KR100545156B1 - Crude Drug Compositions for treating or preventing intestinal disease caused by microorganism - Google Patents

Abstract

The present invention relates to a number of herbal compositions comprising essentially radish ( Raphanus sativus L.) and tea ( Camellia sinensis ), which has no effect on the growth of beneficial bacteria in the intestine in an animal model orally administered the composition of the present invention. By inhibiting the growth of harmful bacteria, it can be used as an effective and safe medicine and health food for improving intestinal function and bacterial intestinal disease.

Intestinal Function Improvement, Microorganism, Bacterial Intestinal Disease, Radish, Tea, Medicine, Health Food

Description

Crude Drug Compositions for treating or preventing intestinal disease caused by microorganism}             

Figure 1a to 1c is a view of the colony morphology in the dilution plate on the day D + 2 of each experimental group, Figure 1a is a normal control group, Figure 1b is a KTG075 treatment group, Figure 1c is a photograph of the loperamide treatment group.

The present invention relates to a combination composition of several herbal medicines having an effective oral activity against bacterial intestinal diseases, and more particularly, to extract and concentrate several herbal medicines such as radish and tea at an appropriate ratio, thereby preventing and improving bacterial intestinal diseases. It relates to a herbal extract having a and a composition containing the same in an appropriate ratio.

Bacterial intestinal diseases include food poisoning caused by E. coli and Salmonella, and chronic / acute gastroenteritis.

Salmonella typhimurium DT104 is different from conventional Salmonella spp. Against ampicillin, chloramphenicol, streptomycin, sulfonamides and tetracycline. The antimicrobial resistance pattern (R-Type ACSSuT) has already been reported in many countries around the world, including developed countries such as the United States, United Kingdom, Canada, Germany, France, Austria and Denmark. In addition to the difficulty in treating antibiotics for infectious diseases, the organism is also at risk of causing the transmission of genes involved in antibiotic resistance to other bacteria through transformation, etc. In the UK, trimethoprim and ciprofloxacin are already present. Salmonella typhimurium DT104 has been reported with additional resistance to. In addition, Escherichia coli 0157: H7 ( Esherichia coli O157: H7) has caused massive food poisoning in the United States and Japan, and shows symptoms such as hemolytic uremic syndrome and hemolytic colitis during infection. Infants and the elderly are known to be deadly. Antimicrobial treatment is the main cause of infection of these organisms, but resistance to various antibiotics due to antibiotic abuse is increasing, so we are looking for useful substances in natural products, and by using natural products to control the intestinal composition of the beneficial flora, potentially gaining significant prevention effects. Attempts are being made.

Radish ( Raphanus sativus L.) used in the present invention belongs to Cruciferae (Cruciferae) and the sugars contained in the roots are mainly glucose and fructose, in addition to kumaric acid, caffeic acid, ferulic acid and phenylpyri. It contains phenylpyruvic acid, gentidinic acid, hydroxy benzoic acid and many amino acids. Alcoholic extracts of radish roots have antimicrobial and antifungal effects (Bo Bo-seop and Shin, Min-Kyo; Hyangjedae, pp582-584, Younglimsa, 1998).

Camellia sinensis belongs to theaceae, and the young and soft leaves of the tea tree mainly contain purine alkaloids and caffeine, and the content is 1 to 5%. In addition, the traces contain theobromine, theophylline, and xanthine. Tannins in tea leaves are mainly galloyl-l-epigallocatechol, which is contained with caffeine. The pharmacological action of tea leaves is mainly caused by xanthine derivatives (caffeine and theophylline), and also contains a large amount of tannic acid, thus acting as antibacterial and vitamin P. Pharmacological action is to excite the central nervous system, promote mental function, increase thinking ability, eliminate fatigue, expand coronary blood vessels, and diuretic (Jung Bo-seop and Shin, Min-Kyo; Hyang-Yangsa Dictionary, pp403-405, Younglimsa, 1998) . The young leaves of the tea tree can produce various kinds of teas with different flavors depending on the processing method. For example, according to the manufacturing methods such as post-fermentation, fermentation, semi-fermentation, and non-fermentation, respectively, tea, black tea, oolong tea, Iron ore, cannon tea, green tea, etc. can be manufactured.

Carrots ( Daucus carota L. var. Sativa DC.) Belong to Umbelliferae, and the roots of carrots include α-, β-, γ-, ε-carotene, lycopene and phytofluene. It contains many carotenoids, vitamins B1, B2 and anthocyanidins. Its pharmacological effect is effective in treating dryness and it is effective in treating indigestion (Jung Bo-seop and Shin Min-kyo; Hyang-Yaksa Dictionary, pp421-422, Yeonglimsa, 1998).

