KR101600884B1 - Composition for improving, treating or preventing constipation comprising Cassia fermented by lactic acid bacteria as an active ingredient - Google Patents

Abstract

The present invention relates to a dietary supplement product for preventing and alleviating constipation by using Senna tora fermented with lactic acid bacteria, a pharmaceutical composition for the same, and a method for preparing the fermented Senna tora in order to increase the weight and moisture content of excrement. The Senna tora fermented with lactic acid bacteria is used for a long time to ensure safety and show little side effects as herbal medicine while showing weight and moisture increasing effects similar to that of Dulcolax-S which uses sennoside, docusate sodium, and bisacodyl as active ingredients and is prohibited from being used as a dietary supplement product due to side effects. Accordingly, the present invention can be used as the pharmaceutical composition for the prevention and treatment of constipation or the dietary supplement product for preventing and alleviating constipation.

Description

[0001] The present invention relates to a composition for improving, treating or preventing constipation, comprising a fermented product of lactic acid bacteria as an active ingredient,

The present invention relates to a health functional food for improving or preventing constipation, a pharmaceutical composition for treating or preventing constipation, and a method for producing a fermented product of a luciferous lactobacillus which increases the moisture content and weight of the liquor.

Constipation is defined as constipation when it is medically less than 3 bowel movements per week or when the bowel movement is 30 g or less per day. Under normal conditions, it is excreted in the stool 24 hours after ingestion of food. However, people with constipation see the stools every few days or even if they see it every day, resulting in a longer stay in the intestinal tract of the resulting food.

The causes of constipation include a decrease in the intestinal motility caused by eating breakfast, a lack of dietary fiber due to westernization of dietary life, a decrease in basic physical strength and abdominal strength due to lack of exercise, dieting, drug addiction due to excessive use of drugs, have.

Constipation, which is said to be the source of all diseases from ancient times, has no appetite and is not only in a state where the abdomen is always bloated, but also toxins of the unexcited side are absorbed into the blood through the intestines, thereby promoting aging of the skin and causing headache, acne, skin rash And when constipation is severe, it breaks the dentition and escapes the dentition when it is defecated, and it causes the hemorrhoids by the toxin in the shift. If constipation persists, cholesterol or fat that should be excreted outside the body may remain in the body, which may lead to arteriosclerosis or gallstone disease. Furthermore, hypertension and cardiac hypertrophy may be exacerbated by heart disease. In addition, constipation is an important disease that requires active prevention and treatment because it is a cause of diverse and serious secondary diseases such as stroke, immunodeficiency, visual disturbance and mental illness (depression).

To date, various medicines and health functional foods have been sold for the treatment or improvement of constipation. However, most of the medicines use stimulant medicines containing anthraquinone derivatives such as senna and rhubarb, and have side effects such as abdominal pain and diarrhea. And there is a problem in taking or continuously taking the food, and in the case of the functional food, the effect of the scientific function is not sufficiently proved. Therefore, the reliability of the product is deteriorating because the fundamental problem of constipation improvement can not be solved.

Cassia obtusifolia L. and Cassia ( Cassia obtusifolia L.) Tora L.) are known. The killer is also called the long-lived Kangnam car. Candidates are cultivated in Central Asia, Japan and elsewhere. Tropical Asia is cultivated in China and Korea.

The shape of the fruit is irregular hexagonal column, one side is pointed. The color of the outer surface is from yellow to black brown, 3 to 6 mm in length, and 2 to 3.5 mm in diameter. The seed coat is firm and shiny, with a narrow striped pattern with polished sides on both sides and a somewhat unusual smell. C. obtusifolia is seed confrontation, C. tora is small.

In case of acute conjunctivitis, ocular hemorrhage, foreign body or pain, and shedding of tears due to the nominal function of cheongryeol in one room, rapid visual acuity due to corneal haze occurring in the eyes of the elderly such as chronic progressive disease, optic nerve, It is the treatment of headache, tinnitus and eye tumor due to decline, obesity, blood pressure, mental stress, treatment of hypertension as coronary artery enlargement, lowering of blood pressure by lowering blood pressure resistance by coronary artery dilation, Mild diarrhea and chronic diseases, cholesterol lowering action is to treat cardiovascular disease, that is, cholesterol level exceeding the normal range or coronary artery stenosis caused by coronary atherosclerosis, and constipation treatment is the treatment of senile constipation, senile hypertension, It is known to treat dry mouth, swelling of the abdomen due to constipation, constipation and restlessness. .

