Example 1

100 mg cornstarch

Lactose 100 mg

2 mg magnesium stearate

After mixing the above ingredients, tablets are prepared by tableting according to a conventional manufacturing method of tablets.

Preparation Example 5-3. capsules

500 mg of fermented deer antler of Example 1

3 mg of crystalline cellulose

Lactose 14.8 mg

0.2 mg magnesium stearate

According to a conventional capsule preparation method, the above ingredients are mixed and filled in a gelatin capsule to prepare a capsule.

Preparation Example 5-4. liquid

100 mg of fermented deer antler of Example 1

10 g isomerized sugar

5 g of mannitol

Purified water appropriate amount

According to a conventional liquid preparation method, each component is added to purified water to dissolve, an appropriate amount of lemon flavor is added, the above components are mixed, purified water is added, the whole is adjusted to 100 by adding purified water, and then filled in a brown bottle and sterilized to prepare a liquid.

Although preferred embodiments of the present invention have been described above, the present invention is not limited thereto, and various modifications can be made within the scope of the technical spirit of the present invention, which also falls within the scope of the appended claims. it is natural

Claims (8)

A food composition for preventing or improving male menopausal syndrome comprising fermented antler as an active ingredient. The method of claim 1,
The fermented antler is a food composition for preventing or improving male menopausal syndrome, characterized in that it is obtained by fermenting the antler extract with lactic acid bacteria or Bacillus subtilis . 3. The method of claim 2,
The antler extract is a food composition for preventing or improving male menopausal syndrome, characterized in that the antler is an extract of water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. 3. The method of claim 2,
The lactic acid bacteria are Lactobacillus genus, Bifidobacterium genus, Leuconostok genus, Pediococcus genus, Weissella genus, Staphylococcus genus, Erococcus genus and Enterococcus genus It is characterized in that any one or more lactic acid bacteria selected from A food composition for preventing or improving male menopausal syndrome. 5. The method of claim 4,
The lactic acid bacteria is Leuconostoc mesenteroides ( Leuconostoc mesenteroides ), Lactobacillus plantarum ( Lactobacillus plantarum ) and Lactobacillus paraplantarum ( Lactobacillus paraplantarum ) Male menopausal syndrome prevention, characterized in that any one or more lactic acid bacteria selected from or a food composition for improvement. The method of claim 1,
The menopausal syndrome is hypogonadism, enlarged prostate, lower urinary tract symptoms, genitourinary function, joint or muscle pain, osteoporosis, dizziness, heat sensation, flushing, hyperhidrosis, decreased physical strength, reduced exercise capacity, muscle strength loss, body hair loss, testosterone Hypothyroidism, decreased bone density, increased arteriosclerosis, increased carotid artery thickness, increased insulin resistance, decreased libido, erectile dysfunction, premature ejaculation, decreased stamina, decreased sperm count and decreased sperm motility, irritability, emotional instability, depression, hot flashes, sleep disturbance , decreased vitality, lethargy, fatigue, decreased work ability, decreased concentration, decreased cognitive ability, decreased spatial perception, and decreased memory. According to claim 1,
The composition is a food composition for preventing or improving male menopausal syndrome, characterized in that increasing the blood testosterone content. A pharmaceutical composition for preventing or treating male menopausal syndrome comprising fermented antler as an active ingredient.