KR102408623B1 - Composition for preventing or treating prostatic diseases comprising Black Ginseng extract or Fermented Black Ginseng extract - Google Patents

Abstract

The present invention relates to a method of extracting an extract containing a functional ingredient for the prevention or treatment of prostate-related diseases from black ginseng and fermented black ginseng.
In detail, the method for producing a black ginseng extract according to the present invention includes the steps of putting dried ginseng into a steamer, raising the temperature to 80°C to 100°C, and steaming the steamed ginseng for 1 hour to 7 hours, and steaming the steamed ginseng at 40°C to 70°C for 3 hours to By repeating the step of drying for 8 hours and then cooling for 10 to 20 hours 3 to 5 times, the yield of total ginsenosides obtainable from black ginseng can be improved.
In addition, by fermenting the black ginseng extract prepared as described above using a Lactobacillus pentosus (ATCC8041) strain, it provides a fermented black ginseng extract excellent in preventing or treating prostate-related diseases.

Description

Composition for preventing or treating prostatic diseases comprising Black Ginseng extract or Fermented Black Ginseng extract, comprising black ginseng extract or fermented black ginseng extract as an active ingredient

The present invention relates to a composition for preventing or treating prostate disease comprising black ginseng or fermented black ginseng extract as an active ingredient. More specifically, it relates to a composition for preventing or treating prostate disease, comprising a black ginseng extract or fermented black ginseng extract with a further increased content of ginsenosides Rh1, Rg2, Rh4, Rg3, Rk1, Rg5, Rh2, and Rh3.

The prostate is an accessory gonad in men composed of glandular tissue and fibromuscular tissue, and although lumps or stones may occur, prostatitis, enlarged prostate, and prostate cancer are the most common diseases. Prostate cancer is one of the most common malignant tumors in men, and it is one of the most common cancers among men in the West. Occurs in people older than three years of age. According to the central cancer registry data, in 2001, male cancer was 2.7% and in 2002, the cancer incidence rate was 2.7%, and in 2002, it ranked 6th. to be. In addition to genetic predisposition, prostate cancer is known to be caused by hormones, diet, and chemicals.

First, in men over 50 years of age, BPH is characterized by bladder storage symptoms such as frequency of urinating 8 or more times a day, nocturia, strong and sudden urge to urinate and unbearable urge to urinate, and delayed urination (when urinating). It is a generic term for symptoms indicating bladder discharge disorders, such as a phenomenon in which urine comes out after moxibustion), interrupted urine (a phenomenon in which the flow of urine is interrupted), and a phenomenon that requires force during urination. Dihydrotestosterone (DHT) converted from (testosterone, T) is known to be involved in prostate enlargement (Urology. 2003 Apr;61(4 Suppl 1):2-7.). Recently, the number of patients with BPH in Korea increased sharply from 458,955 in 2006 to 842,069 in 2011, which is 83.5%. In particular, the number of patients with enlarged prostate appears at a rate of 2 out of 5 men over the age of 60, and more than half of those over the age of 80 will complain of enlarged prostate. The number of patients with benign prostatic hyperplasia is expected to continue to increase in the future due to the entry into an aging society and improvement of the medical examination rate.

In addition, prostatitis is a disease that accounts for about 3 to 12% of all urology patients, but is the most common disease in urology patients under the age of 50. In particular, chronic pelvic pain, the most prominent symptom of prostatitis, is known to be more severe than pain caused by enlarged prostate or sexual dysfunction, and it is common among active young people and adversely affects the quality of life. However, it is difficult to diagnose and treat it because the cause is not clear and there are no characteristic findings on symptoms or tests. The effect of dihydrotestosterone (DHT), which is converted from testosterone (T), a male hormone by 5-alpha reductase, on prostatitis has been reported (The Journal of Urology, Volume 170, Issue 6, Part 1) , December 2003, Pages 2486-2489)

Prostate cancer is a malignant tumor that occurs in the prostate, and prostate cancer is also known to be highly related to the conversion of testosterone to dihydrotestosterone by 5-alpha reductase (J Steroid Biochem Mol Biol. 2002 Nov; 82). (4-5):393-400.). In the West, prostate cancer is the most common cancer among men and shows a high incidence. However, gastric prostate cancer is difficult to detect early because it does not have specific symptoms, so there are many cases of visiting a hospital only after the cancer tissue grows, which interferes with urination, or when prostate cancer metastasizes to other organs, such as bones, and symptoms such as bone pain appear. . Accordingly, there is an increasing demand for prophylactic and therapeutic agents for prostate cancer.