Korean Patent Laid-Open Publication No. 1999-0084271 discloses that a tea bag prepared by mixing a livestock or a livestock lobe with other herbal medicines and simply pulverizing and distributing the livestock tea is effective in improving constipation, and Korean Patent Publication No. 2003-0062493 discloses that a lumberjack First, it is explained that a mixed leach tea prepared by mixing Cassiae japonica extract powder and Cassara sagrada extract powder prepared by extraction, concentration and spray drying has excellent palatability and an effect of improving constipation symptoms. They are merely pulverized and distributed, or extracted and concentrated and dried, or mixed with herbal medicines having different constipation-improving effects.

However, the present inventors have found that fermented product of Lactobacillus lactic acid bacteria obtained by fermenting a herbal medicine, which is a herbal medicine which has been traditionally used for a long time and proved its safety, with lactic acid bacterium is improved or treated with constipation by increasing the number of sides, water content of side, The effect is remarkably improved, and the present invention has been completed.

It is an object of the present invention to provide a health functional food for improving or preventing constipation, which has been used for a long time, has a high safety, and has a significantly improved effect of increasing the moisture content and weight of a side by using a cleanser which is less likely to cause side effects as a herbal medicine.

Another object of the present invention is to provide a pharmaceutical composition for the treatment or prevention of constipation, which has been used for a long time and has a high safety and an effect of increasing the moisture content and weight of the stomach by using a cleanser which is less likely to cause side effects as a herbal medicine.

Another object of the present invention is to provide a method for producing a luciferous fermentation product which is highly effective in increasing the number of feces, the moisture content of the feces and the weight of the feces, Method.

The present invention provides a health functional food for improving or preventing constipation, which comprises a fermented product of a lactic acid fermentor of an active ingredient as an active ingredient.

The fermented product of the Lactobacillus canceller may be obtained by fermenting lactic acid bacteria inoculated with the luciferase extract.

The Cassiae Extract may be an extract of water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The lactic acid bacteria may be any one or more lactic acid bacteria selected from the group consisting of Lactobacillus sp., Bifidobacterium sp., Leuconostoc sp., Pediococcus sp., Enterococcus sp. Lactic acid bacteria.

The health functional food may be in the form of a capsule, tablet, powder, granule, liquid, ring, flake, paste, syrup, gel, jelly or bar.

The present invention also provides a pharmaceutical composition for the treatment or prevention of constipation, comprising a fermented product of a lactic acid fermentor of an active ingredient as an active ingredient.

The fermented product of the Lactobacillus canceller may be obtained by fermenting lactic acid bacteria inoculated with the luciferase extract.

The Cassiae Extract may be an extract of water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The lactic acid bacteria may be any one or more lactic acid bacteria selected from the group consisting of Lactobacillus sp., Bifidobacterium sp., Leuconostoc sp., Pediococcus sp., Enterococcus sp. Lactic acid bacteria.

The present invention also relates to a method for producing a Cassia tumefaciens extract; And a step of inoculating and lyophilizing the Lactobacillus japonica extract with a lactic acid bacterium, wherein the method further comprises increasing the number of sides, the moisture content of the sides, and the weight of the sides.

The fermented product of the lactic acid bacteria of the present invention has been used for a long time and has high safety and is less likely to cause adverse side effects as a herbal medicine. However, sincoside, a drug for the treatment of irritable constipation, or bisacodyl and docusate sodium, , The water content of the side and the weight of the stomach, it can be used as a pharmaceutical composition for the treatment or prevention of constipation or as a health functional food for improving or preventing constipation. .

FIG. 1 is a graph showing the progress of fermentation of lactic acid bacteria through a change in pH measured at 36 and 72 hours after fermentation with a different lactic acid bacterium according to Experimental Example 1. FIG. 1, the extract of Preparation Example 2, and the dilution of Production Example 3, respectively.
2 is a graph showing the number of sides of each test group in Experimental Example 2. In FIG.
3 is a graph showing the weights of the sides of each experimental group in Experimental Example 2. FIG.
FIG. 4 is a photograph of the experimental animal of Experimental Example 2 after sacrificing the experimental animals of each experimental group and then collecting and washing the large intestine.
FIG. 5 is a graph showing a moving distance of Carmine in FIG.
FIG. 6A shows the HPLC-IT / TOP MS pattern of the aqueous solution of the aqueous solution of Cefditor of Preparation Example 2 in Experimental Example 3, and FIG. 6B shows the HPLC-IT / TOP MS pattern of the fermented product of the Cactus Lactobacillus of Preparation 4 in Experimental Example 3 .

