KR20040099670A - A COMPOSITION CONTAINING GENSENOSIDE Rh2 AS AN ACTIVE MATERIAL FOR PREVENTING BRAIN CELL - Google Patents

Abstract

PURPOSE: Provided is a composition containing ginsenoside rh2 as an active ingredient for preventing brain cell. The ginsenoside rh2 is preferably manufactured from the extract of red ginseng in high yields and prevention and treatment effects on stroke. CONSTITUTION: The composition for preventing brain cell is characterized by containing 0.02-90 dried weight % of ginsenoside rh2 as an active ingredient. The ginsenoside rh2 is manufactured by the steps of: washing ginseng with water and acid-containing water; steaming the ginseng at 98-100 deg.C for 1-10 hours, preferably 2-5 hours in a steamer, then drying the steamed ginseng at 60 deg.C for 5 hours in a dryer to give red ginseng; extracting the red ginseng with water or organic solvent to give extract; and inoculating bacterioides sp. and Fusobacterium sp. to the extract of red ginseng.

Description

Brain cell protective composition containing ginsenoside Rｈ2 as active ingredient {A COMPOSITION CONTAINING GENSENOSIDE Rh2 AS AN ACTIVE MATERIAL FOR PREVENTING BRAIN CELL}

The present invention relates to a novel brain cell protective composition containing ginsenoside Rh2 as an active ingredient, in particular a composition for preventing or treating stroke. In addition, ginsenosides Rh2 contained in the composition according to the present invention may preferably be prepared from ginseng, and more specifically, red ginseng prepared from ginseng after water extraction or organic solvent extraction, and then, as a lactic acid bacterium or enteric bacteria, High yield can be obtained by conversion.

Ginseng is a perennial ripening root belonging to the genus Oga ginseng by plant taxonomy and about 11 species are known on earth. Representative species (1) Korean ginseng ( Panax ginseng C.A.Meyer) is native to the Far East of Asia (33-48 ° North Korea, North Manchuria, and parts of Russia), and is very effective. (2) American Ginseng ( Panax quinquefolium L) (3) Panax notoginseng FHChen is wild or grown in southwestern Guangxi province from southeastern Yunnan Province of China, and (4) Panax japonicus CA Meyer is Japan, China. It is known to be distributed in the southwestern region and Nepal (和 漢 藥 百科 圖鑑, Vol. 1, pp. 1-8, Nambatsunehiki, Hoikusa, 1980).

Ginseng is not only stored as a commodity in the new agricultural plant, but has been used as a valuable medicine since ancient times. Many pharmacological experiments so far have revealed that ginseng enhances the nonspecific resistance of the body to stress and has an acidic action. In addition, hypertension, insulin action enhancement, hypoglycemic effect in alloxan diabetic mice, liver RNA synthesis, protein synthesis, sugar and lipid metabolism promoting effect, and anticancer effect were found.

These ginsengs have been used for various diseases such as psychiatric diseases, nervous system diseases and diabetes in the form of herbal medicine mainly in Asian countries such as Korea, China, Japan, and saponins, the main components of the ginseng, are tonic, gangjeong, soothing It is known to have effects on hematopoiesis, antihypertension, etc. (Nambatsuneki et al., 1 , pp. 1-8, 1980).

On the other hand, Korean Patent Publication No. 2000-058997 discloses a manufacturing method of making acid-glycosylated ginseng that is easily absorbed by using sulfuric acid or hydrochloric acid in the decomposition of ginseng by acid-glycosylation.

In addition, Korean Patent No. 1996-017670 discloses a method for producing a ginseng composition containing a large amount of ginsenosides Rg3 and Rg5, unlike conventional ginseng, by treating ginseng at 120 to 180 ° C. to prepare processed ginseng with enhanced efficacy. Is disclosed.

As described above, the prior art is merely a method of preparing ginseng containing a large amount of ginsenosides Rg3 and Rg5 or a low molecular weight ginseng by acid treatment or heat treatment of ginseng. Thus, there is a limit in obtaining a metabolite of saponin, such as ginsenoside Rh2, compound K, which does not exist in nature or is present in very small amounts, but has excellent pharmacological action.

In this regard, Korean Patent No. 1997-061909 discloses a method for mass production of metabolites of saponins centered on compound K from saponins of ginseng using intestinal bacteria.

On the other hand, stroke refers to a syndrome accompanied by sudden consciousness disorder and motor paralysis caused by blood circulation disorders of the brain. There are two main types of strokes: an ischemic stroke that occurs in the ischemic state of the brain tissue due to a decrease or blocking of blood supply to the brain tissue, and the other is a blood vessel that bursts and causes bleeding into the brain tissue. It is a hemorrhagic stroke that occurs. Clinical reports have shown that ischemic stroke accounts for about 80% of all stroke patients.

