KR100555652B1 - Extract of processed Panax Species plant, process for preparing the same, and composition containing the same for preventing and treating cancer and allergy- mediated disease - Google Patents

Abstract

The present invention relates to a processed extract of Panax genus plants with enhanced potency, a method for preparing the same, and a composition for the prevention and treatment of cancer and allergy-related diseases containing the same, and the processed extracts of Panax genus plants according to the present invention. It has excellent anti-cancer or anti-allergic efficacy and can be used as a drug and health functional food useful for the prevention and treatment of cancer and allergy-related diseases.

Panax genus, anticancer, anti-allergic, extract, composition, health functional food.

Description

Extract of processed Panax Species plant, process for preparing the same, and composition containing the same for preventing and treating cancer and allergy- mediated disease}

The present invention relates to a processed extract of Panax genus plants with an excellent anti-cancer or anti-allergic effect, a method for preparing the same, and a composition containing the same.

Panax genus is a perennial root of the genus Araliaceae in the plant taxonomy and is known for over a dozen species on the earth. (1) Panax ginseng is a species in the Far East of Asia (33-48 ° North Korea, It is native to Northern Manchuria and parts of Russia, and has very good efficacy. (2) American Ginseng ( Panax quinquefolia ) grows and grows in the United States and Canada, and (3) Panax Notoginseng is located in southeastern Yunnan, China. Wild or cultivated in the southwestern region, (4) Panax japonica is distributed in Japan, southwestern China and Nepal, (5) Panax trifolia is distributed in eastern North America, (6) Himalayas Ginseng ( Panax pseudoginseng ) is distributed in Nepal, and (7) Vietnamese ginseng ( Panax vietnamensis ) is distributed in Vietnam. (Understanding Korean Ginseng, 9, Korean Ginseng Society, 1995; Advances in Ginseng Research, pp. 127-137, Korean Ginseng Society, 1998). Other Panax genus plants include Panax elegans tee climb (Panax elegatior), Panax Wan Jia Augustine (Panax wangianus), Panax pinra non-typhoid (Panax bipinratifidus).

These panax genus plants have a variety of physiological activity because it is often used as a medicine. In particular, Korean ginseng ( P. ginseng ) has been used as a valuable medicine since ancient times.

Many pharmacological experiments to date have revealed that ginseng strengthens the nonspecific resistance of living organisms to stress and maintains homeostasis. In addition, hypertension, insulin action, hypoglycemic effect in diabetic mice, hepatic RNA synthesis, protein synthesis, sugar and lipid metabolism promoting effect, anti-cancer effect was found. In fact, ginseng is known to have effects on tonic, gangjeong, soothing, hematopoietic and antihypertensive (Understanding Korean Ginseng, Korean Ginseng Society, 1995). On the other hand, American ginseng (or Hwagi ginseng) is a substitute for Korean ginseng and has been used for similar purposes as Korean ginseng.

The most important component in these Panax plants is saponin. The saponin component of the Panax genus plants has 1-4 sugars bound to the Dammarane skeleton, which is different from the saponins of other plants. In particular, about 30 kinds of saponin components are known in Korean ginseng, and high content among them is ginsenosides Rb1, Rb2, Rc, Rd, Rg1, Re, and the like. These saponin components show a variety of drugs, the type and strength of the drug is very different depending on the structure.

Recently, efforts have been made to change the efficacy and usefulness of ginseng by processing ginseng. In particular, the structure of the saponin in such a process may change the drug efficacy.

For example, Korean Patent Publication No. 2000-058997 (October 6, 2000) discloses a method for preparing acid-glycosylated ginseng that is easily absorbed using sulfuric acid or hydrochloric acid.

In addition, Korean Patent Publication No. 1996-017670 (May 23, 1996) shows that ginseng is processed at high temperature to produce processed ginseng with enhanced efficacy, unlike ginseng, which contains large amounts of ginsenosides Rg3 and Rg5. A method of making a composition is disclosed.

The above-mentioned prior art relates to a method of preparing a low molecular weight ginseng composition or ginseng containing a large amount of ginsenosides Rg3 and Rg5 by acid treatment or heat treatment of ginseng, not a method for obtaining a large amount of a specific saponin metabolite.

On the other hand, Korean Patent Publication No. 1996-004217 (February 22, 1996) discloses a method for mass production of metabolites centered on compound K using saponin of ginseng using intestinal bacteria.

Republic of Korea Patent Publication No. 1980-004291 (Method for producing lactic acid bacteria ginseng drink) (1980. 11. 08) separates the intrinsic fragrance component of ginseng and enzymatic decomposition to the organic nitrogen concentration of 0.2 ~ 0.8%, the production content of glucose is lactic acid bacteria When it is more than 3% suitable for fermentation, the method is adjusted to pH 3.8 ~ 4.8 with organic acid, and then the insoluble polymer protein and fiber are removed, the lactic acid bacteria are incubated in the eluate, and then, the intrinsic fragrance component of ginseng is disclosed. have.

