KR102309425B1 - Pharmaceutical Composition Comprising Red Ginseng Oil for Preventing or Treating Prostatic Hyperplasia - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating benign prostatic hyperplasia comprising red ginseng oil, wherein the red ginseng oil of the present invention is prepared by applying a thin film distillation process after supercritical extraction, and the prepared red ginseng oil is Since the improvement effect is excellent, the preparation method of the present invention and the red ginseng oil according to the present invention can be usefully used in the pharmaceutical field and food field for the purpose of improving prostatic hyperplasia.

Description

Pharmaceutical composition for preventing or treating benign prostatic hyperplasia comprising red ginseng oil {Pharmaceutical Composition Comprising Red Ginseng Oil for Preventing or Treating Prostatic Hyperplasia}

The present invention relates to a pharmaceutical composition for preventing or treating benign prostatic hyperplasia comprising red ginseng oil.

Prostatic hyperplasia (prostatic hyperplasia) is a urinary disease in which the prostate is enlarged non-tumorally in men. The exact cause of BPH is not known, but it is known that age and sex hormones influence it.

BPH starts to increase after the age of 50, as the incidence increases with age. When testosterone is excessively present in the blood, testosterone is converted into dihydrotestosterone (DHT) by 5-alpha-reductase (5α-reductase, 5AR), which is bound to androgen receptor (AR). , As a result, smooth muscle (smooth cells) and epithelial cells (epithelial cells) of the prostate transition zone are excessively proliferated and the prostate is enlarged. In addition, as men age, the secretion of male hormones decreases, and as a result, androgen receptors in prostate cells increase in order to maintain the endocrine system balance, DHT binds to more androgen receptor sites and the prostate enlarges. As the prostate enlarges, various urination disorders due to bladder obstruction occur.

These urination disorders are not only unable to hold urine, but also have a thin stream of urine and weak strength, and can't urinate immediately even if they try to urinate. There is a problem in that it affects the kidneys because they are sick, and they urinate frequently, which eventually leads to chronic renal failure due to a decrease in renal function. In addition, such stagnant urine provides good conditions for bacteria to grow in the prostate and urethra, and there is a problem in that complications such as inflammation and stones are easily induced.

Recently, prostatectomy surgery or laser removal is used for the treatment of benign prostatic hyperplasia, but this is only a temporary treatment method and cannot prevent continued enlargement. Drugs used for the treatment of benign prostatic hyperplasia include alpha-adrenergic agonists, which inhibit prostate tension, and drugs that block the expansion of the prostate by decreasing androgen hormone, such as alpha blockers. or androgen inhibitors are used.

As androgen inhibitors, 5-alpha reductase inhibitors are used as therapeutic agents, and finasteride and dutasteride are commercially available. The 5-alpha reductase inhibitor acts to reduce the size of the prostate by inhibiting conversion of testosterone to dihydrotestosterone by 5-alpha reductase. However, it is known that commercial drugs used for such drug therapy can cause side effects such as dizziness, lethargy, and headache. Therefore, there is a need to develop a new natural product that can prevent or treat BPH using a natural product without side effects. As an example of a natural new drug, for example, Patent Document 1 discloses the use of a bokbunja extract to treat benign prostatic hyperplasia.

Among natural products, ginseng is a plant that has been used as a valuable blood tonic in Korea and China from ancient times to the present. It is known to be caused by ginseng saponins, that is, ginsenosides.

Ginseng is divided into fresh ginseng, red ginseng, and white ginseng according to the processing method. Fresh ginseng is fresh ginseng harvested from the field, and contains 70-80% moisture, making it difficult to store for a long time. White ginseng is manufactured by peeling the epidermis of fresh ginseng or drying fresh ginseng in sunlight or hot air, and has a milky white or pale yellow color. Red ginseng is manufactured by washing raw ginseng without peeling off its epidermis and then going through steaming and drying processes, etc., and has a pale yellowish brown or dark red color.

In general, only high-quality fresh ginseng from six-year-old fresh ginseng is selected, steamed, and dried. It has been found that these red ginseng-specific ginsenosides have excellent effects in preventing cancer and protecting brain nerve cells.

The present inventors have reviewed a material capable of improving enlarged prostate, and completed the present invention by confirming that an extract derived from red ginseng improves enlarged prostate.

Korea Registered Patent No. 1,439,859

We aim to provide a natural new drug with fewer side effects while improving prostate enlargement. By applying an additional process to the extract, it was confirmed whether the improvement effect of benign prostatic hyperplasia could be increased.

In order to solve the above problems, the present invention provides the following composition:

1) extracting red ginseng with a supercritical fluid; and

2) thin film distillation of the extract obtained in step 1); a pharmaceutical composition for preventing or treating benign prostatic hyperplasia comprising red ginseng oil prepared by a method comprising a.

The red ginseng oil of the present invention is prepared by additionally introducing a thin film distillation process after supercritical extraction, and the prepared red ginseng oil has an improvement in prostate enlargement compared to red ginseng oil prepared by supercritical extraction or oils prepared by solvent extraction. great.

1 shows the blood DHT measurement results of the red ginseng oil treatment group (experimental group) of Preparation Example 1-2 of the present invention.
Figure 2 shows the DHT measurement results of the prostate tissue of the red ginseng oil treatment group (experimental group) of Preparation 1-2 of the present invention.
3 shows the blood 5-alpha reductase II measurement results of the red ginseng oil treatment group (experimental group) of Preparation Example 1-2 of the present invention.
4 shows the measurement results of 5-alpha reductase I in the prostate tissue of the red ginseng oil treatment group (experimental group) of Preparation Example 1-2 of the present invention.
5 shows the measurement results of 5-alpha reductase II in the prostate tissue of the red ginseng oil treatment group (experimental group) of Preparation Example 1-2 of the present invention.
6 shows the measurement results of prostate-specific antigen (PSA) in the blood of the red ginseng oil treatment group (experimental group) of Preparation Example 1-2 of the present invention.
7 shows the pathological analysis results of the prostate tissue of the red ginseng oil treatment group (experimental group) of Preparation 1-2 of the present invention.
8 shows the results of protein expression (androgen receptor, Bax, Bcl-2 and TGF-β) in the prostate tissue of the red ginseng oil treatment group (experimental group) of Preparation Example 1-2 of the present invention.
9 shows the measurement results of cell activity of BPH-1 cells of the red ginseng oil treatment group (experimental group) of Preparation Example 1-2 of the present invention.
10 shows the protein expression (Bax, Bcl-2 and TGF-β) results of BPH-1 cells of the red ginseng oil treatment group (experimental group) of Preparation Example 1-2 of the present invention.
11 shows the mRNA expression results of androgen receptor in BPH-1 cells of the red ginseng oil treatment group (experimental group) of Preparation Example 1-2 of the present invention.
1 to 6 and 8 to 11 , the result values are expressed as mean±standard deviation. According to Duncan's multiple range test, values that do not have the same letter are significantly different from each other (p<0.05).