The dermis ( Aurantii nobilis Pericapium) is peeled and dried from the fruit of the mature fruit of the Citrus unshiu MARCOR., And the dermis ( Aurantii immaturi Pericapium) is the skin of the immature fruit and the immature fruit of the tangerine. It contains dried essential oils, such as limonene, flavonoid glycosides, hesperidin and citric acid, naringin and aspartic acid. It is known to have bad, Jinhae, expectorant, copulation drug, anti-type I allergic effect (Bo Bo-seop and Shin Min-kyo; Hyang-Yaksa Dictionary, pp783-784, Yeonglimsa, 1998).

Fig ( Ficus carica L.) is a dry fruit of the fig tree of Moraceae, the main ingredient of which contains about 10% of sugars such as glucose and fructose and has a strong sweetness. Organic acids include malic acid and citric acid, as well as benzaldehyde and protease, a protein that is effective in treating cancer, and other enzymes such as lipase, amylase, and oxidase, as well as fiber and protein. Pharmacological action is used for lack of milk, loss of appetite, etc., has the effect of detoxification. The green leaves are applied to hemorrhoids, and the dry juice is used as a medicinal insecticide, and the dried leaves are used as a bath bath for neuralgia (Jung Bo-seop and Shin, Min-Kyo;

Onium ( Allium cepa L.) belongs to the family Liliaceae, with special tinge such as thiol, dimethyl disulfide, diallyl-disulfide and diallyl thioether, thiosulfine acid salt It contains a substance (i) and a small amount of malate. Roots, apertures, and leaves contain coumaric acid, caffeic acid, ferulic acid, and sinapic acid. Onion suppresses the rise of plasma cholesterol, lowers fibrinolytic activity, and can be used for atherosclerosis. It is effective for gastric apathy and enteritis. It has a bactericidal effect and is used as a diuretic and expectorant in folk medicine. ; Hyangjedae Dictionary, pp155-156, Younglimsa, 1998).

Mume Fructus is dried immature fruit of the Prunus mume of the Amygdalaceae. The fruit contains substances such as citric acid, malic acid, fluoride, cytosterol, and oleanolic acid. When immature, it contains clearing. It has antibacterial and antifungal action, and it is an alkaline food for fatigue recovery and constitution improvement. Especially, it has excellent detoxification effect and helps treat stomach upset and food poisoning. Sour taste secretes gastric juice and normalizes digestive system to digest Eliminates insufficiency and gastrointestinal disorders (Bo Bo-seop and Shin Min-kyo; Hyang-Yaksa Dictionary, pp628-629, Younglimsa, 1998).

Apricot ( Prunus armeniaca L.) belongs to the family Prunus armeniaca L. (Amygdalaceae), and the fruit apricot (haengja) contains citric acid and β-carotene, and essential oils include myrcene (myrcene), limonene, p -cymene cymene, terpinolene, and the like. It has the effect of antitussive and expectorant, so it is used for relieving, asthma, bronchitis and acute hepatitis in the private sector, and it is also good for skin beauty and is effective in removing freckles and blemishes. In oriental medicine, the seeds are used as a medicine called algae (杏仁), which is effective for fever, Jinhae, expectoration, and swelling, so it is used for coughing, asthma, bronchitis, sore throat, acute pneumonia and constipation. do. It is distributed in Korea, Japan, China, Mongolia, the United States, and Europe (Bo Bo-seop and Shin Min-kyo); Hyangje Ambassador, pp625-627, Yeonglimsa, 1998).

Anti-cancer activity in radish ( Adv. Exp. Med. Biol. , 289 , pp153-163, 1991: Am. J. Epidermiol. , 144 , pp1015-1025, 1996) and liver detoxification ( Biosci. Biotechnol. Biochem) ., 59, pp1882-1886, 1995: ... Arch Biochem Biophys, 316, pp797-802, 1995) and is known in the United States and Japan, in particular, no extract meantime, various functional foods and in many ways the composition It has been used as a mixture, and various inventions have been proposed for use in health drinks. However, the health beverage composition using such a radish extract is usually only a method used as a beverage composition in vague expectation depending on the digestive function of the radish extract.

As a result of the present inventor's focus on research on foods that can activate intestinal function by acting on beneficial bacteria and harmful bacteria among intestinal bacteria, the extract consisting of several natural herbal medicines and their proper composition ratio inhibits the growth of harmful bacteria in the intestine It was confirmed that there is an excellent improvement effect to complete the present invention.

The present invention provides a pharmaceutical composition for improving bacterial intestinal diseases and health functional foods, which contains several herbal compositions comprising the herbal extracts having intestinal harmful bacterium growth inhibitory activity and their proper combination ratio.

In order to achieve the above object, the present invention provides a pharmaceutical composition for improving bacterial intestinal disease, including a herbal composition comprising essentially a radish and tea extract and a pharmaceutically acceptable carrier.

In the above, radishes include a recent plant such as radish ( Raphanus sativus L. ), turnip ( Brassia Lap.), Tea is Camellia sinensis or green tea, oolong tea, black tea, black tea (China), iron canon (China) And processed tea including unfermented tea and fermented tea.

