WO2015137652A1

WO2015137652A1 - 5-alpha reductase inhibiting fermented camellia japonica seed oil for alleviating benign prostatic hypertrophy and baldness

Info

Publication number: WO2015137652A1
Application number: PCT/KR2015/001925
Authority: WO
Inventors: 정종문; 이승숙
Original assignee: 주식회사 벤스랩; 정종문; 주식회사 코스벤
Priority date: 2014-03-11
Filing date: 2015-02-27
Publication date: 2015-09-17
Also published as: KR101452320B1

Abstract

The present invention relates to a use of fermented Camellia japonica seed oil which is high in the content of free essential fatty acids, and more specifically, to a medicine, a dietary supplement or a cosmetic product for preventing and alleviating benign prostatic hypertrophy and male-pattern baldness, containing fermented Camellia japonica seed oil as an active ingredient since the fermented Camellia japonica seed oil, which is prepared by fermenting Camellia japonica seed oil, is rich in free essential fatty acids and has remarkable 5-α reductase inhibitory activity, thereby preventing and alleviating benign prostatic hypertrophy and male-pattern baldness. According to the present invention, the effects of essential fatty acids can be maximized by increasing the content of essential fatty acids, which are isolated so as to facilitate absorption and in vivo utilization, through the indirect fermentation of Camellia japonica seed oil. In addition, according to the present invention, the fermented Camellia japonica seed oil has a similar activity to finasteride, which prevents and alleviates benign prostatic hypertrophy and male-pattern baldness by having little toxicity as a result of a single-dose toxicity test and cytotoxicity check, having an effect of lowering dihydrotestosterone by inhibiting 5-α reductase, exhibiting an effect of restoring the size of the prostate to a normal level with respect to benign prostatic hypertrophy, promoting the growth of hair follicle cells and exhibiting an effect of alleviating baldness phenomenon of examinees as a result of simple clinical trials, and thus can be applied, prepared, and sold as a cosmetic product, a dietary supplement or a pharmaceutical composition by using a fermented natural oil including fermented Camellia japonica seed oil having identical or similar efficacy and effects to natural product-derived one.

Description

Fermented camellia oil to inhibit 5-alpha reductase to improve prostate hypertrophy and hair loss

The present invention relates to functional foods, pharmaceutical compositions and cosmetic raw materials for inhibiting 5-alpha reductase to improve and prevent prostatic hypertrophy and hair loss, and more particularly, essential unsaturated fatty acids including free oleic acid. The present invention relates to functional foods, pharmaceutical compositions, and cosmetic raw materials for preventing and treating enlarged prostate, and for preventing and improving hair loss, including fermented natural oil including abundant fermented camellia oil (Camellia japonica seed oil) as an active ingredient.

The prostate is an accessory gonad consisting of gland tissue and fibrous muscle tissue. The prostate gland produces and secretes semen and is located just below the bladder and surrounds the urethra. The function of the prostate is reported to be responsible for about 40% of semen and play an important role in the survival and activity of sperm. In addition, the prostate provides sperm with nutrients, maintains proper ionic concentration and acidity, and contains zinc to prevent bacterial infection.

This important function of the prostate gland increases gradually with age. Therefore, the most common prostate disease after middle age is prostatic hyperplasia (BPH).

Prostate hyperplasia is an abnormally enlarged prostate gland that blocks the passage of urine below the bladder or compresses the urethra to prevent urine from draining smoothly and pushes up the bladder to cause urinating more than eight times a day, night urinary, strong and sudden urination Bladder storage symptoms such as urinary urinary urge, delayed urine (the urine comes out of urine when urine is seen), and urinary urine (a break in urine flow) The lower urinary tract symptoms are referred to collectively as symptoms that indicate the discharge of the bladder, such as the need to give strength during urination. As you age, prostate hyperplasia increases and prostate enlargement occurs in more than 50% of men over 60 and in 90% of men over 85.

The cause of enlarged prostate is not yet clear, and as with other chronic diseases, several complex factors are known to work. Prostate hypertrophy occurs mainly in men over 40 who have normal testicles that make androgen, and one of the most important causes is the correlation between testosterone conversion and aging.

Briefly, the mechanism of enlarged prostate is as follows. Excessive testosterone in the blood is converted to dihydrotestosterone (DHT) by the 5-alpha reductase present in the prostate tissue. This DHT stimulates cell division of stromal and epithelial cells in the prostate much more than testosterone, eventually leading to prostatic hypertrophy.

In old age, testosterone levels are gradually lowered but more prostatic hyperplasia occurs. This is due to the dihydrotestosterone, which increases the number of cells in this prostate. Male hormone is present in two forms, testosterone and dihydrotestosterone, which are more important for prostate growth. Although testosterone and dihydrotestosterone bind to the same male hormone receptor in a similar proportion in the prostate, most male hormone receptors in the prostate are more dehydro than testosterone because the dihydrotestosterone-male hormone receptor complex is more stable than the testosterone-male hormone receptor complex. It will remain in combination with testosterone. In addition, unlike testosterone, dihydrotestosterone does not decrease significantly with age, increasing the androgen receptors of prostate cells to maintain the endocrine balance and eventually stimulate other growth factors that lead to prostate proliferation as they age. Prostate enlargement will occur.

5-alpha-reductase, which converts testosterone to dihydrotestosterone, is a microsome's NADPH dependent 3-oxo-5α-steroid-dehydrogenase that reduces double-linked steroids such as testosterone. The enzyme is involved in bile acid synthesis and metabolism of androgen and estrogen in the body. 5-alpha reductase has type 1, type 2, and type 3 subtypes. Type 1 is present in various organs such as skin and liver tissue. Type 2 is mainly present in the urinary tract including the liver and prostate. Testosterone is converted to dihydrotestosterone by at least 90% by 5-alpha reductase in prostate tissue and dihydrotestosterone promotes prostate growth with approximately 10 times more activity than testosterone. Therefore, the efficient inhibition of 5-alpha reductase may reduce the synthesis of physiologically active dihydrotestosterone, which may eventually improve or treat prostatic hyperplasia.

In general, the number of human hair is estimated to be about 100,000 to 150,000. Hair falls from 50 to 100 hairs a day, and hair loss begins when more than 100 hairs fall out a day. The causes of hair loss are many and vary from person to person. Men's hair in particular tends to become thinner at the peak of their 20s. Male pattern baldness (MPB), the most common of the various symptoms of hair loss, has a large genetic characteristic. Another major reason is that male hair abnormalities cause inflammation of the scalp due to weak hair roots and excessive sebum secretion. This falls out.