Hereinafter, the present invention will be described in detail.

The present invention comprises the steps of 1) extracting red ginseng with a supercritical fluid; And 2) thin film distillation of the extract obtained in step 1); provides a pharmaceutical composition for preventing or treating benign prostatic hyperplasia comprising red ginseng oil prepared by a method comprising.

Supercritical fluid extraction is an extraction method in which the principles of solvent extraction and distillation are applied in combination. method of extraction using . In general, since the supercritical fluid has a property of changing solubility and density according to changes in temperature and pressure, the activity of the extract may also vary accordingly. As such, the extraction method using a supercritical fluid enables the setting of high-density conditions by manipulation of pressure and temperature. Therefore, there is an advantage that the loss of natural active ingredients due to heat can be prevented.

In the present invention, as a supercritical fluid, carbon dioxide, which has a critical point close to room temperature, is non-toxic and non-flammable, and is very inexpensive. Supercritical fluid extraction conditions were adjusted in order to enhance the improvement effect of prostatic hyperplasia of red ginseng oil, which is the final product. For example, temperature, time, and pressure were adjusted during supercritical extraction. The conditions are described in detail below.

When extracting with the supercritical fluid, the temperature may be 60° C. to 90° C., preferably 65° C. to 85° C., and more preferably 70° C. to 80° C., but is not limited thereto. If the temperature during extraction is less than the lower limit, the yield of the extract decreases, and when the upper limit is exceeded, the effect of improving BPH is low even if the thin film distillation process is applied to the obtained extract, and the increase in production cost is significant compared to the degree to which the yield is increased. The disadvantage is that the cost of the product increases.

When extracting with a supercritical fluid, the extraction time may be 0.5 to 10 hours, preferably 1 to 8 hours, and more preferably 2 to 5 hours, but is not limited thereto. If the extraction time is less than the lower limit, the yield of the extract is reduced, and if it exceeds the upper limit, the improvement effect of BPH is low even if the thin film distillation process is applied to the obtained extract.

In step 1), the supercritical extraction conditions may maintain the pressure at 50 bar to 500 bar, preferably at 150 bar to 400 bar, more preferably at 200 bar to 350 bar, but is not limited thereto. . When the pressure condition is maintained below the lower limit, the yield of the extract decreases even if the extraction time is extended, and when the pressure condition is maintained above the upper limit, the production cost is consumed more than necessary to maintain the high pressure. In addition, there is a problem in that the purification effect of the obtained extract is lowered, the quality is lowered, and the improvement effect of the enlarged prostate is lowered.

Red ginseng oil of the present invention is obtained by applying a thin film distillation process to the extract obtained by supercritical extraction. Thin-film distillation may use a thin-film distillation generally used. Thin film distillation is a process of increasing the evaporation rate by maximizing the surface area of the mixture in contact with the heat source by making a thin film by physical force of the liquid mixture to be separated. The thin film distiller has a cylindrical heat transfer tube with a heating jacket attached thereto, and a rotating blade inside it, and the extract to be distilled is supplied from the top of the heat transfer cylinder and installed on the rotor It is distributed in the circumferential direction by a distributed distributor and flows down the heat transfer wall while forming a uniform and stable liquid film by the action of flowing the extract by the blades while being stirred by the rotor blades. In the meantime, components not related to the improvement of BPH in the extract are heated by the jacket heating source to vaporize and separate, and condense them to separate and remove them.

In the thin film distillation of step 2), the internal temperature of the distiller may be 150°C to 350°C, preferably 200°C to 300°C, and more preferably 230°C to 270°C, but is not limited thereto. does not If the internal temperature of the distiller is less than the lower limit, the effect of improving the prostate enlargement of red ginseng oil, which is the final product obtained even by applying the thin film distillation process after supercritical extraction, is low. When the internal temperature of the distiller exceeds the upper limit, there is a problem in that the active ingredient is lost.

In the thin film distillation of step 2), the vacuum degree may be 0.001 torr to 100 torr, preferably 0.01 to 50 torr, and more preferably 0.1 to 20 torr, but is not limited thereto. If the degree of vacuum is less than the lower limit, the improvement effect of red ginseng oil in prostate enlargement is low even if a thin film distillation process is applied after supercritical extraction. When the degree of vacuum exceeds the upper limit, there is a problem in that the purification effect is lowered and the quality is lowered, and the improvement effect of the obtained extract is low.

In the thin film distillation of step 2), the distillation time may be 1 hour to 48 hours, preferably 2 hours to 24 hours, and more preferably 4 hours to 10 hours, but is not limited thereto. . If the distillation time is less than the lower limit, the improvement effect of red ginseng oil is low even if the thin film distillation process is applied after supercritical extraction. If the distillation time exceeds the upper limit, the quality is lowered due to the change in the ingredients, so the improvement effect of red ginseng oil is low.

In the thin film distillation of step 2), the input rate of the extract may be 0.1 Hz to 100 Hz, preferably 5 Hz to 60 Hz, and more preferably 10 Hz to 30 Hz, but is not limited thereto. If the input rate of the extract does not satisfy the above range, that is, if the input rate of the extract is less than the lower limit, the thin film is not properly formed, and the red ginseng oil, the final product obtained by applying the thin film distillation process after supercritical extraction, improves prostate enlargement is low If the input rate of the extract exceeds the upper limit, there is a problem in that the purification effect is lowered and the quality is lowered, and the improvement effect of the obtained extract is low.

The red ginseng oil may be obtained by performing a conventional fractionation process in addition to increase the effect of improving the prostatic hyperplasia to obtain a fraction. For example, a fraction obtained by passing red ginseng oil according to the present invention through an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (prepared for separation according to size, charge, hydrophobicity or affinity) The active fractions obtained through various purification methods additionally carried out such as, etc. are also included in the red ginseng oil of the present invention. Red ginseng oil, in which several components are mixed, can be separated according to the properties of the active ingredients through concentration gradient column chromatography, etc. to prepare a specific active fraction having a higher improvement in prostate enlargement. Column chromatography may be performed several times by selecting an appropriate filler as necessary, but is not limited thereto. In using the chromatography, the elution solvent, elution rate, and elution time may be applied to a solvent, rate, or time generally used in the art.

A pharmaceutical composition for preventing or treating benign prostatic hyperplasia of the present invention will be described.

As used herein, the term “prevention” refers to any action that suppresses or delays the onset of BPH by administration of the pharmaceutical composition according to the present invention. As used herein, the term "treatment" refers to any action in which the symptoms of benign prostatic hyperplasia are improved or beneficially changed by administration of the pharmaceutical composition according to the present invention.