In addition, the present invention is a carrot ( Daucus carota var in addition to the essential components extract, that is, radish and tea sativa, dermis ( Aurantii nobilis Pericapium), at least one extract selected from Aurantii immaturi Pericarpium, fig ( Ficus carica L.), onion ( Allium cepa L.), ume ( Mume Fructus ) or apricot ( Prunus armeniaca ) extract To provide a pharmaceutical composition for improving the bacterial enteric disease containing.

Preferred compositions of the present invention include compositions containing radish, tea, carrots, dermis, green skin, figs, onions and ume extracts; Compositions containing radish, tea, carrot, dermis, green skin and onion extracts; Compositions containing radish, tea, carrot, dermis and dermis extracts; A composition containing radish, tea, carrot, dermis, green skin, fig and ume extract, and even more preferably a composition containing radish, tea, carrot, dermis, green skin, fig, onion, ume and apricot extract.

The bacterial intestinal diseases include food poisoning, acute enteritis, acute diarrhea and the like.

In addition, the present invention is a relative blending ratio of radish, tea, carrots, dermis, green skin, figs, onions, buckwheat and apricot extract 1 ~ 2: 0.1 ~ 1: 1-2 ~ 0.01 ~ 1: 0.01 ~ 1: 0.01 ~ 1: It is to provide a pharmaceutical composition of 0.1 ~ 2: 0.01 ~ 1: 0.01 ~ 1.

Such herbal medicines include homogeneous herbal medicines which are obvious in the art and can be used for the same or similar purposes as the prophylactic and therapeutic purposes of the present invention.

For example, the amount of extract of the present invention provides a pharmaceutical composition having a weight ratio of 0.01 to 80%, preferably 1 to 50% of the total composition.

Hereinafter, the present invention will be described in detail.

Herbal extracts for the improvement of bacterial intestinal diseases of the present invention is characterized in that the extraction with water or alcoholic aqueous solution, more specifically,

About 5 to 20 times after washing and drying in a dryer to remove foreign substances present on the surface of individual ingredients such as radish, tea, carrot, dermis, green skin, fig, onion, buckwheat and apricot, dried or undried Preferably, 10 to 12 times the amount of water or lower alcohol is added to the hot water extraction for 1 to 5 hours, preferably 2 to 3 hours at an extraction temperature of 70 to 120 ℃, preferably 90 ℃ or more, lyophilization, vacuum drying Preferably, the first step is a drying step of the individual material extract prepared by the step of making a powder extract by lyophilization;

After filtering the extract obtained from the extraction step, it is concentrated under reduced pressure at 40 to 80 ℃ the extract obtained in a suitable blending ratio, preferably 1 to 2: 0.1 to 1: 1 to 2: 0.01 to 1: 0.01 to 1: 0.01-1: 0.1-2: 0.01-1: 0.01-1, or mixed with other suitable herbal medicines in an appropriate ratio, and then 5-20 times, preferably 10-15 times the volume of these complex herbal medicines. A lower alcohol such as distilled water, methanol, ethanol or the like or a mixed solvent thereof, preferably distilled water alone or a mixed solvent of 10 to 50% water and ethanol, and an extraction temperature of 70 to 120 ° C, preferably 90 ° C or higher 1 to 5 hours, preferably 2 to 3 hours by hot water extraction through freeze-drying, vacuum drying, preferably lyophilized through a second step of drying the compound material extract prepared through the step of making powder extract Get Can.

The present invention basically radishes, tea, carrots, dermis, green skin, figs, onions, plums and apricots 1-80%: 1-80%: 1-80%: 0.1-20%: 0.1-20%: 1 ~ 50%: 1 to 50%: 0.5 to 10%: 1 to 50% by weight of the mixture, in addition, other herbal medicines are characterized in that it is a complex herbal composition extracted from a mixed herbal medicine made by mixing at an appropriate ratio. In particular, radish, carrots, onions are preferably prepared using a juice state.

Meanwhile, the composite herbal composition for improving bacterial intestinal disease of the present invention may be prepared by extracting and concentrating a mixed herbal medicine, and then freeze-drying to prepare a powder extract, and then mixing it with a substrate used for treatment.

The substrate used for oral administration may be used as an alcoholic aqueous solution by adding edible alcohol such as sterile water, saline or drinking water, and soju to the substrate.

Fecal samples were collected from rats orally administered KTG075, the composite herbal composition of the present invention, and the change of viable cell counts in feces was collected, followed by isolation, identification, and in vitro By confirming that KTG075 does not affect the growth of beneficial bacteria through experiments and inhibits the growth of pathogenic enterobacteriaceae, which are harmful bacteria, the composition KTG075 of the present invention can be used as a medicine or health functional food for improving bacterial intestinal diseases. .