Republic of Korea Patent Publication No. 1988-012502 (Method for preparing active lactic acid bacteria beverages using ginseng) (1988. 09. 28), enzymes are applied to fresh juice or extracts treated with ginseng or to increase or increase the ginseng treated dihydrate Subsequently, after culturing the lactic acid bacteria, skim milk, skim milk powder, carbonated water, and vitamins are added and mixed to prepare a carbonated aqueous beverage and ice cream of ginseng lactic acid bacteria with high nutritional value.

Korean Patent Publication No. 1996-23750 (1996. 06. 26) adds 0.001 to 2.39 wt% of ground ginseng or fresh ginseng, ground into 0.1 to 0.6 cm fine particles, and additionally contains water, vitamins, sugars and organic acids. A method for producing a fermented milk composition containing ginseng or ginseng, comprising the step of adding one or two or more ingredients selected from the group consisting of fruits, cereals, and vegetables.

The above-described methods were able to obtain a composition added with ginseng components by adding ginseng fragrance components or added ginseng powder to the existing fermented milk or lactic acid bacteria beverage.

However, the ginseng-containing compositions are merely a composition for beverages to which the ginseng component is added or merely disclose a method for mass production of the metabolite compound (compound K, IH-901) of ginseng saponin.

The present inventors have found that the Panax genus plant extract prepared through special treatment such as lactic acid bacteria or enterobacteriaceae treatment after heat or acid treatment of Panax genus plants is excellent in the treatment effect against cancer and allergic related diseases. Completed.

It is an object of the present invention to provide a process for producing a processed Panax genus plant extract with excellent anticancer or anti-allergic efficacy and an extract of the Panax genus plant produced thereby.

Another object of the present invention to provide a composition for the prevention and treatment of cancer and allergic related diseases containing the extract.

According to the above object, the present invention has a component ratio value ((sum of contents of ginsenosides Rk2, Rh3, PPD and DHPPD) / (sum of contents of ginsenosides Rg3, Rg5 and Rk1)) of 0.1 or more, preferably Provides an extract of the genus Panax plant that is at least 0.2, more preferably at least 0.5.

Wherein PPD is (20S) -protopanaxadiol and (20R) -protopanaxadiol, and DHPPD is 20 (21) -dehydroprotopanaxadiol and 20 (22) -dihydroproto Panaxanadiol is meant to include.

In another aspect, the present invention provides a specially processed Panax genus plant extract characterized in that the lactic acid bacteria or enterobacteriaceae is continuously treated in the extract obtained through acid treatment, heat treatment or a combination thereof.

The raw material of the genus Panax plant of the present invention is not limited, but specifically Panax ginseng , Panax quinquefolia , Panax ginseng ( Panax notoginseng ), which are Panax genus plants containing the dammarane type saponin. ), Panax japonica , Panax trifolia , Himalayan ginseng ( Panax pseudoginseng ), Vietnamese ginseng ( Panax vietnamensis ), or Panax elegatior , Panax wangianus , Panax bipinratifidus , Panax angustifolium , etc. are available, and saponin compounds having a murine skeleton such as roots, stems, leaves, brains, flowers, fruits and their tissue cultures are available. Panax plants containing all can be used as a raw material, the raw materials include Panax plants and processed products of Panax plants and by-products, preferably ginseng, red And it includes 403, misam, ginseng leaves, ginseng extracts and ginseng powder form.

In another aspect, the present invention provides a method for producing a specially processed Panax genus plant extract comprising the step of continuously treating the lactic acid bacteria in the extract obtained through the acid treatment, high temperature heat treatment and a combination thereof.

Hereinafter, the manufacturing process of the specially processed Panax genus plant extract of the present invention will be described.

(1) acid treatment process

Foods containing 0.01 to 50%, preferably 0.1 to 10%, of acid, preferably acetic acid, citric acid, lactic acid, or acidity (for example, Schizandra chinensis) of about 1 to 50 times the weight / volume of the Panax plant raw material Addition is incubated at a culture temperature of 20 to 80 ° C., preferably 40 to 70 ° C., for 1 hour to 2 days, preferably 3 hours to 12 hours.

In order to purify and isolate the culture, an organic solvent, preferably butanol, methanol, ether, and ethyl acetate, is added to the culture, followed by extraction using a solvent extraction method, and the extract is neutralized with a base. An acid treated Panax plant extract is prepared.

Wherein the Panax genus plant raw material and the like processed products and by-products of Panax genus plants, Panax genus plant, preferably a ginseng (Panax ginseng), American ginseng (Panax quinquefolia), jeonchilsam (Panax notoginseng), jukjeol three (Panax japonica , Panax trifolia , Panax pseudoginseng , or Vietnamese ginseng ( Panax vietnamensis ), or Panax elegatior , Panax wangianus , Panax bipinatitipus Panax bipinratifidus ), the roots, stems, leaves, flowers, fruits, brains, their tissue cultures, or extracts thereof of Panax angustifolium .

