KR20140108822A - Composition for improving irritable bowel syndrome - Google Patents

Abstract

The present invention relates to a pharmaceutical composition comprising a wax gourd extract and lactic acid for preventing or treating irritable bowel syndrome, and foods comprising the composition for alleviating irritable bowel syndrome. A formulation comprising the wax gourd extract and lactic acid of the present invention can be widely used for developing formulations for alleviating irritable bowel syndrome by having a relatively excellent effect of alleviating irritable bowel syndrome than a conventional formulation comprising the wax gourd extract and lactic acid.

Description

[Technical Field] The present invention relates to a composition for improving irritable bowel syndrome

The present invention relates to a composition for improving irritable bowel syndrome, more specifically, to a pharmaceutical composition for preventing or treating irritable bowel syndrome including Dong-A extract and lactic acid bacterium, and a food for improving irritable bowel syndrome .

Irritable bowel syndrome is characterized by alternating diarrhea and constipation, chronic or intermittent abdominal pain, insufficient and urgent intermittent bowel movements, bloating, discomfort, gas spots, farts, and unpleasant odors. Although 7 to 15% of people have irritable bowel syndrome, it is difficult to treat them because symptoms appear even when there is no abnormality such as inflammation or tumor in the large intestine. Constipation, which is the main symptom of irritable bowel syndrome, does not excrete harmful toxic substances, which can cause swelling, acne, staining, aging, and atopic dermatitis.

Novartis of Switzerland developed Zelmac (tegaserod hydrogen maleate) as a drug for the treatment of irritable bowel syndrome, but it was banned because of side effects such as ischemic cardiovascular disease (myocardial infarction, stroke, and angina pectoris) Although constipation, which is the main symptom, can be treated with diarrhea but no side effects have been developed, diarrhea is a constipation-inducing agent and constipation is a diarrhea-inducing agent. However, The problem is known. Especially, constipation inducers are known to have side effects such as exhaustion, inactivation, dependence, etc., and diarrheal agents are known to have side effects such as constipation.

The main cause of irritable bowel syndrome is weak or stressful, so it is expected that it will not have a fundamental effect when treating only symptoms such as constipation or diarrhea, and it is expected to have a fundamental effect by relieving stress. In order to develop a therapeutic agent capable of obtaining the active ingredient, there have been many studies to obtain an effective ingredient from a natural product having little or no side effects. However, no results have been reported yet.

Under these circumstances, the present inventors have made intensive researches to develop a therapeutic agent capable of effectively treating irritable bowel syndrome without side effects, and as a result, it has been found that when using a composition containing extracts of Dong-A and lactic acid bacteria, irritable bowel syndrome And the present invention has been completed.

One object of the present invention is to provide a composition for the prevention or treatment of irritable bowel syndrome comprising Dong-A extract and lactic acid bacteria.

Another object of the present invention is to provide a food for improving irritable bowel syndrome comprising Dong-A extract and lactic acid bacteria.

As an embodiment to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating irritable bowel syndrome comprising an East Asian extract and a lactic acid bacterium.

The term " wax gourd (Benincase hispida) "of the present invention means a fruit of a plant belonging to the genus Phytophthora, a dicotyledonous plant, a leaf, a bark, It is said that it passes well and relieves stress and is effective for acne, spots, freckles, atopic skin diseases and so on. For the purpose of the present invention, the Dong-A can be used as an active ingredient of a composition which can be manufactured in the form of Dong-A extract to improve the irritable bowel syndrome.

The term "extract of East Asia" as used herein means an extract obtained by extracting East Asia. In the present invention, the pulverized product of East Asia is extracted with water, acetone, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol or butanol) And the resultant product may be a diluted solution or a concentrated solution of the extract, a dried product obtained by drying the extracted solution, or the adjusted product or the purified product.

The term " lactic acid bacteria "of the present invention means a bacterium which decomposes saccharides such as glucose to produce lactic acid, preferably a probiotic lactic acid bacteria.

