KR20210131131A - Composition for healthy intestine comprising extract from young barley leaves improving enteric microbiota - Google Patents

Abstract

The present invention relates to a prebiotic composition capable of improving the growth of beneficial intestinal bacteria and inhibiting the growth of intestinal harmful bacteria, including a barley sprout or a fraction thereof, and a method for promoting intestinal health comprising a step of administering the composition. The prebiotic composition provided in the present invention promotes the growth of beneficial intestinal bacteria contained in the intestinal flora and exhibits an effect of inhibiting the growth of harmful bacteria in the intestine, thereby being able to be used to improve the intestinal flora and promote intestinal health.

Description

Composition for healthy intestine comprising extract from young barley leaves improving enteric microbiota

The present invention relates to a composition for improving intestinal health according to the improvement of the intestinal flora of the barley sprout extract, and more specifically, the present invention includes a barley sprout extract or a fraction thereof to promote the growth of beneficial intestinal bacteria and inhibit the growth of harmful bacteria in the intestine It relates to a prebiotic composition that can be administered and a method for improving intestinal health comprising administering the composition.

Numerous microorganisms live in a community in the human gastrointestinal track (GI), and such a group is called enteric microbiota. They are essential for normal gastrointestinal physiology and function as they play important roles in immune function, digestion and metabolism while constituting the ecosystem within the intestinal tract. The human gut flora is quite resistant to changes in the environment, but factors such as antibiotics, stress, diet, or certain diseases cause changes in the gut flora. This change in the composition and function of the intestinal flora is called dysbiosis, and the most common definition refers to an imbalance between beneficial bacteria such as Bifidobacteria and Lactobacillus and harmful bacteria such as Escherichia coli. This imbalance is known to be directly or indirectly related to various diseases such as autoimmune disease, inflammatory bowel disease, and obesity. In general, it is believed that it is necessary to maintain the intestinal flora in a state where there are many beneficial bacteria and few harmful bacteria in order to maintain health. The enzymes and metabolites produced are known to be involved in the metabolism of various substances in the human body, and to affect the absorption of nutrients or drugs and the generation of toxic substances.

Probiotics are living microorganisms that can work beneficially by maintaining the balance of the human intestinal flora. The probiotics inhibit the growth of harmful bacteria through a decrease in intestinal pH by production of short chain fatty acids (SCFA) and competition for intestinal nutrients and barrier attachment sites. It is also known to have beneficial functions in the human body, such as immune activation and alleviation of lactose intolerance. Representative microorganisms used as probiotics include bifidobacteria and bacteria belonging to lactic acid bacteria (LAB). Lactic acid bacteria are bacteria that produce lactic acid by fermenting carbohydrates, and include Lactobacillus sp. strains, Streptococcus sp. strains, Leukonostok sp. strains, and the like. On the other hand, bifidobacteria are bacteria that use various oligosaccharides as energy sources, and in the process of converting hexoses into lactic acid or acetic acid, an enzyme called fructose-6-phosphate phosphoketolase acts, and fermentation It has a unique characteristic that it does not generate gas in the process.

The main ingredient used to more effectively proliferate and maintain the probiotics in the intestine is called prebiotics. The prebiotics are not digested by humans, but are selectively fermented by intestinal microbes to control their growth and metabolic changes, and thus may be defined as substances that help the health of the host. As a representative example of the prebiotics, fructo-oligosaccharides (FOS) are known. The fructooligosaccharide is an oligosaccharide in a form in which fructose molecules are combined with sugar by β-1, 2, and since fermentation is possible by Lactobacillus and Bifidobacteria, ingestion of FOS increases Bifidobacterium and Lactobacillus species. It is known that it can be done. However, the FOS can have a beneficial effect on humans by increasing intestinal probiotic strains, but there is a known problem that adverse effects may occur by harmful bacteria and non-probiotics capable of metabolizing intestinal FOS. , there is a demand for the development of a prebiotic composition that can more effectively proliferate and maintain probiotics, and research for developing it is being actively conducted. For example, Korean Patent No. 10-1973514 discloses prebiotics containing grape seed powder.

