KR20130080644A - A composition comprising the extract of red ginseng for treating and preventing atopic disease - Google Patents

Abstract

PURPOSE: A composition containing a black ginseng extract is provided to be effectively used as a composition for preventing and treating an atopic disease since the composition does not have a side effect on a body nor a toxicity and have an atopy inducer excretion reducing effect. CONSTITUTION: A pharmaceutical composition for preventing and treating an atopic disease contains a black ginseng extract as an active ingredient. The black ginseng is obtained through one cycle of the following steps of: a first step of washing Korean ginseng roots of 4 to 6 years old in an ultrasonic cleanser for 10 minutes for 3 times and drying the ginseng roots at a temperature of 35 to 45°C for 22 to 26 hours; a second step of steaming the dried Korean ginseng roots in a steamer at a temperature of 85 to 105°C for 4 to 6 hours, except a pre-heating time; a third step of drying the Korean ginseng roots at a temperature of 55 to 65°C for 10 to 14 hours; a fourth step of steaming the Korean ginseng roots at pressure of 0.1 to 0.20 MPa at a temperature of 110 to 120°C for 5 to 7 hours; and a fifth step of drying the Korean ginseng roots in a dryer for 10 to 14 hours to obtain black ginseng roots.

Description

A composition comprising the extract of red ginseng for treating and preventing atopic disease}

The composition containing the black ginseng extract of the present invention as an active ingredient is useful for treating atopic diseases because it shows an effect of reducing the secretion of atopic inducers.

[Reference 1] Understanding Korean Ginseng, Korean Ginseng Society, p.9, 1995

[Ref. 2] Advances in Ginseng Research, Korean Ginseng Society, p. 127-137, 1998

[Document 3] Korean Unexamined Patent Publication No. 2003-0005089

[Document 4] Republic of Korea Patent Registration No. 10-0754253

[Document 5] Korea Patent Registration No. 10-0781571

[0012] Lee JS, Kim IS, Ryu JS, Yun CY., House dust mite, Dermatophagoides pteronissinus increases expression of MCP-1, IL-6, and IL-8 in human monocytic THP-1 cells. Cytokine. 2008, 42 (3): 365-71. 2008.

The present invention relates to a composition for the prevention and treatment of atopic diseases containing black ginseng extract as an active ingredient and a method for producing the same.

Panax genus is a member of the Orga family, with about 10 species known worldwide. Panax genus plants known to date include Panax ginseng, Panax quinquefolia, Panax notoginseng, Panax japonica, Panax trifolia, Himalaya ginseng, Panax pseudoginseng, Panax vietnamensis, etc. (Understanding Korean Ginseng, p. 9, Korean Ginseng Society, 1995; Advances in Ginseng Research, pp. 127-137, Korean Ginseng Society, 1998). Other Panax plants include Panax elegatior, Panax wangianus or Panax bipinratifidus.

On the other hand, red ginseng is a secondary ingredient conversion during the proper heat treatment process to produce new active ingredients unique to red ginseng (fresh ginseng) or white ginseng, and some active ingredients increase in content. In Korea, after the resale system was abolished in the late 1990s, SMEs entered the ginseng industry since 2001, while large companies dominated the red ginseng market. According to related industry, ginseng products that maximize the efficacy and effect such as strengthening anti-cancer and immunity by using biotechnology technology are released one after another to realize premium ginseng era. This industry situation is also a vital force in terms of new technologies aimed at the global ginseng-related market worth $ 3.5 billion annually.

As mentioned above, the consumption of ginseng, the best health food, is increasing in the era of well-being. However, in the case of SME companies in Geumsan-gun, they are reluctant to share red ginseng processing technology information. In this regard, processing methods and quality of red ginseng concentrate (extract) are different for each processing company and farmer, and there is no standardized and established processing technology. Consumers also prefer large enterprise products, despite their high price, with the bias that they are better in terms of efficacy and quality.

The pathogenesis of atopic dermatitis has not yet been fully understood, but it has been known that atopic dermatitis is caused by a chronic inflammatory response to skin due to a sensitive immune response (allergic reaction) to a common substance (allergen) in the environment. In particular, it is known that the correlation between the amount of house dust mite exposure and the incidence of atopic dermatitis has a significant correlation with the severity of atopic dermatitis. It is also known to reduce the severity of atopic dermatitis by reducing exposure to house dust mites in the environment. In addition, steroids are the most widely used effective therapeutic agents in inflammatory diseases such as bronchial asthma.