The present invention relates to a health functional food for improving or preventing constipation, comprising a fermented product of a lactic acid fermented by a defective lactic acid bacteria as an active ingredient. The present invention also relates to a pharmaceutical composition for treating or preventing constipation, The invention relates to a method for the production of a cetacean extract; And a step of inoculating and lyophilizing the Lactobacillus japonica extract with the lactic acid bacterium, wherein at least one of the number of sides, the moisture content of the sides, To a process for producing fermented lactic acid bacteria.

The loser may be a Cassia obtusifolia L.) or Cassia tora L.), preferably gyeolmyeongja grown in Korea (Cassia tora L.).

The fermented product of the Lactobacillus canceller may be obtained by fermenting lactic acid bacteria inoculated with the luciferase extract.

The extracts include crude extracts as well as crude extracts obtained by treating extraction raw materials with extraction solvents. For example, the Cassiae Extract can be prepared in powder form by an additional process such as vacuum distillation and lyophilization or spray drying. The extract also includes fractions obtained by further fractionating the crude extract.

The Cassiae Extract is an extract of water, an alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, and the extraction method is not particularly limited.

When water is used as the solvent, hot water extraction is preferred. For example, at a temperature of 80 to 105 DEG C, preferably 90 to 100 DEG C for 0.5 to 24 hours, preferably 1 to 6 hours. Even if no hot water is used as a solvent, the components of the cleanser can be extracted from the diluted solution obtained by diluting the cuticle powder with cold water or water at room temperature. When cold water or room temperature water is used to extract the components of the lucifer, it may be extracted separately from the fermentation. In the fermentation by lactic acid bacteria inoculation, the lucifer may be eluted from the lucifer.

An extract of an aqueous solution of an alcohol having 1 to 4 carbon atoms can be used. For example, extraction is carried out with an alcohol aqueous solution such as ethanol, methanol or isopropanol, preferably 20 to 80% by weight aqueous alcohol solution, more preferably 50 to 70% by weight aqueous alcohol solution. When alcohol extracts are used, concentrate or dry the alcohol concentrate or alcohol extract, which has lowered the alcohol content, by evaporating alcohol from the alcohol extract before inoculation of the lactic acid bacteria, and dissolve in water before use.

Also, in the present invention, the Cassiae Extract includes, in a broad sense, a diluted solution in which the Cassiae powder is diluted with water. In the experimental example of the present invention, the fermented product of fermented lactic acid bacteria obtained by fermenting the water-diluted lysate of the luciferase with the fermented product of lactic acid bacteria is somewhat lower than the fermented product of fermented lactic acid bacteria, And the effect was significantly increased compared to the non - Cassiae extract.

A lactic acid bacteria nutrient source such as a protein source, a carbohydrate source, a vitamin or an inorganic source may be further mixed to enhance the growth of lactic acid bacteria before the lactic acid bacteria are inoculated into the Cassiae Extract. The lactic acid bacteria nutrient source may be supplemented with commercially available media or only necessary nutrients.

Also, the above-mentioned Cassiae Extract, or a mixture of the Cassiae Extract and the Lactobacillus nutrient source, may be sterilized by heating before the lactic acid bacteria are inoculated.

The solid content of the Cassiae Radix Extract is not particularly limited, but may be 1 to 15% by weight when there is no other concentration process after extraction, and may be directly used, concentrated or used, concentrated or dried and then diluted.

The lactic acid bacteria may be any one or more lactic acid bacteria selected from the group consisting of Lactobacillus sp., Bifidobacterium sp., Leuconostoc sp., Pediococcus sp., Enterococcus sp. Lactic acid bacteria. Preferably, the lactic acid bacteria are selected from the group consisting of Lactobacillus paracasei, Lactobacillus capryi, Lactobacillus acidophilus, Bifidobacterium longbum, Bifidobacterium brevis, Leuconostomes teneroides, Pediococcus pentosaceus, It's an enterococcus pecalis.

The fermentation conditions are the same as those of general lactic acid bacteria fermentation, so that the fermentation conditions are not particularly limited, but they may be carried out at 25 to 40 DEG C for 24 to 96 hours.

The fermented product of the Lactobacillus canceller may be a fermented product containing lactic acid bacterium cells or a fermented product from which lactic acid bacterium cells have been removed. The fermented product from which the lactic acid bacterium cells have been removed may be sterilized to contain the dead cells, or may be filtrated or centrifuged by removing the cells through filtration or centrifugation. The improvement, treatment, or prevention effect of constipation according to the present invention can be achieved by a composition comprising a component of the Cassiae Extract, which is biologically converted into a component that is easy to use in vivo while the lactic acid bacteria multiply, Components are expected to be produced, and the effect obtained by adding the simple lactic acid bacteria itself is remarkably increased. The fermented product of the Lactobacillus canceller may be used as the fermented product itself or may be dried and pulverized.