 The herbal composition of the present invention may be mixed with a pharmaceutically acceptable carrier as an active ingredient to prepare various bacterial bowel disease improvement compositions.

Compositions comprising extracts according to the present invention are in the form of powders, granules, solutions, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Carriers, excipients and diluents which may be formulated with phosphorus injectables and may be included in the composition comprising the extract 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 have. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are 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, water, and the like. Prepared by mixing sucrose or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The amount of the herbal composition of the present invention may vary depending on the age, sex, and weight of the patient, but is generally administered in an amount of 0.01 to 10 g / kg, preferably 0.1 to 1 g / kg, divided once or several times daily. can do. In addition, the dosage of the extract may be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age and the like, and thus the dosage does not limit the scope of the present invention in any aspect.

The complex herbal composition for improving bacterial intestinal disease produced above may be used in all patients with intestinal diseases and is preferable because it can provide a fundamental treatment.

In addition, the present invention provides a health functional food for improving intestinal diseases, including a herbal composition comprising a radish and tea extract and food supplements that can be added to food.

In addition, the present invention for improving bowel disease containing one or more extracts selected from carrots, dermis, green skin, figs, onions, buckwheat or apricot extract in addition to the essential extract. Provide dietary supplements.

Preferred compositions of the health functional food of the present invention include radish, tea, carrot, dermis, green skin, fig, onion and ume extract; Radish, tea, carrot, dermis, green skin and onion extracts; Radish, tea, carrot, dermis and dermis extracts; It is a functional food containing radish, tea, carrot, dermis, dermis, fig and ume extract, even more preferably radish, tea, carrot, dermis, dermis, fig, onion, ume and apricot extract.

In addition, the present invention is a combination ratio of radish, tea, carrots, dermis, green skin, fig, onion, buckwheat and apricot extract is 1-2: 0.1-1: 1-2: 1: 0.01-1: 0.01-1: 0.01-1: 0.1 It is providing the nutraceutical which is -2: 0.01-1: 0.01-1.

The amount of the herbal composition of the present invention provides a dietary supplement having a weight ratio of 0.01 to 80%, preferably 1 to 50% of the total composition.

The present invention also provides a product for improving bacterial intestinal disease, which contains a herbal composition essentially containing radish and tea and an acceptable food additive.

The product of the present invention is 1 to 80% by weight, tea 1 to 80% by weight, carrot 1 to 80% by weight, dermis 0.1 to 20% by weight, skin peel 0.1 to 20% by weight, fig 1 to 50% by weight, onion 1 It provides a product containing -50% by weight, 0.5-10% by weight, 1-50% by weight apricot extract, 5-30% oligosaccharides and 5-30% dietary components.

Preferably, radish extract 5-30%, tea extract 5-30%, carrot extract 5-30%, dermis extract 1-10%, green skin extract 1-10%, fig extract 5-20%, onion extract 5- It provides a product which is a functional beverage containing 20%, 1-10% mae extract, 1-20% apricot extract, 10-20% oligosaccharide and 10-20% dietary ingredient.

The combination composition according to the present invention is characterized by the synergistic effect of the active ingredient formulated by the combination of the above composition ratio than the sole use for the purpose of providing a bacterial intestinal disease improvement effect.

The composition essentially comprising the radish and tea of the present invention can be used in a variety of food and beverages for the prevention of intestinal diseases.

Examples of foods to which the herbal composition comprising essentially the radish and tea of the present invention may be added include, for example, various foods, beverages, breads, cookies, sachets, candy, gum, tea, vitamin complexes, health functional foods, and the like. There is this.

The composition comprising the radish, tea extract and carrot, dermis, green skin, fig, onion, buckwheat or apricot extract of the present invention, the herbal medicine itself is used as a food, there is little toxicity and side effects, so it is safe even when taken for a long time for the purpose of prevention. Can be used.

The herbal medicine of the present invention may be added to food or beverage for the purpose of improving bacterial intestinal disease. At this time, the amount of the herbal extract in food or beverage is generally added to the functional food composition of the present invention 0.01 to 80% by weight of the total food weight, the health beverage composition is 0.02 to 30g, preferably based on 100ml Preferably at a rate of 0.3 to 5 g.

The health beverage composition of the present invention has no particular limitation on the liquid component except for containing the herbal medicine as an essential ingredient in the indicated ratio, 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, oligosaccharides and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As flavoring agents other than those described above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said natural carbohydrates 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), dietary ingredients, flavoring agents such as synthetic and natural flavoring agents, colorants 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. The present invention can be used as a mixed agent such as lactic acid bacteria preparation beverages or pastes such as yogurt. These components can be used independently or in combination.

The present invention provides a health functional food containing 0.01 to 30% by weight of the vitamins, oligosaccharides, and dietary ingredients as a food supplement.

The proportion of such additives is not so critical but is generally selected in the range of 0 to about 30 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in detail with reference to the following Examples and Experimental Examples, but the present invention is not limited thereto.