In addition, stroke is a greater risk factor than brain damage, including severe brain nerve cell damage caused thereby. Therefore, many studies have been conducted in the art to find the mechanism of occurrence of brain injury due to stroke. However, although the cause of brain nerve cell damage due to stroke has not been specifically identified due to the complexity of the mechanism of brain nerve cell damage, the release of excessive excitatory neurotransmitters, the generation of free radicals, and the inhibition of protein synthesis. , Gene expression abnormalities, and immune response activation.

According to recent reports, contrary to existing theories that the nervous system is an immune privileged organ, multiple sclerosis, trauma, Alzheimer's disease, stroke, especially cerebral ischemic stroke and various brain injuries. Inflammatory reactions may occur even in the central nervous system, and neurodegeneration may be caused by such inflammatory reactions. With this report, it has been speculated that the immune response may be involved in the mechanism of brain cell damage caused by stroke, and as a result, many studies have been known to suppress brain cell damage after stroke through immunosuppression in the nervous system. Is being performed.

For example, many people complain of arthritis or pain, reporting that inhibition of cyclooxygenase-2 (COX-2) in cerebral ischemia has a neuroprotective effect on the inhibition of PGE ₂ production and the resulting inhibition of glutamate. Since COX-2 inhibitors are already being used, it is reported that epidemiological investigations of stroke incidence in these patients could be new targets for the development of prevention and treatment of stroke in the future (Iadecola C. et al., PNAS). ., 30, pp1294-1299, 2001) .

In addition, inflammatory reactions occur in the central nervous system, which has been reported to be one of the leading causes of neurodegeneration. Inflammatory responses after cerebral ischemia are likely to be targets for new stroke, especially cerebral ischemia treatments. It may consist of drugs that inhibit enzymes involved in the free or inhibit the deposition of neutrophils (Dirnagl U. et al., TINS ., 22 , pp391-397, 1999).

In addition, neutrophils penetrated into the brain tissues induce inducible nitric oxide synthase (iNOS) which can release NO, and it is known that NO is released from induced iNOS, leading to brain cell damage, and to the activation of NMDA receptors by glutamate. Neurotoxicity is also reported to be via NO, and development of neuroprotective agents related to inhibition of iNOS expression has been made (Vaughan CJ. Et al., Stroke , 30 , pp1969-73, 1999).

As discussed above, the inflammatory response in the nervous system, which occurs after stroke, especially cerebral ischemia, can lead to brain cell damage and, therefore, can inhibit brain cell damage and treat brain damage by inhibiting inflammation in nerves. Revealed.

The present inventors have conducted a lot of research to find a substance that can protect the brain cells in the event of a stroke while inhibiting the occurrence of a stroke, as a result of the specially processed extract of ginseng, or its lactic acid bacteria fermentation product has a very good brain cell protection effect In particular, it has been found to have a preventive and therapeutic effect on stroke, and based on this finding, on April 8, 2002, as a "composition for the prevention and treatment of stroke containing specially processed ginseng extract with increased efficacy" Filed (2002-18844).

Therefore, the present inventors have conducted a number of studies to find a substance having a protective effect on brain cells, and a stroke prevention and treatment effect, especially in the ginseng extract, or its lactic acid bacteria or enteric bacterial fermented product, which have been specially processed, Senoside Rh2 was found to be an effective ingredient exhibiting the above effects and completed the present invention.

It is therefore an object of the present invention to provide novel brain cell protective compositions.

1 is a diagram illustrating the effect on the brain ischemia of the red ginseng extract bioconversion product and ginsenoside Rh2. (In FIG. 1, Cont represents a control group administered with vehicle only; GB represents an oral administration group of 100 mg / kg of raw white ginseng prepared in Comparative Example 1; HB 100 mg / kg of red ginseng extract prepared in Example 1 HM represents the oral administration group of 100 mg / kg of the red ginseng extract prepared in Example 2 HIM represents the oral administration group of 100 mg / kg of the red ginseng extract prepared in Example 3 Rh2 represents the oral administration group of ginsenoside Rh2 100 mg / kg; the number of animals in the control group is 10 and the number of animals in each experimental group is 5, and * means p <0.05.

In order to achieve the above object, the composition according to the present invention contains ginsenoside Rh2 as an active ingredient of 0.02 to 90% by weight relative to the total dry weight of the composition, thereby protecting the brain cells, in particular stroke, more specifically brain It is characterized by having a prophylactic or therapeutic effect of ischemic stroke.