As used herein, the term " comprising as an active ingredient " is meant to include an amount sufficient to achieve the improvement, treatment, or prevention efficacy or activity of the fermentation product of the deficiency lactic acid bacteria of the present invention.

The health functional food may be a product obtained by formulating the fermented product of the Lactobacillus lucidum in the form of a capsule, tablet, powder, granule, liquid, ring, flake, paste, syrup, gel, jelly or bar, It is produced in the form of general food added to food materials such as tea, spices, gum and confectionery. It means to bring about a certain effect in health when ingested. However, unlike general medicine, There is an advantage that there is no side effect that can occur when taking.

The health functional foods are very useful because they can be ingested routinely. The added amount of the fermented product of the lactic acid bacteria in the health functional food is not uniformly determined depending on the kind of the health functional food to which it is added but may be added within a range that does not deteriorate the original taste of the food, 0.01 to 50% by weight, preferably 0.1 to 20% by weight. In the case of health functional foods in the form of capsules, tablets, powders, granules, liquids, pellets, slices, pastes, syrups, gels, jellies or bars, the amount is usually 0.1 to 100% by weight, preferably 0.5 to 80% %.

The health functional food may contain, as an active ingredient, not only the fermented product of the Lactobacillus lactic acid bacteria but also a component that is ordinarily added at the time of food production, for example, a protein, a carbohydrate, a fat, a nutrient, a seasoning agent and a flavoring agent . Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings. For example, when the health functional food of the present invention is prepared from a drink and a beverage, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, various plant extracts and the like may be further added have.

In a pharmaceutical composition for the treatment or prevention of constipation, wherein the fermented product is a fermented product of a Lactobacillus, for example, 0.001 mg / kg or more, preferably 0.1 mg / kg or more, more preferably 10 mg / kg or more, more preferably 100 mg / kg or more, even more preferably 250 mg / kg or more, and most preferably 0.1 g / kg or more. Since the fermented product of lactic acid bacteria as a natural product has no adverse effect on the human body even when administered in an excessive amount, the quantitative upper limit of the fermented product of the lactic acid bacteria of the present invention can be selected by a person skilled in the art within a suitable range.

The pharmaceutical composition may be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned active ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, A flavoring agent and the like can be used.

The pharmaceutical composition may be formulated to contain at least one pharmaceutically acceptable carrier in addition to the above-described effective ingredients for administration.

The pharmaceutical form of the pharmaceutical composition may be a granule, a powder, a tablet, a coated tablet, a capsule, a suppository, a liquid, a syrup, a juice, a suspension, an emulsion, a drip agent or an injectable liquid agent. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.

Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile water and sterile water suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, And other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added as needed. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.

Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The pharmaceutical composition may be administered orally or parenterally. In the case of parenteral administration, the pharmaceutical composition may be administered by intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration or the like, preferably oral administration.

The appropriate dosage of the pharmaceutical composition will vary depending on factors such as the formulation method, the manner of administration, the age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and responsiveness of the patient, The physician can easily determine and prescribe dosages effective for the desired treatment or prophylaxis. According to a preferred embodiment, the daily dosage of the pharmaceutical composition is 0.001-10 g / kg.

The pharmaceutical composition may be prepared in a unit dose form by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by a person having ordinary skill in the art to which the present invention belongs. Into a capacity container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

Hereinafter, the present invention will be described more specifically with reference to the following examples. However, the following examples are provided only to aid understanding of the present invention, and the scope of the present invention is not limited to the following examples.

Manufacturing example  1: Heat number  Preparation of extract

Cassia tora L., purchased from Jeonnam Agricultural Products Agricultural Cooperative Association (Hwasun Gun), was crushed uniformly with a crusher after washing with water Cristobator powder with an average particle size of 300 ㎛ was prepared and used in refrigeration.

The distilled water, which was 10 times the weight of the crushed killer, was put into a reflux extractor and extracted twice at 100 ° C for 3 hours, followed by cooling and filtration, followed by concentration with a vacuum concentrator and lyophilization to obtain a scalded hot-water extract powder. The yield was 2.2%.

Manufacturing example  2: Preparation of Aqueous Solution Extract of Cereals

Fourfold 50% by weight aqueous ethanol solution of the crude crushed lysate of Preparation Example 1 was added thereto, and the mixture was extracted at room temperature for 3 days. The mixture was filtered, concentrated using a vacuum concentrator, and lyophilized to obtain an aqueous solution of the aqueous ethanol extract of the lucifer. The yield was 9.8%.