On the other hand, ginseng (Panax ginseng C. A. Meyer) is a perennial herb belonging to the Araliaceae family, and the root of ginseng has been known as a tonic or a panacea since ancient times, and various pharmacological effects are recognized by recent studies. Ginseng provides vitality, is used for body weakness, malaise, fatigue, anorexia, vomiting, and diarrhea. As pharmacological effects of ginseng, cerebral cortical excitability and inhibition, balance, anti-fatigue, anti-aging, immune enhancement, heart contraction, gonad stimulation, hyperglycemia suppression, protein synthesis promotion, homeostasis maintenance, anticancer and detoxification effects have been reported. Ginseng is divided into various types according to the processing method. First, ginseng in an unprocessed state is called fresh ginseng or raw ginseng. Fresh ginseng with a high water content has a short shelf life, so it is processed to improve its preservation.

White ginseng refers to fresh ginseng that has not been cooked and dried by sunlight, hot air, or other methods. White ginseng is dried by peeling the skin of fresh ginseng, or drying the skin of white ginseng. At this time, white ginseng dried without peeling is called skin white ginseng.

Taegeuk ginseng is an intermediate product between white ginseng and red ginseng, and refers to fresh ginseng steamed or blanched in boiling water and dried. Usually, fresh ginseng is immersed in water at 80 to 90 degrees Celsius for 10 to 20 minutes and then dried.

Red ginseng is dried by steaming fresh ginseng, and it can be stored for a long time because the moisture content is 15% or less. In the process of steaming ginseng, the components of ginsenosides increase and new components are created, which has the effect of increasing the potency. In addition, compared to unprocessed ginseng, red ginseng not only has better digestion and absorption, but also eliminates side effects and improves taste.

Black ginseng is similar to the method of manufacturing red ginseng, and it is manufactured by repeating the process of steaming and drying fresh ginseng. Black ginseng is ginseng with a higher ginsenoside content compared to fresh ginseng or red ginseng, and it was confirmed that ginsenoside types that did not exist in ginseng exist in black ginseng. That is, as the steroid-sugar bond is broken through the steaming and drying process, new ginsenosides are generated depending on the position of the remaining sugar, thereby exhibiting additional effects. Accordingly, the process of manufacturing black ginseng is studied, but research that can enhance ginsenosides present in trace amounts and enhance its efficacy is active.

Accordingly, the present inventors have tried to develop a natural material that is effective for the treatment of prostate disease without side effects, and the black ginseng extract or fermented black ginseng extract prepared by the manufacturing method of the present invention has a preventive or therapeutic effect on prostate-related diseases, A composition for preventing or treating prostate-related diseases using this was completed.

An object of the present invention is to provide a method for producing a black ginseng extract comprising a functional ingredient for the prevention or treatment of prostate-related diseases.

Another object of the present invention is to provide a black ginseng extract prepared by the method for producing the black ginseng extract.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of prostate-related diseases, including black ginseng extract.

Another object of the present invention is to provide a health functional food composition for preventing or improving prostate-related diseases, including black ginseng extract.

Another object of the present invention is to provide a method for producing a fermented black ginseng extract comprising a functional ingredient for the prevention or treatment of prostate-related diseases.

Another object of the present invention is to provide a black ginseng extract prepared by the method for producing the fermented black ginseng extract.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of prostate-related diseases, comprising a fermented black ginseng extract.

Another object of the present invention is to provide a health functional food composition for preventing or improving prostate-related diseases, including fermented black ginseng extract.

In order to achieve the above object,

The present invention provides a method for producing a black ginseng extract comprising a functional ingredient for the prevention or treatment of prostate-related diseases.

The present invention also provides a black ginseng extract prepared by the method for producing the black ginseng extract.

In addition, the present invention provides a pharmaceutical composition for preventing or treating prostate-related diseases comprising a black ginseng extract.

In addition, the present invention provides a health functional food composition for preventing or improving prostate-related diseases, including black ginseng extract.

In addition, the present invention provides a method for producing a fermented black ginseng extract comprising a functional ingredient for the prevention or treatment of prostate-related diseases.