Example 1. Preparation of Radish Extract

After cutting radish (Hungryong seedling) and drying it, 100 g of 10 g of water was added to 1 l of water and extracted for 2 hours at 100 ° C. The extract was concentrated under reduced pressure using a vacuum condenser (Buchi R020, Germany) and freeze dried. , Or radish was finely squeezed or radish was concentrated under reduced pressure as described above, and then lyophilized radish extract was used as a sample.

Example 2. Preparation of Tea Extract

Purchasing green tea (Shinheung Dawon, Hwagae), oolong tea (Taiwanese), Iron Gwaneum (Chinese), Boicha (Chinese) and Black tea (Sri Lanka) were carried out in the same manner as in Example 1 to prepare and obtain an extract. And each was used as a sample.

Example 3. Preparation of Carrot Extract

Washed and dried carrots (heungjong seedlings) 100g or raw carrots were cut or obtained by performing the same method as in Example 1, the juice was used as a sample.

Example 4. Preparation of Dermis Extract

100 g of dried dermis (Gyeongdong Market) was obtained in the same manner as in Example 1, and used as a sample.

Example 5. Preparation of Peel Extract

100 g of dried Cheongpi (Gyeongdong Market) was obtained by performing the same method as Example 1, which was used as a sample.

Example 6 Preparation of Fig Extract

100 g of dried figs (Samho Agricultural Cooperative) were obtained by the same method as Example 1, and used as a sample.

Example 7. Preparation of Onion Extract

100 g of onion (heungjong seedlings) or raw onion was cut or obtained by performing the same method as in Example 1 by taking out juice, which was used as a sample.

Example 8. Preparation of Ome Extract

100 g of ume (Gyeongdong Market) was obtained by the same method as Example 1, and used as a sample.

Example 9 Preparation of Apricot Extract

100 g of dried apricots (Samho Agricultural Cooperative) was obtained by the same method as Example 1, and used as a sample.

Example 10 Preparation of Herbal Composition I

Of the herbal extracts prepared in Examples 1 to 9, radishes, green tea, carrots, dermis, green skin, figs, onions, and ume were combined in a weight ratio as shown in Table 1 below and used as samples.

group Herbal medicine Composition ratio (%)    I radish 30 green tea 10 carrot 20 dermis 5 Cheongpi 10 FIG 10 onion 10 Hawk 5

Example 11 Preparation of Herbal Composition II (KTG075)

Among the herbal extracts prepared in Examples 1 to 9, radishes, oolong tea, carrots, dermis, green skin, fig, onion, ume, and apricot were named KTG075 by combining them in a weight ratio as shown in Table 2 below. It was used as a sample in the following experimental example.

group Herbal medicine Composition ratio (%)     Ⅱ radish 20 Oolong tea 15 carrot 20 dermis 10 Cheongpi 10 FIG 10 onion 10 Hawk 5 apricot 10

Example 12 Preparation of Herbal Composition III

Of the herbal extracts prepared in Examples 1 to 9, radishes, black tea, carrots, dermis, blue skin, onions were used in combination by weight composition ratio as shown in Table 3 below.

group Herbal medicine Composition ratio (%)   Ⅲ radish 20 black tea 20 carrot 20 dermis 10 Cheongpi 10 onion 20

Example 13 Preparation of Herbal Composition IV

Radish, carrot, tea (iron guanyin), dermis and green skin were combined in the weight ratio as shown in Table 4 below. The extract was concentrated under reduced pressure and freeze-dried to use it as a sample.

group Herbal medicine Composition ratio (%)   Ⅳ radish 30 Tea (iron Kannon) 20 carrot 30 dermis 10 Cheongpi 10

Example 14 Preparation of Herbal Composition V

Combining radish, black tea, carrot, dermis, green skin, fig, and ume with the weight composition as shown in Table 5, 20 times purified water was added, and the extract obtained by extracting at 100 ° C. for 2 hours was concentrated under reduced pressure, and then freeze-dried. Used as.

group Herbal medicine Composition ratio (%)    Ⅴ radish 15 black tea 15 carrot 30 dermis 5 Cheongpi 2 FIG 20 Hawk 3

Example 15 Preparation of Herbal Composition VI

The radish, onion, and carrots were concentrated in a composition as shown in Table 6 below, added to the combined weight ratio of the remaining herbal medicines, added to 5 times purified water, and extracted at 100 ° C. for 2 hours. It was dried and used as a sample.

group Herbal medicine Composition ratio (g)    Ⅵ radish 20 Boycha 10 carrot 15 dermis 10 Cheongpi 10 FIG 10 onion 20 Hawk 5

Example 16. Preparation of herbal composition VII

Of the herbal extracts prepared in Examples 1 to 9, radishes, green tea, carrots, dermis, green skin, figs, onions, plums and apricots were combined in the weight ratio as shown in Table 7 below and used as a sample in the following experimental example. It was.