Hereinafter, the present invention will be described in more detail.

Ginsenoside Rh2 contained as an active ingredient in the composition according to the present invention can be prepared using ginseng as a raw material. That is, ginsenoside Rh2 can be obtained by extracting the product (red ginseng) obtained by heating ginseng at 98-100 degreeC in water or an organic solvent, and then bioconverting the obtained extract liquid to lactic acid bacteria or enterobacteriaceae. The ginsenoside Rh2 thus obtained may be any of 20 (R) -ginsenoside Rh2, 20 (S) -ginsenoside Rh2 or a mixture thereof.

More specifically, it may be produced by a method comprising the following steps (1) to (4):

(1) washing ginseng raw materials with water and then further washing with water containing optionally acid to remove foreign matter from ginseng raw materials, especially fresh ginseng (optional process);

(2) The washed ginseng obtained through the above step (1) was steamed at 98-100 ° C. for 1-10 hours, preferably 2-5 hours, in a conventional red ginseng manufacturer, and then the obtained product was about 60 Preparing red ginseng by drying in a conventional dryer for about 5 hours at &lt; RTI ID = 0.0 &gt;

(3) obtaining water extract or organic solvent extract from red ginseng obtained in step (2);

(4) bioconversion of the red ginseng water extract or organic solvent extract using lactic acid bacteria or intestinal bacteria.

On the other hand, the step (1) is an optional step, the step (2) to (4) may be an essential continuous manufacturing step. In addition, the process (4) may not be cultured by mixing the red ginseng extract and the strain, it can be prepared to encapsulate the reaction in the human intestine, all these processes are selectively depending on the form of the final ginseng product Applicable

On the other hand, ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc and the like contained in the raw materials are converted to ginsenoside Rg3 during the manufacturing process of red ginseng, and ginsenoside Rg3 by bioconversion Etc. are bioconverted through lactic acid bacteria or enteric bacteria and converted to the final metabolite, ginsenoside Rh2. Those skilled in the art can make appropriate modifications to the method including the above processes so that a large amount of ginsenoside Rh2 is contained as a final product, and other processes other than the above processes may be included. It will be understood that modifications and the like apparent to those skilled in the art are within the scope of the present invention.

Hereinafter, each process is demonstrated concretely.

In the step (1), the ginseng raw materials include ginseng and processed ginseng products and by-products, and preferably include ginseng, red ginseng, white ginseng, rice ginseng, ginseng leaves, ginseng extract and ginseng in powder form. In addition, the washing step of the step (1) can be washed only with water, but after washing with water, especially with an aqueous solution containing a physiologically acceptable acid, it is possible to remove the contaminated microorganisms attached to the ginseng raw materials. have. Ingredients that can be used for the acid wash include acetic acid, citric acid, lactic acid or food with acidity and the like.

According to the present invention, when washing only with water, the total bacteria count was about 13 × 10 ³ / g, but when washing with an acid-containing aqueous solution was further performed after washing, the total bacteria count was about 6 × 10 ³ / g. The total bacterial count was counted, and it was shown that washing with an acid-containing aqueous solution could remove more than about 50% of contaminating microorganisms (counting method: about 5 g of ginseng was accurately weighed and placed in a sterile pack (Korea 3M Co., Ltd.). (9 times), finely grind, dilute 10 times and 100 times again, of which 1 ml is inoculated vertically into petrifilm (Three Korea Inc.), cover the top film without foaming, and then press down on the pressure plate. After 2 seconds to 60 seconds, when gelation is complete, incubate for about 24-48 hours to count red colonies). On the other hand, as the acid-containing aqueous solution used therein, acetic acid-containing aqueous solution, specifically, 0.1% acetic acid-containing aqueous solution can be used.

Red ginseng is produced through the process (2) (Commercial ginseng products are commercially available ginseng harvested immediately; white ginseng dried at room temperature; red ginseng prepared by heating the ginseng at 98 to 100 ℃ There is this). In order to manufacture the ginsenoside Rh2 desired in the present invention, it is preferable to prepare red ginseng from fresh ginseng.

On the other hand, a method for producing red ginseng from ginseng raw materials is commonly known in the art, for example, washed ginseng obtained through the above step (1) is 1-10 at 98-100 ° C with a conventional red ginseng manufacturer It can be prepared by steaming for an hour, preferably 2-5 hours, and then drying in a conventional dryer at 60 ° C. for 5 hours, but obvious modifications in the art are carried out for the steaming temperature and time, drying process after steaming, and the like. Can be. Such modifications are also within the scope of the present invention.