The pharmaceutical composition of the present invention may further contain, in addition to the red ginseng oil, other ingredients that can give a synergistic effect to the effect of the red ginseng oil, preferably within a range that does not impair the effect of the present invention. have. For example, it may contain conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances, or carriers.

The route of administration of the pharmaceutical composition includes oral, intravenous, intramuscular, intraarterial, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual or rectal, for example, topical application by application. method can be applied. The parenteral includes subcutaneous, intradermal, intravenous, intramuscular, intralesional injection or infusion techniques.

The pharmaceutical composition of the present invention may be administered in a pharmaceutically effective amount.

In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level includes the subject type and severity, age, sex, type of disease, The activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the rate of excretion, the duration of treatment, factors including concurrent drugs, and other factors well known in the medical field can be determined according to factors. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. and may be administered single or multiple. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, and can be easily determined by a person skilled in the art.

The pharmaceutical composition of the present invention is not particularly limited as long as it is an individual for the purpose of improving the symptoms of benign prostatic hyperplasia, and any one is applicable. For example, non-human animals such as monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cattle, sheep, pigs, goats, etc., humans, birds and fish can be used.

The pharmaceutical composition may contain one or more active ingredients having the same or similar function in addition to the red ginseng oil. The composition may be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, and syrups, external preparations, suppositories, and sterile injection solutions according to conventional methods, respectively.

Solid preparations for oral administration include powders, granules, tablets, capsules, soft capsules, pills, and the like. Liquid formulations for oral administration include suspensions, solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have.

Formulations for parenteral administration include powders, granules, tablets, capsules, sterilized aqueous solutions, solutions, non-aqueous solutions, suspensions, emulsions, syrups, suppositories, aerosols, etc. It can be formulated and used in the form, and preferably a pharmaceutical composition for external use on the skin of a cream, gel, patch, spray, ointment, warning agent, lotion, liniment agent, pasta agent or cataplasma can be prepared and used. , but is not limited thereto. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used.

The composition may further contain adjuvants such as preservatives, stabilizers, wetting agents or emulsification accelerators, salts and/or buffers for regulating osmotic pressure, and other therapeutically useful substances, and can be mixed, granulated or It can be formulated according to the coating method.

The dosage of the pharmaceutical composition may vary depending on various factors including the age, weight, general health, sex, administration time, administration route, excretion rate, drug formulation, and severity of a specific disease of the individual.

In addition, the pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers. In addition, the pharmaceutical composition may be administered at a dosage within the range of 0.001 to 200 mg/kg based on an adult, and when the pharmaceutical composition is for external use, once a day to 5 times a day in an amount of 1.0 to 3.0 ml based on an adult It is preferable to apply it once and continue it for 1 month or more, but the dosage is not limited to the scope of the present invention.

When the pharmaceutical composition is formulated in a unit dosage form, the red ginseng oil of the present invention as an active ingredient is preferably contained in a unit dose of about 0.01 to 1,500 mg, and the dosage required for adult treatment depends on the frequency and intensity of administration. Therefore, the range of about 1 to 500 mg per day is usually, but not limited to, a higher daily dose may be desirable for some patients.

In one embodiment of the present invention, it was confirmed that, after supercritical extraction, when red ginseng oil to which a thin film distillation process was applied was treated to an enlarged prostate cell model, the proliferation of prostate cells was inhibited (see Table 1). In addition, when the same sample is treated in an enlarged prostate animal model, the weight of the prostate is reduced and the content of DHT involved in prostate cell proliferation is reduced in blood and prostate tissue without increasing ALT and AST in blood, 5- Alpha reductase I and II are decreased in the blood and prostate tissue, prostate-specific antigen in the blood is decreased, the thickening of prostate epithelial cells is decreased, the size of the lumen is maintained normally, androgen receptor in the prostate tissue is decreased and apoptosis is decreased. It was confirmed that the expression of proteins involved in the phenomenon is regulated (reduction of Bcl-2 and TGF-β, increase of Bax) (see Table 2, Table 3, and FIGS. 1 to 8). In addition, when the red ginseng oil of the present invention is treated with enlarged prostate cells (BPH-1), the cell activity of BPH-1 is reduced, the expression of proteins involved in apoptosis shows results consistent with animal experiments, and androgen It was confirmed that the mRNA expression of the receptor was also reduced (see FIGS. 9 to 11 ).

In addition, the present invention comprises the steps of 1) extracting red ginseng with a supercritical fluid; And 2) thin film distillation of the extract obtained in step 1); provides a health food composition for improving prostate enlargement comprising a red ginseng oil prepared by a method comprising.

The manufacturing method of red ginseng oil is the same as described in the "pharmaceutical composition for preventing or treating enlarged prostate including red ginseng oil ", and thus description is omitted.

The red ginseng oil is preferably contained in an amount of 0.005 to 20.0% by weight based on the total weight of the health functional food, and more preferably contained in an amount of 0.01 to 10.0% by weight, but is not limited thereto. When the effective content of the red ginseng oil is less than 0.005% by weight, it is not possible to alleviate the symptoms of benign prostatic hyperplasia, and when it exceeds 20% by weight, the apparent synergistic effect according to the content increase is lowered, which is uneconomical.

The health functional food may further contain, in addition to the red ginseng oil of the present invention, other ingredients that can give a synergistic effect to the effect of the red ginseng oil, preferably within the range that does not impair the effect of the present invention. have. For example, it may contain conventional adjuvants such as stabilizers, solubilizers, vitamins, pigments and fragrances, or carriers.

In addition, it may include ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. Examples of the carbohydrate include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent, a natural flavoring agent taumatin, a stevia extract, and a synthetic flavoring agent may be used. For example, when the health functional food of the present invention is manufactured as a drink, citric acid, high fructose, sugar, glucose, acetic acid, malic acid, fruit juice, natural extract, etc. may be additionally included in addition to the red ginseng oil of the present invention.

There is no particular limitation on the type of the health functional food. Examples of foods to which the red ginseng oil of the present invention can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea , drinks, alcoholic beverages, and vitamin complexes, and includes all health functional foods in the ordinary sense.

Hereinafter, the present invention will be described in detail by way of Preparation Examples and Examples.

However, the following Preparation Examples, Experimental Examples, and Examples are only for illustrating the present invention, and the content of the present invention is not limited by the following Preparation Examples and Examples.

<Preparation Example 1> Preparation of extract

<1-1> Red ginseng oil (after supercritical extraction, thin film distillation process is not applied)

After putting 1.0 kg of red ginseng (Panax ginseng CA Meyer, root) powder into a supercritical extraction tank (Natex), close the extraction tank lid, add liquid carbon dioxide (CO ₂ ), and extract 10.3 g of red ginseng oil at 250 bar and 75° C. for 3 hours. was obtained.