The term " probiotic lactic acid bacteria "of the present invention refers to a single or complex strain of live lactic acid bacteria that improves the properties of intestinal microorganisms upon ingestion and has a beneficial effect on the host. Unlike antibiotics that kill off harmful bacteria, While suppressing the growth or proliferation of the harmful microorganisms, resulting in the multiplication of the host. Such probiotic lactic acid bacteria or foods containing the same can prevent abnormal fermentation by germs in the intestines and improve the intestinal environment to aid digestion and absorption of food, improve the function of the large intestine, improve the constipation, diarrhea, And treatment, reduction of blood cholesterol, prevention of cancer, and suicidal action can improve the health of the recipient.

The lactic acid bacterium which can be used as such a probiotic microorganism is not particularly limited, but may be a microorganism belonging to the genus Lactobacillus sp., Bacillus sp., Or Bifidobacterium bifidum sp. Preferably Lactobacillus brevis, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus spp., Lactobacillus spp., Lactobacillus spp. Lactobacillus delbrueckii subsp bulgaricus, Bifidobacterium bifidum, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus helveticus, Lactobacillus delbrueckii subsp bulgaricus, Bifidobacterium bifidum, Bifidobacterium longum, Bifidobacterium breve, , Bifidobacterium adolescentis, Bifidobacterium < RTI ID = 0.0 > infantis, Bifidobacterium psendolongum and Bifidobacterium animalis may be used singly or in combination.

Foods containing the probiotic lactic acid bacteria are mainly dairy products such as fermented milk, syrup, milk powder, and ice cream, but the range of other foods including confectionery and bakery products is increasing. Currently, the products containing probiotic lactic acid bacteria include probiotic products, Products, and genetically modified products are being actively developed. The probiotic microorganism uses the microbial cell itself, the microbial agent uses the antimicrobial substance secreted by the microbial cells, the feed additive uses the probiotic agent, and the human body drug includes the formulant and the lactic acid bacterial agent.

In the Functional Food Act, the efficacy of probiotic lactic acid bacteria is recognized as "inhibition of harmful bacteria, smooth activity of bowel movements" and classified as belonging to the fermented microbial probiotics as a functional raw material. Only Lactobacillus, Lactococcus, Enterococcus, Streptococcus, and Bifidobacterium are recognized as probiotic lactic acid bacteria in the functional functional foods. These microorganisms are cultured and dried, containing 100,000,000 CFU / g or more of live cells and 100,000,000 ~ 10,000,000,000 CFU / g.

In addition, the pharmaceutical compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. The pharmaceutical composition for the prevention or treatment of irritable bowel syndrome according to the present invention may be administered orally or parenterally in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations, suppositories, And the like. In the present invention, the carrier, excipient and diluent which can be contained in the pharmaceutical composition for the prevention or treatment of irritable bowel syndrome include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin , Calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a commonly used filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant or the like is used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, ), Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use may include various excipients such as wetting agents, sweetening agents, fragrances, preservatives, etc. in addition to water and liquid paraffin, which are simple diluents commonly used in suspension, liquid solutions, emulsions and syrups have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

As another embodiment for achieving the above object, the present invention provides a food for improving irritable bowel syndrome comprising an East Asian extract and a lactic acid bacterium.

The Dong-A extract or lactic acid bacterium of the present invention has been conventionally used as a medicinal or edible food, and its safety has been proved, so it can be used as a food composition.