Subsequently, in step (3), water extract or organic solvent extract is obtained from red ginseng. For example, about 1 to 50 times weight / volume of water in dried red ginseng, alcohol as organic solvent, preferably ethanol, in particular 70% alcohol, or 20 to 80 ° C. in a mixture of water and alcohol, preferably Is extracted and concentrated for 1 hour to 2 days, preferably 3 hours at a temperature of 50 to 60 ℃, to obtain a water extract or organic solvent extract from red ginseng. However, there is no particular limitation on the kind of extraction solvent, extraction temperature and extraction time, and if ginsenoside Rh 2 is obtainable, it can be used arbitrarily.

The step (4) is a step of bioconversion of red ginseng extract, especially water extract or organic solvent extract using enterobacteriaceae or lactic acid bacteria, microorganisms used for bioconversion, conditions for bioconversion using the microorganisms, and bioconversion The period is not particularly limited as long as ginsenoside Rh2 can be obtained in sufficient yield after bioconversion. For example, the red ginseng water extract or organic solvent extract prepared in step (3) may be cultured at 20-50 ° C., preferably 25-40 ° C. for 6 hours to 5 days, preferably 24 hours to 48 hours. In, by incubating with lactic acid bacteria or enterobacteria, bioconversion process can be performed.

On the other hand, the enteric bacteria that can be used in the process (4) can be any one that can metabolize ginseng saponin to produce the ginsenoside Rh2, preferably, genus Bacterioides, Fusobacte Enteric bacteria of the genus Fusobacterium, more preferably bacteroid JY-6 ( Biol. Pharm. Bull. , 23 , pp1481-1485, 2000), Fusobacterium K-60 ( Biol. Pharm. Bull. , 23 , pp1481-1485, 2000).

In addition, the lactic acid bacteria used in the bioconversion process of the red ginseng extract of step (4) can be any one that can metabolize ginseng saponin to produce the ginsenoside Rh2, preferably Bifidobacterium (Bifidobacterium) Lactic acid bacteria, more preferably Bifidobacterium infantis, Lactobacillus lactis, Bifidobacterium H-1 (KCCM-10493), Bifidobacterium C-13 (Kyung Hee University) Pharmacy Kim Dong-hyun)), Bifidobacterium C-34 (Kyung Hee Univ.

Among them, Bifidobacterium H-1 is a microorganism developed by the present inventors to obtain a bioconversion product of red ginseng extract containing a large amount of ginsenoside Rh2, wherein Bifidobacterium H-1 is a deposit number. It has been deposited with the Korean Culture Center of Microorganisms as KCCM-10493 (Deposit Date: 2003.05.06).

Bifidobacterium H-1 strain according to the present invention is Gram-positive, bacillus, anaerobic strain, fructose 6-phosphate phosphoketolase (fructose 6-phosphate phosphoketolase) positive, sugar availability as follows Has

Kinds Sugar soluble (+, positive;-negative) Kinds Sugar soluble (+, positive;-negative) Amygdalin arabinose cellobiose dextrin escalin fructose galactose gluconate glucose glycogen inositol inulin lactose -+-+ /-++-+-+ /-+ Maltose mannitol mannose mannose +++-+++ /-+-+-+

On the other hand, the Bifidobacterium H-1 has the same general taxonomic characteristics as compared to the existing Bifidobacterium, but when used in lactic acid bacteria fermentation of ginseng, a higher content of ginsenoside Rg3 to ginsenoside Rh2. The conversion can provide a bioconversion product containing a large amount of ginsenoside Rh2.

The ginseng extract of the present invention obtained through the above manufacturing steps (1) to (4) has a ratio of (ginsenoside Rh2 / (ginsenoside Rg3 + Rc + Rd + Rb1 + Rb2) of 0.1 or more, which is the above production process It contains 10 times more ginsenoside Rh2 than red ginseng extract obtained through (1) to (3).

In addition, the bioconversion product of the red ginseng which passed through the said manufacturing process (1)-(4) may be used as it is, and may be used by freeze-drying it. Alternatively, centrifugation of the extract of the red ginseng bioconversion product in water or organic solvent extraction, and then filtered the supernatant, concentrated under reduced pressure to remove impurities and sediment in the bioconversion red ginseng extract and concentrated at the same time, and then freeze-dried and It can also be used in the form of concentration through the same drying process.

As a suitable solvent for extracting ginsenoside Rh2 as an active ingredient from the extract of the bioconversion product, a lower alcohol solution such as water, methanol and ethanol can be used, and as an extraction method, a special extraction method such as supercritical extraction can be used. The active ingredient may be separated, and then a drying process by a lyophilization method and an agitation process or a dilution process for manufacturing a processed beverage or a processed food may be further performed.