(2) heat treatment process

In the present invention, the high temperature heat treatment of the genus Panax plants is performed at 110 to 180 ° C. for 0.5 to 20 hours, preferably at 110 to 140 ° C. for 2 to 5 hours. The heating time depends on the heating temperature, which must be heated for a long time at low temperature and only a relatively short time at high temperature. At this time, the heating is performed using hot air or steam, nitrogen, helium, carbon dioxide, oxygen or a mixture of these, it is preferable to perform in a closed container such as a heat sterilizer to increase the efficiency. If desired, it may be desirable to add a small amount of water into the container, or to immerse the Panax plant itself in water to heat it in a closed container. The heat-treated Panax genus plants are extracted with water, methanol, ethanol, or a mixed solvent thereof to obtain a heat-treated extract. If necessary, the extract can be further purified. That is, by extracting an organic solvent such as butanol, ethyl acetate, ethanol in the extract to obtain a partially purified heat treatment extract.

Wherein the Panax genus plant raw material and the like processed products and by-products of Panax genus plants, Panax genus plant, preferably a ginseng (Panax ginseng), American ginseng (Panax quinquefolia), jeonchilsam (Panax notoginseng), jukjeol three (Panax japonica , Panax trifolia , Panax pseudoginseng , or Vietnamese ginseng ( Panax vietnamensis ), or Panax elegatior , Panax wangianus , Panax bipinatitipus Panax bipinratifidus ), the roots, stems, leaves, flowers, fruits, brains, their tissue cultures, or extracts thereof of Panax angustifolium .

(3) Bioconversion Process

Lactic acid bacteria or enteric bacteria were added to the Panax genus plant extract obtained by the acid treatment of (1) or the heating treatment of (2) for 8 hours to 8 days, preferably 24 hours to 3 days, and 20 to 50 ° C. Preferably it is cultured at a culture temperature of 25 to 40 ℃ to obtain a strain-treated Panax genus plant extract.

Lactic acid bacteria that can be used for this purpose include the metabolism of ginsenosides Rg3, Rg5, Rk1 to ginsenosides Rh2, Rh3, Rk2, protofanaxadiol (PPD), 20-dihydroprotopanaxadiol (DHPPD). Any one that can be produced is possible, preferably, the Bifidobacterium genus lactic acid bacteria, more preferably Bifidobacterium infantis, Bifidobacterium bifidum, lactose Bacillus lactis, Clostridium butyricum, Bifidobacterium K-103, Bifidobacterium K-506, Bifidobacterium K-513, Bifidobacterium K-525 ( Arch. Pharm. Res. , 21 , p54-61, 1988) can be used one or a mixed strain thereof selected from lactic acid bacteria.

Intestinal bacteria that can be used for this purpose also metabolize ginsenosides Rg3, Rg5 and Rk1 in panax plant saponins, and also ginsenosides Rh2, Rh3, Rk2, protopanaxanadiol, and 20-dihydroprotopanaxadiol. As long as it can produce any one, preferably in the genus Bacterioides (Bacterioides), Fusobacterium (Fusobacterium), Eubacterium (Eubacterium) intestinal bacteria, more preferably bacterioid JY- 6 ( Biol. Pharm. Bull. , 23 , pp1481-1485, 2000), Bacterioides stercoris, Fusobacterium K-60 ( Biol. Pharm. Bull. , 23 , pp1481-1485, 2000 ), Eubacterium L-8 ( Biol. Pharm. Bull. , 23 , pp1481-1485, 2000), or mixed strains thereof may be used.

The following steps (4) to (6) can be selectively applied depending on the form of the final Panax plant product.

(4) a process of concentrating the panax plant extract of step (3) as it is under reduced pressure, lyophilization or spray drying.

(5) a process of centrifuging the Panax genus plant extract of step (3) to remove impurities and precipitates in the extract, followed by concentration of the supernatant under reduced pressure, lyophilization, or spray drying.

(6) An extraction process for extracting only an active ingredient contained in the panax plant extract of step (4) or (5) with an appropriate solvent, and suitable solvents include water, methanol, ethanol, and the like. The same lower alcohol solution may be used, and the active ingredient may be separated using a special extraction method such as supercritical extraction method, followed by a drying process by freeze drying and a stirring process for manufacturing a processed beverage or processed food. (Agitation Process) or dilution process (Dilution Process) may be further performed.

Through the above process, the ginsenosides Rb1, Rb2, Rc, and Rd, which are compounds contained in the raw materials, are converted to ginsenosides Rg3, Rg5, and Rk1 by acid treatment or heat treatment, followed by continuous lactic acid bacteria or enterobacterial treatment. The sugar at position 3 of these saponin structures breaks off and is converted to ginsenosides Rk2, Rh2, Rh3, protopanaxadiol (PPD) and DHPPD (20-dehydro-protopanaxadiol) to substantially constitute constituent saponins (ginsenoside Rk2 + Rh3 + PPD + DHPPD) / (ginsenoside Rg3 + Rg5 + Rk1) ratio of more than 0.1 to produce a genus Panax plant extract.

The present invention also provides all related Panax genus plant products commercially available containing the specially processed Panax genus plant extracts produced by the above-described manufacturing process.