Under this background, the present inventors have made intensive research efforts to develop a prebiotic composition that can more effectively propagate and maintain probiotics. As a result, it is confirmed that barley sprout extract can be used as an active ingredient of the prebiotic composition, and the present invention was completed.

The main object of the present invention is to provide a prebiotic composition comprising a barley sprout extract or a fraction thereof.

Another object of the present invention is to provide a method for improving intestinal health comprising administering the composition.

One embodiment of the present invention for achieving the above object provides a prebiotic composition comprising a barley sprout extract or a fraction thereof.

As used herein, the term "sprout barley" refers to a young plant that sprouts from barley and grows to about 15 cm. Typically, the sprouted barley can be obtained by immersing barley in water for about a day, removing moisture, sown under conditions that prevent moisture loss, germinate, and then grow for 7 to 10 days.

As used herein, the term "extract" refers to a liquid component obtained by immersing a target substance in various solvents and then extracting it for a certain time at room temperature or a heated state, and a solid obtained by removing the solvent from the liquid component. it means. In addition, in addition to the result, it can be comprehensively interpreted as including all of the diluent of the result, the concentrate thereof, the prepared product thereof, the purified product, and the like.

In the present invention, the extract means a barley sprout extract. The sprouted barley extract can be obtained by extracting the whole plant of the sprouted barley, its dried products, processed products, etc., alone or in combination, with water or various organic solvents. At this time, the organic solvent used is not particularly limited as long as an extract containing rutonarin or saponarin, which exhibits an effect of inhibiting differentiation into adipocytes, can be obtained, but is preferably water, a polar solvent or a non-polar solvent. More preferably, water, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol or butanol, etc.), a mixed solvent thereof, etc. may be used, and most preferably, ethanol or a mixed solvent thereof may be used. have. In addition, the method for obtaining the extract is also not particularly limited as long as an extract containing rutonarin or saponarin, which exhibits an effect of inhibiting differentiation into adipocytes, can be obtained, but is preferably a plant of the sprouted barley. Cold extraction method in which the whole, its dried product, processed product, etc. are immersed in the solvent and extracted at room temperature of 10 to 25 ° C., heat extraction method in which extraction is performed by heating to 40 to 100 ° C. Methods such as reflux extraction can be used.

As used herein, the term "fraction" refers to a result obtained by a fractionation method for separating a specific component or a specific group from a mixture containing various components.

In the present invention, the fraction can be interpreted as a fraction containing rutonarin or saponarin, obtained by applying the sprout barley extract to various fractionation methods. The fraction of the extract can be obtained by applying the extract to various fractionation methods, the fractionation method is not particularly limited thereto, but a solvent fractionation method performed by treating various solvents, an ultrafiltration membrane having a constant molecular weight cut-off value It may be an ultrafiltration fractionation method performed by passing through a chromatographic fractionation method, and a chromatographic fractionation method performing various chromatography (prepared for separation according to size, charge, hydrophobicity or affinity). In particular, the solvent used in the solvent fractionation method is not particularly limited thereto, but a polar solvent or a non-polar solvent may be used, and preferably a non-polar solvent may be used. The solvent fractionation method may be performed in a manner of sequentially fractionating the extract from a solvent with a high level of non-polarity to a solvent with a low level. For example, a method of sequentially fractionating the extract using hexane or ethyl acetate may be used. can

The term "prebiotics" of the present invention is used in the same meaning as a prebiotic composition, and is mainly used by useful microorganisms in the large intestine to promote the growth or activity of microorganisms, thereby exhibiting a beneficial effect on health. It means food ingredients from Mars.

In the present invention, the prebiotic composition, including barley sprout extract or a fraction thereof, can be interpreted as exhibiting an effect of promoting the growth of beneficial intestinal bacteria and inhibiting the growth of harmful intestinal bacteria.

As used herein, the term "intestinal beneficial bacteria" refers to beneficial bacteria that previously inhabit the host's intestine and strains that have a beneficial effect on the intestinal environment when ingested and reach the intestine. These are bacteria that survive in gastric and bile acids, reach the small intestine, multiply and settle in the intestine, exhibit useful effects in the intestine, and satisfy the non-toxic and non-pathogenic conditions. That is, it may be understood as the same meaning as probiotics.