The most common cause of masculine alopecia (more than about 90%) is an abnormality of androgen alopecia. As with prostatic hyperplasia, over-synthesis of dihydrotestosterone by 5-alpha reductase causes dihydrotestosterone to bind to androgen receptors around the hair follicles, shrinking and devastating the hair follicles and eventually reducing the hair growth period. Because of the reduced growth phase, the roots of the hair, the hair roots, do not develop sufficiently and at the same time there is not enough pigment accumulation. This immature hairless power often comes out of the scalp and eventually begins to lose hair. Continued hair follicles eventually die and permanent hair loss occurs, which makes it difficult to treat other than hair transplantation. Dihydrotestosterone provides the factors that make the hair shorter and longer during the growth cycle of the hair and eventually make the hair smaller and smaller as the growth cycle continues. As hair loss progresses, the hair follicles contract and instead, the sebaceous glands become larger and the hair becomes thinner and smaller, usually oily, causing rashes or sores. The thinner head turns into a downy hair and becomes invisible to our eyes, and the scalp inside the head begins to look into it. After 20 repetitions of the process of thinning and falling off, the hair follicles will die forever and no remedy will ever recover. While testosterone primarily affects the growth of armpit hair and pubic hair, dehydrotestosterone is known to affect beards and baldness, but it is not clear.

Many drugs are known for treating prostatic hyperplasia and masculine alopecia. However, it is most important to inhibit 5-alpha reductase for fundamental treatment.

Inhibitors of 5-alpha reductase synthesis include Finasteride and Dutasteride. Testosterone is converted to dihydrotestosterone by 5-alpha reductase in prostate tissue. Therefore, taking a 5-alpha reductase inhibitor that effectively blocks the conversion of dihydrotestosterone can reduce the size of the prostate gland. After six months of use, it is known that the size of the prostate is reduced to the maximum. Finasteride, a competitive inhibitor of 5-alpha reductase, developed by Merck, also inhibits type 1 but mainly reduces type 2 5-alpha reductase, reducing dihydrotestosterone in the prostate and blood. Dutasteride has a longer half-life than Finasteride, and is 45 times stronger for type 1 enzymes and 2.5 times stronger for type 2 enzymes. Most of the metabolism in the liver, hepatic failure should be limited to use. Synthetic inhibitors of these 5-alpha reductases can be regarded as a fundamental treatment for prostatic hyperplasia, but patients who take 5 mg / day for 4 to 6 months for long periods of time will be effective. . Side effects include erectile dysfunction, decreased libido, gynecomastia, and ejaculation disorders.

Finasteride is also used to treat androgenetic alopecia. Finasteride was first used to treat prostatic hyperplasia, but has been recognized by the US Food and Drug Administration to treat hair loss at a daily dose of 1 mg per day. As mentioned above, this drug also has a hair loss treatment effect through the mechanism of inhibiting 5-alpha reductase present in the scalp. However, at least 3-5 months is effective, and if you stop taking it back to its original condition within 12 months. In addition, because the structure of finasteride itself is similar to sex hormones such as testosterone and progesterone hormones, side effects are often found in women, and in particular, strictly limit the use of mothers. Even factories producing finasterides prevent women from participating directly in production.

Minoxidil was also developed for the treatment of hypertension and observed to inhibit hair loss in hypertensive patients. The mechanism of action of minoxidil is not clear, but it can be seen to facilitate the production and development of hair by expanding the blood vessels of the scalp and easily supplying oxygen and nutrients to the hair follicles. There are few side effects, but the maximum effect should be applied for a long time more than 6 months, the effect disappears when stopped. It is also not an inhibitor of 5-alpha reductase and therefore does not lower the concentration of dihydrotestosterone.

Recently, many herbal remedies have emerged to compensate for the side effects and disadvantages of conventional drug therapies for the treatment of enlarged prostate and masculine alopecia. Although herbal remedies show other possibilities for the treatment of enlarged prostate and masculine alopecia, there are currently no long-term clinical results compared to placebo controls.

Saw palmetto, reported to be used by more than 2 million people in the United States alone, is an extract from the fruit of the saw palm tree and is a representative natural-derived functional food most commonly used for prostatic hyperplasia. In the past, Indians have used saw palmetto fruit as a remedy for the genitourinary system. The main components of saw palmetto are TG-type fatty acids and phytosterols, which act as 5-alpha reductase inhibitors to improve prostate hyperplasia and hair loss. In Korea, saw palmetto has been recognized as effective in improving prostate hyperplasia and is now marketed as an individually recognized product. Recently, it is produced and sold as a therapeutic agent for enlarged prostate by using coucurbite seed oil as an active ingredient. In fact, this seed oil is an European pumpkin seed extract and is imported from European pharmaceutical companies. As a mechanism of action, pharmaceutical companies explain that coucurbite seed oil suppresses blood cholesterol causing prostatic hypertrophy and prevents changes in male hormones to treat urination disorder caused by hypertrophy.

Camellia is also called winter rose and grows in Shandong Province of China, Taiwan, South Korea, Japan, etc. It grows mainly at 300-1,100m above sea level and is usually 1.5-6m in height. Flowers bloom from January to March. In Korea, it is grown on Tongyeong and Jeju Island. In particular, camellia oil, which is produced in Tongyeong, is imported from Japan and specialized in atopic dermatitis. Camellia oil is a fatty oil derived from the seeds of Camellia japonica L. of theaceae and its related plants.The fatty acid is similar to olive oil and contains up to 82-86% of oleic acid. Similar uses are used in creams, emulsions, etc., especially for hair. Does not deteriorate even at high temperatures of 160-220 degrees Celsius. Oleic acid, which is found in camellia oil, lowers LDL cholesterol, a low-density cholesterol that is harmful to the body, while raising high levels of HDL cholesterol, a high-density cholesterol that is good for the body, preventing high blood pressure and heart disease. It is known to be effective in preventing cancer and reducing memory due to aging.

In nature, however, it does not exist as free oleic acid, but is usually covalently attached to glycerol in ester form. Commercial oleic acid is either K + or Na + attached to the saponification of oil, ethyl ester form by ethanolysis, or manufactured through several industrial processes.