In addition, according to the present invention, the bioconversion product of red ginseng prepared by the preparation method comprising the steps (1) to (4) was extracted with BuOH, and then concentrated and silica gel column (5 x 50 cm, elution solvent: chloroform : Methanol = 20: 1) can be isolated to isolate ginsenoside Rh2. Its chemical properties are as follows:

20 (R) -ginsenoside Rh2: white crystals, mp 208-210 ° C. (dec.) FAB-MS ( m / z ) 623 [M + 1] ⁺ .

In addition, the results of ¹³ C-NMR spectrum analysis thereof are as shown in Table 1 below:

Chemical Displacement of 20 (S) -ginsenoside Rh2 Isolated from ¹³ C-NMR Spectrum carbon Chemical displacement (δ / ppm) carbon Chemical displacement (δ / ppm) C-1 39.12 3-Glc1 106.92 C-2 27.05 2 75.76 C-3 88.78 3 78.72 C-4 39.66 4 71.65 C-5 56.35 5 78.34 C-6 18.43 6 63.05 C-7 35.15 C-8 40.00 C-9 50.38 C-10 36.94 C-11 32.02 C-12 70.96 C-13 49.54 C-14 51.69 C-15 31.32 C-16 26.70 C-17 54.77 C-18 16.77 C-19 15.61 C-20 72.94 C-21 26.83 C-22 35.88 C-23 22.97 C-24 126.30 C-25 130.73 C-26 25.78 C-27 17.66 C-28 28.14 C-29 16.34 C-30 17.65

Ginsenoside Rh2 obtained by the manufacturing method of the said ginsenoside Rh2 may be any of 20 (R) -ginsenoside Rh2, 20 (S) -ginsenoside Rh2, or a mixture thereof.

According to the present invention, the ginsenoside Rh2 isolated from the bioconversion product of red ginseng may be used alone to further improve the protection of brain cells, in particular, the treatment or prevention of stroke, but as a pharmacologically active ingredient contained in the red ginseng extract. Panaxitridol, Panaxidol, Panaxinol, Ginsenoside Rc, Ginsenosides Rb1, Rb2, Compound K, 20 (R) -Protopanaxadiol, 20 (S) -Protopanaxadiol, 20 ( It may further contain one or more saponin derivatives selected from the group consisting of S) -ginsenoside Rhl, 20 (S) -protopanaxatriol. Preferably, it is used in the form of a bioconversion product (hereinafter referred to as 'bioconversion product of red ginseng extract') by lactic acid bacteria or enterobacteriaceae of the red ginseng extract.

On the other hand, the present invention can provide a variety of processed ginseng products using the bioconversion product of the red ginseng extract.

For example, the processed ginseng (or red ginseng) products include ginseng dry powder, extract, ampoule preparations, teas, tablets and the like.

In addition, the present invention provides a dietary supplement comprising a brain cell protective composition according to the present invention exhibiting a protective effect against brain cells and stroke, and a food acceptable additive. The food-acceptable food supplement additives are commonly known in the art, and those skilled in the art will obviously recognize the method for preparing the health supplement food. In addition, the health supplement may be any of a variety of forms, such as health drinks, jelly, candy, sweets. The present invention is not limited by these forms.

The effect on the prevention and treatment of stroke of the composition according to the present invention, which contains ginsenoside Rh2 as an active ingredient, ischemic properties of ugonin and its derivatives in vivo using a midbrain aortic occlusion model, in which the test method has already been established. This can be confirmed by the effect of inhibiting stroke.

The method involves inserting nylon filament into the internal carotid artery to occlude the midbrain aorta and then reperfusion by removing the filament after 120 minutes. At this time, the samples are orally administered to confirm the effect on ischemic stroke. It is.

On the other hand, the composition according to the present invention containing ginsenoside Rh2 as an active ingredient may further comprise suitable carriers, excipients and diluents commonly known in the art.

As carriers, excipients and diluents which may be included in the composition according to the invention containing ginsenoside Rh2 as an active ingredient, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia Rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil have.

Compositions according to the present invention containing ginsenoside Rh2 as an active ingredient, oral formulations, external preparations, suppositories, and sterilization, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, respectively, according to conventional methods It can be formulated and used in the form of injectable solutions.