The present invention also provides a black ginseng extract prepared by the method for producing the fermented black ginseng extract.

In addition, the present invention provides a pharmaceutical composition for preventing or treating prostate-related diseases comprising a fermented black ginseng extract.

In addition, the present invention provides a health functional food composition for preventing or improving prostate-related diseases comprising a fermented black ginseng extract.

When black ginseng is prepared by the method for producing black ginseng extract according to the present invention, the yield of total ginsenosides obtainable from black ginseng can be improved.

In addition, by fermenting the black ginseng extract prepared as described above using a Lactobacillus pentosus (ATCC8041) strain, the ginsenoside yield is further improved, and a method for producing a fermented black ginseng extract excellent in preventing or treating prostate-related diseases and It provides a fermented black ginseng extract prepared by the above manufacturing method.

1 is a schematic diagram for manufacturing black ginseng of the present invention.
2 is a diagram showing the comparison of the contents of ginsenoside components Rh1, Rg2, Rh4, Rg3, Rk1, Rg5, Rh2, Rh3 of white ginseng, manufactured black ginseng and fermented black ginseng before processing.
Figure 3a is a view showing the prostate cancer proliferation inhibitory effect of saw palmetto (Saw palmetto) as a control.
Figure 3b is a diagram showing the prostate cancer proliferation inhibitory effect of black ginseng hot water extract (BGH).
Figure 3c is a diagram showing the prostate cancer proliferation inhibitory effect of black ginseng 30% alcohol extract (BGH).
Figure 3d is a view showing the confirmation of the prostate cancer proliferation inhibitory effect of black ginseng 70% alcohol extract (BGH).
Figure 3e is a diagram showing the prostate cancer proliferation inhibitory effect of fermented black ginseng hot water extract (BGH).
Figure 3f is a diagram showing the prostate cancer proliferation inhibitory effect of fermented black ginseng 30% alcohol extract (BGH).
Figure 3g is a view showing the confirmation of the prostate cancer proliferation inhibitory effect of fermented black ginseng 70% alcohol extract (BGH).
Figure 4a is a view showing the confirmation of the prostate cancer proliferation inhibitory effect of compound Rh1 isolated from fermented black ginseng extract.
Figure 4b is a view showing the confirmation of the prostate cancer proliferation inhibitory effect of the compound Rh4 isolated from the fermented black ginseng extract.
Figure 4c is a view showing the confirmation of the prostate cancer proliferation inhibitory effect of compound Rg3 isolated from fermented black ginseng extract.

Hereinafter, preferred embodiments of the present invention will be described in more detail. However, it should be understood that the present invention may be embodied in many different forms and is not limited to the described embodiments. It should be noted that the embodiments of the present invention described below are intended to sufficiently convey the spirit of the present invention to those skilled in the art.

In addition, terms used in this specification are terms used to properly express preferred embodiments of the present invention, which may vary depending on the intention of a user or operator or customs in the field to which the present invention belongs. Therefore, definitions of these terms should be made based on the content throughout this specification. Throughout the specification, when a part "includes" a certain element, it means that other elements may be further included, rather than excluding other elements, unless otherwise stated.

Throughout this specification, '%' used to indicate the concentration of a specific substance is (w/w) % for solid/solid, (w/v) % for solid/liquid, and Liquid/liquid is (v/v) %.

According to an embodiment of the present invention, a) heating the dried ginseng from room temperature to 80°C to 100°C and steaming for 1 hour to 7 hours; b) drying the steamed ginseng as in step a) at 40° C. to 70° C. for 3 hours to 8 hours, followed by cooling for 10 hours to 20 hours; c) repeating steps a) and b) 3 to 5 times to prepare black ginseng; and d) drying at 25° C. to 40° C. for 1 to 5 days using a dryer and then naturally drying for 5 to 20 days so that the total moisture content of black ginseng is 15% or less; e) including the step of extracting the black ginseng extract from the black ginseng using an extraction solvent, it is possible to prepare a black ginseng extract.

In an embodiment of the present invention, step a) may be characterized in that the dried ginseng is steamed at 90° C. to 95° C. for 3 to 5 hours, but is not limited thereto.

In one embodiment of the present invention, step b) may be characterized in that the steamed ginseng is dried at 50° C. to 60° C. for 5 to 6 hours and then cooled for 15 hours, but is not limited thereto.