<1-2> Red ginseng oil (after supercritical extraction, thin film distillation process is applied)

After putting 1.0 kg of red ginseng (Panax ginseng CA Meyer, root) powder into a supercritical extraction tank (Natex), close the lid of the extraction tank, add liquid carbon dioxide (CO ₂ ), and extract 10.3 g of oil at 250 bar and 75° C. for 3 hours. obtained. The oil extracted through the supercritical extractor was distilled at a vacuum degree of 1 torr for 6 hours at a temperature of 240° C. inside the distiller using a distillation device (VDL 70-4), and the input rate of the extract was set to 15 Hz, and then purified red ginseng oil 5.5 g was obtained.

<1-3> Red Ginseng Oil (Hexane Extract)

Put 1.0 kg of red ginseng (Panax ginseng CA Meyer, root) powder in a stirring tank (10L), add 10L of hexane (Daejeonghwageum, 95%), stir for 4 hours (HT-DX), and extract 3 times, After collecting the extract extracted in each round, the extract was filtered using a 10 μm filter. The filtrate was concentrated at 40-50° C. using a rotary vacuum concentrator (BUCHI, R-220) to obtain 18.3 g of red ginseng oil.

<1-4> Ginseng seed (Hexane extraction)

Put 100 g of ginseng seed (Panax ginseng CA Meyer, fruit) powder in a flask (2L), add 1L of hexane (Daejeonghwageum, 95%), stir for 4 hours (HT-DX), and extract 3 times, After collecting the extract extracted in each round, the extract was filtered using a 10 μm filter. The filtrate was concentrated at 40-50° C. using a rotary vacuum concentrator (BUCHI, R-220) to obtain 21.4 g of ginseng seed oil.

<Example 1> Confirmation of the improvement of benign prostatic hyperplasia - In vitro confirmation

<1-1> Cell culture and cell proliferation inhibitory ability measurement

Testosterone produced in the testis moves into prostate cells and is converted to dihydrotestosterone (DHT) by 5-alpha-reductase, and the converted DHT binds to androgen receptor (AR) and moves into the nucleus. and induce cell proliferation. Therefore, since BPH develops through this series of processes, in this example, a change in the inhibition of proliferation of prostate cells in relation to BPH was confirmed. Inhibition of the DHT-induced PSA secretion by Verbascum xanthophoeniceum and Serenoa repens extracts in human LNCaP prostate epithelial cells. J Ethnopharmacol. 2014 8;155 reported by Marcoccia D et al. (1):616-25) was applied and the experiment was conducted as follows.

LNCaPFGC (Androgendependent Prostate Cancer) cells, a prostate cancer cell line with androgen receptors, were purchased from the Korea Cell Line Bank, and RPMI1640 (Gibco, Grand Island, NY, USA) medium was used, 5% CO ₂ , and cultured at 37° C. in a cell incubator.

LNCaPFGC ^{The cell line was diluted to a concentration of 2×10 4} cells/well in a 96-well plate, aliquoted, and cultured for 24 hours. LNCaPFGC cells untreated with the sample were treated as a normal group, and DHT (Tokyo chemical, Tokyo, Japan) was added to a concentration of 10 nM. LNCaPFGC cells treated with was set as a negative control, saw palmetto extract (Daehan chemtech co., LTD, Seoul, Korea) was set as a positive control. Each of the samples prepared in the above preparation examples were analyzed by concentration (0, 50, 100, 200). μg) to the cells for 48 hours. After that, the culture medium is removed, and the Cyto X (Cell viability assay kit, LPS solution, Daejeon, Korea) solution is diluted 10% in the culture medium, 100 μl of each is added, and then reacted in a cell incubator at 37° C. for 30 minutes with a multi-plate reader (Multiskan). Go, Thermo Scientific, MA, USA) and the absorbance was measured at 450 nm. Cell proliferation inhibitory ability was compared with the absorbance after sample treatment of the corresponding sample treatment group when the absorbance after sample treatment of the negative control group, that is, the DHT treatment group was 100%, [cell proliferation rate (%) = (sample after DHT treatment) Absorbance of treatment group)/(absorbance of negative control after DHT treatment)×100]. A value of 100 or more indicates an increase in the proliferation of the prostate cancer cell line after sample treatment, and a value of 100 or less indicates that the proliferation of the prostate cancer cell line is suppressed after the sample treatment.

<1-2> Confirmation of efficacy of the extracts prepared in Preparation Examples 1-1 to 1-4 - In vitro confirmation

The effects of treatment with the extracts prepared in Preparation Examples 1-1 to 1-4 alone at a concentration of 100 μg/ml were confirmed. The degree of cell proliferation compared to the negative control is shown in Table 1 below.

normal group Negative control group (DHT-treated group) Positive control group (saw palmetto extract treatment group) Red ginseng oil of Preparation Example 1-1 (thin film distillation not applied) Red ginseng oil of Preparation Example 1-2 (thin film distillation applied) Red ginseng oil of Preparation Example 1-3 (Hexane extraction) Ginseng seed oil of Preparation Example 1-4 (Hexan extraction) Cell proliferation rate (%) 76.6 100 91 116 79 103 118

As a result, in the case of the negative control group treated with DHT, cell proliferation was increased compared to the normal group. Among the extracts of Preparation Examples 1-1 to 1-4 obtained by different extraction methods of ginseng-derived raw materials, the extract that significantly inhibited cell proliferation compared to cell proliferation after DHT treatment of the negative control group was the red ginseng of Preparation Example 1-2. It was an oil (applied by thin film distillation after supercritical extraction), and it was found to be superior to the effect of saw palmetto extract, which is used to improve prostate enlargement. Due to such proliferation inhibition, the cell proliferation of the group treated with the red ginseng oil (applied by thin film distillation after supercritical extraction) of Preparation Example 1-2 showed a tendency to be inhibited to the extent of cell proliferation of the normal group not treated with DHT.

On the other hand, the red ginseng oil of Preparation Example 1-1 (thin film distillation not applied after supercritical extraction) had more than 16% cell proliferation compared to the negative control, and the red ginseng oil of Preparation Example 1-2 (after supercritical extraction) in which cell proliferation was inhibited. Thin film distillation applied) showed an increase in cell proliferation by more than 37% compared to the treated group, indicating that it would not be suitable for use for the purpose of improving BPH. And, when the extract obtained by using hexane as a solvent, such as the red ginseng oil of Preparation Example 1-3 and the ginseng seed oil of Preparation Example 1-4, was treated, cell proliferation was increased compared to the negative control.