In the present invention, the beneficial intestinal bacteria are not particularly limited thereto, but as an example, Lactobacillus sp., Leuconostoc sp., Lactococcus sp., Bifidobacter It may be a Bifidobacterium sp., Enterococcus sp., or Streptococcus sp. strain, and as another example, it may be a Bifidobacterium sp. or Lactobacillus sp. strain.

As used herein, the term "intestinal harmful bacteria" refers to harmful bacteria existing in the intestine of the host and strains that are harmful to the intestinal environment when ingested and reach the intestine.

In the present invention, the intestinal harmful bacteria are not particularly limited thereto, but as an example, Pseudomonas-aeruginosa sp., Vibrio sp., Staphylococcus sp., Clostridium sp., Eubacterium sp., Escherichia sp. or Bacteroides sp. may be a strain, as another example, Clostridium It may be a genus, Staphylococcus genus, or Escherichia genus strain, and as another example, Clostridium perfringens ) It may be a strain or E. coli ( E. coli ).

Sprout barley included as an active ingredient in the prebiotic composition provided in the present invention has been used as a food resource for a long time, and its extract has been used for a long time to prove its safety, so the prebiotic composition is in the form of a pharmaceutical composition or food composition It can be prepared, and for this purpose, it may further include an appropriate carrier, excipient or diluent commonly used in the preparation of pharmaceutical compositions or food compositions.

First, the pharmaceutical composition may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., oral dosage forms, external preparations, suppositories, and sterile injection solutions according to conventional methods, respectively. In the present invention, carriers, excipients and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, it is prepared using diluents or excipients, such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the extract and its fractions, for example, starch, calcium carbonate, It is prepared by mixing sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, solutions, emulsions, syrups, etc., which are commonly used simple diluents such as water and liquid paraffin, as well as various excipients, such as wetting agents, sweeteners, fragrances, and preservatives. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used.

The content of the extract or its fraction of sprouted barley contained in the pharmaceutical composition is not particularly limited thereto, but may be included in an amount of 0.0001 to 50% by weight, more preferably 0.01 to 20% by weight, based on the total weight of the final composition. .

Next, the food composition may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. . In addition, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu). In addition, it may include amino acids such as lysine, tryptophan, cysteine, and valine. In addition, preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder and high bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxytoluene ( BHT), etc.), colorant (tar pigment, etc.), color developer (sodium nitrite, sodium nitrite, etc.), bleach (sodium sulfite), seasoning (MSG sodium glutamate, etc.), sweetener (dulcin, cyclmate, saccharin, sodium, etc.) Food additives such as flavorings (vanillin, lactones, etc.), expanding agents (alum, D-potassium hydrogen tartrate, etc.), strengthening agents, emulsifiers, thickeners (flavors), film agents, gum base agents, foam suppressants, solvents, and improving agents ) can be added. The additive is selected according to the type of food and used in an appropriate amount.

The content of the extract or its fraction of sprouted barley contained in the food composition is not particularly limited thereto, but may be included 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, In particular, when the food is a beverage, it may be included in a ratio of 1 to 30 g, preferably 3 to 20 g, based on 100 ml.

Another embodiment of the present invention provides a method for improving intestinal health of an individual comprising administering the prebiotic composition.

As described above, a prebiotic composition comprising a barley sprout extract or a fraction thereof may exhibit an effect of promoting the growth of beneficial intestinal bacteria and inhibiting the growth of harmful bacteria in the intestine, so that the prebiotic composition is administered to an individual In this case, it is possible to enhance the intestinal health of the subject.

In the present invention, the term "individual" includes, without limitation, mammals including mice, livestock, humans, etc., which are intended to promote intestinal health.