Manufacturing example  Three: Dilute  Produce

The water-diluted lysate was prepared by mixing water so that the crushed killer of Preparation Example 1 was 7% by weight.

Manufacturing example  4: Lactic acid bacteria Fermentation product of  Produce

The lactic acid bacteria of the Cassia tora extract, or Cassia tora water diluted solution of Production Example 3 diluted Cassia tora extract powder of Preparation Example 1 or 2 to 5% by weight in distilled water, sterilized 121 ℃, 1.5 atm, for 15 minutes, Table 1, each MRS broth and GAM broth for 24 hours at 37 ° C to activate the culture. Lactic acid bacteria were inoculated into the Cassiae Extract at a concentration of 5% by weight (1 × 10 ⁷ CFU / ml) And cultured at 150 rpm with stirring to obtain a culture solution.

The culture broth was centrifuged at 6,000 rpm for 15 minutes at 4 ° C, and the supernatant, from which the cells were removed, was concentrated and lyophilized to prepare a fermented product of the cultivar Lactobacillus.

division Strain name Access route Use badge L1 Lactobacillus kefiri MJ90 From Tibetan mushrooms
detach MRS L2 Lactobacillus paracasei Separated from aged Kimchi for 2 years MRS L3 Lactobacillus acidophillus 128 Korea Food Research Institute MRS L4 Lactobacillus plantarum 144 Korea Food Research Institute MRS B1 Bifidobacterium longum Korea Food Research Institute RCM  / GAM P1 Pediococcus pentosaceus Korea Food Research Institute MRS

Experimental Example  1: Lactic acid bacteria Fermented  Identification of fermentation characteristics

In order to confirm the fermentation characteristics of the fermented product of the Lactobacillus koreans in Preparation Example 4, the pH and viable cell count of the fermented product were checked at 36 and 72 hours after inoculation.

Lactobacillus spp., Pediococcus spp. Were plated on MRS agar, and Bifidobacterium spp. Was blended with 5% horse blood containing 5% horse blood. The viable cell count was determined by diluting 1 ml of the sample with 9 ml of sterile physiological saline, agar and anaerobic jar (BBL Gas Pak Anaerobic System). After incubation at 37 ° C for 24-48 hours, the number of colonies formed was counted, and the average number of colonies was multiplied by the dilution factor Colony per culture was calculated.

end. pH  change

The types of lactic acid bacteria and pH changes with time of culture are shown in Figs. 1A, 1B and 1C. Fig. 1 (a) is the extract of Preparation Example 1, Fig. 1 (b) is the extract of Preparation Example 2, and Fig. 1 (c) is the fermentation of the dilution of Production Example 3.

The initial pH was 6.4, decreased from 5.2 to 5.8 at 36 hours, and decreased from 4.9 to 5.5 at 72 hours.

Comparing changes in pH by lactic acid bacteria Lactobacillus kefiri MJ90 (L1) and Lactobacillus paracasei (L2) Lactobacillus acidophilus 128 (L3) , Lactobacillus plantarum 144 (L4), Bifidiobacterium longum (B1) , Pediococcus pentosaceus (P1).

When the lactic acid bacteria were the same, there was almost no difference in the pH of the cultured medium using the scalded hot water extract of Preparation Example 1, the Casser's ethanol extract of Preparation Example 2, The pH at the time and 72 hours was somewhat higher.

I. Viable cell count  change

Two kinds of lactic acid bacteria (L1 and L2) which had relatively high pH change among 6 kinds of lactic acid bacteria were re-preincubated and inoculated with 1% by weight of lactic acid bacteria in a 6% And the number of viable cells was measured after fermentation at 37 ° C for 24 hours. The results are shown in Table 2.

division Viable cell count ( CFU / ml ) Before fermentation After fermentation ( Production Example 1 ) After fermentation ( Production Example 2 ) After fermentation ( Production Example 3 ) L1 5.0 × 10 ⁵ 7.2 x 10 ⁸ 6.9 x 10 ⁸ 3.3 x 10 ⁸ L2 5.0 × 10 ⁵ 2.6 x 10 ⁸ 2.2 x 10 ⁸ 1.0 × 10 ⁸

As shown in the above Table 2, there was almost no difference between the use of the hydrothermal extract of Cercillator in Preparation Example 1 and the ethanol aqueous solution of Preparation Example 2 as the substrate, but in the case of using the dilution liquid of Cercillation Water of Preparation Example 3, Which was slightly less than half of the viable cell counts of Production Examples 1 and 2.