<Example 2> Confirmation of the improvement of benign prostatic hyperplasia - in vivo confirmation

<2-1> Induction of benign prostatic hyperplasia in experimental animals

In order to confirm the improvement effect of prostate enlargement in the prostate enlargement induction animal model of the red ginseng oil of the present invention obtained in Preparation Example 1-2, the literature reported to Shin IS et al. (Ursolic acid reduces prostate size and dihydrotestosterone level in a rat model of benign) Prostatic hyperplasia, Food Chem Toxicol., 2012, 50, 884-888) by applying the method disclosed in the experiment as follows. The rats were castrated to exclude the effect of internal testosterone, and testosterone propionate (TP, T0028, TCI, JAPAN) was injected subcutaneously for 8 weeks in the castrated rats to induce an enlarged prostate, TP After oral administration of the red ginseng oil of Preparation Example 1-2, and finasteride used as a treatment for benign prostatic hyperplasia, respectively, along with the administration of , the effect of improving prostatic hyperplasia was confirmed.

7-week-old male Sprague Dawley rats weighing 260 to 300 g were supplied from Coretech (Pyeongtaek, Korea) and used after acclimatization for 7 days. 55 ± 5%) and light-dark cycle (12 h) were automatically adjusted. Castration was performed to rule out the effect of internal testosterone in the rats. After inhalation anesthesia, male SD rats were fixed with the back facing the operator except for the normal group, and then the skin at the end of the scrotum was incised to cut off the left and right testicles and epididymis, and the incision was sutured to perform surgery. After castration was performed and stabilization for 10 days, the wound at the surgical site was healed, and then the experiment was started by administering TP to the castrated rat. Six animals were assigned to each group with almost the same weight for each group based on body weight. Except for the normal group, the remaining experimental animals (negative control group, positive control group, and experimental group) were administered subcutaneous injection of TP at 3 mg/kg/day at 3 mg/kg/day according to the change in body weight until the end of the experiment for 8 weeks. did. In order to minimize the error of the experiment, the total injection volume based on the weight measured every 3 days was dissolved in the vehicle corn oil so that 100 μl was used, and the normal group was injected with only corn oil 100 μl without TP.

The negative control group is a group administered external testosterone, excluding the effect of internal testosterone. The positive control group is a group administered with finasteride (Finasteride, F1293, Sigma, USA), which excludes the influence of internal testosterone, externally administers testosterone, and exhibits a therapeutic effect on benign prostatic hyperplasia. Finasteride was dissolved in corn oil as a vehicle at a concentration of 10 mg/kg/day and administered orally with TP administration for 8 weeks. The experimental group excludes the effect of internal testosterone, administers testosterone from the outside, and administers the red ginseng oil of Preparation Example 1-2. In order to confirm the effect of the red ginseng oil of the present invention, red ginseng oil was administered in an amount of 25 mg/kg/day, 50 mg/kg/day, 100 mg/kg/day and 200 mg/kg/day for 8 weeks of TP administration, Oral administration was performed along with TP administration for 8 weeks by dissolving in corn oil as a vehicle. After 8 weeks administration, each group was sacrificed to measure prostate weight, blood ALT and AST measurement, blood and prostate tissue DHT measurement, 5-alpha reductase I and II measurement, prostate-specific antigen measurement, prostate histological analysis, Protein expression analysis of prostate tissue was performed.

<2-2> Prostate weight measurement

The prostate was removed by incision of the abdomen of the sacrificed rat, washed once in PBS, and then lightly drained to measure the weight. The prostate ratio is calculated as prostate weight (mg)/weight (100g) × 100 Thus, it is shown in Table 2 below.

army Prostate weight (g) Prostate Ratio
(Prostate weight (mg) / Weight (g) * 100) normal group 0.52±0.03 ^d 0.11±0.01 ^d negative control 0.96±0.05 ^a 0.22±0.01 ^a Experimental group 25 0.94±0.07 ^a 0.21±0.02 ^a Experimental group 50 0.90±0.04 ^ab 0.21±0.01 ^a Experimental group 100 0.79±0.03 ^bc 0.18±0.01 ^b Experimental group 200 0.76±0.04 ^c 0.18±0.01 ^b positive control 0.59±0.03 ^d 0.14±0.01 ^c

In Table 2, the results are expressed as mean±standard deviation. According to Duncan's multiple range test, values that do not have the same letter are significantly different from each other (p<0.05).

The prostate weight of the negative control group, which induced BPH, was significantly increased compared to the normal group, and the prostate weight of the positive control group was reduced by 39% to the level of the normal group. In the case of the experimental group, the weight of the prostate was significantly reduced by about 20% in the experimental group 100 and the experimental group 200. In the prostate ratio, which represents the weight of the prostate to body weight, a significant decrease of 19% was observed in the experimental group 100 and the experimental group 200.

Prostate weight is used as a major indicator in the development of benign prostatic hyperplasia. Therefore, from the above results, the experimental group 100 and the experimental group 200 cause a significant decrease in the size of the prostate, and it can be inferred that this is the result of alleviating the development of enlarged prostate.

<2-3> Blood ALT and AST measurement

The liver index (mg/weight 100g) was calculated by calculating the weight of the liver extracted by incision of the abdomen of the sacrificed rat per 100 g of body weight.

In addition, to check the blood levels of aspartate aminotransferase (AST) and alanine aminotrasferase (ALT) as indicators of liver damage, rat blood was collected, centrifuged at 3,000 rpm for 20 minutes, and the serum fraction was collected and used for analysis. . Analysis of AST and ALT in serum was measured using a kit from Asan Pharmaceutical (Seoul, Korea), and is shown in Table 3 below in Karmen units according to a standard curve.

army liver index
(mg/weight 100g) ALT
(karmen) AST
(karmen) normal group 3.82±0.14 ^a 67.01±3.81 ^a 96.30±5.44 ^b negative control 4.06±0.16 ^a 69.53±5.47 ^a 110.11±3.98 ^b Experimental group 25 3.68±0.08 ^a 62.83±4.70 ^ab 133.67±9.02 ^a Experimental group 50 3.99±0.14 ^a 60.23±1.98 ^ab 98.58±3.80 ^b Experimental group 100 3.93±0.23 ^a 59.57±6.26 ^ab 97.87±4.68 ^b Experimental group 200 3.90±0.24 ^a 63.73±7.77 ^ab 107.11±9.39 ^b positive control 3.59±0.16 ^a 50.41±2.31 ^b 95.71±4.31 ^b

In Table 3 above, the results are expressed as mean ± standard deviation. According to Duncan's multiple range test, values that do not have the same letter are significantly different from each other (p<0.05).

From the above results, it was confirmed that there was no significant difference in liver index between the normal group, the negative control group, the positive control group and the experimental group. In addition, there was no significant difference between the normal group, negative control group, positive control group, and experimental group in AST and ALT indicators indicating liver toxicity. However, AST was significantly increased in experimental group 25, but it is within a safe range, so it does not give much significance to safety.