The composition comprising the extract of Dong-A and the lactic acid bacterium is contained in an amount of 0.01 to 100% by weight, more preferably 1 to 80% by weight based on the total weight of the food composition. When the food is a beverage, it may be contained in a proportion of 1 to 30 g, preferably 3 to 20 g based on 100 ml. In addition, the composition may contain additional ingredients which are commonly used in food compositions and which can improve odor, taste, vision and the like. For example, vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid and the like. It is also possible to use zinc (Zn), iron (Fe), calcium (Ca), phosphorus (P), magnesium (Mg), manganese (Mn), copper (Cu) ), And the like. It can also contain more than 20 natural amino acids. It is also possible to add preservatives (antiseptics, potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate), disinfectants (such as bleaching powder and highly bleached white powder, sodium hypochlorite), antioxidants (butylhydroxyanisole (BHA), butylhydroxytoluene (Sodium saccharinate, aspartame, stevioside, etc.), coloring agent (sodium nitrite, sodium nitrate), bleaching agent (sodium sulfite), seasoning (sodium MSG glutamate) (Food additive) such as flavorings (vanillin, lactones, etc.), swelling agents (alum, potassium hydrogen D-tartrate), reinforcing agents, emulsifiers, thickeners, encapsulating agents, gum bases, foam inhibitors, The additives may be selected according to the type of food and used in an appropriate amount.

A functional food for improving irritable bowel syndrome can be prepared using the food composition containing the Dong-A extract and the lactic acid bacterium.

As a specific example, the above-mentioned composition can be used to produce a processed food having improved storage properties while utilizing the characteristics of agricultural products, livestock products or aquatic products. The processed food may be, but is not limited to, margarine, fat continuous or water continuous or bicontinuous spread, fat reduced spread, chocolate, or chocolate coating or chocolate fillings, Such as bakery products, ice cream, ice cream coatings, ice cream containing products, dressings, mayonnaise, cheese, cream substitutes, dried soup, drinks, cereal bars, sauces, snack bars, dairy products,

Specific examples of the processed foods include confectionery, drinks, liquor, fermented foods, canned foods, processed milk products, processed marine foods, and noodles. The sweets include biscuits, pies, cakes, breads, candies, jellies, gums, cereals (including dinner utensils such as cereal flakes). Drinks include carbonated beverages, functional ionic beverages, juices (such as apples, pears, grapes, aloes, citrus fruits, peaches, carrots, tomato juices, etc.) and sikhye. The mainstream includes sake, whiskey, shochu, beer, liquor, and fruit wine. Fermented foods include soy sauce, miso, and kochujang. Canned products include canned goods (eg, tuna, mackerel, saury, canned fish, etc.), canned products (beef, pork, chicken, turkey canned food, etc.) and canned products (corn, peach and canned pineapple). Milk processed foods include cheese, butter, yogurt and the like. Meat processed foods are pork cutlet, beef cutlet, chicken cutlet, sausage. Sweet and sour pork, nuggets, and nubani. And noodles such as sealed packaging raw noodles. In addition, the composition may be used in retort food, soup and the like.

In another embodiment, the composition may be used alone or as a core active ingredient to produce a processed food that can directly exhibit the effect of the composition. Such processed food may be a tablet, a powder, a granule, a capsule, a ring, a liquid agent, and the like, but is not particularly limited thereto.

The term " functional food "in the present invention is the same term as a food for special health use (FoSHU). In addition to the nutritional supply, the term " functional food " The term " health food "in the present invention refers to a food having an active health promotion or promotion effect as compared to a general food, and a health supplement food refers to a food . In some cases, the terms functional foods, health foods, and health supplements are commonly used. The food may be prepared in various forms such as tablets, capsules, powders, granules, liquids, and circles in order to obtain effects useful for the improvement and prevention of irritable bowel syndrome.

The formulation containing the Dong-A extract and the Lactobacillus bacteria according to the present invention can exhibit a relatively superior irritable bowel syndrome improving effect than the conventional Dong-A extract or the formulation containing the Lactobacillus bacterium, and thus can be widely used for the development of an agent for improving irritable bowel syndrome will be.