In one embodiment of the present invention, step c) may be characterized in that the steps a) and b) are repeated three times, but is not limited thereto.

In one embodiment of the present invention, step d) is dried at 30 ° C. to 35 ° C. for 1 to 3 days using a dryer, and then dried naturally for 10 to 15 days so that the total moisture of black ginseng is 15% or less. It may be characterized as such, but is not limited thereto.

According to another embodiment of the present invention, a) heating the dried ginseng from room temperature to 80°C to 100°C and steaming for 1 hour to 7 hours; b) drying the steamed ginseng as in step a) at 40° C. to 70° C. for 3 hours to 8 hours, followed by cooling for 10 hours to 20 hours; c) repeating steps a) and b) 3 to 5 times to prepare black ginseng; and d) drying at 25° C. to 40° C. for 1 to 5 days using a dryer and then naturally drying for 5 to 20 days so that the total moisture content of black ginseng is 15% or less; e) extracting the black ginseng extract from the black ginseng using an extraction solvent; and f) mixing the black ginseng extract with a Lactobacillus pentosus (ATCC8041) culture solution and fermenting it for 5 to 15 days at pH 4 to 8, 25 to 50 ° C. Black ginseng extract can be prepared.

In one embodiment of the present invention, in step f), the black ginseng extract prepared as described above is mixed with a Lactobacillus pentosus (ATCC8041) culture solution and fermented at pH 5 to 7, 30 to 45° C. for 10 days. It may be characterized in that, but is not limited thereto.

As defined in the present invention, "culture solution" means a culture solution obtained by culturing a specific microorganism in a culture medium, a concentrated culture solution, a dried product of a culture solution, a culture filtrate, a concentrated culture filtrate, or a dried product of the culture filtrate, and the strain After inoculating and culturing the containing or strain, it may be a culture filtrate from which the bacteria are removed, that is, the strain. In addition, the formulation of the culture is not limited, and may be, for example, a liquid, an emulsion, or a solid.

As another embodiment of the present invention, it is possible to provide a black ginseng extract or fermented black ginseng extract prepared as described above.

The extract according to the present invention may be obtained by extraction and separation from nature using extraction and separation methods known in the art, and "extract" as defined in the present invention is obtained from black ginseng or fermented black ginseng using an appropriate solvent. It is extracted, and includes, for example, a crude extract, a polar solvent-soluble extract, or a non-polar solvent-soluble extract. As a suitable solvent for extracting the extract from the black ginseng or fermented black ginseng, any pharmaceutically acceptable organic solvent may be used, and water or an organic solvent may be used, but is not limited thereto, for example, purified water , methanol (methanol), ethanol (ethanol), propanol (propanol), isopropanol (isopropanol), alcohols having 1 to 4 carbon atoms, including butanol (butanol), acetone (acetone), ether (ether), benzene (benzene) Various solvents such as , chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane may be used alone or in combination.

As the extraction method, any one of methods such as hot water extraction, cold extraction, reflux cooling extraction, solvent extraction, steam distillation, ultrasonic extraction, elution, and compression may be selected and used. In addition, the desired extract may be further subjected to a conventional fractionation process, and may be purified using a conventional purification method.

There is no limitation on the method for preparing the extract of the present invention, and any known method may be used. For example, the extract included in the composition of the present invention may be prepared in a powder state by an additional process such as distillation under reduced pressure and freeze-drying or spray-drying the primary extract extracted by the hot water extraction or solvent extraction method described above.

In another embodiment of the present invention, it is possible to provide a pharmaceutical composition for preventing or treating prostate-related diseases, including the black ginseng extract or fermented black ginseng extract prepared as described above.

As used herein, the term "prostate-related disease" refers to a disease occurring in the prostate, which is the male reproductive and urinary tract, for example, one selected from the group consisting of enlarged prostate, prostatitis, prostate cancer, prostate abscess, and prostate calcification. It may be the above diseases, but is not limited thereto, and may include all of various diseases caused by problems with the prostate. Prostate abscess is a disease in which bacteria enter the prostate through the urethra to form a sac of pus followed by acute inflammation and rapidly necrosis of the surrounding prostate tissue. Inability to urinate causes severe pain in urination. Prostate calcification refers to a disease in which limestones settle in the prostate. The stones refer to crystallization of calcium, oxalate, phosphate, etc. remaining without being all liquid when too many components are present in body fluids including urine.