group Herbal medicine Composition ratio (%)    VII radish 30 green tea 10 carrot 25 dermis 5 Cheongpi 5 FIG 5 onion 10 Hawk 5 apricot 5

Example 17 Preparation of Healthy Beverages

3000 mg of radish extract of Example 1, 3000 mg of tea extract of Example 2, 2000 mg of carrot extract of Example 3, 100 mg of dermis extract of Example 4, 100 mg of dermal extract of Example 5, fig of Example 6 Purified water was added to 2000 mg of the extract, 200 mg of the onion extract of Example 7, 50 mg of the mae extract of Example 8, 100 mg of the apricot extract of Example 9, 100 mg of citric acid, 100 g of oligosaccharide, and 1 g of the dietary ingredient, and a total volume of 900 ml. made. After mixing the above components according to the conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution was filtered and obtained in a sterilized 2 L container, sealed sterilization and refrigerated.

Reference example. Strain culture medium

The BCP agar medium (Eiken, Japan) used in the present invention is 2.5 g of yeast extract, 5 g of peptone, 1 g of glucose, 0.1 g of L-cysteine, 0.04 g of bromcresol purple, and agar. Prepared by adding 15 g / l.

NB (nutrient broth, Difco) medium was prepared by adding 3 g of beep extract (peptone), 5 g / L peptone and adjusted to pH 6.8.

NA (nutrient agar, Difco) medium was prepared by adding 3 g of beef extract, 5 g of peptone, 15 g / L of agar.

MRS (Mann-Rogosa-Sharpe, Difco) liquid medium is 10 g of proteose peptone (No. 3), 10 g of beef extract, 5 g of yeast extract, 20 g of dextrose, 1 g of Twin-80 , 2 g of triammonium citrate, 5 g of sodium acetate, 0.1 g of MgSO _{4 ·} 7H ₂ O, 0.05 g of MnSO _{4 ·} 4H ₂ O and 2 g / L of K ₂ HPO ₄ It prepared by adjusting to pH 6.0-6.5.

MRS agar medium was prepared by adding 1.2-1.5% agar to the MRS liquid medium. 0.05% L-cysteine was added when MRS medium was used in anaerobic conditions.

Experimental Example 1. Effects of KTG075 on intestinal virulence

1-1. Animal testing

Male Sprague Dawley rats (220-240 g, 5-8 rats / group) were acclimated in metabolic cages for 6 days, then from day 7 (D day) A feed containing 1 mg of feramide (loperamide, Sigma) was supplied for free intake. Only normal water was supplied to the normal control group and the loperamide control group, and the KTG075-administered group was supplied with a sample solution prepared to contain KTG075 (7 mg / ml) in the bottled water. The feces obtained on the day during the experiment were received in aluminum foil which was previously laid under the cage, and the viable cell count was measured within 18 hours.

1-2. Measurement of Viable Cell Count

1 g of each fecal sample was ground and suspended in 100 ml of physiological saline (containing 0.05% L-cysteine in the case of anaerobic bacteria), diluted in succession (range: 10 ⁴ to ⁸ ), and spread on NA plates. Colonies growing during 7-day incubation in miracle and anaerobic conditions (H ₂ -5%: CO ₂ -15%: N ₂ -80% / Mart AJ9028 chamber and Anoxomat WS8000, Netherlands) were counted. Viable counts were expressed as cfu (colony forming units / g wet feces) per gram of feces.

1A to 1C are photographs of colonies grown on a plate diluted with 10 ⁷ days of feces of D + 2 days in each experimental group, FIG. 1A is a colony form of a normal control group, FIG. 1B is a colony of a KTG075 treatment group, and FIG. 1C Is a picture of colonies of loperamide treatment group.

On a solid medium for measuring viable cell counts, after one week, colonies of transparent lactic acid bacteria of 1 mm or less in diameter were laid on the surface, and other various colonies, opaque milky white with a diameter of 2 to 5 mm, grew. Colonies were observed (see FIGS. 1A-1C). Much fewer colonies were observed on the plate of KTG075 treated group than other normal control and loperamide treated groups.

The results are shown in Table 8 as c.f.u. by colony counts grown in aerobic and anaerobic conditions.

No significant change in total counts of viable cells was found between the normal control group, the loperamide treated group, the KTG075-administered group, or before and after the loperamide administration. However, after the KTG075 administration, the number of viable cells measured in aerobic conditions was slightly decreased from 2 days (D + 2) to 4 days (D + 4 days). It means that it can be suppressed.