Such processable related Panax plant products include Panax plant dry powders, extracts, ampoules, teas, tablets, capsules, solutions, ointments and the like.

The specially processed Panax genus plant extract of the present invention is a conventional unprocessed Panax genus plant or acid treatment in the cytotoxicity test on cancer cells and the antiallergic effect test in Rat Mast Cell using RBL-2H3 cells. Only process or heat treatment showed excellent efficacy compared to the coarse Panax plant extract.

Accordingly, the present invention provides a pharmaceutical composition for preventing and treating cancer diseases, including the Panax genus plant extract, which has been specially processed by the above-described manufacturing process.

Specifically, the cancer includes cancers such as liver cancer, lung cancer, skin cancer, breast cancer, ovarian cancer, blood cancer, uterine cancer, and the like.

In another aspect, the present invention provides a pharmaceutical composition for the prevention and treatment of anti-allergic related diseases, including Panax genus plant extract, which has been specially processed by the manufacturing process.

Specifically, the allergy-related diseases include diseases such as rhinitis, dermatitis, asthma and autoimmune deficiency.

The composition comprising Panax genus plant extract of the present invention may further comprise a suitable carrier, excipient and diluent according to conventional methods.

Carriers, excipients and diluents that may be included in the composition comprising the Panax genus plant extract of the present invention include 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. .

Compositions comprising Panax genus plant extract according to the present invention, respectively, according to a conventional method of oral formulations, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injectable solutions It can be formulated and used in the form.

The amount of Panax genus plant extract of the present invention may vary depending on the age, sex, and weight of the patient, but may be administered once to several times in an amount of 0.1 to 100 mg / kg. In addition, the dosage of the Panax genus plant extract 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 comprising the plant extract of Panax genus of the present invention can be used in various formulations, such as drugs, foods and beverages for the prevention and treatment of cancer and allergic diseases in the above formulation. Examples of the food to which the Panax genus plant extract can be added include various foods, beverages, gums, teas, vitamin complexes, and health functional foods.

Panax genus plant extract of the present invention has little toxicity and side effects, so can be used with confidence even for long-term use for the purpose of prevention.

Panax genus plant extract of the present invention may be added to food or beverages for the purpose of preventing cancer or allergic related diseases. At this time, the amount of the Panax plant extract in the food or beverage may be added at 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.02 to 5 g, preferably 0.3 to 1g based on 100 ml Can be added.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for having the extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates, etc. as additional ingredients, as in general 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 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 proportion of said 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), synthetic flavors and natural flavoring agents, colorants and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, 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. 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.

Comparative Example 1. Preparation of crude Panax plant extract

20 g each of Korean ginseng, Hwagi ginseng, and whole chile were sliced, and 60% ethanol was added thereto, followed by extraction under reflux for 3 hours, followed by concentration under reduced pressure, to obtain 4.5 g, 4.0 g, and 3.7 g of crude white ginseng, Hwagi ginseng, and chile extract, respectively.

Comparative Example 2. Preparation of Acid-treated Panax plant extract

Chop 20g each of Korean Ginseng, Hwagisam and Whole Chil, add 1000 ml of water containing 0.1% lactic acid, and incubate at 60 ℃ for 5 hours, extract this culture with butanol, and extract 2.5 g of Korean Ginseng, Hwagi and Whole Chil Extract. , 2.8 g, 3.2 g each were obtained.

Comparative Example 3 Preparation of Heat Treatment Panax Plant Extract

100 g each of Korean ginseng, Hwagi ginseng and whole chicks were heated with steam at 130 ° C. for 2 hours, and 60% ethanol was added thereto to reflux for 3 hours. The extract was concentrated under reduced pressure to obtain 42 g, 35 g, and 37 g of Korean ginseng, Hwagi ginseng, and Ginseng extract, respectively.

Example 1 Preparation of Processed Panax Plant Extract

Dissolve 1 g of each of the plants in the Panax genus plant extract prepared in [Comparative Example 3] as a raw material in 20 ml of water, and then add 100 mg (wet weight) of Fusobacterium K-60 for 72 hours at 37 ° C. After incubation, the culture was centrifuged, and the supernatant was concentrated under reduced pressure and dried to prepare 550 mg, 530 mg, and 430 mg of the processed Panax genus extract, respectively.

Example 2 Preparation of Processed Panax Plant Extract

1 g of each of the heat-treated Panax genus plant extracts prepared in [Comparative Example 3] was dissolved in 20 ml of water, respectively, and then Bifidobacterium K-506 (Kim Dong-hyun et al . , Arch. Pharm. Res. , 21 , pp54-61, 1988) 50 mg and Bifidobacterium K-103 (Kim, et al . , Arch. Pharm. Res. , 21 , pp54-61, 1988) 50 mg, Bifidobacterium KK-1 50 mg ( Wet weight) was added thereto, followed by incubation at 37 ° C. for 72 hours, followed by centrifugation of the culture and concentration of the supernatant under reduced pressure, thereby preparing 580 mg, 450 mg, and 410 mg of processed Panax genus extracts, respectively.