Recently, a lot of mechanisms for steroids have been revealed, but the mechanisms are still not fully understood. Some patients, such as bronchial asthma and rheumatoid arthritis, have steroid resistance that does not respond to steroids. Although there are not many patients showing such steroid resistance, it is usually serious and suffers from complications, and the cost of treatment is a big problem. That is, in a chronic inflammatory response, various cytokines such as tumor growth factor (β, TGF-β), IL-4, IL-13, and the like involved in fibrosis are secreted through various cells. This increases the secretion of IL-6, which activates fibroblasts, resulting in differentiation and proliferation of many fibroblasts, and overproducing the production of extra cellular matrix, resulting in cell and tissue transformation and fibrosis. Cause.

Atopic dermatitis is treated with steroid-based drugs, but this is why it becomes resistant to steroid preparations due to tissue degeneration and fibrosis as it becomes chronic. Monocyte chemoattractant protein-1 (MCP-1) binds to the chemokine receptor (CCR2), and mice lacking the MCP-1 gene damage the chemotaxis of monocytes and infect certain bacteria. Weakening of resistance was observed, a phenomenon similar to that seen in animals with the CCR2 gene removed. In addition, MCP-1 has been reported to convert Th0 cells into cells that secrete Th2 cytokines in vitro. Intravenous injection of MCP-1 decreased IL-12 production and increased IL-4 production. This indirectly means that IgE-dependent allergic inflammation can be exacerbated.

IL-8 is an important inflammatory chemokine secreted by airway epithelial cells, which is important for early inflammatory reactions and is caused by bronchial hypersensitivity by IL-8, increased in allergic rhinitis or bronchial asthma, and IL-8 is inhibited by steroids.

Recently, as a result of efforts to further increase the active ingredient content of ginseng, black ginseng using the principle of Gujeungpo, which is the most representative method of using water and fire among the processing methods of herbal medicine, which is one of steaming methods, has been developed ( Korean Patent Application Publication No. 2003-0005089; Manufacturing method of black ginseng and black rice ginseng) In this laboratory, patents have been registered for a standardized manufacturing method of black ginseng that can be shortened in time and mass-produced compared to this method (Korean patent registration) No. 10-0754253; Novel black ginseng manufacturing method and composition containing the same, Patent registration No. 10-0781571; Pharmaceutical composition containing the extract of the novel black ginseng).

However, there is no mention or suggestion about the therapeutic effect of black ginseng extract prepared by the method developed by the present inventors for atopic diseases.

Accordingly, the present inventors confirmed that the black ginseng extract showed the effect of reducing the secretion of inflammatory factors such as MCP-1, IL-6, IL-8 in EoL-1 and THP-1 cells treated with mite extract The present invention has been completed.

In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of atopic diseases containing black ginseng extract as an active ingredient.

Black ginseng root extract of the present invention can be prepared through the following process.

Black ginseng root as defined herein is 3 to 7 years old, preferably 4 to 6 years old Korean ginseng after washing three times for 10 minutes with an ultrasonic cleaner, 10 to 70 ℃, preferably 12 to 36 at a temperature of 30 to 50 ℃ A first step of drying time, preferably 20 to 28 hours; A second step of putting dried Korean ginseng root into a steamer for 1 to 8 hours, preferably 4 to 6 hours at a temperature of 60 to 120 ° C., preferably 85 to 105 ° C. except for a preheating time; A third step of drying at a temperature of 40-80 ° C., preferably 50-70 ° C. for 6-18 hours, preferably 9-15 hours; 4 to 8 hours, preferably 5 at a temperature of 95 to 155 ° C., preferably at a temperature of 100 to 120 ° C., under a pressure of 0.05 to 0.45 MPa, preferably 0.10 to 0.20 MPa, more preferably 0.10 to 0.14 MPa A fourth step of steaming for 7 to 7 hours; The black ginseng root (hereinafter referred to as BPG) of the present invention can be obtained by a fifth step of drying in a drier for 8 to 16 hours, preferably 10 to 14 hours.