The amount of ginsenoside Rh2 contained in the composition according to the present invention may vary depending on the age, sex, and weight of the patient, but the amount of 0.1-100 mg / kg may be administered once to several times daily. In addition, the dosage of the extract or compound may be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age and the like. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

The composition according to the present invention containing ginsenoside Rh2 as an active ingredient can be used in various forms such as pharmaceuticals, foods and beverages for the protection and treatment of brain cells and stroke. Examples of foods to which the ginseng extract or compound can be added include various foods, beverages, gums, teas, vitamin complexes, and health supplements.

Ginsenoside Rh2 itself contained in the composition according to the present invention has little toxicity and side effects, so it is a drug that can be used safely even for long-term administration for the purpose of prevention.

The composition according to the present invention containing ginsenoside Rh2 as an active ingredient may be added to food or beverage for the purpose of protecting brain cells and preventing stroke. At this time, the amount of the extract or compound in the food or beverage is generally added to the health food composition of the present invention to 0.01 to 15% by weight of the total food weight, the food health beverage composition is 0.02 to 0.1g based on 100 ml Preferably, it can add in the ratio of 0.3-1 g.

The health beverage composition of the present invention has no particular limitation on the liquid component except for containing the extract as an essential ingredient in the indicated proportions, and may contain various flavors or natural carbohydrates, etc. as additional ingredients, like ordinary drinks. 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 conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The ratio of said natural carbohydrate is generally about 1-20 g, preferably about 5-12 g per 100 m1 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. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

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

EXAMPLE

Example 1 Preparation Example of Processed Ginseng Extract (1)

1 kg of 5 year old ginseng purchased from Geumsan Ginseng Market was washed with water and finally washed with acetic acid containing 0.1%, steamed at 98-100 ℃ for 4 hours with steaming machine (KI-HT01; Doosung Engineering), and dried at 60 ℃. It is dried for 5 hours with KU-DR99; International Corporation. This dried red ginseng was extracted with 70% alcohol to obtain a red ginseng extract. This was concentrated, suspended in water, extracted with BuOH and concentrated to obtain a red ginseng extract (hereinafter referred to as HB).

Example 2 Preparation Example of Processing Ginseng (Red Ginseng) Extract (2)

1 kg of 5 year old ginseng purchased from Geumsan Ginseng Market was washed with water and finally washed with acetic acid containing 0.1%, steamed at 98-100 ℃ for 4 hours with steaming machine (KI-HT01; Doosung Engineering), and dried at 60 ℃. It is dried for 5 hours with KU-DR99; International Corporation. This dried red ginseng was extracted with 70% alcohol to obtain a red ginseng extract. The red ginseng extract was suspended in sterile distilled water to 5%, added Bifidobacterium H-1 strain (wet weight 10 grams), incubated for 24 hours at 37 ° C, extracted with BuOH, concentrated and processed. 4 g of ginseng was obtained (hereinafter referred to as HM).

Example 3 Preparation of Processed Ginseng Extract (3)

1 kg of 5 year old ginseng purchased from Geumsan Ginseng Market was washed with water and finally washed with acetic acid containing 0.1%, steamed at 98-100 ℃ for 4 hours with steaming machine (KI-HT01; Doosung Engineering), and dried at 60 ℃. It is dried for 5 hours with KU-DR99; International Corporation. This dried red ginseng was extracted with 70% alcohol to obtain a red ginseng extract. Suspend the red ginseng extract in sterile distilled water to 5% and add intestinal bacterium (10 grams of wet weight obtained by suspending human feces in tryptic soy broth) at 37 ° C. After incubation for 72 hours, the extract was extracted with BuOH and concentrated to obtain 4 g of processed ginseng (hereinafter referred to as HIM).

Example 4 Production of Ginsenoside Rh2 from Processed Ginseng Extract Preparation (9)

1 kg 5 years old ginseng purchased from Geumsan Ginseng Market was washed with water and finally washed with 0.1% acetic acid and steamed at 98-100 ℃ for 4 hours with steaming machine (KI-HT01; Doosung Engineering), and dried at 60 ℃. It is dried for 5 hours with KU-DR99; International Corporation. This dried red ginseng was extracted with 70% alcohol to obtain a red ginseng extract. Suspend the red ginseng extract in sterile distilled water to 5%, add intestinal bacterium (10 g of cell weight obtained by suspending human feces in soy broth) and incubate for 24 hours at 37 ℃ Extracted with BuOH and concentrated to obtain a processed ginseng 12g, which was eluted with silica gel curl (5 x 50 cm, eluting solvent: chloroform: methanol = 20: 1) to separate 0.5g ginsenoside Rh2.