Therefore, essential fatty acids including oleic acid and linoleic acid, which are free from glycerol skeletal molecules, are fermented to facilitate absorption and use of camellia oil containing abundant essential fatty acids oleic acid and linoleic acid for easy absorption and use. The present invention was completed by preparing a fermented natural oil having the effect of preventing and improving prostatic hyperplasia and male hair loss by increasing the content of 5-alpha reductase.

It is an object of the present invention to separate natural oils including camellia oil into fatty acids (Fatty acid) and glycerol (Glycerol) through the most relaxed process of pre-microbial culture and post-enzyme fermentation, and thus new biochemical functions It aims to manufacture by adding.

In addition, an object of the present invention is to select a microbial strain having a high function of lipase that removes essential unsaturated fatty acids from glycerol skeletal molecules of camellia oil and incubate it in a lipase-induced medium, the culture medium containing a high titer of lipase is ammonium sulfate Precipitates, concentrates and dialysates enzyme proteins to eliminate various metabolic final substances that may occur during microbial cultivation and contaminate the final fermented product, and eliminates the fermentation odor produced by direct fermentation. Ferment the camellia oil using lipase concentrated fermentation solution to block the source.

It is also an object of the present invention to provide a use for preventing and improving prostatic hyperplasia and hair loss by preparing a fermentation oil containing a large amount of free essential fatty acids to inhibit 5-alpha reductase.

In addition, an object of the present invention is to produce a fermented natural oil for oral use for the enlargement of the prostate gland, and for oral and skin application for the prevention and treatment of hair loss to have a synergistic effect.

In order to achieve the above object, the present invention provides a fermented camellia oil containing abundant essential fatty acids, including oleic acid by fermenting camellia oil.

The present invention also provides a method for producing fermented camellia oil comprising the step of fermenting from a camellia oil using a fermentation enzyme or fermentation enzyme composition.

In addition, the present invention is fermented vegetable oil containing one or more of olive oil, canola oil, olive oil, rapeseed oil, moringa oil or sunflower seed oil in addition to camellia oil, and contains abundant essential essential fatty acids such as oleic acid. To provide fermented natural oils.

The present invention also provides a method for producing fermented natural oil comprising the step of fermenting using a fermentation enzyme or fermentation enzyme composition from the various vegetable oils.

In addition, the present invention provides a pharmaceutical composition or health functional food that inhibits 5-alpha reductase, improves and prevents prostatic hypertrophy and hair loss by using fermented camellia oil containing abundant essential fatty acids as an active ingredient.

In another aspect, the present invention provides a skin external preparation, cosmetic composition, or other quasi-drugs to inhibit 5-alpha reductase, improve and prevent hair loss by using fermented camellia oil containing abundant essential fatty acids as an active ingredient.

In addition, the present invention provides a pharmaceutical composition or a health functional food that inhibits 5-alpha reductase, improves and prevents prostatic hypertrophy and hair loss by using a fermented natural oil containing abundant essential fatty acids as an active ingredient.

In addition, the present invention provides a skin external preparation, cosmetic composition, or other quasi-drugs for inhibiting 5-alpha reductase, improving and preventing prostatic hypertrophy and hair loss by using fermented natural oil containing abundant essential fatty acids as an active ingredient. .

According to the present invention, fermented camellia oil contains a large amount of essential fatty acids including oleic acid as compared to other pre-fermentation oils and shows an effective effect.

In addition, according to the present invention, the microorganism producing lipase is first induced in culture, and then the culture medium containing the lipase is precipitated, concentrated and dialyzed using ammonium sulfate to physically emulsify the fermentation broth and camellia oil of high titer. After mixing, the fermentation process is carried out under optimum conditions, which can eliminate microbial waste contamination and odors peculiar to direct fermentation. The production efficiency of free unsaturated fatty acids can be greatly improved.

In addition, according to the present invention fermented camellia oil has the ability to inhibit 5-alpha reductase to lower blood dihydrotestosterone (dihydrotestosterone) to treat, improve or prevent the enlarged prostate.

In addition, according to the present invention, rich free essential fatty acids containing fermented camellia oil have the effect of preventing or improving male hair loss by inhibiting 5-alpha reductase.

1 is a graph showing the results of analyzing the prostate weight reduction effect of fermented camellia oil.

Figure 2 is a graph showing the results of analyzing the inhibitory effect of 5-alpha reductase of fermented camellia oil.

3 is a graph showing the results of analyzing the growth promoting effect of the fermented camellia oil.

Figure 4 is a graph showing the results of measuring the cytotoxicity of mouse-derived macrophages of fermented camellia oil.

Figure 5 is a graph showing the results of measuring the long-term weight change of female mice according to a single toxicity test for fermented camellia oil mice.

Figure 6 is a graph showing the results of measuring the long-term weight change of male mice according to a single toxicity test for fermented camellia oil mice.

Figure 7 is a graph showing the results of measuring the weight change of female and male mice according to a single toxicity test for fermented camellia oil mice.

The present invention fermented camellia oil to provide a fermented camellia oil containing abundant essential fatty acids, including oleic acid.

Hereinafter, the present invention will be described in detail.

The present invention provides a fermented camellia oil containing fermented camellia oil in abundance.

Camellia japonica seed oil is known as Camellia japonica seed oil, and the raw material is fat oil obtained from seed removed from Camellia japonica Linne ( Theaceae ). Extraction and manufacturing methods of camellia oil include cold pressing obtained by pressing at room temperature, heat extraction by pressing at high temperature, and solvent extraction using ethanol or hexane at high temperature and high pressure. extraction methods. Camellia oil contains 80% or more of oleic acid and contains palmitic acid, linoleic acid and linolenic acid. It has good skin absorption and has a soothing effect. In addition, it is known to inhibit the growth of Staphylococcus aureus and is effective in atopic dermatitis, skin itching and the like.

The active ingredient of fermented camellia oil used in the present invention refers to an essential unsaturated fatty acid, especially an oil containing a large amount of oleic acid.

The essential fatty acid preferably contains a large amount of free oleic acid, linoleic acid, gamma linolenic acid and the like. In addition, the fermented camellia oil containing a large amount of free essential fatty acids is preferably effective to inhibit the 5-alpha reductase.

The oleic acid does not readily oxidize and produce lipid peroxides compared to other fatty acids. Oleic acid is known to prevent arteriosclerosis and heart disease by removing LDL-cholesterol in the blood like linoleic acid and alpha-linolenic acid. It is also the most abundant fatty acid in breast milk, helping the baby grow and develop.