Approximately 3 to 7 times, preferably 4 to 6 times mass water of black ginseng root prepared by the high temperature and high pressure method was added thereto, and the mixture was left at room temperature for 3 to 7 hours, and then water containing purified water and lower carbon atoms having 1 to 4 carbon atoms. Alcohol or a mixed solvent thereof, preferably 80% ethanol, added 5 to 20 times the mass, preferably 8 to 12 times the mass for 2 to 8 hours, preferably 3 to 4 hours, 1 to The black ginseng root crude extract of the present invention (hereinafter referred to as BPGE) may be obtained by cooling, filtration and concentration steps of the extract obtained by reflux extraction five times, preferably three times.

Suspending the crude black ginseng root crude extract obtained by suspending water, and then adding a nonpolar solvent such as ether or dichloromethane to remove the nonpolar material; Black ginseng root butanol soluble extract of the present invention containing a large amount of ginseng saponin (hereinafter referred to as BPGBE) through the final step of extracting saponin present in ginseng 2 to 6 times and concentrating under reduced pressure by adding saturated butanol to the remaining aqueous layer. Can be obtained.

Black ginseng extract as defined herein includes black ginseng root crude extract (BPGE) and black ginseng root butanol soluble extract (BPGBE) as described above, and specifically, water of black ginseng, lower alcohols such as methanol, ethanol, butanol or the like Mixed solvent, preferably water or 1-100% water and ethanol mixed solvent, more preferably about 1-70% water and ethanol mixed solvent.

The present invention also provides a health functional food for the prevention and improvement of atopic diseases containing black ginseng extract as an active ingredient.

Hereinafter, a method of obtaining the black ginseng extract of the present invention will be described in more detail.

For example, the black ginseng of the present invention is 3 to 8 years old ginseng, preferably 5 to 7 years old ginseng, preferably Geumsan ginseng at 10 to 80 ℃, preferably 20 to 60 ℃ about 6 hours to 7 days, Preferably, the first step of naturally drying for 12 to 48 hours; A second step of washing the naturally dried ginseng and naturally drying the second only to remove moisture; A third step of steaming the dried ginseng at 60 to 120 ° C., preferably 80 to 110 ° C., for about 1 to 48 hours, preferably 1 to 24 hours, and more preferably 2 to 12 hours; The first steamed ginseng is dried at 30 to 80 ° C., preferably at 40 to 70 ° C. for about 1 to 72 hours, preferably 2 to 48 hours, more preferably 6 to 24 hours. Step 4; The dried ginseng is about 1 to 14 hours, preferably 2 to 12 hours at a pressure of 0.01 to 0.50 MPa, preferably 0.05 to 0.20 MPa, at about 60 to 150 ° C, preferably 80 to 120 ° C. A fifth step of steaming secondary during; The second steamed ginseng is third to dry at about 30 to 80 ℃, preferably 40 to 70 ℃ for about 1 to 72 hours, preferably 2 to 48 hours, more preferably 6 to 24 hours The black ginseng dry powder of the present invention (hereinafter referred to as BPG) can be obtained through a process including six steps.

In addition, the black ginseng extract of the present invention after washing and drying the black ginseng, about 1 to 50 times the sample weight, preferably about 5 to 25 times (w / v) volume of water, C ₁ to C ₄ lower alcohol Or a mixed solvent thereof, preferably water or ethanol, more preferably water or 5 to 80% ethanol as an extraction solvent, at a reaction temperature of about 70 to 120 캜, preferably 80 to 110 캜. A second step of extracting a conventional extraction method such as a heat extraction method, a hot water extraction method, an ultrasonic extraction method, a reflux extraction method, and preferably a hot water extraction method for 1 to 21 days, preferably 3 to 7 days; Extraction of the primary extracted extract for about 1 to 6 hours, preferably 3 to 5 hours at a reaction temperature of about 70 to 120 ℃, preferably 80 to 110 ℃ conventional extraction such as heat extraction, ultrasonic extraction, reflux extraction A third step of extracting 1 to 10 times, preferably 2 to 7 times by a method, preferably by heat extraction; It is possible to obtain black ginseng root ethanol soluble extract (BPGE) of the present invention through a process comprising a method of preparing a fourth step of filtering and extracting the extract obtained in the above step under reduced pressure.

