KR20200125206A - Composition for improvementing, preventing or treating inflammatory bowel disease or leaky gut syndrome comprising fractions or extract of pepper leave - Google Patents

Abstract

The present invention relates to a composition for ameliorating, preventing or treating inflammatory bowel disease or leaky gut syndrome. The present invention, more specifically, relates to a composition for ameliorating, preventing or treating inflammatory bowel disease or leaky gut syndrome, containing a red pepper leaf extract as an active ingredient.

Description

Composition for improving, preventing or treating inflammatory bowel disease or leaky gut syndrome comprising fractions or extract of pepper leave}

The present invention relates to a composition for improving, preventing or treating inflammatory bowel disease or leaky gut syndrome, and by containing red pepper leaf extract as an active ingredient, it is useful for functional foods and drugs for the prevention, improvement or treatment of inflammatory bowel disease or leaky gut syndrome. Can be used.

In addition to the basic functions of digestion, absorption, and excretion of food, the intestinal tract of the human body performs an immunological function as a barrier to the intestinal mucosa from entering the bloodstream of microorganisms, their by-products, antigens, and toxins. That is, the merit film is a barrier that blocks foreign substances and has a dual function of passing and absorbing them.

Intestinal health is very important because if there is a problem with these intestinal functions, not only does it cause problems in the supply of nutrients to the body itself, and symptoms such as repetitive constipation, diarrhea, and abdominal pain greatly interfere with daily life.

Inflammatory bowel disease is a disease in which the intestine is chronically inflamed, and the cause of the disease and the cure method are not yet known. In general, it refers to Crohn's disease and ulcerative colitis. Crohn's disease and ulcerative colitis may show symptoms such as diarrhea, abdominal pain, nausea, fever, loss of appetite, weight loss, and fatigue. Crohn's disease occurs mainly in the small and large intestine, but ulcerative colitis is invented only in the large intestine. In addition to this, long-type Behcet's disease, which is relatively common in Korea, also belongs to this.

Drugs used to treat such inflammatory bowel disease include steroidal immunosuppressants, 5-aminosalicylic acid (5-ASA) drugs that block the production of prostaglandins (e.g., sulfasalazine), Mesalazine, etc., are used, and they cause serious side effects such as fullness, headache, rash, liver disease, leukopenia, agranulocytosis, and male infertility, as well as having insignificant therapeutic effect on inflammatory bowel disease. Use is restricted.

Therefore, if it is possible to develop a therapeutic agent for inflammatory bowel disease that does not cause side effects, it is expected that it will be possible to treat patients with inflammatory bowel disease more safely and effectively. However, a treatment without side effects has not yet been developed.

Under this background, the present inventors have made diligent efforts to develop a therapeutic agent for inflammatory bowel disease derived from natural products that do not cause side effects, and as a result, a composition containing red pepper leaf extract is effective in the treatment of not only inflammatory bowel disease but also intestinal leak syndrome, The present invention was completed by confirming that it enhances the intestinal immune function.

Korean Patent Application Publication No. 10-2012-0067405

It is an object of the present invention to provide a food composition for improving or preventing inflammatory bowel disease or leaky intestinal syndrome using red pepper leaf extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of inflammatory bowel disease or leaky intestinal syndrome, using red pepper leaf extract as an active ingredient.

In order to achieve the above object, the present invention provides a food composition for improving or preventing inflammatory bowel disease or leaky intestinal syndrome, comprising red pepper leaf extract as an active ingredient.

The red pepper leaf extract may be extracted with water, alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The composition may increase immune activity, suppress an inflammatory response, and reduce activation of inflammatory cells.

The composition may be one that enhances IgA expression and inhibits or reduces the expression of IL-6 and LTB ₄ .

The inflammatory bowel disease may be a disease selected from the group consisting of Crohn's disease, ulcerative colitis, bowel Behcet's disease, infectious enteritis, ischemic bowel disease, and radiation enteritis.

The leaky gut syndrome may be induced by a high fat diet.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating inflammatory bowel disease or leaky intestinal syndrome, comprising red pepper leaf extract as an active ingredient.

The red pepper leaf extract may be extracted with water, alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The composition may increase immune activity, suppress an inflammatory response, and reduce activation of inflammatory cells.

The composition may be one that enhances IgA expression and inhibits or reduces the expression of IL-6 and LTB ₄ .

The inflammatory bowel disease may be a disease selected from the group consisting of Crohn's disease, ulcerative colitis, bowel Behcet's disease, infectious enteritis, ischemic bowel disease, and radiation enteritis.

The leaky gut syndrome may be induced by a high fat diet.

The composition of the present invention includes a red pepper leaf extract, thereby suppressing inflammation and activating immunity to improve, prevent or treat inflammatory bowel disease. In addition, the composition of the present invention has the effect of improving, preventing or treating intestinal leak syndrome by enhancing intestinal permeability.