The fermentation enzyme is preferably a lipase derived from the genus Rizopus or Candida, or a fermentation enzyme concentrate for fermentation of natural plants concentrated through extraction, ammonium sulfate, and dialysis in plants and animals.

The present invention also provides a method for preparing fermented camellia oil.

Fermented camellia oil used in the present invention can be prepared as follows. A method of preparing fermented camellia oil for achieving the purpose is obtained by separating fatty acids from glycerol, which is a skeletal molecule constituting oil, using a fermentation concentrate containing lipase produced by microorganisms. Fermentation results in a richer free essential fatty acid content than before fermentation.

For oil fermentation, microbial culture, a strain that produces large amounts of lipase, is added directly to the oil, and a small amount of artificial emulsifier is added to emulsify the oil so that it can be easily fermented. Add various nutrients for this and incubate it for a long time under optimum conditions. At the end of the incubation, centrifugation is usually performed to separate the microbial cells and the oil culture. The microbial direct fermentation oil thus produced usually has a characteristic smell of microbial fermentation, and it is troublesome to separate cultured microorganisms from fermentation oils. It is also likely that the final fermented oil product is contaminated. To this end, the final fermentation oil must be passed through the adsorption column or a distillation or filtration process must be added to remove odors and contaminants. In particular, the high fat-soluble substance as a compound that is released as the microorganism grows is easily contaminated with the final fermentation oil, and it is difficult to remove it later from the fermentation oil.

Therefore, in the present invention, the fermentation microorganisms are pre-cultured in a lipase-inducing medium, and then only the culture medium is recovered and concentrated, and then emulsified and fermented by the high titer of lipase-containing culture solution and oil by physical method. Second, to maximize the production of free unsaturated fatty acids, third, to minimize the microbial contamination of the final fermented oil product, fourth, to harm the body or consumers because it does not depend on artificial emulsifiers or other chemical processes Fifth, there is no contamination of chemicals that can make an unfavorable impression. Fifth, it is possible to produce the maximum amount of essential unsaturated fatty acids freed in the most natural way, thus increasing competitiveness over other fermented products. Sixth, concentrate fermentation filtrate with ammonium sulfate. To the buffer solution By removing the fat-soluble and low molecular weight compounds made by the microorganisms produced as by-products during fermentation, only pure fermented oil can be prepared.

Fermentase used in the present invention is preferably a lipase (Lipase), the fermentation enzyme composition is preferably a composition comprising a lipase, in particular, pre-cultivation of microorganisms capable of producing lipase and enrich the enzyme filtrate obtained One is preferable. Lipases derived from plants or animals can be included in the preparation of the present invention, without being limited to lipase fermentation concentrates derived from microorganisms.

Hereinafter, a method of preparing fermented camellia oil will be described in detail.

The method for preparing fermented camellia oil used in the present invention comprises the steps of: i) mixing the fermented enzyme or fermented enzyme composition in a weight ratio of 1: 8 to 8: 1 in camellia oil; ii) blending the mixture of step i) in a blender for 5-10 minutes and then physically emulsifying by sonicating for 30-60 minutes; iii) fermenting the mixture of step ii) with mixing at 300-400 rpm for fermentation temperature of 25-45 degrees Celsius, pH 5.0-8.0 and fermentation time 48-120 hours; And iv) separating the fermentation oil from the fermentation concentrate by centrifugation and ultrafiltration of the fermentation product of step iii).

Method for producing a fermentation enzyme used in the present invention comprises the steps of a) culturing a microorganism (including genus lypus or Candida microorganisms) under specific induction conditions to obtain a fermentation broth with a large amount of lipase enzyme added to a high titer; b) concentrating and dialysis the enzyme filtrate obtained from the culture; c) adding the microbial fermentation broth and vegetable oil and fermenting at optimum conditions (physical emulsification, optimal pH, optimal temperature, optimal mixing rpm and fermentation time); And d) recovering the fermented oil.

In the method, step a) is a step of incubating the microorganisms to produce as much as possible a high titer of primary lipase under aerobic conditions.

In the above method, step b) is a step of separating the cells from the culture medium after the primary culture and concentrated dialysis of the enzyme.

In the above method, each cultured strain culture was centrifuged at 4 ℃, 10,000 rpm conditions to isolate the cells from the culture medium, the supernatant was treated with ammonium sulfate at 4 ℃ to precipitate proteins After recovering the protein containing the enzyme, the concentrated enzyme can be dissolved in pH 7.0 (sodium phosphate buffer, 50 mM) and dialyzed in the same buffer to obtain a concentrated lipase.

In the method, step c) is a step of fermentation by adding the obtained lipase concentrated solution and vegetable oil.

Herein, the temperature of the oil is preferably 40 ° C. The pH of the fermentation enzyme solution to be added is preferably pH 7.0. The mixing ratio of the vegetable oil and the fermented enzyme solution is preferably a ratio of 50% by weight of vegetable oil to 50% by weight of the fermented enzyme solution. When mixing vegetable oil and fermented enzyme solution, blend with a blender for about 10 minutes, sonicate for 10-60 minutes, and put into fermenter. 350 rpm is preferable for the mixing speed. The fermentation time is preferably 120 hours. As for the temperature to ferment, 40 degreeC is preferable. This is because the lipase and the vegetable natural oil must be fermented enough to allow the production of the free essential fatty acids.

The present invention also fertilizes vegetable oils including one or more of olive oil, canola oil, olive oil, rapeseed oil, moringa oil or sunflower seed oil in addition to camellia oil to enrich free essential fatty acids such as oleic acid. It provides fermented natural oil containing.

The essential fatty acid contains a large amount of free oleic acid, linoleic acid or gamma linolenic acid, and the fermented camellia oil containing a large amount of the free essential fatty acid is preferably effective to inhibit the 5-alpha reductase.

In addition, the present invention is a glass such as oleic acid comprising the step of fermenting one or more of olive oil, canola oil, olive oil, rapeseed oil, moringa oil or sunflower seed oil by treating the fermentation enzyme or fermentation enzyme composition It provides a method for producing a fermented natural oil containing abundant essential fatty acids.

The present invention also provides a pharmaceutical composition for the prevention and treatment of enlarged prostate, containing fermented camellia oil according to the present invention, and a dietary supplement, cosmetic composition or quasi-drug for preventing and improving prostatic hyperplasia.

Such compositions are based on fermented camellia oil as a main ingredient and may comprise as an additional ingredient a combination selected from a pharmaceutically, food or cosmetically acceptable carrier such as vitamins and the like.