As used herein, the term “prevention” may refer to any action of inhibiting or delaying the onset of prostate-related diseases by administering the pharmaceutical composition for preventing or treating prostate-related diseases according to the present invention to an individual.

As used herein, the term “treatment” may refer to any action for improving or benefiting symptoms of a prostate-related disease by administering the composition of the present invention to a subject suspected of having a prostate-related disease.

The pharmaceutical composition of the present invention may further include an adjuvant in addition to the active ingredient. The adjuvant may be used without limitation as long as it is known in the art, for example, Freund's complete adjuvant or incomplete adjuvant may be further included to increase the immunity thereof.

The pharmaceutical composition according to the present invention may be prepared in a form in which the active ingredient is incorporated into a pharmaceutically acceptable carrier. Here, the pharmaceutically acceptable carrier includes carriers, excipients and diluents commonly used in the pharmaceutical field. Pharmaceutically acceptable carriers that can be used in the pharmaceutical composition of the present invention include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The pharmaceutical composition of the present invention may be formulated in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, or sterile injection solutions according to conventional methods, respectively. .

In the case of formulation, it can be prepared using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, surfactant, etc. commonly used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations include at least one excipient in the active ingredient, for example, starch, calcium carbonate, sucrose, lactose, gelatin. It can be prepared by mixing and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid formulations for oral administration include suspensions, solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. can Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As the base of the suppository, witepsol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used.

The pharmaceutical composition according to the present invention may be administered to an individual by various routes. Any mode of administration can be envisaged, for example, by oral, intravenous, intramuscular, subcutaneous, intraperitoneal injection.

The pharmaceutical composition may be formulated in various oral or parenteral dosage forms.

Formulations for oral administration include, for example, tablets, pills, hard, soft capsules, solutions, suspensions, emulsifiers, syrups, granules, and the like. crose, mannitol, sorbitol, cellulose and/or glycine), lubricants (eg silica, talc, stearic acid and its magnesium or calcium salts and/or polyethylene glycol). In addition, the tablet may contain a binder such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidine, and optionally starch, agar, alginic acid or a disintegrant such as its sodium salt or a boiling mixture and/or absorbent, coloring, flavoring and sweetening agent. The formulation may be prepared by conventional mixing, granulating or coating methods.

In addition, a representative formulation for parenteral administration is an injection formulation, and examples of the solvent for the injection formulation include water, Ringer's solution, isotonic saline, or suspension. The sterile, fixed oil of the injectable preparation can be used as a solvent or suspending medium, and any non-irritating fixed oil including mono- and di-glycerides can be used for this purpose.

In addition, the injection preparation may use a fatty acid such as oleic acid.

In another embodiment of the present invention, it is possible to provide a health functional food composition for preventing or improving prostate-related diseases, including the black ginseng extract or fermented black ginseng extract prepared as described above.

In addition to containing black ginseng extract or fermented black ginseng extract as an active ingredient, the "health functional food composition" of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients like a conventional food composition.

Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. The above-mentioned flavoring agents can advantageously use natural flavoring agents (Taumatin), stevia extract (eg rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.). The food composition of the present invention may be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes and health supplements. There is this.

In addition, the health functional food composition contains various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavoring agents, colorants and thickeners (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice beverages and vegetable beverages.