Lactobacillus It was confirmed that the number of viable cells of the culture prepared by using the kefiri MJ90 (L1) strain was three times higher than the number of viable cells of the culture prepared using the Lactobacillus paracasei (L2) strain.

Experimental Example  2: Confirmation of effect through animal experiment

end. Experimental animals and Experimental group

Male, 4-week-old SD (Sprague Dawley) rats were purchased from Multiscience (Daejeon) and used for breeding at a temperature of 20 ± 2 ° C and a humidity of 55 ± 5% for 12 hours. Experimental animals were purified for 1 week after purchase, and 5 mice per group were placed in the experimental group as shown in Table 3.

division Experimental group NOR Normal feed consumed in normal control LOP Negative controls were administered subcutaneously at 4 mg / kg for 5 days to induce constipation. CON1 Sennoside 50 mg / kg After one week oral administration,
Lipheramide 4 mg / kg subcutaneously for 5 days Positive control CON2 After oral administration of Dulcolax S 36.25 mg / kg per week,
Lipheramide 4 mg / kg subcutaneously for 5 days Positive control Comparative Example 1 After oral administration of 200 mg / kg of the water-extract of Cercariae of Preparation Example 1 for 1 week,
4 mg / kg subcutaneous administration of rofemide for 5 days Comparative Example 2 After 200 mg / kg of a water extract of Cercariae of Preparation Example 2 for 1 week orally,
4 mg / kg subcutaneous administration of rofemide for 5 days Example 1-1 Production Example 1 Fermentation product of strain L1 (Preparation Example 4) After oral administration of 100 mg / kg for 1 week,
4 mg / kg subcutaneous administration of rofemide for 5 days Examples 1-2 Production Example 1 Fermentation product of strain L1 (Preparation Example 4) After oral administration of 200 mg / kg for 1 week,
4 mg / kg subcutaneous administration of rofemide for 5 days Example 2-1 Production Example 2 Fermentation product of strain L1 (Preparation Example 4) After oral administration of 100 mg / kg for 1 week,
4 mg / kg subcutaneous administration of rofemide for 5 days Example 2-2 Production Example 2 Fermentation product of strain L1 (Preparation Example 4) After oral administration of 200 mg / kg for 1 week,
4 mg / kg subcutaneous administration of rofemide for 5 days Example 3-1 Production Example 3 After the oral administration of 100 mg / kg of the fermentation product of the strain L1 (Production Example 4) for 1 week,
4 mg / kg subcutaneous administration of rofemide for 5 days Example 3-2 Production Example 3 After the oral administration of 100 mg / kg of the fermentation product of the strain L1 (Production Example 4) for 1 week,
4 mg / kg subcutaneous administration of rofemide for 5 days Example 4-1 Production Example 2 Fermentation of strain L2 (Preparation Example 4) After oral administration of 100 mg / kg for 1 week,
4 mg / kg subcutaneous administration of rofemide for 5 days Example 4-2 Production Example 2 Fermentation product of strain L2 (Preparation Example 4) After oral administration of 200 mg / kg for 1 week,
4 mg / kg subcutaneous administration of rofemide for 5 days Example 5 Loperamide was subcutaneously administered at 4 mg / kg for 5 days,
Production Example 2 Fermentation product of strain L1 (Preparation Example 4) 200 mg / kg per 1 week oral administration

Each of the experimental groups except for the normal control (NOR), the negative control (LOP) and the Example 5 were orally administered the respective positive control drugs, the samples of the comparative examples and the examples for 5 days before the administration of the looperamide.

All but the negative control (LOP), the positive control (CON1, CON2) except for the normal control (NOR), and the Example 5 resulted in constipation by subcutaneous administration of rofelamide for 5 days, Lt; RTI ID = 0.0 > 6 < / RTI >

In Example 5, Fermented Lactobacillus Lactobacillus strain Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus (LOP) mg / kg After 5 days of oral administration, the last day of the addition of the final fermentation animals were sacrificed.