Such compositions may further contain one or more active ingredients which exhibit the same or similar function in addition to the above ingredients.

Specifically, the composition is not limited to camellia oil but may include a vegetable oil containing a large amount of oleic acid or linoleic acid in essential fatty acids. This may include olive oil, canola oil, rapeseed oil, sunflower seed oil, pecan oil, tea tree oil, and the like. When fermented to include a large amount of free essential unsaturated fatty acids can have a similar effect to fermented camellia oil.

Such compositions may be formulated in a variety of oral and parenteral formulations, which may be formulated using conventional diluents or excipients, such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, and the like.

Dosages of such compositions vary depending on the patient's weight, age, sex, health condition, diet, time of administration, method of administration, rate of excretion and severity of the disease, with the daily dosage being approximately based on the amount of the composition. It may be from 0.0001 to 10 g / kg, may be administered 1 to 6 times a day.

In the present invention, the composition for treating, improving and preventing prostatic hyperplasia is to effectively inhibit 5-alpha reductase to improve prostatic hyperplasia by dihydrotestosterone (dihydrotestosterone) made by excess of 5-alpha reductase or It means treatment and prevention, and its clinical effect means restoring the enlarged prostate to normal size due to enlargement of the prostate.

In addition, the present invention provides a pharmaceutical composition for preventing and treating hair loss, and a health functional food, cosmetic composition or quasi-drug for preventing and improving hair loss, containing fermented camellia oil as an active ingredient according to the present invention.

In the present invention, the pharmaceutical composition for treating, improving and preventing hair loss refers to male hair loss caused by dihydrotestosterone, which is excessively produced by 5-alpha reductase by effectively inhibiting 5-alpha reductase. Means to improve, treat, and prevent hair loss when the composition is orally administered or applied directly to the scalp.

In particular, the present invention means that the fermented camellia oil is quickly absorbed into the scalp by using it for cosmetic compositions or coatings to have a more effective male-type hair loss improving effect.

In addition, the present invention is fermented vegetable oil containing one or more of olive oil, canola oil, olive oil, rapeseed oil, moringa oil or sunflower seed oil in addition to camellia oil, and contains abundant essential essential fatty acids such as oleic acid. Provided is a pharmaceutical composition for preventing and treating prostatic hyperplasia, which contains fermented natural oil as an active ingredient, and a dietary supplement, cosmetic composition, or quasi-drug for preventing and improving prostatic hyperplasia.

In addition, the present invention is fermented vegetable oil containing one or more of olive oil, canola oil, olive oil, rapeseed oil, moringa oil or sunflower seed oil in addition to camellia oil, and contains abundant essential essential fatty acids such as oleic acid. The present invention provides a pharmaceutical composition for preventing and treating hair loss, and a dietary supplement, cosmetic composition, or quasi-drug for preventing and improving hair loss.

Hereinafter, the present invention will be described in detail with reference to the following examples, but the scope of the present invention is not limited by the following examples.

<실시예 1> 리파아제를 생성하는 미생물과 배지 조성 및 침전, 농축, 투석과정을 거친 고농축 발효효소액 제조Example 1 Preparation of Highly Concentrated Fermentation Enzyme Solution After Microbial and Lipid Formation and Precipitation, Concentration, and Dialysis

Candida rugosa or other Candida microorganism culture medium that produces lipase for fermenting camellia oil contains 2% olive oil, 0.3% yeast extract, 0.3% malt extract, 0.5 A composition of% peptone, 1% dextrose was used, incubated at pH 7.0, 30 degrees Celsius, and culture medium of Rizopus oryzae or other Rizopus microbial culture was 2% olive oil. , 4% potato starch, 1% dextrose composition was used, was cultured at pH 6.0, 30 degrees Celsius was used to prepare a high titer fermentation concentrate with a large amount of lipase production secretion. The culture time of the microorganism was cultured for 70-90 hours in the case of Candida strain, and 100-120 hours in the case of Rizopus strain. When the microbial culture was terminated, the microbial cells were allowed to settle by centrifugation at 8,000 rpm, and only the supernatant was recovered. Then, 60-80% ammonium sulfate was added slowly at 4 degrees Celsius to stir the proteins in the culture medium. The precipitated enzyme protein was centrifuged at 12,000 rpm at 4 degrees Celsius to settle and remove the supernatant, and then dissolved in 50 mM sodium phosphate buffer (pH 7.0). This concentrated enzyme protein was repeatedly dialyzed 2-4 times at 4 degrees Celsius with the same buffer to remove substances with low molecular weight. In order to measure the lipase titer of the high titer of concentrated fermentation enzyme solution, color development through saponification was used. Preparation of the copper acetate-pyridine reagent was made to 5% (w / v) cupric acetate (Sigma) and prepared by adjusting the pH to 6.1 using pyridine (Merck). The substrate was made to contain 10% olive oil in 1% polyvinylalcohol and 2.5 ml of the substrate and 1 ml of the concentrated fermentation enzyme solution were reacted for 30 minutes at 37 degrees Celsius. After the reaction, the reaction was terminated with 5 ml acetone (acetone). The produced free fatty acid was dissolved by adding 2.5 ml of isooctane, and 1 ml of copper-acetate-pyridine was added to develop a copper reaction. At this time, the degree of color development was measured for absorbance at 715nm. The lipolytic activity of lipase was expressed in units of units and 1 unit was defined as 1 μmol of the fatty acids produced.

Table 1

	Cell weight (g / 100 ml)	Lipase Activity (U / ml)
Candida rugosa	2.7	4.8
Rhizopus oryzae	2.9	3.2

<실시예 2> 고농축 발효효소액을 이용한 동백 오일의 발효Example 2 Fermentation of Camellia Oil Using Highly Concentrated Fermentation Enzyme Solution

In order to set the optimum temperature condition for the enzyme reaction, the temperature of camellia oil was set to 37 ° C, and the fermented enzyme solution produced by microbial culture and highly concentrated was dialyzed in sodium phosphate buffer (50 mM) at pH 7.0. After mixing and using each other. That is, after mixing at a ratio of 50% by weight of oil to 50% by weight of the highly concentrated fermentation enzyme solution (including lipase), blended in a blender for 10 minutes and then sonicated for 30 minutes to complete the physical emulsification process, the rotation speed 350 rpm, Fermentation was carried out at 37 ℃ for 72 hours. After fermentation, the oil was centrifuged at 10,000 rpm for 20 minutes to separate the fermentation enzyme solution and the enzyme residue, followed by ultrafiltration to obtain only fermented natural oil.