1 is a graph showing the measurement of the concentration of IgA in serum for the normal feed group (NC), the positive control group (GC), the first experimental group (WEPL50), and the second experimental group (WEPL100).
2 is a measurement of the expression level of cytokine IL-6 in serum for the normal feed group (NC), the high fat diet group (HF), the positive control group (GC), the first experimental group (WEPL50), and the second experimental group (WEPL100). This is the graph shown.
Figure 3 is a normal feed group (NC), high fat diet group (HF), positive control group (GC), the first experimental group (WEPL50) and the second experimental group (WEPL100) in serum cytokine leukotriene B4 (LTB ₄ ) expression amount It is a graph showing by measuring.
Figure 4 shows the detection concentration of FITC-dextran in the normal feed group (NC), the high fat diet group (HFD), the positive control group (GC), the first experimental group (WEPL50) and the second experimental group (WEPL100) over time. It is a graph.
5 is a graph showing the area under the curve of the result measured from FIG. 4.

The present invention relates to a composition for improving, preventing or treating inflammatory bowel disease or leaky intestinal syndrome, using red pepper leaf extract as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention relates to a composition for improving, preventing or treating inflammatory bowel disease or leaky intestinal syndrome, using red pepper leaf extract as an active ingredient.

Pepper ( Capsicum annuum ) is a tropical plant, classified as an annual plant belonging to the Solanaceae family , and is one of the most consumed vegetables worldwide. In Korea, it is cultivated mainly from late spring to summer, and has been used as a food ingredient for a long time as the main ingredient of kimchi. Currently, there are about 20 to 30 varieties of pepper known worldwide, of which Capsicum annuum, Capsicum frutescens, Capsicum chinense, Capsicum baccatum, and Capsicum pubescens are the most cultivated. Pepper is not only rich in vitamins and minerals, but it has been reported that it contains a large amount of phenolic compounds such as carotenoid, quercetin, luteolin and capsaicinoids, but studies on pepper leaves are insufficient.

In the present invention,'leaky gut syndrome' or'leaky gut syndrome'; LGS' refers to a phenomenon in which intestinal membrane cells maintain a certain gap between cells, and during the process of digestion and absorption, any stimulation or damage is applied, increasing the permeability of the intestinal membrane through which a polymer material can reciprocate into the gap between cells. It collectively refers to symptoms caused by a phenomenon in which a polymer material in the blood leaks into the intestinal lumen due to improper functioning of the intestinal wall or a phenomenon in which a polymer material in the lumen directly enters the blood (leaky gut). These symptoms appear in various clinical conditions such as aging, allergy, multiple trauma, rheumatoid arthritis, inflammatory bowel disease, chronic fatigue syndrome, and irritable syndrome. In addition, pathogens, antigens, and decay substances are introduced into the intestinal mucosa due to an increase in permeability of the intestinal mucosa or damage to the intestinal mucosa. It causes various inflammatory reactions and immune reactions.

Such leaky gut syndrome complains of various, extensive and ambiguous symptoms without specific symptoms. Specifically, it has a peculiarity in that it has symptoms that complain in common in various diseases.

The exact cause of the leaky gut syndrome has not been identified, but the use of non-steroidal antiin ammatory drugs (NSAIDs) used as pain relievers for a long time, antibiotics and steroids, anticancer chemistry as an anticancer therapy In the case of therapy or radiation therapy, when a change in the composition of the normal intestinal flora occurs, the growth of fungi in the intestine, ingestion of excessive food, ingestion of spoiled food, ingestion of heavy metals or toxic substances, excessively irritating food Ingestion of or hypersensitivity to certain foods, excessive use of alcohol, multiple trauma, exposure to acute chronic mental stress, chronic intestinal bacteria, parasites, and yeast infections can be cited.

Specifically, the composition for improving, preventing, or treating leaky gut syndrome may be referred to as a composition for improving, preventing, alleviating, suppressing and treating that the permeability of the intestine is increased due to the various causes. This has the effect of improving, preventing, and treating intestinal leak syndrome, which causes various diseases, as the intestinal permeability increases, so that polymeric substances in the blood leak into the intestinal lumen, or polymeric substances in the lumen enter the branch blood. The improvement, prevention, or treatment of leaky gut syndrome may mean suppressing a phenomenon in which the intestinal membrane cell gap is loosened and thus the permeability of the intestinal membrane through which a polymer material can reciprocate is increased.

The inflammatory bowel disease may be a disease selected from the group consisting of Crohn's disease, ulcerative colitis, bowel Behcet's disease, infectious enteritis, ischemic bowel disease, and radiation enteritis.

According to the experimental results of the present invention, it can be seen that when a high-fat diet is administered for a long time, inflammatory factors such as cytokine IL-6 are increased and activation of inflammatory cells is increased. This induces inflammation in the intestine over time, and soon enteritis is induced.

At this time, it can be seen that the combined administration of the red pepper leaf extract increases the immune activity, suppresses the inflammatory response, and reduces the activation of inflammatory cells. Therefore, it can be seen that the composition comprising the red pepper leaf extract according to the present invention as an active ingredient has an effect of improving, preventing or treating inflammatory bowel disease.