I. Weight, feed intake and Quantity of water

All experimental groups except for Example 5 were divided into two groups: from the start of ropelamide administration to the sacrifice of experimental animals, Example 5 shows the change in body weight (g) of experimental animals from the start of administration of ferulamide fermented product to sacrifice of experimental animals, Feed intake (g) and water volume (mL) are shown in Table 4.

division Weight change (g) Feed intake (g) Inhalation volume (mL) NOR 288 ± 11 14.1 ± 1.8 42.4 ± 5.6 LOP 290 ± 8 13.2 ± 1.9 39.2 ± 4.5 CON1 291 ± 9 13.8 ± 1.9 41.1 ± 5.8 CON2 289 ± 13 13.6 ± 1.6 39.5 ± 6.4 Comparative Example 1 288 ± 12 13.8 ± 1.8 38.4 ± 5.2 Comparative Example 2 290 ± 13 13.5 ± 1.7 40.1 ± 4.8 Example 1-1 286 ± 11 13.4 ± 1.7 39.2 ± 5.9 Examples 1-2 289 ± 10 14.1 ± 1.8 38.6 ± 4.8 Example 2-1 287 ± 13 13.2 ± 1.9 41.1 ± 4.1 Example 2-2 290 ± 6 14.2 ± 1.9 40.5 ± 5.5 Example 3-1 285 ± 12 13.5 ± 1.5 39.5 ± 5.8 Example 3-2 288 ± 10 13.8 ± 1.8 39.9 ± 5.4 Example 4-1 286 ± 14 14.2 ± 1.8 41.2 ± 5.8 Example 4-2 290 ± 10 14.1 ± 1.8 40.8 ± 6.1 Example 5 288 ± 9 13.9 ± 2.0 40.2 ± 5.8

There was no significant difference in feed intake and insulin intake during the experiment and no significant difference in body weight gain.

All. Number of sides and weight

In Example 5, the sides of the cage were removed from the cage for 4 days from the start of fermentation, and the rest were removed from the cage for 4 days from the start of ropelamide administration. The sides were collected for 1 day after the bed was changed, 2 and 3.

The number of stools was reduced to 23.1 ± 3.6 compared with the control group (NOR) of 47.2 ± 5.8, and the ropeferramide negative control group (LOP) was reduced to 23.1 ± 3.6, It was confirmed that the effect of increasing the number of sides was better than that of Dulcolax S (CON2).

In Comparative Examples 1 and 2, the water extract of the Cassiae and the aqueous solution of the aqueous ethanol solution of the categorizer showed little effect of increasing the number of sides, and in Examples 1-2 and 2-2 and 4 -2 showed the effect of increasing the number of sides equivalent to sennoside (CON1). In Example 3-2, the number of sides equivalent to Dulcolax S was increased although it was somewhat lower than that of senoside.

The effect of increasing the number of sides was increased in the dose-dependent manner in Examples 1-1, 1-2, 2-1, 2-2, 3-1, 3-2, 4-1 and 4-2, In Examples 2-1 and 2-2 in which fermentation was carried out, the effect of increasing the number of sides was higher, and also in the case of fermentation with the L1 strain in the same manner, in Examples 2-1 and 2-2 in which the aqueous solution of the aqueous ethanol solution of the cutter was fermented, 1 and 1-2, respectively.

In Example 5, in which the L1 fermented product of the aqueous solution of the aqueous ethanol solution of the lysimeter of Preparation Example 2 was administered after inducing constipation, the number of the stools was 26.2 ± 3.1, which was significantly increased again compared with the negative control (LOP).

(LOP) was reduced to 2.6 ± 0.32 compared with 4.8 ± 0.89 in the normal control group (NOR), and the induction of constipation by ropelamide was confirmed. There was no significant difference in the weight of the sides of Dulcolax S (CON2).

Comparative Example 1 showed no effect of increasing the weight of the stomach in the water-extract-treated group of Comparative Example 1, but increased the weight of the stomach in the group treated with the aqueous solution of the aqueous ethanol solution of the Criterion of Comparative Example 2.

As in Comparative Example 2, the weight of the stool was significantly increased in Examples 1-2 and 2-2 administered with 200 mg / kg, as compared with Comparative Example 2, and a significant difference was observed in Comparative Example 2 in Example 4-2 But the weight of the stool was significantly increased as compared with Comparative Example 1. In Example 3-2, the effect of weight on the sides equivalent to sennoside and Dulcolax S was shown.

The effect of increasing the weight of the stomach was increased in a dose-dependent manner in Examples 1-1, 1-2, 2-1, 2-2, 3-1, 3-2, 4-1 and 4-2, In Examples 2-1 and 2-2 in which the aqueous solution of the aqueous ethanol solution of the cutter was fermented, the effect of increasing the weight of the stool was more excellent than those in Examples 1-1 and 1-2, 2-1 and 2-2, the weight increase effect of the stool was higher than those of Examples 4-1 and 4-2 fermented using the strain L2.