<2-4> Measurement of DHT in blood and prostate tissue

In order to measure changes in blood DHT, the level of DHT was measured in the serum of sacrificed rats using an ELASA kit (BioVendor, Brno, Czech Republic). Briefly, 50 μl of serum was incubated with HRP-conjugated DHT and DHT-specific antibody in 96-well microplates pre-coated with goat-anti-rabbit antibody for 1 hour at room temperature with agitation. After incubation, the substrate solution was added, and the reaction was stopped with sulfuric acid solution after 15 minutes, and the difference was measured at 450 nm within 10 minutes, and the results are shown in FIG. 1 . The measurement value was calculated using a standard graph drawn with the DHT standard solution, and the unit was expressed as pg/mL.

In addition, for DHT measurement in prostate tissue, PBS with a pH of 7.4, 4 times the tissue weight, was added and homogenized, centrifuged at 4 ° C. 5000 rpm for 5 minutes, and then the supernatant was taken and DHT was measured in the same manner as in blood. 2 is shown.

In the results of Figure 1, compared with the normal group, the negative control group showed a significant increase as 1893.9 ± 109.3 pg/mL, and in the case of the experimental group, the concentration-dependent decrease was 1506.7 ± 120.9 pg/mL and 1506.7 ± 120.9 pg/mL in the experimental group 100 and 200, respectively. It was 1507.1 ± 177.1 pg/mL, which was significantly different from the negative control group (p<0.05). In the case of the positive control group, it was 1028.4 ± 139.5 pg/mL, which was significantly different from the negative control group (p<0.05), which was significantly lower than that of the experimental groups 100 and 200.

In addition, as a result of comparing the DHT level in the prostate tissue from FIG. 2 , compared with the normal group, the negative control group was significantly higher as 1005.9 ± 62.3 pg/mL (p<0.05), and the experimental group 100 and the experimental group 200 were each 698.9 ± 77.9 and 712.6 ± 89.9 pg/mL, which was significantly lower than that of the negative control group (p<0.05), which was not significantly different from that of the positive control group of 736.0 ± 51.8 pg/mL.

Such a decrease in DHT is considered to be a result of reducing the size of the prostate by inhibiting the proliferation of prostate cells, and the results of the decrease in the size of the prostate can be confirmed in Table 2 above.

<2-5> Measurement of 5-alpha reductase I and II

In the serum of sacrificed rats, 5-alpha reductase II was measured using a 5-alpha reductase II ELISA kit (Cusabio, Wuhan, Hubei, China), and the results are shown in FIG. 3 .

In addition, the prostate tissue was treated to reduce 5-alpha using the 5-alpha reductase I ELISA kit (Cusabio, Wuhan, Hubei, China) and the 5-alpha reductase II ELISA kit (Cusabio, Wuhan, Hubei, China), respectively. Enzymes I and 5-alpha reductase II were measured, and the results are shown in FIGS. 4 and 5, respectively. For the treatment of the prostate tissue, 100 mg of prostate tissue was homogenized in 1 mL of PBS, and then frozen at -20 ° C for overnight, and after two freeze-thaw cycles to obtain a homogenate in which the cell membrane was destroyed, centrifuged at 5000 x g for 5 minutes. One supernatant was used for analysis.

3, the negative control group showed a significant increase in serum 5-alpha reductase II compared to the normal group, and the experimental group 25 and the experimental group 50 had no significant difference from the negative control group, but the experimental group 100 (1760.3 ± 229.1 pg/ mL) and experimental group 200 (1693.2 ± 304.4 pg/mL) showed a significant difference (p<0.05), which was similar to the positive control group (1778.0 ± 164.7 pg/mL).

In FIG. 4, 5-alpha reductase I in the prostate tissue was increased to 613.3 ± 65.6 pg/mL in the negative control group compared to the normal group at 458.5 ± 66.5 pg/mL, but no significant difference was observed, except for the low concentration of 25. and 400.6 ± 35.3, 395.4 ± 42.1, and 379.6 ± 36.3 pg/mL in experimental group 50, experimental group 100, and experimental group 200, respectively, which were significantly lower than in the negative control group (p<0.05).

In FIG. 5 , 5-alpha reductase II in the prostate tissue increased with a significant difference to 351.5 ± 36.3 pg/mL in the negative control group compared to the normal group (75.5 ± 12.7 pg/mL), and 240.8 in the positive control group. It showed a significant decrease compared to the negative control group at ± 24.8 pg/mL (p<0.05). In the case of the experimental group, except for the experimental group 25 and the experimental group 50, which were low concentrations, the experimental group 100 and the experimental group 200 showed a significant decrease to 213.0 ± 36.5 and 221.1 ± 31.8 pg/mL, respectively (p<0.05).

In benign prostatic hyperplasia, 5-alpha reductase I and II are overexpressed, of which 5-alpha reductase II is the dominant type. In benign prostatic hyperplasia, 5-alpha reductase II is an enzyme that promotes the conversion of testosterone to DHT. From the above results, the experimental group reduced 5-alpha reductase I in the prostate tissue to the level of the positive control group from the experimental group 50 to the experimental group 200, and the 5-alpha reductase II in the blood and prostate tissue was effectively reduced in the experimental group 100 and the experimental group 200. , which appears to be the result of decreased prostate weight and decreased DHT levels.

<2-6> Measurement of Prostate Specific Antigen

After 200-fold dilution of the serum separated from the sacrificed rats, a PSA ELISA kit (Cusabio) was used to measure the prostate-specific antigen (PSA), and the results are shown in FIG. 6 . After the analysis was performed, the values measured at 450 nm were calculated based on the standard curve and a graph was prepared.

6, the negative control group (15.96 ± 2.35 pg/mL) was significantly higher than the normal group (9.76 ± 0.90 pg/mL), and in the case of the experimental group, 11.45 ± 0.88 pg/mL in the experimental group 50 and 9.59 in the experimental group 100, respectively. ± 0.90 pg/mL, 8.30 ± 1.03 pg/mL in experimental group 200, which was significantly lower than that of the negative control group (p<0.05). No significant difference was found.

PSA is a glycoprotein produced in small amounts by prostate cells and is used as a semi-quantitative indicator of prostate volume in BPH or prostate cancer, and elevated blood PSA levels are often observed in BPH patients.

Although the exact mechanism between BPH and blood PSA level is not known, PSA level tends to rise according to prostate size, and correlation between prostate disease including BPH and blood PSA level has been steadily reported. The elevated blood PSA level is thought to be due to the reduction in the size of the enlarged prostate due to the enlarged prostate.