In the method of promoting intestinal health of the present invention, the administration route of the prebiotic composition may be administered through any general route. The prebiotic composition of the present invention is not particularly limited thereto, but may be administered intraperitoneally, intravenously, intramuscularly, subcutaneously, intradermally, oral administration, intranasal administration, intrapulmonary administration, or rectal administration as desired. have. However, during oral administration, since the extract or its fractions of sprouted barley may be denatured by gastric acid, the oral composition should be formulated to coat the active agent or to protect it from degradation in the stomach. In addition, the composition may be administered by any device capable of transporting the active agent to a target cell.

The prebiotic composition provided in the present invention promotes the growth of beneficial intestinal bacteria contained in the intestinal flora and exhibits the effect of inhibiting the growth of harmful intestinal bacteria, so it can be utilized to improve the intestinal flora and promote intestinal health .

1 is a graph showing the results of measuring changes in the activity level of β-glucosidase derived from intestinal microorganisms according to the treatment concentration of barley sprout extract.

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

Example 1: Preparation of sprout barley extract

The sprouted barley harvested in about 7-10 days after sowing the large grain barley No. 1 was dried and powdered. 80% fermented alcohol was added to the powder, extracted with stirring at room temperature for about 24 hours, and the solvent was concentrated with a concentrator and then freeze-dried to prepare a sprouted barley extract.

Example 2: Effect of improving intestinal flora of barley sprout extract

Example 2-1: Measurement of β-glucosidase activity of intestinal microbial culture

β-glucosidase secreted from intestinal microbes decomposes glycosides to separate sugars and non-glycosides. It is known that when glycosides are decomposed, reactivity increases and absorption in the body increases. In particular, it is known that cyanide glycosides such as amygdalin, coumarin glycosides, and phenolic glycosides are released after decomposition to have side effects in the living body. The purpose of this study was to examine the effect of barley sprout extract on the activity of β-glucosidase derived from intestinal microorganisms.

First, barley sprout extract was added in various amounts to 5 mL of GAM (General Anaerobic Medium) medium, and then the pH of the medium was adjusted to 7.2 and sterilized.

Then, fresh feces of healthy Koreans were diluted 10-fold with an anaerobic medium, and then 5 μL of the sterilized medium was treated, and the sample was obtained by culturing at 37° C. in an anaerobic incubator.

100 μL of the obtained sample, 300 μL of 0.1 M phosphate buffer (pH 6.0) and 200 μL of 2 mM PNP β-D-glucoside were mixed and reacted at 37°C. 400 μL of 0.5 N NaOH was added to the reaction mixture to terminate the reaction, 1 mL of distilled water was added, and then centrifuged (3000 rpm, 20 minutes) to obtain a supernatant. The absorbance (optical density) of the obtained supernatant was measured at 405 nm, and the content of the free PNP was calculated using this, and this was used as the activity level of β-glucosidase (FIG. 1). At this time, it was determined that the higher the content of the free PNP, the higher the activity level of β-glucosidase.

1 is a graph showing the results of measuring changes in the activity level of β-glucosidase derived from intestinal microorganisms according to the treatment concentration of barley sprout extract.

As shown in FIG. 1 , as the treatment concentration of the barley sprout extract increased, the content of free PNP decreased, confirming that the activity level of β-glucosidase was lowered.

Therefore, it was found that the barley sprout extract can improve the intestinal environment in a concentration-dependent manner.

Example 2-2: Analysis of the effect of barley sprout extract on microorganisms in the intestinal flora

To analyze the effect of barley sprout extract on the proliferation of beneficial intestinal bacteria (Bifidobacterium sp. strain and Lactobacillus sp. strain) and intestinal harmful bacteria (Clostridium sp. strain, E. coli and Staphylococcus sp. strain) contained in the intestinal flora did.

First, EG liquid medium (Beef extract 2.0 g, Proteose peptone No.3 10.0 g, Yeast extrat 5.0 g, Na2HPO4 4.0 g, Soluble starch 0.5 g, Glucose 1.5 g, L-cystein 0.4 g, Silicon antifoamer 0.25 mL, Tween 80 0.5 g, 1000 mL of distilled water) was added to 5 mL of barley sprout extract in an amount of 1.0% (w/v), and then sterilized under anaerobic conditions.