<비교예 1> 발효한 올리브오일Comparative Example 1 Fermented Olive Oil

Olive oil fermented in the same manner as in <Example 2> was obtained.

<비교예 2> 발효한 카놀라오일Comparative Example 2 Fermented Canola Oil

Canola oil fermented in the same manner as in <Example 2> was obtained.

<비교예 3> 발효하지 않은 동백오일<Comparative Example 3> Uncambed Camellia Oil

<비교예 4> 발효하지 않은 올리브오일Comparative Example 4 Unfermented Olive Oil

<비교예 5> 발효하지 않은 카놀라오일Comparative Example 5 Canola Oil Not Fermented

<실험예 1> 발효한 동백 오일의 유리된 필수지방산 생산 수율 측정Experimental Example 1 Measurement of Free Essential Fatty Acid Production Yield of Fermented Camellia Oil

Essential fatty acid content was determined using HPLC. At this time, in order to quantify the content of each essential fatty acid, a standard reagent for each fatty acid was obtained, a standard curve was first obtained by HPLC, and fermented camellia oil was analyzed and compared with the standard curve. HPLC column (C18,250 x 4.6 mm ID) was used, and the mobile phase was mixed with 0.005% TFA in acetonitrile: 0.005% TFA in water in a volume ratio of 8: 2 with a flow rate of 1.0 ml / min. , The sample injection amount was used 0.01 ml and measured at a wavelength of UV 210 nm. The slope of the mobile phase was started at 20:80 (v / v) and maintained for 30 minutes, after which the fatty acids were analyzed with increasing polarity.

Shown in Table 3 below shows the content of fatty acids described in the certificate of camellia oil (Tongyeong, Korea), canola oil (Quebec, Canada), olive oil (greek acid) used for the present invention. These fatty acids are more than 99% ester-bonded to glycerol backbone molecules and the total content of free fatty acids is less than 1%.

TABLE 2

Total fatty acid content of natural oil purchased for use in the present invention

	Oleic acid	Linoleic acid	Linolenic acid	Palmitic acid	Stearic acid
Camellia oil (Tongyeong)	85-87%	4.5%	0.3%	8.5%	1.1%
Olive oil (Henry Lamotte)	72-75%	5.0-6.0%	0.6%	3.7%	11.5%
Canola oil (from Canada)	63.4-66.0%	17.0-21.2%	8.7-9.2%	4.2-4.5%	1.6%

TABLE 3

Fatty acid content before and after fermentation of camellia oil, olive oil and canola oil

Fermentation	content (%)
Fermentation	Oleic acid (OA)	Linoleic acid (LA)	Gamma-linolenic acid (GLA)
Example 2 (fermented camellia oil)	38.76	2.06	0.10
Comparative Example 1 (fermented olive oil)	25.35	1.02	0.02
Comparative Example 2 (fermented canola oil)	19.25	0.95	0.05
Comparative Example 3 (camellia oil before fermentation)	Not detected	Not detected	Not detected
Comparative Example 4 (Olive Oil before Fermentation)	Not detected	Not detected	Not detected
Comparative Example 5 (Canola oil before fermentation)	Not detected	Not detected	Not detected

As can be seen from Table 3 above, free essential fatty acids were hardly measured in the non-fermented oils, and essential fatty acids liberated only in the fermented vegetable oils were detected. It can be seen that the highest. In addition, it can be confirmed that the most free oleic acid among essential fatty acids.

<실험예 2> 생체내(Experimental Example 2 In vivo in vivoin vivo ) 실험을 통한 동백 발효오일의 전립선 무게 변화 측정Measurement of Prostate Weight of Camellia Fermented Oil by Experiment

The animals to be used for the experiment were the following experiments using 260-300g 7-week-old male Sprague Dawley rats (Semtaco, Osan, Korea). The rats were allowed to acclimate for one week, and the rats were tested under conditions of 18-22 degrees Celsius, 40-50% humidity, 12 hours dark and 12 hours bright. In the comparison group, finasteride was used to treat prostatic hypertrophy. In order to rule out the effects of internal testosterone, the male SD rats except for the normal group were anesthetized with ether, and then the abdominal medial incision was made through the abdominal cavity, and the testicles were placed into the abdominal cavity. The testicles were fixed to the side abdominal wall. After being allowed to rest for one week, the group of each individual was administered the composition according to the present invention (fermented camellia oil) after the induction of prostate hypertrophy by hormonal stimulation (control), prostate hypertrophy by hormonal stimulation. Experiment group 1, the comparison group administered with finasteride after prostate hypertrophy induced by hormone stimulation, and the experimental group 2 administered the composition (fermented camellia oil) of the present invention to confirm the other effects and toxicity of the composition. Each group was assigned the same weight of six dogs in each group based on weight, and propionate testosterone (TP) is injected subcutaneously at 5 mg / kg / day for three weeks and the composition of the present invention In order to see the effect of fermented camellia oil was administered orally to the experimental group in the amount of 100 mg / kg / day or 400 mg / kg / day, the control group was orally administered 2 mg / kg / day for 3 weeks.

Table 4

Experimental group	Sample and Daily Dose (21 days)
normal		Vehicle
Control	TP (5 mg / kg / day)	Vehicle
Experimental group
		1	Example 2 Fermented Camellia (100 mg / kg / day)
Example 2 Fermented Camellia (400 mg / kg / day)
Comparison	Finasteride (2 mg / kg / day)
Experiment group 2		Example 2 Fermented Camellia (100 mg / kg / day)
		Example 2 Fermented Camellia (400 mg / kg / day)

As shown in Table 4 above, oral administration was performed for 3 weeks, and the results were confirmed by sacrifice of mice. The results are shown in Table 5 and FIG. 1.

Table 5

Experimental group	Prostate Weight (g / kg)
normal	0.534 ± 0.075
Control	0.890 ± 0.053
Experimental group 1	0.685 ± 0.081
Experimental group 1	0.595 ± 0.045
Comparison	0.572 ± 0.039
Experiment group 2	0.525 ± 0.031
Experiment group 2	0.553 ± 0.047

As a result of the experiment, as shown in Figure 1, the weight of the control group was increased by about 60% than the weight of the prostate in the control group, and

experimental groups

1 and 2 administered <Example 2> inhibited hypertrophy of 23.1% and 33.2%, respectively. The effect was shown. In addition, experimental group 2, which did not induce prostate hypertrophy, showed a similar level to the normal group, and fermented camellia oil did not seem to inhibit any side effects or prostate development in healthy prostate.