Comparative Example 1. Preparation of Raw Ginseng Extract

5 g of fresh ginseng, purchased from Geumsan Ginseng Market, is chopped and added 5 times the amount of water, extracted at 60 ℃ for 5 hours, concentrated under reduced pressure with a concentrator (Eyella, KN-IN Evaporator, Japan), and freeze-dried (three-way cold heat SFDSM24L model , Korea) to obtain 1 g of a dry white ginseng dry powder.

Example 5. Content Analysis Experiment

2 g each of the raw ginseng (Comparative Example 1) and the ginseng powders of Examples 1, 2 and 3 of Comparative Example 1 were taken, and 100 ml of methanol was extracted three times. The extract was concentrated under reduced pressure, and then suspended in 100 ml of water, extracted three times with 100 ml of ether, and concentrated under reduced pressure. Then, the resultant was extracted three times with 100 ml of butanol and concentrated under reduced pressure, and then the obtained concentrate was dissolved in 100 ml of methanol to obtain 100 mg of saponin fraction, which was then purified by TLC (developing solvent: CHCl ₃ : MeOH: H ₂ O = 65: 35: 10). , Coloring reagent: 5% methanol sulfate solution, Detector: Shimadzu TLC Scanner CS-9301PC) was analyzed to obtain the saponin content analysis results as shown in Table 2.

ingredient Content in Saponin Fraction (%) Comparative Example 1 Example 1 Example 2 Example 3 Ginsenoside Rb1 13.4 5.1 2.7 2.3 Ginsenoside Rb2 8.9 3.5 2.1 1.9 Ginsenoside Rc 9.2 1.2 2.5 2.0 Ginsenoside Rd 3.3 1.1 2.1 1.2 Ginsenoside Rg3 <1 6.8 2.5 2.3 Δ ²⁰ -ginsenoside Rg3 <1 2.5 1.2 <1 compound K <1 <1 2.6 2.8 Ginsenoside Rh2 <1 <1 3.2 3.9 Protopananacidio Diol <1 <1 <1 <1

As a result, the content of ginsenoside Rg3 increased rapidly when ginseng was removed, and the content of ginsenoside Rh2 metabolized by ginsenoside Rg3 in steamed ginseng (red ginseng) rapidly increased when ginseng was treated with lactic acid bacteria. Could confirm.

Experimental Example 1 Experiment of Effect on Brain Cell Protection and Stroke

The therapeutic effect of the processed ginseng extract of the present invention and the compounds isolated therefrom against brain cell protection and stroke are compared with the pharmacological effects of the raw white ginseng dry powder of Comparative Example 1 and the acidic white ginseng dry powder of Comparative Example 2 In order to perform the following experiment.

The rat ischemic animal model using rats (Biogenomeics, Korea) was administered at 1, 3, 10, and 50 mg / kg approximately 5 minutes before the start of reperfusion and reperfusion. Were extracted, a brain slice was made to a thickness of 2 mm using a brain matrix, and then stained using a 2, 3, 5-triphenyltetrazolinum chloride (TTC) staining method. The cerebral infarction was analyzed using an image analysis system. As a result, as shown in Figure 1, the biotransformation products of the red ginseng extract prepared in Examples 2 and 3, and ginsenoside Rh2 obtained in Example 4 showed significant brain cell protective activity. It has never been reported in the prior art that ginsenoside Rh2 has a protective effect on brain cells. Meanwhile, in FIG. 1, Cont is a control group (vehicle administration group), GB is Comparative Example 1, HB is Example 1, HM is Example 2, HIM is Example 3, and ginsenoside Rh2 represents a sample of Example 4. , * Indicates significant significance at 95% significance level compared to control.

In conclusion, the treatment of lactic acid bacteria or intestinal bacteria with ginseng shows the enhanced efficacy of preventing and treating the brain cells, especially the stroke. It has been found that it can be effective in manufacturing.

Hereinafter, an example of the preparation of the pharmaceutical composition will be described, but it is not intended to limit the present invention but merely to explain in detail.

Formulation Example 1 Preparation of Powder

According to the preparation method of powder in the pharmacopeia formulation, it is prepared in the following ingredient content per one packet.

Dry powder of Example 3 50 mg

Lactose · ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Talc ㆍ ················ 10 mg

Formulation Example 2 Preparation of Tablet

According to the preparation method of tablets in the pharmacopeia formulation, it is prepared in the following component content per tablet.

Dry powder of Example 3 50 mg

Corn starch ㆍ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Lactose · ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Magnesium stearate 2 mg

Formulation Example 3 Preparation of Capsule

According to the preparation method of capsules in the pharmacopeia formulation, it is prepared in the following component content per capsule.