Or the black ginseng extract of the present invention after washing and drying the black ginseng, about 1 to 50 times the weight of the sample, preferably about 2 to 25 times (w / v) volume of water, C ₁ to C ₄ lower alcohol Or a mixed solvent thereof, preferably water or ethanol, more preferably water or 5 to 80% ethanol as an extraction solvent, at a reaction temperature of about 70 to 120 캜, preferably 80 to 110 캜. A second step of extracting by a conventional extraction method such as a heat extraction method, a hot water extraction method, an ultrasonic extraction method, a reflux extraction method, preferably a reflux extraction method for from about 24 hours, preferably 1 to 7 hours; Suspending the extract with distilled water and then fractionating with a nonpolar solvent such as ether, chloroform and hexane to remove the nonpolar solvent solubles and obtaining an aqueous layer; Black ginseng root butanol soluble extract containing ginseng saponin of the present invention through a process comprising a fourth step of preparing a saturated butanol layer obtained by performing fractionation by adding saturated butanol to the aqueous layer and filtering under reduced pressure. Hereinafter referred to as BPGBE.

Accordingly, the present invention provides a method for producing black ginseng dry powder and black ginseng extract as described above.

Atopic diseases as defined herein include, but are not limited to, atopic dermatitis, ecstasy, psoriasis, and the like.

In another aspect, the present invention provides a pharmaceutical composition and health functional food for the treatment and prevention of atopic diseases containing the black ginseng extract prepared by the method as an active ingredient.

The composition of the present invention comprises 0.1 to 50% by weight of the herbal extract based on the total weight of the composition.

However, the composition is not limited thereto, and may vary depending on the condition of the patient, the type of disease, and the progress of the disease.

Compositions comprising the extract of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions.

The composition containing the extract according to the present invention may be formulated in the form of powders, granules, tablets, capsules, oral preparations such as suspensions, emulsions, syrups and aerosols, external preparations, suppositories and sterilized injection solutions, Examples of carriers, excipients and diluents that can be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium Silicates, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose, or the like. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol gelatin and the like can be used.

Preferred dosages of the extracts of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the extract is preferably administered at 0.01 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg per day. The administration may be carried out once a day or divided into several doses. Therefore, the dose is not intended to limit the scope of the present invention in any aspect.

The composition of the present invention may be administered to mammals such as rats, mice, livestock, humans, and the like in various routes. All modes of administration can be expected, for example by oral and rectal or intravenous methods.

In another aspect, the present invention provides a health functional food for the improvement of forgetfulness and degenerative brain disease containing black ginseng dry powder or black ginseng extract as an active ingredient.

Health functional foods containing the extract of the present invention can be used in a variety of drugs, foods and beverages for the prevention and improvement of allergic inflammatory diseases. Examples of the foods to which the extract of the present invention can be added include various foods, beverages, gums, tea, vitamin complexes, health supplements and the like, and they can be used as powders, granules, tablets, capsules or beverages have.

Accordingly, the present invention also provides a food and food additive containing black ginseng extract as an active ingredient having an effect of preventing and improving atopic diseases.

Extract of the present invention may be added to food or beverage for the purpose of preventing and improving atopic diseases. At this time, the amount of the extract in the food or beverage is generally the health food composition of the present invention can be added to 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.02 to 10g, preferably 0.3 based on 100ml To 1 g.

At this time, in the case of the food additive containing the black ginseng extract of the present invention, it can be used to add 0.01 to 10% by weight based on the total weight of the food, the amount of the black ginseng extract in the health functional food or beverage is 0.01 of the total food weight To 30% by weight, the health beverage composition may be added in a ratio of 0.02 to 5g, preferably 0.3 to 1g based on 100ml.

The health functional beverage composition of the present invention has no particular limitation on the other ingredients other than the above-mentioned extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. 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 sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As natural flavors other than those described above, natural flavors (such as tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin)) and synthetic flavors (saccharin, aspartame, etc.) have. The proportion of the natural carbohydrate is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof. , Organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected in the range of 0.1 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

The composition containing the black ginseng extract of the present invention has no side effects or toxicity to the human body, MCP-1, IL-6, IL-8, etc. in EoL-1 and THP-1 cells treated with mite extracts causing atopy Since it shows the effect of reducing the secretion of atopy-inducing factors of can be usefully used as a composition for the prevention and treatment of atopic diseases.