FIG. 1 is a graph and photograph showing the anticancer activity of each tablet against human lung cancer A549 cells.
2 is a graph and photograph showing the anticancer activity of each tablet against human colon cancer HCT-116 cells.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example  1: Diarrheal improvement effect of Dong-A extract and lactic acid bacteria

From August 1, 2012 to January 30, 2013, 36 people with diarrhea symptoms of food poisoning, 44 people with diarrhea symptoms of pathogenic diseases, and 40 people with diarrhea symptoms of alcohol or other causes were grouped And 100 ml of Dong-A solution or 10 g of lactic acid bacteria every 12 hours was taken as an improvement completion point. At this time, the extracts of Dong-A were dried at 65 ° C for 48 hours, hot-extracted for 14 hours, and then filtered to obtain Lactobacillus casei and Bifidobacterium bifidum The live mixed culture culture was used (Table 1). The lactic acid bacteria contained in the culture dried product are maintained at a certain level of their physiological activities.

Long-term improvement effect of diarrhea of Dong-A extract and Lactobacillus (Unit: number / total number) Diarrhea cause ingredient Time to 80% improvement 3h 6h 12h 24h 48h 72h sum χ2 (p) food poisoning Lactobacillus
Dong-A Extract 2
8 6
10 8
11 7
4 6
3 3
0 32/36
36/36 33.257
(0.000) Pathogenic disease Lactobacillus
Dong-A Extract 5
9 6
14 7
10 9
5 9
2 6
One 42/44
41/46 Alcohol, other Lactobacillus
Dong-A Extract 2
9 4
12 7
9 10
5 7
3 4
2 34/40
40/40 Sum Lactobacillus
Dong-A Extract 9
26 16
36 22
30 26
14 22
8 13
3 108/120
117/120

*: p < 0.05, **: p < 0.01, ***: p < 0.001

As shown in Table 1, 120 patients suffering from diarrhea were treated with lactic acid bacteria for 72 hours, and 80% of food poisoning-type diarrhea, 100% of pathogenic disease-type diarrhea, 85% of alcohol and other diarrhea were improved , And 108 of the total number of participants were improved, showing a 90% improvement rate. Food-borne diarrhea, alcohol and other diarrhea that did not improve 100% until 72 hours continued to improve after 72 hours. The severity of food poisoning diarrhea was 12 to 24 hours, pathogenic diarrhea was 24 to 48 hours, and alcohol and other forms of diarrhea were the most improved at 24 hours.

In addition, 100% of food poisoning type diarrhea, 91% of pathogenic disease type diarrhea, and 100% of alcohol and other type of diarrhea were obtained when Dong - ah extract was applied to 120 people for 72 hours. Of the total number of participants, an improvement of 117% was achieved, with a 98% improvement. The improvement rate of food poisoning type diarrhea was 6 ~ 12 hours, the pathogenic disease type diarrhea was 6 hours, and that of alcohol and other type of diarrhea was 6 ~ 12 hours faster than that of lactic acid bacteria.

Therefore, it was confirmed that lactic acid bacteria or Dong - A extract had significant effects (p <0.001) on the improvement of food poisoning diarrhea, pathogenic disease diarrhea, liquor and other diarrhea.

The results were analyzed by severity and age (Table 2).

Effect of age on diarrhea of Dong-A extract and Lactobacillus (Unit: number / total number) Age (years) 2-15 16-30 31 or more sum χ2 (p) Lactobacillus food poisoning
Pathogenic disease
Alcohol
total 8/12
14/14
4/8
26/34 12/12
16/16
14/16
42/44 12/12
14/14
14/16
40/42 32/36
44/44
32/40
108/120 3.885
(0.422) Dong-A Extract food poisoning
Pathogenic disease
Alcohol
total 12/12
16/18
6/6
34/36 12/12
14/14
16/16
42/42 12/12
10/12
18/18
40/42 16/16
40/44
40/40
116/122 7.150
(0.128)

*: p < 0.05, **: p < 0.01, ***: p < 0.001

As shown in Table 2, there was no significant difference according to age, but lactic acid bacteria showed 66.7% improvement in food poisoning type diarrhea aged 2-15 years and 100% improvement in ages higher than that , Pathogenic diarrhea was 100% in all ages, diarrhea of 2 ~ 15 years of age and other diseases showed 50% improvement, and 100% of the age group was higher.