<실험예 3> 발효 동백 오일의 5-알파 환원효소의 억제효과 측정Experimental Example 3 Inhibition of 5-alpha Reductase of Fermented Camellia Oil

5-alpha reductase inhibitory effect was performed according to Hirosumi's method. In a micro tube, 40 mM potassuim phosphate, 0.2 mM NADPH buffer solution was mixed with the enzyme and the fermented camellia oil sample prepared in Example 2, 1 mM DTT, and 120 nCi [1.2.6.7-3H] T. After adjusting to 500 μl and mixing well, it was reacted at 37 ℃ 60 minutes. Then, 1 ml of ethyl acetate was added to terminate the reaction, and centrifuged at 1500 rpm for 5 minutes to obtain a supernatant. The supernatant was completely dried at 60 ° C., dissolved in 50 μl ethyl acetate, and developed in TLC. The solvent used for the separation conditions of TLC was 50% cyclohexane, 50% ethyl acetate. Testosterone, a product appearing on TLC, was identified by UV irradiator at 254 nm, and dihydrotestosterone was confirmed by spraying 10% sulfuric acid. For quantitative analysis, testosterone and dihydrotestosterone sites were cut out and 10 ml of Ultima GoldTM cocktail (PerkinElmer, Massachusetts, USA) was added to the liquid scintillation analyzer (Packard Bioscience, Meriden, USA). The conversion rate of testosterone by 5-alpha reductase was calculated by the following equation.

5-alpha converting enzyme inhibitory activity (%)

= {1- (dihydrotestosterone / testosterone + dihydrotestosterone)} × 100

As a result of the experiment, as shown in Figure 2, in this experiment to see whether the fermented camellia oil inhibits the 5-alpha reductase finasteride used as a positive control group was able to inhibit about 98% at a concentration of 0.5 ppm. IC ₅₀ values reported in Finasteride's paper are 69 nM (type II 5-alpha reductase) and 360 nM (type I 5-alpha reductase). Given that the molecular weight is 372.5 daltons, it has an IC ₅₀ value at 0.03-0.13 ppm. On the other hand, the IC ₅₀ value of fermented camellia oil is about 80 ppm considering the value of negative control of camellia oil before fermentation.

<실험예 4> 모낭세포의 성장 촉진 효과 측정Experimental Example 4 Measurement of Growth Promotion Effect of Hair Follicle Cells

Dermal papilla cells are cells that give hair growth orders in the lower part of the hair follicles below the pores. Therefore, it was tested whether fermented camellia oil promotes the growth of dermal papilla cells.

Human dermal papilla cells were used as cell lines used in the experiment. Cell culture was incubated at 37 ℃, 5% CO ₂ using DMEM medium added with 10% FBS. Cells were dispensed at 1 x 10 ⁵ cells / mL for each well of 96 wells and then incubated for 24 hours. After 24 hours, the medium used for the previous culture was removed, the sample was dissolved in DMSO, diluted with medium so that the final concentration of DMSO was 1%, and the concentration of fermented oil (μg / mL) was 37 ° C. and 5% CO ₂ , respectively. Each for 24 hours. As a positive control, finasteride was treated with 0.5 ppm. After the treatment, 20 μl of 0.2% MTT solution was added to each well, followed by reaction for 3 hours at 37 ° C. in a 5% CO ₂ incubator. After the reaction, all supernatants were removed, DMSO 150 μl was added, and all formazan produced at room temperature was dissolved for 10 minutes, and the change in absorbance was measured at 570 nm using an ELISA reader. In order to determine the effect on DMSO, the experiment was divided into a control group and a control group for 1% DMSO, and cell growth rate (%) was expressed using the following equation.

Cell growth rate (%) = (absorbance of sample treatment group / absorbance of control group) × 100

As a result of the experiment, as shown in FIG. 3, the growth and proliferation effect of the dermal papilla cells was excellent at 400 ppm of fermented camellia oil, and the dermal papilla cells were grown by 120 ± 4.5% compared to the control group. 400 ppm of fermented camellia oil was more effective than 104.5 ± 3.3% of 0.5 ppm finasteride.

<실험예 5> 세포 안전성 측정Experimental Example 5 Cell Safety Measurement

MTT assay was performed to determine the toxicity of Raw 264.7 cells, a mouse-derived macrophage line of fermented camellia oil of the present invention.

Toxicity of macrophages was analyzed by MTT [(3- (4,5-dimethyl thiazol-2-yl) -2,5-diphenyl tetrazolium bromide) (Sigma, MO, USA)]. Raw264.7 cells were dispensed at 1 × 10 ⁴ cells / mL for each well of 96 wells and then incubated at 37 ° C., 5% CO ₂ for 24 hours. After 24 hours, the medium used for the previous culture was removed, the sample was dissolved in DMSO, diluted with medium so that the final concentration of DMSO was 1%, and the concentration of fermented oil (μg / mL) was 37 ° C. and 5% CO ₂ , respectively. Were treated for 5 hours and 24 hours, respectively. After the treatment, 20 μl of 0.2% MTT solution was added to each well, followed by reaction for 3 hours at 37 ° C. in a 5% CO ₂ incubator. After the reaction, all supernatants were removed, DMSO 150 μl was added, and all formazan produced at room temperature was dissolved for 10 minutes, and the change in absorbance was measured at 570 nm using an ELISA reader. In order to determine the effect on DMSO, the experiment was divided into a control group and a control group for 1% DMSO, and cell survival rate (%) was expressed using the following equation.

Cell viability (%) = (absorbance of sample treatment group / absorbance of control group) × 100

As shown in Figure 4, it can be seen that the cytotoxicity to the fermented camellia oil prepared by the present invention is not toxic even at high concentration of 1000 ppm.