The present invention will be described in more detail based on the following examples, but the present invention is not limited thereto.

Example  1. Preparation of Black Ginseng Root

After purchasing five-year-old ginseng produced and distributed in Geumsan, 1kg of ginseng root was put in a screw washer (Jungwoo Machinery, Korea) and washed three times for 10 minutes. At this time, care was taken not to peel during the cleaning process.

When steamed steamed ginseng without drying, ginseng roots are often bursting, so to prevent this, first dried at 40 ℃ for 24 hours. Ginseng roots dried at low temperature for 24 hours were put into a steamer and steamed for 1 hour at 95 ° C except for the preheating time. The preheating time is the time from the steamer to water vapor, which normally took about 30 minutes. After keeping the internal temperature of the dryer with a heating wire at 60 ℃, spin the pen and dry the second ginseng evenly for 12 hours, then steam it again at 115 ℃ for 6 hours under the pressure of 0.12MPa 250 g of black ginseng root of the present invention was prepared by time drying 3 times (hereinafter, referred to as BPG).

Example  2. Black Ginseng Root Crude extract  Produce

20 g of BPG prepared by the method of Example 1 was taken and pulverized, 100 ml of water was added thereto, and the mixture was left at room temperature for 5 hours, followed by 1 liter of 80% ethanol (20% water). It was. The extracted solution was cooled to room temperature, filtered and the ethanol solvent was removed using a vacuum condenser to obtain 7.4 g of black ethanol soluble extract of black ginseng root (hereinafter referred to as BPGE).

Example  3. Black Ginseng Root Butanol  Preparation of Soluble Extracts

200 ml of water was suspended in 3.2 g of BPGE obtained in Example 2, and 200 ml of ether was added to remove the nonpolar material. 200 ml of saturated butanol was added to the remaining aqueous layer to extract saponins present in ginseng, and then the solvent was removed under a reduced pressure concentrator to obtain 0.7 g of black ginseng root butanol soluble extract containing a large amount of ginseng saponin (hereinafter referred to as BPGBE). .

The BPGBE was used as a sample for component analysis and high pathogenic avian influenza virus experiments.

Example  4. Analysis of Black Ginseng Saponin Components

In order to analyze the components of ginseng saponin present in the black ginseng root extract, 20 mg of BPGBE obtained in Example 3 was dissolved in 1 ml of methanol, and the filtrate was filtered through a 0.45 μm filter. It was. As a result of component analysis, Rb1 2.3 mg, Rb2 2.1 mg, Rc 1.3 mg, Rd 1.4 mg, Re 0.5 mg, Rg1 1.2 mg, Rk1 3.7 mg, Rg5 4.7 mg, Rg3 per 1 g of BPGBE. It was confirmed that (S) was 5.4 mg and Rg3 (R) was 4.5 mg.

Column: Shiseido C18 column (5, 1504.6 mm) Flow rate 1 / min Detector ELSD-LT detector Solvent A; CH ₃ CN: H ₂ O: 5% CH ₃ COOH = 15: 80: 5, B; CH ₃ CN: H ₂ O = 80:20 0 min (0% B), 0-10 min (30% B), 10-25 min (50% B), 25-40 (100% B), 40- 50 minutes (100% B), 50-55 minutes (0% B), 55-58 minutes (0% B) Temperature 40 degrees

Experimental Example  1. Black Ginseng Atopy Inhibition  activation

In order to measure atopic inhibition activity in the sample obtained in the above example, MCP-1, IL-6, IL- was added by adding a mite extract known to cause atopy to THP-1 cells by modifying the method disclosed in the literature as follows. After promoting the secretion of 8, black ginseng extract was added to observe the decrease (Lee JS, Kim IS, Ryu JS, Yun CY., House dust mite, Dermatophagoides pteronissinus increases expression of MCP-1, IL-6, and IL-). 8 in human monocytic THP-1 cells.Cytokine. 2008, 42 (3): 365-71. 2008.).