Dry powder of Example 3 50 mg

Corn Starch ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Lactose · ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Magnesium stearate 2 mg

Formulation Example 4 Preparation of Injection

According to the preparation method of injection in the pharmacopeia formulation, it is prepared in the following component content per ampoules (2 ml).

Dry powder of Example 3 50 mg

Sterile distilled water for injection ㆍ ···············

pH regulator ㆍ ·······················

Formulation Example 5 Preparation of Liquid

According to the preparation method of the liquid formulation in the Pharmacopoeia General Formulation, it is prepared in the following component content per 100 ml of the liquid formulation.

Dry powder of Example 3 50 mg

Heterosaccharides ·················· 10 g

Mannitol ····················· 5 g

Purified water ㆍ ······················

In addition, the health beverage is prepared as follows.

0.1 to 80% of dry powder of Example 3, 5 to 10% of sugar, 0.05 to 0.3% of citric acid, 0.005 to 0.02% of caramel, 0.1 to 1% of vitamin C, and then mixed with 79 to 94% of purified water A syrup was made, and the syrup was sterilized at 85 to 98 ° C. for 20 to 180 seconds, mixed with cooling water at a ratio of 1: 4, and then injected with 0.5 to 0.82% of carbon dioxide gas to give a high content of ginsenoside Rh2. A carbonated beverage containing a bioconversion product was prepared.

Instant sterilization by homogeneously mixing the dry powder of Example 3 with subsidiary materials such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%) and water (75%) This was packaged in small packaging containers such as glass bottles and plastic bottles to prepare a healthy beverage.

Although the composition ratio is a composition suitable for a preferred beverage in a preferred embodiment, the compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and use purpose.

As described above, the bioconversion product of the red ginseng extract and the ginsenoside Rh2 isolated therefrom have a very excellent stroke treatment effect, therefore, the novel brain cells containing the ginsenoside Rh2 as an active ingredient Protective compositions can be very usefully used.

Claims (10)

Ginsenoside Rh2 as an active ingredient, based on the total weight of the composition, the brain cell protective composition, characterized in that it contains 0.02 to 90% by weight dry weight. 2. The brain cell protective composition according to claim 1, which protects the brain cells from brain cell damage induced by stroke. 3. The brain cell protective composition according to claim 2, wherein the stroke is an ischemic stroke. The brain cell according to any one of claims 1 to 3, wherein the ginsenoside Rh2 is prepared by water extraction or organic solvent extraction of red ginseng obtained by steaming ginseng, followed by bioconversion to enterobacteriaceae or lactic acid bacteria. Protective composition. The brain cell protective composition according to claim 4, wherein the ginsenoside Rh2 is prepared by a method comprising the following steps (1) to (4): (1) washing ginseng raw materials with water, and then further washing with water containing optionally acid to remove foreign substances from ginseng raw materials, especially fresh ginseng; (2) The washed ginseng obtained through the above step (1) was steamed at 98-100 ° C. for 1-10 hours, preferably 2-5 hours, in a conventional red ginseng manufacturer, and then the obtained product was about 60 Preparing red ginseng by drying in a conventional dryer for about 5 hours at &lt; RTI ID = 0.0 &gt; (3) obtaining water extract or organic solvent extract from red ginseng obtained in step (2); (4) bioconversion of the red ginseng water extract or organic solvent extract using lactic acid bacteria or intestinal bacteria. The brain cell according to claim 5, wherein the enteric bacteria that can be used in the step (4) are at least one enteric bacterium selected from the group consisting of Bacterioides and Fusobacterium strains. Protective composition. The method according to claim 5, wherein the lactic acid bacteria that can be used in the step (4) are Bifidobacterium infantis, Lactobacillus lactis, Bifidobacterium H-1 (KCCM 10493), BP Brain cell protective composition, characterized in that it is selected from one or more strains of lactic acid bacteria selected from the group consisting of gambakterium C-13 (Kyung Hee University Pharmacy Kim Dong-hyun) and Bifidobacterium C-34 (Kyung Hee University Pharmacy Kim Dong-hyun). The process according to any one of claims 1 to 3, wherein panaxitriol, panaxidol, panaxinol, ginsenoside Rc, ginsenosides Rb1, Rb2, compound K, 20 (R) -protopanaxa Further contains one or more saponin derivatives selected from the group consisting of diols, 20 (S) -protopanaxadiol, 20 (S) -ginsenoside Rh1, 20 (S) -protopanaxatriol Brain cell protective composition. Lactobacillus Bifidobacterium H-1 (KCCM 10493) A dietary supplement comprising a brain cell protective composition according to any one of claims 1 to 3 and a food supplement acceptable food additive which exhibit a protective effect on brain cell protection and stroke.