<2-7> Histological analysis of prostate

After immersing the prostate tissue in 10% formalin solution, making a paraffin section with a thickness of 4 μm, dewaxing and dehydration, and staining and analyzing the tissue with hematoxylin and eosin (H&E) for general histopathological examination. It was photographed at a magnification of 200, and the results are shown in FIG. 7 .

7, it can be observed that the epithelial cells of the prostate tissue of the negative control group (B) are thickened and the lumen is narrowed compared to the normal group (A). In the case of the positive control group (G), compared with the negative control group, the epithelial cell thickening was decreased, and it was confirmed that recovery was similar to that of the normal group (A). Experimental group 25(C), Experimental group 50(D), Experimental group 100( E), in the experimental group 200 (F), the epithelial cell thickening was decreased compared to the negative control group (B), and it was confirmed that the lumen was maintained to a similar degree to that of the normal group.

<2-8> Protein expression analysis of prostate tissue

DHT binds to AR present in the nuclei of prostate epithelial and strut cells and induces structural changes and dimerization. It binds to and induces prostate growth. Elevated levels of nuclear AR have been reported in thickened prostate tissue compared to normal prostate.

In addition, apoptosis is essential for maintaining homeostasis, and apoptosis occurs more frequently in the prostate epithelium than in the case of enlarged prostate in normal prostate. In the case of an enlarged prostate, it is known that apoptosis occurs 4 times less than in the normal prostate, and it is known that Bcl-2, which inhibits apoptosis, increases and causes prostate growth. Bax is a pro-apoptotic protein that stimulates the release of cytochrome c from mitochondria and promotes apoptosis. Normal prostate tissue exhibits little Bcl-2 expression in epithelial cells, but Bcl-2 expression increases and Bax expression tends to decrease in an enlarged prostate.

Western blot was performed to analyze the expression of androgen receptor (AR) and Bax, Bcl-2 and TGF-β in prostate tissue. carried out. The prostate tissue was homogenized and centrifuged at 3000 rpm for 6 minutes to collect the supernatant, and the protein was quantified by the Bradford method. Electrophoresis was performed on 8, 10, 15% SDS gel based on 50 μg of protein between each group, and Western blot was performed. After transfer to the nitrofibrin membrane, blocking was performed at 4° C. with 5% skim milk for 16 hours, and each antibody was used and printed on an X-ray film, and the results are shown in FIG. 8 s. The antibodies used were as follows; Bcl-2 (Santa Cruz Biotechnology, Dallas, Texas, United States), Bax (Cell Signaling Technology, Danvers, Massachusetts, United States), TGF-β1 (Santa Cruz Biotechnology, Dallas, Texas, United States), androgen receptor (Sigma) -Aldrich, St. Louis, MO, United States) were all diluted 1:1000 and used.

8 , in the case of Bax, the expression of the experimental group 100 and the experimental group 200 was higher than that of the negative control group. In the case of Bcl-2, the expression in the negative control group was significantly increased compared to the normal group, and it showed a tendency to decrease again in the experimental group 25, the experimental group 50, the experimental group 100 and the experimental group 200.

TGF-β, which is regulated by Bcl-2 and Bax apoptosis genes, is balanced in normal prostate by inhibition through apoptosis mechanism. A theory has been proposed that the proliferation of cells is promoted, and the abnormal imbalance between the proliferation and apoptosis of these prostate cells is thought to be the main cause of inducing prostate growth. In FIG. 8 , it can be seen that TGF-β was increased in the negative control group, but the expression was significantly decreased in the experimental group.

In addition, AR increased in BPH, and its expression tended to decrease in a concentration-dependent manner.

<Example 3> Confirmation of the improvement of benign prostatic hyperplasia - In vitro confirmation

<3-1> Cell activity of enlarged prostate cells

BPH-1, an enlarged prostate cell line, was treated with RPMI1640 (Gibco, Grand Island, NY, USA) medium containing 10% FBS (Fetal Bovine Serum) and antibiotics (Penicillin/Streptomycin 100U) at 37°C, 5% CO ₂ , Cultured under 95% humidity atmosphere conditions.

BPH-1 cells were seeded into 96 wells at 8500 cells/well, and after the cells were implanted, a sample was added and incubated for 24 hours, followed by 5 mg/mL MTT reagent (3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide, Sigma) was added. After 2 hours, the medium was removed, washed once with PBS, and then 100 μl of DMSO was added to dissolve the formed formazan, followed by measurement at 570 nm. Cell activity was shown in FIG. 9 by calculating the relative value with 100% of the control group.

9 shows the concentration-dependent proliferation inhibitory effect of red ginseng oil administration in BPH-1 cells. At a concentration of 25 μg/ml, there was a significant decrease of about 15%, and at a concentration of 100 μg/ml, a decrease of about 50% (p<0.05). If it can have an effect on the control of BPH, it does not necessarily mean that BPH-1 cell proliferation is inhibited, but it can be predicted to be involved in the cycle regulation related to the cell cycle.

<3-2> Protein expression analysis of enlarged prostate cells

In order to analyze the expression of Bax, Bcl-2, and TGF-β in BPH-1 cells, Western blot was performed in the same manner as described in "<2-8> Protein expression analysis of prostate tissue" during the animal experiment. The results are shown in FIG. 10 .

In the case of BPH-1 cells, the expression of Bax protein was increased in a concentration-dependent manner from 50 μg/ml after treatment with red ginseng oil compared to the control group. In the case of Bcl-2 protein, a marked decrease in expression was observed from 100 μg/ml. In the case of TGF-β, a tendency to decrease expression was observed from 200 μg/ml. These results have the same tendencies as the results seen in the animals previously induced for enlarged prostate, confirming that red ginseng oil can control the main cause of inducing prostate growth through abnormal imbalance of apoptosis.

<3-3> Confirmation of mRNA expression of androgen receptor (AR)

Real-time PCR was performed in BPH-1 cells to confirm the mRNA expression of androgen receptor. First, RNA was extracted from BPH-1 cells using RNA iso (Takara, Ohtsu, Japan) and reverse transcribed with PrimeScript™ reverse transcriptase (Takara, Ohtsu, Japan) to synthesize cDNA. Next, real-time PCR was performed using a light cycler 96 (Roche, Mannheim, Germany), and the results are shown in FIG. 11 . The primer sequences used are as follows. AR (sense; CTC ACC AAG CTC CTG GAC TC (SEQ ID NO: 1), antisense; CAG GCA GAA GAC ATC TGA AAG (SEQ ID NO: 2)), GAPDH (sense; ATG TTC GTC ATG GGT GTG AAC (SEQ ID NO: 3), antisense: GCA TGG ACT GTG GTC ATG AGT (SEQ ID NO: 4)).