Treatment of atopic dermatitis extract on THP-1 cells increases the amount of atopy-inducing cytokine, MCP-1, IL-6, and IL-8. Therefore, we tried to observe whether the amount of MCP-1, IL-6, IL-8 increased when black ginseng was treated in this pathological condition.

1-1. THP -1 cell culture

Human acute monocytic leukemia cell (THP-1), a monocyte, was 2.0x10 ⁵ / m in RPMI 1640 medium, antibiotics (penicillinm 10 ⁴ U / ml, streptomycin 10mg / ml, amphotericin B 25µg / Ml) and 10% FBS, and incubated for 3 days in a 37 ℃ CO ₂ incubator.

1-2. MCP -One, IL -6, IL -8 measurements

THP-1 cells were dispensed in 24 well plates at 2.0x10 ⁶ / m in RPMI containing 0.5% FBS and incubated in an incubator (37 ° C, 5% CO ₂ ) for 16 hours. After incubation, black ginseng (final concentration 100 μg / ml) was treated for 14 hours, followed by HDE (1 μg / ml) for 24 hours, respectively, followed by monocyte chemoattractant protein-1 (monocyte) in supernatant using ELISA. chemoattractant protein-1 (MCP-1), IL-6 and IL-8 were measured.

1-3. MCP -1 secretion inhibitory activity

In order to measure the inhibitory activity of MCP-1 secretion in the sample obtained in the above example, the method disclosed in the literature was modified to add a mite extract known to cause atopy to THP-1 cells to promote secretion of MCP-1. Black ginseng extract was added and observed to decrease (Lee JS, Kim IS, Ryu JS, Yun CY., House dust mite, Dermatophagoides pteronissinus increases expression of MCP-1, IL-6, and IL-8 in human monocytic THP -1 cells.Cytokine. 2008, 42 (3): 365-71. 2008.).

As a result of the experiment, as shown in Table 2, the treatment of mite extracts that induce atopy on THP-1 cells can be seen that the secretion of MCP-1 rapidly increased (582pg / ml). Treatment of black ginseng extract (BPGE) of the present invention in this pathological condition was reduced to 486 pg / ml. On the other hand, the treatment of black ginseng butanol soluble extract (BPGBE) containing a large amount of black ginseng saponin was weaker than the dexamethasone used as a positive control group, but it was reduced to almost normal level at 86 pg / ml. In other words, black ginseng was determined to be effective in inhibiting atopic dermatitis and the main activity was black ginseng saponin.

1-4. IL -6 secretion inhibitory activity

In order to measure IL-6 secretion inhibitory activity in the sample obtained in the above example, the method described in the literature was modified to add a tick extract known to cause atopy to THP-1 cells to promote secretion of IL-6. Black ginseng extract was added and observed to decrease (Lee JS, Kim IS, Ryu JS, Yun CY., House dust mite, Dermatophagoides pteronissinus increases expression of MCP-1, IL-6, and IL-8 in human monocytic THP -1 cells.Cytokine. 2008, 42 (3): 365-71. 2008.).

As a result of the experiment, as shown in Table 2, the treatment of mite extracts inducing atopy on THP-1 cells showed that the secretion of IL-6 increased rapidly (803 pg / ml). In this pathological condition, black ginseng extract (BPGE) of the present invention was reduced to 697 pg / ml. On the other hand, the treatment of black ginseng butanol soluble extract (BPGBE) containing a large amount of black ginseng saponin was weaker than dexamethasone used as a positive control group, but showed a 50% inhibitory activity at the level of 394 pg / ml. In other words, black ginseng was determined to be effective in inhibiting atopic dermatitis and the main activity was black ginseng saponin.

2-5. IL -8 secretion inhibitory activity

In order to measure IL-8 secretion inhibitory activity in the sample obtained in the above example, the method disclosed in the literature was modified to add a mite extract known to cause atopy to THP-1 cells to promote secretion of IL-8. Black ginseng extract was added and observed to decrease (Lee JS, Kim IS, Ryu JS, Yun CY., House dust mite, Dermatophagoides pteronissinus increases expression of MCP-1, IL-6, and IL-8 in human monocytic THP -1 cells.Cytokine. 2008, 42 (3): 365-71. 2008.).