The number of viable cells observed in the feces of rats during the experiment Experimental group Viable cell count conditions <D + 2> <D + 4> Normal control Exhalation 2.9 ㅧ 10 ⁸ -2.7 × 10 ⁹ 2.2 × 10 ⁸ -1.3 × 10 ⁹ anaerobe 2.5-4.2 × 10 ⁹ 1.7-3.3 × 10 ⁹ KTG075 treatment group Exhalation 2.2-7.5 × 10 ⁸ 4.7-8.7 × 10 ⁸ anaerobe 3.4-6.9 × 10 ⁹ 5.1-6.9 × 10 ⁹ Loperamide Treatment Group Exhalation 2.9 × 10 ⁸ -1.2 × 10 ⁹ 3.2 × 10 ⁸ -1.5 × 10 ⁹ anaerobe 4.3-4.9 × 10 ⁹ 3.2-9.1 × 10 ⁹

This inhibition was also observed on the solid medium for measuring the viable cell count at D + 2 days. In other words, the small size of the colony and transparent lactic acid bacteria were growing well, but the number of other opaque milky colonies decreased (see FIGS. 1A to 1C). In fact, the small and transparent colonies of lactic acid bacteria are invisible, and other enterobacteriaceae ( Klebsiella pneumoniae RF-8, Dermacoccus nishinomiyaensis RF-17, etc.) with large colonies can be distinguished.

Dominant bacteria among these were in vitro culture experiments of Experimental Examples 1-3, ie, in vitro effects of KTG075 components on the growth of enterobacteriaceae. It was isolated, purified, and identified as a MIDI system for use in impact experiments.

1-3. Isolation of Strains and Identification of Strains

Colonies growing predominantly from each plate at the time of viable cell counts were transplanted into new NA and BCP plates and purified again to isolate the dominating bacteria. Culture Microorganisms were incubated for 3 days in NA plates or for 7 days in BCP agar plates. Identification of the isolated strain was carried out by the method of microbial identification system (Microbial Identification System, MIDI; Microbial ID, Inc., Newark, Delaw-are) using the type and distribution of cell fatty acids.

Methyl esters of fatty acids were prepared according to the standard test method of MIDI, and the extracts were prepared using GC (Hewlett-Packard, 5890A), methyl phenyl silicone fused silica capillary column (HP 19091B-102, 25 m × 0.2 mm) was used to analyze the distribution of fatty acids using a Chromatopac CR 4A Data Analyzer.

The results are shown in Table 9. Fatty acid analysis results obtained here are summarized in Table 10, RF number refers to the number of strains isolated from the feces of the rat during the experiment.

Identification of RF Isolators Using a MIDI System RF number Colony morphology Cellular fatty acids Identification result One 2 mm diameter colony Milky-white, irregular, convex, undulate, dull on NA medium Table 10 Streptococcus sanguis 5 Punctiform, raised, intact, smooth, translucent on BCP agar medium  Table 10 Lactobacillus del brewek (Lactobacillus delbrueckii) 8 Colony 5 mm in diameter on NA medium, white, irregular, convex, wavy, concentric, opaque Table 10 Klebsiella pneumoniae 11 3 mm diameter colonies, white, round, flat, whole, smooth, opaque on NA media Table 10 Brevibacillus brevis 12 Punctiform, raised, intact, smooth, translucent on BCP agar  Table 10 Lactobacillus helveticus 17 Colony 3 mm diameter on NA, yellow, round, pulvinate, intact, contoured, opaque Table 10 Dermacoccus nishinomiyaensis

Cellular Fatty Acid Profiles of RF Separators RF number 10: 0 12: 0 13: 0 13: 0 iso 14: 0 14: 0 iso 14: 1 w5c 15: 0 15: 0 iso 15: 0 ante -iso 15: 1 w6c 16: 0 16: 0 iso 16: 1 isoH 17: 0 17: 0 iso One - 1.02 - - 22.85 - - - - - - 42.61 - - - - 5 0.84 6.28 0.67 - 18.82 - - - - - - 10.31 - - - - 8 - 5.25 0.37 - 10.95 - - 1.33 - - - 27.99 - - 0.39 - 11 - - - 0.29 0.28 2.00 - - 30.17 52.24 - 1.02 2.29 - - 5.34 12 2.57 3.81 - - 3.11 - 1.75 - - - - 14.59 - - - - 17 - - - - 0.50 - - 1.56 7.34 0.33 2.09 2.83 2.23 0.82 2.83 15.15 RF number 17: 0 cyclo 17: 0 ante -iso 17: 0 10- met 17: 1 w8c 17: 1 iso w9c 17: 1 ante -iso w9c 18: 0 18: 0 iso 18: 1 w7c 18: 1 w9c 19: 0 19: 0 iso 19: 0 cyclo w8c 20: 0 20: 1 w9c One - - - - - - 2.31 - 9.11 - - - 16.44 - - 5 - - - - - - 1.76 - 1.60 56.48 - - - - 3.25 8 18.94 - - - - - 0.18 - 12.16 - - - 2.26 - - 11 - 4.08 - - - - 0.83 0.20 - - - 0.63 - 0.43 - 12 - - - - - - - - - 49.50 - - - - - 17 - 2.41 2.26 26.74 14.43 0.91 - - 0.64 6.24 - - - - -

Experimental Example 2 Effect of KTG075 on the Growth of Enterobacteriaceae

In order to investigate the effect of KTG075 on the growth of enterobacteria, in vitro culture experiments were performed on the optimal medium containing KTG075 at a certain concentration against bacteria known to be beneficial and harmful to enterobacteria. .