Next, fresh feces of healthy Koreans are diluted (100-1000 times) with an anaerobic medium, and then added to the sterilized medium at a content of 2.5% (v/v), and then in an anaerobic incubator (Gas- pak jar) at 37° C. for 60 hours.

After the culture is completed, the culture solution is sequentially diluted 10 times up to 10^7 times to obtain a culture diluent, and 50 μL of the culture diluent is plated on each agar plate medium suitable for intestinal microbiota, Incubated at 37°C. At this time, in order to culture the Bifidobacterium sp. strain, it is cultured in anaerobic conditions using BS agar medium, and in order to culture the Lactobacillus sp. strain, MRS (Man-Rogosa-Sharpe) agar medium is used in anaerobic conditions. In order to culture the Clostridium sp. strain, it was cultured in anaerobic conditions using SPS (Sulfite Polymyxin Sulfadiazine) agar medium, and to culture Clostridium perfringens strain, it was cultured in anaerobic conditions using Differential Reinforced Clostridial agar medium. In order to culture E. coli, McConkey agar medium was used for culturing under aerobic conditions, and for culturing Staphylococcus sp. strains, it was cultured under aerobic conditions using Mannitol salt agar medium. In addition, as a control, a sample cultured in a medium not containing sprouted barley extract was used.

After the culture was completed, the proliferation level of the strain was compared by measuring the number of colonies of the bacteria under a microscope (Table 1).

Change in the culture medium by barley sprout extract (log CFU=colony forming unit)/mL intestinal flora strain control Barley sprout extract (1.0%) Bifidobacterium genus strain
Lactobacillus genus strain
Clostridium genus strain
Clostridium perfringens strain
coli
Staphylococcus genus strain 8.5
6.1
8.6
5.1
7.4
3.4 9.3
7.4
8.5
4.4
6.4
3.3

As shown in Table 1 above, in the case of beneficial intestinal bacteria (Bifidobacterium sp. strain and Lactobacillus sp. strain) included in the intestinal flora, the proliferation level of each strain was increased when the barley sprout extract was treated, and In the case of some intestinal harmful bacteria (Clostridium sp. strain and Staphylococcus sp. strain) included, there was no change according to the treatment of sprouted barley extract, and some of the intestinal harmful bacteria (Clostridium perfringens strain and E. In this case, it was confirmed that the proliferation level of each strain was reduced when treated with sprouted barley extract.

As such, it can be seen that the barley sprout extract promotes the growth of beneficial intestinal bacteria contained in the intestinal flora and exhibits the effect of inhibiting the growth of harmful bacteria in the intestine, so it can be used as an active ingredient in a prebiotic composition that can improve the intestinal flora. could

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention, rather than the above detailed description, all changes or modifications derived from the meaning and scope of the claims described below and their equivalents.

Claims (7)

A prebiotic composition comprising a barley sprout extract or a fraction thereof.
According to claim 1,
The extract is a composition obtained by extracting sprouted barley with a solvent selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, and mixed solvents thereof.
According to claim 1,
The fraction is a fraction obtained by subjecting the sprouted barley extract to a method selected from the group consisting of solvent fractionation, ultrafiltration fractionation, chromatography fractionation, and combinations thereof.
According to claim 1,
The prebiotic composition promotes the growth of beneficial intestinal bacteria included in the intestinal flora, and exhibits an effect of inhibiting the growth of intestinal harmful bacteria.
5. The method of claim 4,
The intestinal beneficial bacteria are Lactobacillus sp., Leuconostoc sp., Lactococcus sp., Bifidobacterium sp., Enterococcus sp. ) or Streptococcus genus ( Streptococcus sp.) will be a strain, the composition.
5. The method of claim 4,
The intestinal harmful bacteria are Pseudomonas-aeruginosa sp., Vibrio sp., Staphylococcus sp., Clostridium sp., Eubacterium genus ( Eubacterium sp.) Escherichia genus ( Escherichia sp.) or bacteroid genus ( Bacteroides sp.) strain, the composition.
A method for promoting intestinal health of an individual comprising administering the prebiotic composition of any one of claims 1 to 6 to an individual other than a human.

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