As a result of the experiment, as shown in Table 2, the treatment of mite extracts that induce atopy on THP-1 cells can be seen that the secretion of IL-8 is sharply increased (2013pg / ml). In this pathological condition, black ginseng extract (BPGE) of the present invention was reduced to 1656 pg / ml. On the other hand, the treatment of black ginseng saponin soluble extract of black ginseng butanol (BPGBE) is weaker than the dexamethasone used as a positive control group, but was found to be reduced to the level of about 50% to 1103g / ml level. In other words, black ginseng was determined to be effective in inhibiting atopic dermatitis and the main activity was black ginseng saponin.

matter Volume of MCP-1 (pg / ml) Amount of IL-6 (pg / ml) IL - 8 of  Sheep ( pg Of ml ) Normal group 31 19 76 Tick treatment group 582 803 2013 Dexamethasone treatment group 29 21 132 Black Ginseng Extract (BPGE) 486 697 1656 Black ginseng Butanol  Soluble extract ( BPGBE ) 86 394 1103

Taken together, black ginseng irradiated with atopy inhibits the activity, especially MCP-1 secretion was reduced to almost normal levels.

Experimental Example 2. Acute Toxicity Test

The acute toxicity test was carried out as described below using specific pathogen-free (SPF) SD rats of 6 weeks old supplied from the experimental supply center.

After the oral administration of the black ginseng root crude extract of Example 2 at a dose of 1 g / kg to two animals in each group, the mortality, clinical symptoms, and weight changes of the animals were observed. An autopsy was performed, and autopsy was performed to visually observe the abnormalities of the organs and thoracic organs.

As a result of the experiment, there were no clinical symptoms or dead animals in all animals treated with the test substance, and no toxic change was observed in weight change, blood test, blood biochemical test, and autopsy findings. As a result, the extract of the present invention did not show a change in toxicity even in rats up to 1 g / kg, respectively, it was confirmed that the minimum lethal dose (LD ₅₀ ) is determined to be a safe substance more than 1 g / kg.

It is merely to illustrate the preparation of the pharmaceutical composition comprising the black ginseng root extract of the present invention as follows, it is not intended to limit the present invention by this only intended to explain in detail.

Preparation Example 1. Preparation of powder

BPGE (Example 2) 20 mg

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

Formulation Example 2. Preparation of tablets

BPGBE (Example 3) 10 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

Formulation Example 3. Preparation of capsules

BPGE (Example 2) 10 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Formulation example  4. Preparation of injections

BPGBE (Example 3) 10 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO ₄ 12H ₂ O 26 mg

(2 ml) per ampoule in accordance with the usual injection method.

Formulation example  5. Liquid  Produce

BPGE (Example 2) 20 mg

10 g per isomer

5 g mannitol

Purified water

Each component was added to purified water in accordance with the usual liquid preparation method and dissolved, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was adjusted to 100 ml with purified water, And sterilized to prepare a liquid preparation.

Formulation example  6. Manufacture of health food

BPGBE (Example 3) 1000 mg

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

0.13 mg of vitamin B ₁

0.15 mg of vitamin B ₂

0.5 mg of vitamin B ₆

Vitamin B ₁₂ 0.2 g

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

50 μg folic acid

Calcium Pantothenate 0.5mg

Mineral mixture quantity

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

Formulation example  7. Manufacture of health drinks

BPGE (Example 2) 100 mg

15 g of vitamin C

100 g of vitamin E (powder)

Iron lactate 19.75 g

3.5 g of zinc oxide

Nicotinamide 3.5 g

0.2 g of vitamin A

Vitamin B ₁ 0.25 g

Vitamin B ₂ 0.3 g

Water quantification

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 ° C for about 1 hour. The resulting solution was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, It is used in the production of the health beverage composition of the invention.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Claims (12)