In the case of food poisoning type diarrhea, 100% of food poisoning type diarrhea, 88.9% of 2 ~ 15 year old cases, 100% of 16 ~ 30 years old and 83.3% And 100% for other forms of diarrhea.

Therefore, it was confirmed that Lactobacillus was effective against pathogenic diarrhea and Dong - A extract showed excellent therapeutic effect against other diarrhea.

Example  2: Tablet  Manufacturing and Characterization

14 kg of Dong-A were dried at 65 ° C for 48 hours, and then hydrothermally extracted for 14 hours. Then, 14 L of 26 brix extract was obtained by filtration, which was freeze-dried to obtain 3.6 kg of dry extract powdery extract.

The present invention also relates to a method for producing a lactic acid bacterium, which comprises using the above-mentioned extract of Dong-A, lactobacillus strains as a mixed culture culture of Lactobacillus casei and Bifidobacterium bifidum, The mixture containing the magnesium stearate was pushed through a Hanli Phamatec HLT-30 tablet machine (100,000 tablets per hour) to prepare an oval tablet of 1.3 x 0.7 x 0.5 cm (0.45 g). At this time, the ratios of the respective components are shown in Table 3 below.

Material cost of Dong-A extract and Lactobacillus tablet (%) Tablet Dong-A Extract Lactobacillus Lactic acid bacteria and Dong-A extract Dong-A Extract
Lactobacillus
Crystalline cellulose
Lactose
Magnesium stearate 33.2
-
66.4
-
0.5 -
49.8
-
49.8
0.5 33.2
33.2
33.2
-
0.5

The carbohydrate, crude protein, crude fat, ash and moisture of the tablet were analyzed according to the method of food revolution (KFIA 2000) (Table 4) and the disintegration was analyzed according to the function of the functional food (KFDA 2009a).

General composition of Dong-A extract and lactic acid bacteria preparation (%) Tablet Dong-A Extract Lactobacillus Lactic acid bacteria and Dong-A extract carbohydrate
Crude protein
Crude fat
Ash
moisture 72.8
13.7
5.3
3.5
4.7 73.6
17.4
4.1
1.1
3.7 66.5
16.3
7.2
4.4
5.6

As shown in Table 4, both the Dong-A extract and the Lactobacillus preparation prepared by the tablet, the Dong-A extract and the Lactobacillus preparation, which were compared with each other, contained the most carbohydrate because the crystalline cellulose and lactose were added as excipients .

In East Asia, when the content of concentrated components is calculated, the monosaccharides are 8.8% of fructose, 2.5% of sucrose, 8.0% of glucose, and 2.5% of other sugars, totaling 21.2%. Organic acid is 12.9%, while citric acid is 10.2%, malic acid is 2.1%, succinic acid is 2.5% and fumaric acid is 0.005%. Crude protein was 13.7% and crude fat was 5.3%.

In addition, the disintegration of the extract of Dong-A and Lactococcus tablets with water as a solvent was suitable for the requirement that it should be disintegrated within 30 minutes after 18 minutes (KDFA 2009a). 38 minutes for the Dong-A extract tablet and 7 minutes for the lactobacillus tablet.

Example  3: Tablet Irritable Bowel Syndrome  Improvement effect

The improvement effect of each tablet prepared in Example 2 on irritable bowel syndrome from August 1, 2012 to January 30, 2013 was evaluated in 36 cases of diarrhea predominant type, 40 cases of constipation type predominant type, (44%), and others (abdominal pain, tortoise type), were assessed as improvement completion points (Table 5). At this time, significance was crossovered (χ2) using statistical analysis program SPSS 20.0.