Example 3 Preparation of Panax Plant Extract

1 g of Panax genus plant extract prepared in [Comparative Example 2] was dissolved in 20 ml of water, respectively, as a raw material, and then bacterioid JY-6 50 mg, Eubacterium L-8 50 mg, and bacterioides stericis 50 After adding mg (wet weight) and incubating at 37 ° C. for 2 days, the solution was centrifuged and the supernatant was concentrated under reduced pressure to obtain 580 mg, 630 mg, and 450 mg of processed ginseng extract, 630 mg, and 450 mg, respectively.

Example 4 Preparation of Korean Ginseng Extract

1 g of Korean ginseng extract obtained in [Comparative Example 2] was dissolved in 20 ml of water as a raw material, and then Bifidobacterium K-506 ( Arch. Pharm. Res. , 21 , pp54-61, 1988) 50 mg and 50 mg (wet weight) of Bifidobacterium K-103 ( Arch. Pharm. Res. , 21 , pp54-61, 1988) were added and incubated at 37 ° C. for 72 hours. After centrifugation of this solution, the supernatant was concentrated under reduced pressure to obtain 430 mg of processed ginseng extract.

Example 5 Preparation of Korean Ginseng Extract

1 g of the Korean ginseng extract obtained in [Comparative Example 1] was taken as an original ingredient, 20 ml of water containing 1% citric acid was added thereto, and incubated at 60 ° C. for 5 hours, followed by neutralization with sodium hydroxide or calcium gluconic acid ( pH 6.8-7.0) and centrifuged to take the supernatant. 50 mg of Bifidobacterium K-506 and 50 mg of Bifidobacterium KK-2 (wet weight) were incubated at 37 ° C. for 2 days, and the culture solution was centrifuged and the supernatant was concentrated under reduced pressure. Ginseng extract 350 mg was obtained.

Example 6 Preparation of Processing Korean Ginseng Leaf Extract

1 g of dried dry ginseng leaves were extracted with reflux with methanol, the solvent was concentrated under reduced pressure, the residue was suspended in water, extracted with ether solvent, the remaining aqueous layer was extracted with butanol, and the butanol layer was concentrated under reduced pressure. Soluble fractions were obtained. The fractions were heated at 130 ° C. for 3 hours, and then dissolved in 20 ml of water. Human fresh intestinal flora (Kim Hee University, Professor Dong-Hyun Kim, Biol. Pharm. Bull. , 25 , pp58-61, 2002) 100 mg (wet weight) Incubated at 37 ℃ for 2 days and centrifuged to take the supernatant. This solution was extracted with 50 ml of saturated butanol, and concentrated under reduced pressure and dried to obtain 100 mg of Korean ginseng leaf extract.

Experimental Example 1. Content Analysis Experiment

The extracts obtained in Comparative Examples 1, 2 and 3 and Examples 1, 2 and 3 were each taken in an amount corresponding to 500 mg as a raw plant, suspended in water, and extracted with n-BuOH. The BuOH layer was concentrated under reduced pressure, and the remaining residue was dissolved in 5 ml of methanol, and the solution was injected into HPLC by membrane filtration. At this time, HPLC was carried out as follows by slightly modifying the measuring method described in the literature (Kwon et al., J. Chromatography A , 921 , pp335-339, 2001), and summarized the analysis results in the following [Table 1].

[Measuring conditions]

Column: LiChrosorb RP-18

Solvent: A = H ₂ O, B = CH ₃ CN gradient elution

      0 min (B 15%); 10 minutes (B 34.5%); 25 minutes (B 47.5%);

      40 minutes (B 80%); 50 minutes (B 100%)

Flow rate: 1 ml / min

Detector: Vaporized Light Scattering Detector (ELSD)

Changes in Relative Content of Saponin Components by Processing Methods sample Rk2 + Rh3 PPD ¹⁾ DHPPD ²⁾ Rg3 Rg5 + Rk1 Ratio ³⁾ Comparative Example 1 Korean ginseng - - - - - Fire ginseng - - - - - Prepainted - - - - - Comparative Example 2 Korean ginseng - - - 28 10 - Fire ginseng - - - 32 12 - Prepainted - - - 25 8 - Comparative Example 3 Korean ginseng - - - 34 47 - Fire ginseng - - - 25 42 - Prepainted - - - 22 35 - Example 1 Korean ginseng 14 5 One 13 20 0.61 Fire ginseng 10 3 2 10 17 0.56 Prepainted 9 2 3 8 12 0.70 Example 2 Korean ginseng 12 8 3 11 20 0.74 Fire ginseng 10 5 3 10 22 0.56 Prepainted 7 3 2 6 14 0.60 Example 3 Korean ginseng 2 2 One 13 5 0.28 Fire ginseng 3 2 One 15 4 0.32 Prepainted 2 One One 14 3 0.23 1) PPD: Protopanaxadiol 2) DHPPD: 20-dihydroprotopanaxadiol 3) Ratio: Ratio of (Rk2 + Rh3 + PPD + DHPPD) / (Rg3 + Rg5 + Rk1) * * Concentrations are expressed as the ratio of individual peaks to total saponin peak area.