In Example 5, in which the L1 fermented product of the aqueous solution of the aqueous ethanol solution of the Criterion of Preparation Example 2 was administered after inducing constipation, the weight of the stool was 3.18 ± 0.28, which was significantly increased again compared with the negative control (LOP).

la. Carmine  Coloring matter movement distance

The carmine pigment was transferred to saline at a concentration of 1.5% (w / v) in physiological saline. Oral administration of carmine dye was performed at 3 mL for 6 days. After 20 minutes of oral administration, FIG. 4 shows a photograph of the large intestine after ligating both the cecum to the rectum and washing the organs with PBS. The distance traveled by the Carmine pigment shown in FIG. 4 was measured and shown in FIG. .

The translocation distance of Carmine pigment was 13.4 ± 0.94 cm in the normal control group (NOR), and the negative control group (LOP) was decreased to 6.6 ± 0.75 cm in the ropeferramide administration, Side lengths (CON1) and Dulcolax es (CON2) increased significantly in the length of the bowel movement compared to the negative control (LOP).

In Comparative Example 1, there was no significant difference from the negative control group (LOP) in the long movement distance, but in the Comparative Example 2, the short movement distance was significantly increased as compared with the negative control group (LOP).

In Examples 3-1 and 2-1, although they were administered in ½ amount of Comparative Example 2, they showed a similar long-distance travel distance as Comparative Example 2, and in Examples 3-2 and 3-2 in which the same amount as in Comparative Examples 1 and 2 was administered, It was confirmed that the long travel distance of 2-2 was remarkably increased as compared with Comparative Example 2.

Examples 3-2 and 2-2 showed a field migration distance equal to or greater than that of the positive control selenoside and Dulcolax S, and in particular, Example 2-2 fermented with the L1 strain was equivalent to the normal control (NOR) And the distance of the longest movement.

Experimental Example  3: Before and after fermentation HPLC - IT / TOP MS  Pattern comparison

IT / TOP MS device (Shimadzu, Japan) at the same concentration as that of Preparation Example 3, which was obtained by fermenting the aqueous solution of the aqueous solution of the aqueous solution of the decanter of Preparation Example 2 and the aqueous solution of the aqueous solution of the aqueous ethanol solution of the decanter of Preparation Example 2, Were used to perform HPLC-IT / TOP MS to compare patterns. Acquity UPLC C-18, 1.7 m 2.1 x 100 mm column, with a gradient of 70% aqueous solution in a 10% aqueous solution of acetonitrile at a flow rate of 0.3 mL / min.

The pattern of the aqueous solution of the aqueous solution of the lysate of Preparation Example 2 is shown in Fig. 6a, and the pattern of the fermented product of the Lactobacillus of Preparation Example 3 is shown in Fig. 6b. Through the above pattern comparison, it was confirmed that a large number of peaks were lowered or disappeared through fermentation in the aqueous solution of the aqueous ethanol solution of the liquor of Production Example 2, and a new peak was produced instead.

Claims (10)

A health functional food for improving or preventing constipation, characterized by increasing at least one of the number of sides, the moisture content of the sides and the weight of the stomach with fermented product of the Lactobacillus fermentor obtained by fermenting the luciferase inoculum with lactic acid bacteria. delete [Claim 3] The health food according to claim 1, wherein the Cassiae Extract is an extract of water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The composition according to claim 1, wherein the lactic acid bacterium is any one or more lactic acid bacteria selected from the group consisting of Lactobacillus sp., Bifidobacterium sp., Leuconostoc sp., Pediococcus sp., And Enterococcus sp. Lactic acid bacteria. Health functional foods. The composition according to claim 1, wherein the health functional food is a capsule, tablet, powder, granule, liquid, ring, flaky, paste, syrup, gel, jelly or bar form. Health functional food for. A pharmaceutical composition for the treatment or prevention of constipation characterized by increasing at least one of the number of sides, the moisture content of the sides, and the weight of the stomach with fermented product of the Lactobacillus fermented product obtained by fermenting the Lactobacillus japonica extract with the lactic acid bacteria. delete [Claim 7] The pharmaceutical composition for treating or preventing constipation according to claim 6, wherein the Cassiae Extract is an extract of water, an alcohol having 1 to 4 carbon atoms or a mixed solvent thereof. The method according to claim 6, wherein the lactic acid bacterium is any one or more lactic acid bacteria selected from the group consisting of Lactobacillus genus, Bifidobacterium genus, Leuconostoc genus, Pediococcus genus, and Enterococcus genus. A pharmaceutical composition. Producing a Cercerium extract; And a step of inoculating lactic acid bacteria into the cuttlefish extract and fermenting the cuttlefish extract, wherein the number of sides, the moisture content of the sides, and the weight of the sides are increased.

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