Improvement of irritable bowel syndrome over time in tablet (unit: number / total number) Symptom type Tablet Time to 80% improvement 3h 6h 12h 24h 48h 72h sum χ2 (p) Diarrhea Lactobacillus
Dong-A Extract
Lactic acid bacteria and Dong-A extract 2
8
19 6
10
12 8
11
4 7
4
One 6
3
0 3
0
0 32/36
36/36
36/36 37.392
(0.000) Constipation Lactobacillus
Dong-A Extract
Lactic acid bacteria and Dong-A extract 2
10
16 4
14
18 5
10
6 6
4
0 4
0
0 3
0
0 24/40
38/40
40/40 42.518
(0.000) Typical Lactobacillus
Dong-A Extract
Lactic acid bacteria and Dong-A extract 3
12
26 4
18
14 6
12
4 6
2
0 5
0
0 2
0
0 26/44
44/44
44/44 55.890
(0.000) Etc Lactobacillus
Dong-A Extract
Lactic acid bacteria and Dong-A extract 3
12
26 6
14
10 7
15
8 8
One
0 4
0
0 3
0
0 31/44
42/44
44/44 55.345
(0.000) Sum Lactobacillus
Dong-A Extract
Lactic acid bacteria and Dong-A extract 10
42
87 19
56
54 25
48
22 27
11
One 21
3
0 11
0
0 113/164
160/164
164/164 37.392
(0.000)

*: p < 0.05, **: p < 0.01, ***: p < 0.001

As shown in Table 5, in the case of the lactic acid bacteria tablet, the diarrhea symptom improved by 89%, the constipation symptom by 60%, the general symptom by 59% and the other symptoms by 70.5% . In addition, the Dong-A Extract Tablet improved diarrhea, constipation and general symptoms by 100% and other symptoms by 95.5%. Finally, Dong-A Extract + Lactobacillus tablets were all 100% improved.

In addition, the lactobacillus tablets showed the highest improvement efficiency at 12 to 24 hours for diarrhea, constipation, general and other irritable bowel syndrome symptoms, while the Donga extract tablet had the highest level at 3 to 12 hours, Tablets were highest at 3 to 6 hours.

Therefore, all of the above tablets showed significant effects (p < 0.001) on the improvement of irritable bowel syndrome, and Donga extract + lactic acid bacteria tablet showed the most excellent effect.

On the other hand, the results were classified according to age and age (Table 6).

Improvement of irritable bowel syndrome by age of each tablet (unit: number / total number) Age (years) 2-15 16-30 31 or more sum χ2 (p) Lactobacillus diarrhea
Constipation
Typical
Etc 12
4
6
9 10
10
12
12 10
10
8
10 32/36
24/40
26/44
31/44 3.880
(0.693) Dong-A Extract diarrhea
Constipation
Typical
Etc 12
10
14
14 12
14
14
12 12
14
16
16 36/36
38/40
44/44
42/44 0.972
(0.987) Lactic acid bacteria +
Dong-A Extract diarrhea
Constipation
Typical
Etc 12
12
16
17 12
12
14
14 12
16
14
16 36/36
40/40
44/44
44/44 0.801
(0.992)

*: p < 0.05, **: p < 0.01, ***: p < 0.001

As shown in Table 6, there was no significant difference between the tablets.

Example  4: Tablet  Analysis of physiological activity

The physiological activities (Angiotensin converting enzyme inhibitory activity, Xanthine oxidase inhibitory activity, Superoxide dismutase (SOD) -like activity, Antioxidative activity, α-Glucosidase inhibitory activity and Tyrosinase inhibitory activity) of each tablet prepared in Example 2 were compared (Table 7).

First, the Angiotensin converting enzyme inhibitory activity was measured by modifying a known method. 50 mL of the sample was dried under reduced pressure, and the same amount of ethyl acetate was added to remove the supernatant. To the filtrate (60 μL) obtained by filtration, 30 μL of ACE solution (100 mU / mL) extracted from rabbit lung powder and a substrate solution (HHL) dissolved in 500 mM NaCl and 6.5 mM Hip-His-Leu in 100 mM boric acid buffer solution (pH 8.3) were mixed and reacted at 37 ° C. for 30 minutes. The reaction was stopped with 0.5 N HCl to obtain 228 nm The amount of hippuric acid was measured by the absorbance of. 1 unit was the amount that produced 1 μmol of hippuric acid from HHL at 37 ° C for 1 min.