Lactobacillus such as Lactobacillus and Bifido bacterium were cultured in MRS liquid medium (containing 0.05% L-cysteine) and other strains in NB liquid medium. Inoculated with 100 μl of the preculture of each strain in a 15-ml Corning culture tube containing 10 ml of NB or MRS liquid medium, and cultured at 37 ° C. to measure the viable cell count after 72 hours, without adding KTG075. Compared with. KTG075 was added to the test group to contain 7,000 ㎍ / mL component, and the lactic acid bacteria were cultured by filling a mixed gas of H ₂ -5%: CO ₂ -15%: N ₂ -80% in the anaerobic phase of Mart AJ9028. It was. After incubation, the number of viable cells was measured and compared, and the number of viable cells was expressed as the number of viable microorganisms per 1 ml of culture medium.

Test strains include Bifidobacterium bifidum ( KCTC 3442), Bifidobacterium longum ( KCTC 3128), Lactobacillus casei 160 ( Lactobacillus casei 160), Lactobacillus delbruecki RF-5 ( Lactobacillus delbrueckii RF-5), Lactobacillus helveticus RF-12 (Lactobacillus helveticus RF- 12), Escherichia coli (Escherichia coli), Proteus vulgaris (Proteus vulgaris), Salmonella typhimurium (Salmonella typimurium), Staphylococcus Staphylococcus aureus , Yersinia enterocolityca , Klebsiella pneumonia RF-8 were used.

As shown in Table 11, the results of the growth experiments, KTG075 is Lactobacillus casei, Lactobacillus helveticus and Bifidobacterium bifidem, Bifidobacterium longgum known as beneficial bacteria among the disclosed strains. It did not affect the growth of the back. However, it was known as a harmful bacterium, and food poisoning and acute gastritis, Salmonella typhimurium, infectious pathogen Proteus vulgaris, food poisoning and acute enterococci, Yersinia enterocholica, food poisoning and acute enterococci, and infectious pathogen Klepsi Ella pneumoniae, etc. suffered from growth.

Inhibitory Effects of KTG075 on Various Intestinal Bacterial Strains Treatment / antagonist effect Salmonella Typhimurium Prodeus Vulgaris Yersinia Enterocholicica Escherichia collie Klebsiella pneumoniae RF-8 Lactobacillus casei 160 Lactobacillus Helveticus RF-12 Bifidobacterium Bifiderm KCTC3442 Bifidobacterium longgum KCTC3128 Control 1.9-3.8 × 10 ⁸ 3.0-5.7 × 10 ⁸ 3.2-4.5 × 10 ⁸ 1.4 × 10 ⁸ 5.6 × 10 ⁸ 1.8-6.1 × 10 ⁸ 1.1 × 10 ⁸ 2.0 × 10 ⁷ 3.8 × 10 ⁸ KTG0757,000 µg / ml 1 × 10 ⁶ or less 8.8 × 10 ⁷ 1 × 10 ⁴ or less 1.5 × 10 ⁸ 3.5 × 10 ⁷ or less 6.5 × 10 ⁸ 1.1 × 10 ⁸ 1.0 × 10 ⁷ 1.4 × 10 ⁸ Antagonistic effect Growth inhibition  Growth inhibition Growth inhibition none Growth inhibition none none none none

Fecal samples were collected from rats orally administered KTG075 of the present invention to observe changes in viable cell counts, and then isolated, identified and in vitro Through experiments, KTG075 does not affect the growth of beneficial bacteria and inhibits the growth of pathogenic intestinal bacteria, which are harmful bacteria, the composition KTG075 of the present invention can be used as a medicine or health functional food for improving bacterial intestinal diseases. .

Claims (9)

The blending ratio of radish, tea, carrot, dermis, green skin, fig, onion, buckwheat and apricot extract is 1-2: 0.1-1: 1-2: 0.01-1: 0.01-1: 0.01-1: 0.1-2: 0.01 1: A pharmaceutical composition for the prevention and treatment of bacterial intestinal diseases, including extract of 0.01 to 1 and a pharmaceutically acceptable carrier The pharmaceutical composition of claim 1, wherein the radish is Raphanus sativus L. or Turnip ( Brassia Lap.). The pharmaceutical composition of claim 1, wherein the tea is Camellia sinensis , green tea, oolong tea, black tea, black tea, or iron Guanyin. delete delete delete delete delete delete

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