After treatment with red ginseng oil in BPH-1 cells, the mRNA expression of AR was decreased, and it showed a significant decrease compared to the control group not treated with red ginseng oil at all concentrations. This is a result consistent with the tendency for AR expression to decrease after treatment with red ginseng oil in animals previously induced by prostate enlargement, and shows that red ginseng oil can regulate the expression level of AR inducing prostate growth by binding with DHT.

<Preparation Example 2> Preparation of pharmaceutical formulations

<2-1> Preparation of powder

2 g of red ginseng oil of the present invention

1 g lactose

The above ingredients were mixed and filled in an airtight cloth to prepare a powder.

<2-2> Preparation of tablets

100 mg of red ginseng oil of the present invention

Corn Starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets were prepared by tableting according to a conventional method for manufacturing tablets.

<2-3> Preparation of capsules

100 mg of red ginseng oil of the present invention

Corn Starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above ingredients, the capsules were prepared by filling in gelatin capsules according to a conventional manufacturing method of capsules.

<2-4> Preparation of the ring

1 g of red ginseng oil of the present invention

1.5 g lactose

1 g of glycerin

0.5 g of xylitol

After mixing the above components, it was prepared so as to be 4 g per ring according to a conventional method.

<2-5> Preparation of granules

150 mg of red ginseng oil of the present invention

Soybean extract 50 mg

glucose 200 mg

Starch 600 mg

After mixing the above ingredients, 100 mg of 30% ethanol was added and dried at 60° C. to form granules, and then filled in a bag.

<Production Example 3> Preparation of health food

<3-1> Manufacturing of wheat flour food

0.5 to 5.0 parts by weight of the red ginseng oil of the present invention was added to wheat flour, and breads, cakes, cookies, crackers and noodles were prepared using this mixture.

<3-2> Preparation of soups and gravies

By adding 0.1 to 5.0 parts by weight of the red ginseng oil of the present invention to soup and broth, processed meat products for health promotion, soup and broth of noodles were prepared.

<3-3> Preparation of ground beef

Ground beef for health promotion was prepared by adding 10 parts by weight of red ginseng oil of the present invention to ground beef.

<3-4> Production of dairy products

5-10 parts by weight of red ginseng oil of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<3-5> Manufacture of wire type

Brown rice, barley, glutinous rice, and barley radish were pregelatinized by a known method and dried, and then roasted and prepared as a powder having a particle size of 60 mesh with a grinder.

Black soybeans, black sesame, and perilla were also steamed and dried by a known method, and then roasted and prepared as a powder having a particle size of 60 mesh with a grinder.

The red ginseng oil of the present invention was concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by drying with a spray and hot air dryer was pulverized to a particle size of 60 mesh with a pulverizer to obtain a dry powder.

It was prepared by mixing the grains, seeds and red ginseng oil of the present invention prepared above in the following ratios.

Grains (30 parts by weight of brown rice, 15 parts by weight of barley, 20 parts by weight of barley), seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame), red ginseng oil of the present invention (3 parts by weight), reishi (0.5 parts by weight), Rehmannia (0.5 parts by weight)

<Production Example 4> Preparation of beverage

<4-1> Manufacturing of health drinks

Instant sterilization is performed by homogeneously mixing 5 g of red ginseng oil of the present invention with auxiliary materials such as high fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%), and water (75%). Then, it was prepared by packaging it in a small packaging container such as a glass bottle or a plastic bottle.

<4-2> Preparation of vegetable juice

Vegetable juice was prepared by adding 5 g of red ginseng oil of the present invention to 1,000 ml of tomato or carrot juice.

<4-3> Preparation of fruit juice

Fruit juice was prepared by adding 1 g of red ginseng oil of the present invention to 1,000 ml of apple or grape juice.

<110> KOREA GINSENG CORP. <120> Pharmaceutical Composition Comprising Red Ginseng Oil for Preventing or Treating Prostatic Hyperplasia <130> 2019-DPA-3453 <150> KR 10-2018-0110636 <151> 2018-09-17 <160> 4 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Androgen Receptor primer sense <400> 1 ctcaccaagc tcctggactc 20 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Androgen Receptor primer anti-sense <400> 2 caggcagaag acatctgaaa g 21 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> GAPDH primer sense <400> 3 atgttcgtca tgggtgtgaa c 21 <210> 4 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> GAPDH primer anti-sense <400> 4 gcatggactg tggtcatgag t 21

Claims (13)

1) Supercritical extraction of red ginseng using carbon dioxide as a supercritical fluid; and
2) thin film distillation of the extract obtained in step 1); contains red ginseng oil prepared by a method comprising;
A pharmaceutical composition for preventing or treating benign prostatic hyperplasia that reduces the expression of androgen receptors. delete The method according to claim 1,
A pharmaceutical composition for preventing or treating benign prostatic hyperplasia, wherein the temperature is 60° C. to 90° C. when extracted with a supercritical fluid. The method according to claim 1,
A pharmaceutical composition for the prevention or treatment of benign prostatic hyperplasia, wherein the extraction time is 0.5 to 10 hours when extracted with a supercritical fluid. The method according to claim 1,
In the thin film distillation of step 2), the internal temperature of the distiller is 150° C. to 350° C. A pharmaceutical composition for preventing or treating benign prostatic hyperplasia. 6. The method of claim 5,
In the thin film distillation of step 2), the internal temperature of the distiller is 200° C. to 300° C. A pharmaceutical composition for preventing or treating benign prostatic hyperplasia. The method according to claim 1,
In the thin film distillation of step 2), the degree of vacuum is 0.001 torr to 100 torr of a pharmaceutical composition for preventing or treating benign prostatic hyperplasia. 8. The method of claim 7,
In the thin film distillation of step 2), the degree of vacuum is 0.01 torr to 50 torr of a pharmaceutical composition for preventing or treating benign prostatic hyperplasia. The method according to claim 1,
In the thin film distillation of step 2), the distillation time is 1 hour to 48 hours, a pharmaceutical composition for preventing or treating benign prostatic hyperplasia. 10. The method of claim 9,
In the thin film distillation of step 2), the distillation time is 2 hours to 24 hours, a pharmaceutical composition for preventing or treating benign prostatic hyperplasia. The method according to claim 1,
In the thin film distillation of step 2), the input rate of the extract is a pharmaceutical composition for preventing or treating benign prostatic hyperplasia carried out in a range of 0.1 Hz to 100 Hz. 12. The method of claim 11,
In the thin film distillation of step 2), the input rate of the extract is 5 Hz to 60 Hz, a pharmaceutical composition for preventing or treating benign prostatic hyperplasia. 1) Supercritical extraction of red ginseng using carbon dioxide as a supercritical fluid; and
2) thin film distillation of the extract obtained in step 1); contains red ginseng oil prepared by a method comprising;
A health food composition for improving enlarged prostate that reduces the expression of androgen receptors.

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