<실험예 6> 단회 독성 측정Experimental Example 6 Single Toxicity Measurement

The experimental animals were rats of 6-week-old Sprague Dawley (SD) strain (Samtako, Korea). These animals did not show any general symptoms at the time of acquisition and during adaptation. Breeding conditions were carried out after setting the temperature 22 ± 2 ℃, relative humidity 45 ± 10%, lighting time 12 hours (6 am to 6 pm), solid feed for experimental animals (Samtako, Korea) and water purification system Tap water was used freely. In the single toxicity test of fermented camellia oil, the experimental group dissolved and administered fermented camellia oil in distilled water, the control group administered distilled water, and rats were fasted 12 hours before sample administration and used as a single toxic oral dose of normal functional foods. Observation was carried out for 14 days after oral administration at a concentration of 2 g / kg. Clinical signs observations were performed in accordance with the Food and Drug Administration's Toxicity Test Standards and OECD Test Guideline 420 (TG 420). That is, all the experimental animals were observed every hour for 6 hours after administration on the day of administration, and the change of general condition of the animal, the expression of intoxication symptoms and the presence of death were observed once a day from the following day to 14 days. In addition, the weight change on the day and 14 days of administration of the test substance was measured. After the end of the test, the animals were anesthetized and killed, and then visually observed for abnormalities in appearance and internal organs.

As a result of the experiment, as shown in Figures 5 to 7, there were no individuals who died in males and females according to the administration of the sample, four observations on the day and once a day for 14 days from the next day, camellia fermentation compared to the control group There was no specific behavioral change in the oil group. In addition, as can be seen in Figures 5 to 7 body weight change after 14 days of sample administration and the change in organ weight after autopsy also did not change significantly compared to the control group. Therefore, it can be seen that the fermented camellia oil has no animal single toxicity.

<실험예 6> 간이임상 시험Experimental Example 6 Simple Clinical Trial

A male clinical subject with alopecia was selected using the fermented camellia oil of the present invention and a simple clinical trial was conducted.

Subjects were set up in groups of six for 12 adult males with mild hair loss from 33 to 55 years of age (approximately more than 50 hairs falling off per day and self-aware of their hair loss). One group applied 5 ml of fermented camellia oil to the hair once a day (before bedtime) and wrapped the hair the next day. The other group was placebo group once a day before fermenting camellia oil, 5 The ml was applied to the hair for the same period (4 weeks) and the results were observed. To determine the effect, the average number of hair loss, visual evaluation, and the individual's personal opinions were selected as the evaluation scale after the head was closed, and evaluated in five stages. . More details are shown in Table 6 below. The overall test results are shown as the average of each score.

Table 6

Clinical trial evaluation index of the present invention

Score	Visual evaluation	Personal opinion	Hair loss
One	weakening	weakening	increase
2	No change	No change	No change
3	Slightly improved	Slightly improved	Slightly reduced
4	Moderate improvement	Moderate improvement	Usually reduced
5	Much improvement	Much improvement	Much reduced

TABLE 7

Clinical Trial Results of the Invention

Score	Example 2 (fermented camellia oil)	Comparative Example 3 (camellia oil before fermentation)	Remarks
Visual evaluation	3.4	2.7	21%
Personal opinion	4.1	3.2	28%
Hair loss	3.9	2.5	36%
Comprehensive improvement	3.8	2.8	27%

The results of the simplified clinical trials of the present invention showed improvement of 21 to 36% in visual evaluation, individual findings, and hair loss compared to <Comparative Example 3>, which was not fermented as shown in Table 7 above. It can be seen that there is an improvement effect.

Claims

Fermented camellia oil fermented by treating fermented enzyme or fermented enzyme composition to camellia oil.
The method of claim 1, wherein the fermented camellia oil contains a high content of free essential unsaturated fatty acids (Oleic acid (OA), linoleic acid (LA) or gamma- linolenic acid (gamma- Linolenic acid, GLA) Fermented camellia oil characterized by.
The method of claim 1, wherein the fermentation enzyme is a lipase derived from the genus Rizopus or Candida microorganism, or a fermentation enzyme concentrate for fermentation of natural plants concentrated through extraction, ammonium sulfate treatment and dialysis in plants and animals. Fermented camellia oil.
A fermented natural oil obtained by fermenting one or two or more vegetable oils of olive oil, canola oil, olive oil, rapeseed oil, moringa oil or sunflower seed oil by treating a fermentation enzyme or fermentation enzyme composition.
i) mixing the fermenting enzyme or fermenting enzyme composition in camellia oil in a weight ratio of 1: 8 to 8: 1;

ii) blending the mixture of step i) in a blender for 5-10 minutes and then physically emulsifying by sonicating for 30-60 minutes;

iii) fermenting the mixture of step ii) with mixing at 300-400 rpm for fermentation temperature of 25-45 degrees Celsius, pH 5.0-8.0 and fermentation time 48-120 hours; And

iv) separating the fermentation oil from the fermentation concentrate by centrifugation and ultrafiltration of the fermentation product of step iii).
The method according to claim 5, wherein the fermentation enzyme or fermentation enzyme composition used in step i)

a) culturing the lipase producing microorganism in an induction medium; And

b) removing the microbial cells from which the fermentation is completed in the culture through centrifugation and filtration, and then concentrating the protein in the culture by precipitation with ammonium sulfate; And

c) a method for preparing fermented camellia oil, comprising the step of recovering the enzyme fermentation solution after dialysis of the precipitated and concentrated protein with a buffer solution (50 mM, sodium phosphate buffer, pH 7.0).
The method according to claim 5, wherein the fermentation enzyme of step 1) is a lipase isolated from the genus Rizopus strain including Rhizopus oryzae or the Candida genus strain including Candida rugosa, or plants and animals. A method for producing fermented camellia oil, characterized in that the lipase enzyme extracted from the pancreas.
A pharmaceutical composition for preventing and treating enlarged prostate or hair loss, comprising the fermented camellia oil of claim 1 as an active ingredient.
Prostate hypertrophy or hair loss prevention and functional health food containing the fermented camellia oil of claim 1 as an active ingredient.
Claim 1 cosmetic composition for preventing and improving enlarged prostate gland or hair loss containing the fermented camellia oil as an active ingredient.
Prophylactic enlargement or hair loss prevention and improvement qualitative product containing the fermented camellia oil of claim 1 as an active ingredient.
A pharmaceutical composition for preventing and treating prostatic hypertrophy or hair loss, comprising the fermented natural oil of claim 4 as an active ingredient.
Prostate hypertrophy or hair loss prevention and functional health food containing the fermented natural oil of claim 4 as an active ingredient.
Claim 4 cosmetic composition for preventing and improving enlarged prostate or hair loss containing the fermented natural oil as an active ingredient.
A quasi-drug for preventing and improving enlarged prostate or hair loss, comprising the fermented natural oil of claim 4 as an active ingredient.