ACE inhibitory activity (%) = (CT / CB) x 100

Next, Xanthine oxidase inhibitory activity was measured according to a known method. That is, 100 μL of a solution obtained by dissolving 1 mg / mL of a sample in 600 μL of a 0.1M potassium phosphate buffer solution (pH 7.5) was added, and 200 μL of a substrate solution in which 1 mM xanthine was dissolved was added thereto, and xanthine oxidase (0.2 unit / mL) The reaction was stopped by adding 200 μL of 1 N HCI. The reaction was stopped by centrifugation at 12,000 rpm for 10 minutes to remove proteins, and the generated uric acid was measured at 292 nm.

Xanthine oxidase inhibitory activity (%) = [1- {A (Samples) -B (Blank) / C (Control)}]

Next, the superoxide dismutase (SOD) -like activity was homogenized by adding 55 mM Tris-cacodylic acid buffer (TCB, pH 8.2) to each tablet according to a known method and centrifuged to adjust the supernatant to pH 8.2. , 50 μL of 24 mM pyrogallol was added to 950 μL of the sample solution, and the absorbance was measured at 420 nm for the initial 2 minutes.

SOD-like activity (%) = (AB) / A × 100

Next, the antioxidant activity was measured using a reducing power of 1,1-diphenyl-2-picrylhydrazyl (DPPH) according to a known method. That is, 0.8 mL of DPPH solution (dissolved in 12.5 mg of DPPH) was added to 0.2 mL of the sample, reacted for 10 minutes, and the absorbance at 525 nm was measured and compared with the control.

Antioxidant activity (%) = [1- {T10min- (T0min-Control) / control}] x100

Next, the? -Glucosidase inhibitory activity was measured according to a known method. 50 μL of a 0.2 unit / ml α-glucosidase solution was added to 50 μL of the concentrated sample, and the mixture was preliminarily reacted at 37 ° C. for 5 minutes. Then, 50 μL of 1.5 mM p-NPG was added thereto and reacted at 37 ° C. for 25 minutes. The reaction was stopped by adding 100 μL of M Na ₂ CO ₃ and the inhibition rate was calculated by measuring the absorbance at 405 nm

α-Glucosidase inhibitory activity (%) = {(Control-Test) / Control} × 100

Finally, tyrosinase inhibitory activity was measured according to known methods. In other words, a mixture of 200 μL of 5 mM L-DOPA and 800 μL of 0.1 M phosphate buffer (pH 6.0) was added to 500 μL of the sample, and 11 units of tyrosinase was added thereto. The reaction was carried out at 35 ° C. for 2 minutes. Absorbance was measured at 475 nm, .

Tyrosinase inhibitory activity (%): {Control- (Test-Balk) / Control} x 100

Physiological activity (%) of each tablet Tablet Dong-A Extract Lactobacillus Dong-A Extract
+ Lactic acid bacteria ACE inhibitory activity
XOD inhibitory activity
SOD-like activity
Antioxidant activity (DPPH)
α-Glucosidase inhibitory activity
Tyrosinase inhibitory activity 47.9 (+ - 0.2)
5.4 (+/- 0.5)
nd
23.4 (+ - 0.3)
nd
13.2 (+/- 0.3) 21.4 (+/- 0.7)
nd
nd
4.4 (± 0.3)
nd
3.5 (+/- 0.9) 27.1 (+/- 0.1)
nd
nd
20.3 (+/- 0.3)
nd
3.6 (+ - 0.2)

1) ACE; angiotensin converting enzyme, XOD; xanthine oxidase, SOD; superoxide dismutase 2) nd; not detected.

As shown in Table 7, the physiological activity of the Dong-A extract tablet showed 47.9% inhibition of angiotensin converting enzyme (ACE), which means antihypertensive, and 13.2% inhibition of tyrosinase, which indicates a whitening effect. Was 23.4%. Next, the lactobacillus tablet inhibited the activity of angiotensin converting enzyme by 21.4% and inhibited the remainder by less than 5%. Finally, the extracts of Dong-A extract and Lactobacillus acidus inhibited the activity of angiotensin converting enzyme by 27.1% and antioxidant activity by 20.3%, but the inhibitory activity against other enzymes was less than 5% or not inhibited.