Pharmaceutical composition for the prevention and treatment of atopic diseases containing black ginseng extract as an active ingredient. The method of claim 1,
The black ginseng, the first step of washing 4 to 6 years old Korean ginseng root three times for 10 minutes with an ultrasonic cleaner, and drying for 22 to 26 hours at a temperature of 35 to 45 ℃; Putting the dried Korean ginseng root into a steamer and steaming for 4 to 6 hours at a temperature of 85 to 105 ° C except for a preheating time; A third step of drying at a temperature of 55 to 65 ° C. for 10 to 14 hours; A fourth step of steaming for 5 to 7 hours at a temperature of 110 to 120 ° C. under a pressure of 0.1 to 0.20 MPa; Pharmaceutical composition, characterized in that the black ginseng obtained through a process comprising the step of obtaining a black ginseng root made by performing a first step of drying for 10 to 14 hours in a drier. The method of claim 1,
The black ginseng extract is a pharmaceutical composition comprising black ginseng root crude extract (BPGE) and black ginseng root butanol soluble extract (BPGBE). The method of claim 3, wherein
The black ginseng root crude extract (BPGE) is added to the water of about 3 to 7 times, preferably 4 to 6 times the mass of the black ginseng root prepared by the high temperature and high pressure method, and left at room temperature for 3 to 7 hours, including purified water. Water, lower alcohols having 1 to 4 carbon atoms or mixed solvents thereof, preferably 80% ethanol, added 5 to 20 times the mass, preferably 8 to 12 times the mass, 2 to 8 hours, preferably 3 Pharmaceutical composition, characterized in that the crude extract of black ginseng root of the present invention through the cooling, filtration and concentration step of the extract obtained by refluxing 1 to 5 times, preferably 3 times during the time to 4 hours. The method of claim 3, wherein
The black ginseng root butanol soluble extract (BPGBE) is suspended by adding water to the black ginseng root crude extract, and then adding non-polar substance by adding ether or dichloromethane; Pharmaceutical composition, characterized in that the ginseng saponin-containing black ginseng root butanol soluble extract obtained through the final step of extracting saponin present in ginseng 2-6 times and concentrated under reduced pressure by adding saturated butanol to the remaining water layer. The method of claim 1,
The atopic disease is a pharmaceutical composition characterized in that it comprises atopic dermatitis, eczema, psoriasis and the like. Health functional food for the prevention and improvement of atopic diseases containing black ginseng extract as an active ingredient. 8. The method of claim 7,
The black ginseng, the first step of washing 4 to 6 years old Korean ginseng root three times for 10 minutes with an ultrasonic cleaner, and drying for 22 to 26 hours at a temperature of 35 to 45 ℃; Putting the dried Korean ginseng root into a steamer and steaming for 4 to 6 hours at a temperature of 85 to 105 ° C except for a preheating time; A third step of drying at a temperature of 55 to 65 ° C. for 10 to 14 hours; A fourth step of steaming for 5 to 7 hours at a temperature of 110 to 120 ° C. under a pressure of 0.1 to 0.20 MPa; Health functional food characterized in that the black ginseng obtained through a process comprising the step of obtaining a black ginseng root made by performing a fifth step of drying for 10 to 14 hours in a dryer once. 8. The method of claim 7,
The black ginseng extract is a health functional food comprising black ginseng root crude extract (BPGE) and black ginseng root butanol soluble extract (BPGBE). The method of claim 9,
The black ginseng root crude extract (BPGE) is added to the water of about 3 to 7 times, preferably 4 to 6 times the mass of the black ginseng root prepared by the high temperature and high pressure method, and left at room temperature for 3 to 7 hours, including purified water. Water, lower alcohols having 1 to 4 carbon atoms or mixed solvents thereof, preferably 80% ethanol, added 5 to 20 times the mass, preferably 8 to 12 times the mass, 2 to 8 hours, preferably 3 Health functional food, characterized in that the black ginseng root crude extract of the present invention through the cooling, filtration and concentration step of the extract obtained by refluxing 1 to 5 times, preferably 3 times for a time to 4 hours. The method of claim 9,
The black ginseng root butanol soluble extract (BPGBE) is suspended by adding water to the black ginseng root crude extract, and then adding non-polar substance by adding ether or dichloromethane; Health functional food characterized in that the ginseng saponin-containing black ginseng root butanol soluble extract obtained by the final step of extracting saponin present in ginseng 2-6 times and concentrated under reduced pressure by adding saturated butanol to the remaining water layer. 8. The method of claim 7,
The atopic disease is a health functional food, characterized in that it includes atopic dermatitis, eczema, psoriasis.