As a result of the experiment, the sample of [Comparative Example 1] did not detect any nonpolar saponin components such as ginsenosides Rg3, Rg5, Rk1, Rk2, Rh3, PPD and DHPPD, which were not contained in the raw ginseng. Because it is. Panax genus plants obtained in [Comparative Example 2] was found to be mainly high in the content of ginsenoside Rg3. In the Panax genus plants obtained in [Comparative Example 3], the content of ginsenosides Rg3, Rg5, and Rk1 is mainly high, and ginsenosides Rk2, Rh3, PPD, and DHPPD are not detected or very low. Extracts of the present invention obtained in 2, 3 was found to increase the content of ginsenosides Rk2, Rh3, PPD, DHPPD.

Experimental Example 2: Drug potentiation confirmation experiment (anticancer effect)

In order to compare the anticancer effect of the processed Panax genus plant extract of the present invention, the anticancer experiment described in Carmichael et al. (Carmichael, J. et al; Cancer Res., 47 , pp936-940, 1987) The experiment was carried out as follows.

Liver cancer cell line (HepG2; Korea Cell Line Bank, Cat.No. 58065), human-derived cancer cell line, was 10% FBS, 1% antibiotic-antimycotics (GIBCO, Cat.No. 15240-062) and 2.2g, respectively. Incubated with RPMI 1640 medium solution (Sigma, R-6504) to which the / L sodium bicarbonate was added (5% CO ₂ ) as follows.

Each of the cancer cell lines was treated with 0.25% trypsin to remove the cells from the flask, and the number of cells was adjusted to 3 × 10 ⁴ cells / well, and 180 μl per well was put into a 96 well culture plate for 24 hours at 37 ° C. and 5% CO _2. Cells were attached by culturing in a conditional saturated CO ₂ incubator. After the sterilization of Panax genus plant extract samples of Comparative Examples 1, 2, 3 and Examples 1, 2, 3 with high-pressure steam, 20 μl of each well was added at different concentrations, followed by 5% CO ₂ . Incubated for 48 hours in saturated incubator. After 48 hours, 50 μl of 20 mg / ml MTT reagent (Cig. Cat. No. M-5655, Sigma Co., Ltd.) was added to each well, followed by reaction in a CO ₂ incubator for 4 hours, and then 100 μl of DMSO was added to the remaining precipitate. In addition, the absorbance was measured at 580 nm using an ELISA analyzer (Bio. TeK Instrument Inc., USA) to determine the extent of cell growth inhibition.

Table 2 shows the 50% cancer cell growth inhibition concentration (GI ₅₀ ). As shown in Table 2, compared with Comparative Examples 1 and 2, the case of Comparative Example 3 showed a strong anticancer effect. However, it was confirmed that the processed Panax genus plant extracts of Examples 1, 2, and 3 exhibited more potent anticancer effects than Comparative Example 3.

Cancer Cell Growth Inhibitory Activity Test Results (GI ₅₀ , Raw Ginseng ㎍ / mL) Korean ginseng Fire ginseng Prepainted Comparative Example 1 > 5000 > 5000 > 5000 Comparative Example 2 1500 2040 2580 Comparative Example 3 720 1200 1320 Example 1 240 300 390 Example 2 310 370 400 Example 3 480 580 600

Experimental Example 3: Anti-allergic effect experiment using RBL-2H3 cell line

Inagaki et al . ; Int. Arch. Allergy Appl. Immunol., 87 , pp254-259, 1988 to compare the antiallergic effects of the processed Panax genus plant extracts of the present invention . According to the anti-allergy effect test method described in the following experiment was performed.

DBL (Dulbeccos' modified Eagle's medium, Sigma, 22256) containing 10% FBS (fetal bovine serum) and L-glutamine from RBL-2H3 cell line (rat basophil cell line, Korea Cell Line Bank, Cat.No. 22256) (Sigma D-5648) was incubated for 2 hours in a humidified 5% CO ₂ incubator at 37 ° C., and the adherent cells were suspended using trypsin-EDTA solution. It was used for this experiment.

The RBL-2H3 cells were dispensed into 5 wells of 5 × 10 ⁵ cells / well in 24 wells, followed by incubation for 12 hours with 0.5 μg / ml of mouse monoclonal IgE. The cells were washed with 0.5 ml of siraganian buffer (119 mM NaCl, 5 mM KCl, 0.4 mM MgCl ₂ , 25 mM PIPES, 40 mM NaOH, pH7.2) and then again 0.16 ml of siraguanian buffer (5.6 mM). Glucose, 1 mM CaCl ₂ , 0.1% BSA was added) and then incubated at 37 ° C. for 10 minutes. 40 μl of the processed Panax genus plant extract samples of Comparative Examples 1, 2, 3 and Examples 1, 2, and 3 were added thereto, and then, after 20 minutes, 0.02 ml of antigen (DNP-BSA 1 μg / ml Cells were activated at 37 ° C. for 10 minutes and then centrifuged at 2000 rpm for 10 minutes to transfer 0.025 ml of the supernatant to 96 wells. To this was added 0.025 ml of substrate solution 1 mM p-NAG (a solution of p-nitrophenyl-N-acetyl-β-D-glucoamide) in 0.1 M citrate buffer to pH 4.5, which was then heated to 60 ° C. at 37 ° C. After incubation for 10 minutes, 0.2 ml of 0.1 M Na ₂ CO ₃ / NaHCO ₃ was added to stop the reaction, and the absorbance was measured at 405 nm by ELISA analyzer.