The health functional food composition of the present invention may be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, and the like. In the present invention, the term "health functional food composition" refers to food manufactured and processed using raw materials or ingredients useful for the human body according to Health Functional Food Act No. 6727, and contains nutrients for the structure and function of the human body. It refers to ingestion for the purpose of obtaining useful effects for health purposes such as regulation or physiological action. The health functional food of the present invention may contain normal food additives, and unless otherwise specified, whether it is suitable as a food additive is related to the item according to the general rules and general test method of food additives approved by the Food and Drug Administration. It is judged according to the standards and standards. The items listed in the 'Food Additives Code' include, for example, chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, high pigment, and guar gum; Mixed preparations, such as a sodium L-glutamate preparation, a noodle-added alkali agent, a preservative preparation, and a tar color preparation, etc. are mentioned. For example, a health functional food in tablet form is granulated by a conventional method by mixing the active ingredient of the present invention with an excipient, a binder, a disintegrant and other additives, followed by compression molding with a lubricant, etc., or The mixture can be compression molded directly. In addition, the health functional food in the form of tablets may contain a corrosive agent or the like, if necessary. Among health functional foods in the form of capsules, hard capsules can be prepared by filling a mixture of the active ingredient of the present invention with additives such as excipients in ordinary hard capsules. It can be prepared by filling the mixture mixed with the capsule base such as gelatin. The soft capsules may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary. The health functional food in the form of a ring can be prepared by molding a mixture of the active ingredient of the present invention with an excipient, a binder, a disintegrant, etc. by a known method, and if necessary, it can be coated with sucrose or other skinning agent, Alternatively, the surface may be coated with a material such as starch or talc. The health functional food in the form of granules can be prepared in a granular form by a conventionally known method by mixing a mixture of an excipient, a binder, a disintegrant, etc. of the active ingredient of the present invention, and may contain a flavoring agent, a flavoring agent, etc. can

Hereinafter, the present invention will be described in detail by way of embodiments of the present invention with reference to the accompanying drawings. However, the following examples are presented as examples of the present invention, and when it is determined that detailed descriptions of well-known techniques or configurations known to those skilled in the art may unnecessarily obscure the gist of the present invention, the detailed description may be omitted, and , the present invention is not limited thereby. Various modifications and applications of the present invention are possible within the scope of equivalents interpreted therefrom and the description of the claims to be described later.

Example 1. Preparation of black ginseng extract

Black ginseng extract was prepared in the same manner as shown in FIG.

More specifically, the first washed ginseng is dried in a hot air dryer at 35° C. for 10 to 15 hours and then naturally dried for 2-3 days. Put the pre-dried ginseng in the steamer as described above, preheat it at 90°C for 40 minutes, and then steam it at 90-95°C for 3-5 hours. After that, the first steamed ginseng is dried at 50-60° C. for 5-6 hours using a dryer, and then cooled and dried for 15 hours. Then, after repeating the above-mentioned steaming and drying three times, using a dryer again, drying at 30-35° C. for 1 to 3 days, and then natural drying for 10 to 15 days to obtain black ginseng with a moisture content of 15% or less.

Using the black ginseng obtained as described above, the brain heads and fine roots (fine roots) of black ginseng are removed, and the black ginseng main ginseng is used and powdered using a grinder. After adding 80% methanol to the obtained powdered black ginseng to 100 g/1L, reflux extraction was performed three times for 12 hours each, followed by precipitation, filtration with filter paper, concentration under reduced pressure and freeze-drying at 45° C. or lower to obtain 45 g of a freeze-dried product.

Example 2. Preparation of fermented black ginseng extract

100 mg to 200 mg of glucose was added as a carbon source based on 10 g of the freeze-dried product of black ginseng prepared as in Example 1, and 10 mg - 20 mg of peptone was additionally added to sterilize. Thereafter, the Lactobacillus pentosus (ATCC8041) strain culture solution was inoculated to a concentration of 50 g/L and cultured. The strain was cultured using a fermenter (10 L) for 10 days, maintained at 37° C., pH 5-7. After culturing, the culture medium was centrifuged to remove the culture medium and then sterilized (120° C., 30 minutes, 1.5 atm) so that no further culture was possible. After fermentation, the sterilized black ginseng fermented product was finally extracted with 30% (V/V) fermented alcohol under reflux conditions at 70° C. for 4 hours and precipitated, followed by filtration with filter paper. After combining the two extracts with the filtrate, the fermented black ginseng extract concentrated under reduced pressure at 40° C. or less was obtained and freeze-dried.

Example 3. Preparation of fractions of fermented black ginseng extract and separation of compounds

The black ginseng fermentation extract obtained as in Example 2 was partitioned and extracted three times with EtOAc (1 L) / H ₂ O (1 L), and the H ₂ O layer was again partitioned with n-BuOH (0.8 L) four times. extracted. Each layer was concentrated under reduced pressure to obtain an EtOAc fraction, an n-BuOH fraction, and a H ₂ O fraction.

8 kinds of compounds, Rh1, Rg2, Rh4, Rg3, Rk1, Rg5, Rh2 and Rh3 were obtained.