Example  5: Tablet  Analysis of antitumor activity

To each well of 24-well plate, 500 mL of cell culture medium suspension containing 0.1 x 10 ⁴ human lung cancer A549 cells and colon cancer HCT116 cells was added and cultured overnight at 37 ° C in a cell incubator. On the next day, the cell culture medium was removed and the cell culture medium was removed from the A549 cells for 24 hours by adding Dong-A &lt; / RTI &gt; extract + Lactobacillus, Dong-A extract and Lactobacillus acid preparations with different concentrations of 0, 100, 250 and 500 占 퐂 / A549 and HCT116 cells were inoculated with trypsin-EDTA solution in 24 wells to prepare a cell suspension. Trypsin was added to about 10 mL cell suspension to count the number of living cells without dying under a microscope. The experiment was repeated three times. The photographs of A549 and HCT116 cells were exposed for 24 hours as described above, and then the cell culture medium was removed and taken under a microscope (FIGS. 1 and 2).

FIG. 1 is a graph and photograph showing the anticancer activity of each tablet against human lung cancer A549 cells. As shown in FIG. 1, treatment of A549 cells of human lung cancer with concentrations of 100, 250 and 500 / / ml of Dong-A extract tablet did not affect cancer cell proliferation (A), but the lactobacillus tablet was treated at a concentration of 250 / / (B), and the concentration of the extracts of L. acidophilus and L. acidophilus was 67%, 500 ㎍ / ㎖ at 100 ㎍ / ㎖ and 250 ㎍ / ㎖, respectively, (C).

2 is a graph and photograph showing the anticancer activity of each tablet against human colon cancer HCT-116 cells. As shown in FIG. 2, treatment of human colon cancer HCT116 cells with the concentrations of 100, 250 and 500 μg / ml did not affect the proliferation of cancer cells (A, B), but the extracts of Dong- Tablets were inhibited by about 70% when treated at a concentration of 100 μg / ml, 68% when treated at a concentration of 250 μg / ml, and 58% when treated at a concentration of 500 μg / ml.

Claims (9)

A pharmaceutical composition for the prevention or treatment of irritable bowel syndrome comprising a lactic acid bacterium and an East Asian extract.
The method according to claim 1,
Wherein the extract is obtained by extracting Dong-A using a solvent selected from the group consisting of water, acetone, lower alcohols having 1 to 4 carbon atoms, and mixed solvents thereof.
The method according to claim 1,
Wherein said lower alcohol is selected from the group consisting of methanol, ethanol, propanol and butanol.
The method according to claim 1,
Wherein the lactic acid bacterium is selected from the group consisting of Lactobacillus sp., Bifidobacterium bifidum sp. Microorganisms and mixed strains thereof.
The method according to claim 1,
Lt; RTI ID = 0.0 &gt; pharmaceutically &lt; / RTI &gt; acceptable carrier, excipient or diluent.
A health-functional food showing the effect of preventing or ameliorating irritable bowel syndrome comprising the composition of any one of claims 1 to 5.
The method according to claim 6,
Wherein the composition is added or a food containing the composition alone.

8. The method of claim 7,
Foods in the form of adding such compositions may be used in a variety of forms including margarine, fat continuous or water continuous or bicontinuous spreads, fat reduced spreads, chocolate, or chocolate coatings or chocolate fillings or bakery Selected from the group consisting of confectioneries such as buckwheat, ice cream, ice cream coatings, ice cream inclusions, dressings, mayonnaise, cheese, cream substitutes, dried soup, drinks, cereal bars, sauces, snack bars, dairy products, Health functional food.
8. The method of claim 7,
Wherein the food comprising the composition alone is selected from the group consisting of tablets, powders, granules, capsules, pills and liquids.

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