Table 3 shows the 50% inhibitory concentration (IC ₅₀ ), compared to Comparative Examples 1, 2, and 3 compared to Comparative Examples 1, 2 and 3 of the present invention, Panax plants extracts allergic to RBL-2H3 cell line. Inhibitory effect experiments showed that the allergic effect is more excellent.

Results of anti-allergic effect in RBL cells (IC ₅₀ ; raw ginseng ㎍ / mL) sample Korean ginseng Fire ginseng Prepainted Comparative Example 1 > 50 > 50 > 50 Comparative Example 2 > 50 > 50 > 50 Comparative Example 3 > 50 > 50 > 50 Example 1 12 17 22 Example 2 15 19 21 Example 3 24 28 29

Experimental Example 4: Passive transdermal anaphylaxis experiment Experiment using animal model

Passive Cutaneous Anaphylaxis Experiments The following experiment was performed to confirm the effects of the Korean ginseng extract samples of Comparative Examples 1 and 2 and Examples 4, 5 and 6 on allergy using an animal model.

 First, the back of the mouse (ICR-based Korean animal) in which 10 g of a solution of IgE serum of DNP-BSA (Dinitrophenol-human serum albumin (Sigma, A-6661) diluted with physiological saline was anesthetized with ether. (50 µl), and manually sensitized. After 48 hours, 0.2 ml of saline solution containing 0.2 mg of DNP-HSA and 1.6 mg of Evans blue was injected into the tail vein and killed by cervical dislocation 30 minutes later. The amount of Evans blue pigment leaked by the back and the like was measured. A portion of the back of the mouse (1 cm 2 of the site where IgE was injected) was cut out into a test tube, and 0.7 ml of 1 M-KOH was added thereto and incubated overnight at 37 ° C. 4 ml of 0.6 M phosphate-acetone mixed solution (5:13) was added to the test tube, shaken, filtered, and the extracted pigment was subjected to colorimetric determination at 620 nm.

The Korean ginseng extract of Comparative Examples 1 and 2 and the processed ginseng extract samples of Examples 4, 5 and 6 were dissolved or suspended in physiological saline and administered orally one hour before administration of the antigen (DNP-HSA).

As shown in Table 4, when the raw treatment of Comparative Example 1 did not have an anti-allergic effect, the acid treatment of Comparative Example 2 had a low anti-allergic effect, but processed as in Examples 4, 5, 6 The anti-allergic effect of ginseng extract was excellent.

Experimental Results of Passive Transdermal Anaphylaxis Effect of Korean Ginseng Treatment Sample Capacity (mg / kg) % Inhibition Comparative Example 1 50 0 Comparative Example 2 50 20 Example 4 50 52 Example 5 50 62 Example 6 50 54 DSCG (positive control) 100 37

Based on the above results, chemical treatment and biological treatment of Panax genus plants show enhanced efficacy of preventing and treating diseases such as cancer and allergy. It has been found that it can be effective in preparing health functional foods.

Experimental Example 5. Acute Toxicity Test

1. Oral administration

 ICR-based mice and Sprague Dawley (cod hyochang science) were divided into four groups of 10 mice each, orally administered with the extracts of Example 1 of the present invention at doses of 500, 725, 1000 and 5000 mg / kg, respectively. After two weeks of toxicity, none of the four groups died and no symptoms were found.

2. Intraperitoneal administration

 ICR mice (25 ± 5 g) and Sprague Dawley (cod hyochang science) were divided into four groups of 10 animals each, and the extracts of Example 1 of the present invention were 25, 250, 500 and 725 mg, respectively. Toxicity was observed for 24 hours after intraperitoneal administration at / kg. There were no deaths in all four groups and no symptoms were apparent in the control group.

From the above results, it was confirmed that the extract of the present invention has little acute toxicity.

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.

Example 1 Dry Powder 50 mg

Lactose ......................................... 100 mg

Talc ......................................... 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.

Example 1 Dry Powder ............... 50 mg

Corn starch ..................... 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.

Example 1 Dry Powder 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).

Example 1 Dry Powder ............. 50 mg

Sterile Distilled Water for Injection ...

pH adjuster ...............

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.

Example 1 Dry Powder ......................................... 50 mg

Isomerized sugar ......................................... 10 g

Mannitol ......................................... 5 g

Purified water .............................

In addition, the health beverage is prepared as follows.