Example 4. Rh1, Rg2, Rh4, Rg3, Rk1, Rg5, Rh2 and Rh3 content comparison

The content of 8 kinds of compounds obtained in Example 3 was confirmed by using HPLC.

As a result, as shown in FIG. 2 , no compounds other than Rh1 and Rg2 were detected in white ginseng before processing, and eight compounds Rh1, Rg2, Rh4, Rg3, Rk1, Rg5, Rh2 and Rh3 in the black ginseng prepared according to the present invention. were all detected. In particular, it was confirmed that the content of Rh1, Rh4, Rg3, Rk1, Rg5, Rh2, Rh3 increased in the manufactured fermented black ginseng than in the manufactured black ginseng.

Experimental Example 1. Confirmation of cytotoxicity and prostate cancer proliferation inhibitory effect of black ginseng extract and fermented black ginseng extract

In order to confirm the cytotoxicity and prostate cancer proliferation inhibitory effect of the black ginseng extract and the fermented black ginseng extract of the present invention, the cancer growth inhibitory effect was confirmed using human prostate stromal cells (WPMY-1) and human prostate cancer cell line (LNCap).

Human prostate stromal cells (WPMY-1) were cultured in RPMI 1640 medium supplemented with 10% FBS and 1% penicillin/streptomycin (100 u/ml) at 37° C., 5% CO ₂ using a cell incubator. Cytotoxicity of black ginseng extract and fermented black ginseng extract to human prostate stromal cells (WPMY-1) and human prostate cancer cell line (LNCap) was measured using MTT assay. Cultured WPMY-1, LNCap cells were dispensed in a 96 well plate at a concentration of 2×10 ⁴ cells/well, and after pre-incubation for 24 hours, black ginseng water, 30% alcohol, 70% alcohol extract, fermented black ginseng black ginseng water, 30% Alcohol, 70% alcohol extract was treated by concentration (50, 100, 150, 200 μg/ml) and further cultured for 24 hours. In order to measure cell viability, MTT solution was treated and reacted at 37°C for 4 hours, and the absorbance was measured at 570 nm by dissolving purple-stained formazan by adding DMSO. The group not treated with black ginseng extract or fermented black ginseng extract showed a cell viability of 100%.

3 is a human prostate stromal cell line and cancer cell line black ginseng hot water extract (BGH), black ginseng 30% alcohol extract (BG30), black ginseng 70% alcohol extract (BG70), fermented black ginseng hot water extract (FBGH), fermented black ginseng 30% alcohol extract (FBG30) and fermented black ginseng 70% alcohol extract (FBG70) were treated for each concentration, respectively, and it is a graph showing the survival rate after 24 hours.

As shown in Figures 3b and 3c, black ginseng hot water extract (BGH) and black ginseng 30% alcohol extract (BG30) were not cytotoxic to human prostate stromal cells (WPMY-1), 200 ug/ml and 148 ug/ml, respectively. At a concentration of ml, the growth of prostate cancer cells was inhibited by 50%. Black ginseng 70% alcohol (BG70) was toxic to prostate stromal cells, as shown in FIG. 3d , and inhibited cancer cell growth at about 110 ug/ml.

As shown in FIGS. 3e and 3f, in the case of fermented black ginseng hot water extract (FBGH) and fermented black ginseng 30% alcohol extract (FBG30), there is no cytotoxicity to human prostate stromal cell lines, and there is a superior effect compared to unfermented black ginseng extract was confirmed.

As shown in FIG. 3g , it was confirmed that the 70% alcohol extract of fermented black ginseng (FBG70) was toxic to prostate stromal cells and inhibited cancer cell growth at about 145 ug/ml.

Experimental Example 2. Confirmation of cytotoxicity and prostate cancer proliferation inhibitory effect of compounds isolated from fermented black ginseng extract

In order to confirm the cytotoxicity and prostate cancer proliferation inhibitory effect of the compound isolated from the fermented black ginseng extract of the present invention, the cancer growth inhibitory effect was confirmed using human prostate stromal cells (WPMY-1) and human prostate cancer cell line (LNCap).