Example 1 Syrup mixed with 0.1 to 80% of dry powder, 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 mixed with 79 to 94% of purified water The syrup is sterilized at 85 to 98 ° C. for 20 to 180 seconds, mixed with cooling water at a ratio of 1: 4, and then contains a specially processed Panax plant dry extract which is injected with 0.5 to 0.82% of carbon dioxide gas. To produce a carbonated beverage.

Example 1 Homogeneous sterilization by homogeneous mixing of dry powder and subsidiary materials such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%) and water (75%) Health drinks are prepared by packing them in small packaging containers such as glass bottles and plastic bottles.

Formulation Example 6 Preparation of Health Functional Food

Extract of Example 1 ................. 1000 mg

Vitamin Blend .....................

Vitamin A Acetate ............... 70 μg

Vitamin E ............... 1.0 mg

Vitamin B1 ............... 0.13 mg

Vitamin B2 ............... 0.15 mg

Vitamin B6 ............... 0.5 mg

Vitamin B12 ........................ 0.2 μg

Vitamin C ............... 10 mg

Biotin ............ 10 μg

Nicotinic Acid Amide ... 1.7 mg

Folic acid ............... 50 ㎍

Calcium Pantothenate ......................................... 0.5 mg

Mineral mixture .....................

Ferrous Sulfate ............... 1.75 mg

Zinc Oxide ............... 0.82 mg

Magnesium Carbonate ............ 25.3 mg

Potassium monophosphate ......................................... 15 mg

Dicalcium 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 mixtures is mixed with a component suitable for a health functional food in a preferred embodiment, the composition ratio may be arbitrarily modified, and the above components may be mixed according to a conventional health food manufacturing method. Next, the granules may be prepared and used for preparing the health functional food composition according to a conventional method.

Formulation Example 7 Preparation of Health Functional Drink

Extract of Example 1 ..... 1000 mg

Citric Acid ......................................... 1000 mg

Oligosaccharide ............. 100 g

Plum concentrate ........................... 2 g

Taurine ......................................... 1 g

Purified water is added ............ 900 ml total

After mixing the above components in accordance with the conventional health functional beverage manufacturing method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained by sterilization in a sterilized 2 ℃ container, sealed sterilized and then refrigerated It is used to prepare a health functional beverage composition of the present invention.

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

Compositions containing specially processed Panax plant extracts, which have undergone physicochemical treatments such as heat treatment and acid treatment, and biological treatments such as lactic acid bacteria and enterobacteriaceae, have the effect of effectively inhibiting cancer or allergic diseases. It can be usefully used as a prophylactic and therapeutic agent for cancer and allergy-related diseases.

Claims (26)

The composition ratio of constituent saponin ((sum of contents of ginsenosides Rh2, Rh3, PPD (protopanaxadiol) and DHPPD (20-dehydroprotopanxadiol)) / (sum of contents of ginsenosides Rg3, Rg5, Rk1) is 0.2 or more. Processed extract of the genus Panax plant having an antiallergic effect. delete delete delete The genus Panax ginseng , Panax quinquefolia , Panax notoginseng , Panax japonica , Panax trifolia , Panax pseudoginseng , Vietnam three one member selected from (Panax vietnamensis), or Panax elegans are tees climb (Panax elegatior), Panax Wan Jia Taunus (Panax wangianus), Panax non pinra typhus (Panax bipinratifidus), Panax eye styryl poly Titanium (Panax angustifolium) Extracts that are plants above. The extract of claim 1, wherein the genus Panax plants are roots, stems, leaves, brainstems, flowers, fruits and their tissue cultures. delete delete Method for producing panax genus plant extracts with enhanced anti-allergic efficacy with a specially treated anti-allergic effect comprising the step of culturing Bifidobacterium spp. . delete A first step of producing a panax genus plant extract by subjecting the panax genus plant material to a high temperature heat treatment at 110 to 180 ° C. for 0.5 to 20 hours; Panax plants with increased anti-allergic efficacy comprising a second step of adding Bifidobacterium lactic acid bacteria or intestinal bacteria to the high temperature heated Panax genus plant extract to obtain a biologically treated Panax genus plant extract Method of Preparation of Extracts. delete delete delete delete delete The method of claim 11, wherein the lactic acid bacteria are selected from the group consisting of Bifidobacterium K-103, Bifidobacterium K-506, Bifidobacterium KK-1, Bifidobacterium KK-2 and mixed strains thereof. Way. 12. The method according to claim 9 or 11, wherein the enterobacteriaceae are selected from the group consisting of Bacterioides strains, Fusobacterium strains, Eubacterium strains and mixed strains thereof. . 12. The enteric bacterium is selected from the group consisting of bacterioid JY-6, Fusobacterium K-60, Eubacterium L-8, Bacteroides stercoris, and mixed strains thereof. Way. delete delete A pharmaceutical composition for preventing and treating allergic diseases comprising the ginseng extract of claim 11 as an active ingredient and a pharmaceutically acceptable carrier. 23. The composition of claim 22, wherein the allergic related diseases include rhinitis, dermatitis, asthma disease. The pharmaceutical composition according to claim 22, wherein the pharmaceutically-used formulation form is any one selected from powders, granules, tablets, capsules, solutions, and injections. delete delete