Human prostate stromal cells (WPMY-1) were cultured in RPMI 1640 medium supplemented with 10% FBS and 1% penicillin/streptomycin (100 u/ml) at 37° C., 5% CO ₂ using a cell incubator. The cytotoxicity of the compounds isolated from the fermented black ginseng extract to human prostate stromal cells (WPMY-1) and human prostate cancer cell lines (LNCap) was measured using MTT assay. Cultured WPMY-1, LNCap cells were dispensed into 96 well plates at a concentration of 2×10 ⁴ cells/well, and after pre-incubation for 24 hours, the compounds isolated from the fermented black ginseng extract were separated by concentration (25, 50, 75, 100, 125 μg/ml) and further incubated for 24 hours. In order to measure cell viability, MTT solution was treated and reacted at 37° C. for 4 hours, and the absorbance was measured at 570 nm by dissolving purple-dyed formazan by adding DMSO. The group not treated with black ginseng extract or fermented black ginseng extract showed a cell viability of 100%.

4 is a graph showing the survival rate after 24 hours by treating each concentration of the compound isolated from the fermented black ginseng extract in human prostate stromal cell lines and cancer cell lines.

As shown in Fig. 4a, it was confirmed that Rh1 had no cytotoxicity to human prostate stromal cell lines and had an excellent inhibitory effect on cancer cell growth. Rh4 (Fig. 4b) and Rg3 (Fig. 4c) also had no cytotoxicity to human prostate stromal cell lines, and it was confirmed that the effect of inhibiting cancer cell growth was excellent. In the case of Rh1, it was confirmed that cancer cell growth was inhibited at about 78.11 ug/ml. In addition, it was confirmed that Rh4 and Rg3 were excellent in inhibiting cancer cell growth at about 64.12 ug/ml and about 78.12 ug/ml, respectively.

Although the preferred embodiments of the present invention have been described in detail above, the present invention is not limited to the specific embodiments described above, and it is common in the technical field to which the present invention pertains without departing from the gist of the present invention as claimed in the claims. Various modifications and improvements are possible by those having the knowledge of, of course, and these modifications and improvements should not be individually understood from the technical spirit or prospect of the present invention.

Claims (14)

a) heating the dried ginseng from room temperature to 80°C to 100°C and steaming for 1 hour to 7 hours;
b) drying the steamed ginseng as in step a) at 40° C. to 70° C. for 3 to 8 hours and then cooling the steamed ginseng for 10 to 20 hours;
c) repeating steps a) and b) 3 to 5 times to produce black ginseng; and
d) extracting the black ginseng extract using an extraction solvent from the black ginseng main ginseng except for the head and fine roots (root) of the black ginseng;
A method for producing a black ginseng extract with an increased content of ginsenoside Rh4, comprising a. The method of claim 1,
Black ginseng extract comprising a step of drying so that the total moisture of black ginseng becomes 15% or less by drying naturally for 5 to 20 days after drying at 25° C. to 40° C. for 1 to 5 days using a dryer after step c) manufacturing method. delete The method of claim 1,
The solvent used for the extraction is methanol, ethanol, acetone, ethyl acetate, hexane, butanol, methylene chloride, water, or a method for producing a black ginseng extract that is extracted with any one solvent selected from the group consisting of mixed solvents thereof. A black ginseng extract prepared by the method of claim 1. A pharmaceutical composition for preventing or treating prostate cancer, comprising the black ginseng extract prepared by the method of claim 1. delete A health functional food composition for preventing or improving prostate cancer comprising a black ginseng extract prepared by the method of claim 1 . a) preparing a black ginseng extract by the method of claim 1; and
b) mixing the black ginseng extract with a Lactobacillus pentosus (ATCC8041) culture solution and fermenting it for 5 to 15 days at pH 4 to 8 and 25 to 50° C. to prepare a fermented black ginseng;
A method for producing a fermented black ginseng extract with an increased content of ginsenoside Rh4, comprising a. A fermented black ginseng extract prepared by the method of claim 9 . 10. The method of claim 9, wherein the fermented black ginseng extract is ginsenosides Rh1, Rg2, Rh4, Rg3, Rk1, Rg5, any one component selected from the group consisting of Rh2 and Rh3 is increased than the black ginseng extract, fermented black ginseng extract manufacturing method. A pharmaceutical composition for preventing or treating prostate cancer, comprising the fermented black ginseng extract prepared by the method of claim 9 . delete A health functional food composition for preventing or improving prostate cancer, comprising the fermented black ginseng extract prepared by the method of claim 9.

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