The red pepper leaf extract is more specifically characterized in that it enhances IgA expression and inhibits or reduces the expression of IL-6 and LTB ₄ .

In the present invention, the'extract' includes not only the crude extract obtained by treating the extraction raw material with the extraction solvent, but also the processed product of the crude extract. For example, the red pepper leaf extract may be prepared in a powder state by additional processes such as distillation under reduced pressure and freeze drying or spray drying.

In addition, the'extract' in the present invention also includes a fraction obtained by further fractionating the crude extract. That is, the red pepper leaf extract includes not only those obtained using an extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, a polymer fraction obtained by passing through an ultrafiltration membrane having a certain molecular weight cut-off value, separation by various chromatography (made for separation according to size, charge, hydrophobicity or affinity), etc. It can be purified through a method.

The polymer fraction may be a fraction obtained by separating only a polymer material having a molecular weight of 10 kDa or more after obtaining a red pepper leaf extract or a red pepper leaf extract using an extraction solvent.

In addition, in the present invention, the'pepper leaf extract' is an extract of the red pepper leaf using water, alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, and the extraction method does not need to be particularly limited. Preferably, the red pepper leaf extract may be a hot water extract of red pepper leaf or an ethanol solution of red pepper leaf, and may be, for example, a red pepper leaf alcohol extract. Most preferably, it may be a hot water extract of red pepper leaves.

In addition, the red pepper leaf extract may be processed by the red pepper leaf extract, and then repeatedly extracted one or more times with water, alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The red pepper leaves may be cut into a certain size, dried using a shade or a dryer, and then crushed or used as it is. The drying period is not particularly limited as long as the moisture content in the red pepper leaves is less than 20%.

Compared to other parts of pepper, using pepper leaves enhances IgA expression when administered on a high-fat diet for a long period of time, inhibits or reduces IL-6 expression and LTB ₄ expression by more than 1.5 to 2.5 times, and has intestinal permeability. Since it is also inhibited by 2 to 5 times or more, red pepper leaf extract is remarkably excellent in preventing, improving, or treating inflammatory bowel disease as well as intestinal leak syndrome compared to other parts of pepper.

In the present invention, "containing as an active ingredient" or "including as an active ingredient" means including an amount sufficient to improve, prevent or treat inflammatory bowel disease or leaky intestinal syndrome of the red pepper leaf extract of the present invention. Since the red pepper leaf extract is a natural product and does not have side effects on the human body even if it is administered in an excessive amount, the upper and lower limits of the quantity of the red pepper leaf extract included in the composition of the present invention can be selected and carried out by a person skilled in the art within an appropriate range.

In the present invention, when the composition containing the red pepper leaf extract as an active ingredient is treated, the composition promotes IgA expression when administered on a high fat diet for a long period of time, and inhibits or reduces the expression of IL-6 and LTB ₄ , It was confirmed that there is an effect of preventing, improving or treating inflammatory bowel disease, and it was confirmed that it has an effect of inhibiting and reducing intestinal permeability.

The composition of the present invention may be variously used as a food composition for improving and preventing inflammatory bowel disease or leaky gut syndrome using the red pepper leaf extract of the present invention as an active ingredient.

The food composition according to the present invention may be formulated in the same manner as the pharmaceutical composition described below and used as a functional food or may be added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gum, ice cream, vitamin complexes, health supplements. Etc.

The food composition of the present invention may include not only red pepper leaf extract as an active ingredient, but also ingredients that are commonly added during food production, and include, for example, proteins, carbohydrates, fats, nutrients, flavoring agents, and flavoring agents. Examples of the aforementioned carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is made of drinks and beverages, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts, in addition to the red pepper leaf extract of the present invention, may be further included.

The present invention provides a health functional food comprising a food composition for improving, suppressing or preventing inflammatory bowel disease or leaky gut syndrome comprising the red pepper leaf extract as an active ingredient. Health functional foods are foods prepared by adding red pepper leaf extract to food materials such as beverages, teas, spices, chewing gums, confectionery, or encapsulating, powdering, or suspending them. However, unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long time by using food as a raw material. The health functional food of the present invention obtained in this way is very useful because it can be consumed on a daily basis. The amount of red pepper leaf extract added in such health functional foods cannot be uniformly regulated depending on the type of health functional food to be targeted, but can be added within the range that does not damage the original taste of the food. To 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of health functional foods in the form of pills, granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.

In another example, as described above, it may be used as a pharmaceutical composition for preventing or treating inflammatory bowel disease or leaky intestinal syndrome using red pepper leaf extract as an active ingredient. In addition to the red pepper leaf extract, which is an active ingredient, it can be prepared using a pharmaceutically suitable and physiologically acceptable auxiliary agent, and the auxiliary agent includes excipients, disintegrants, sweeteners, binders, coating agents, expanding agents, lubricants, lubricants or flavoring agents. Etc. can be used.

For administration, the pharmaceutical composition may include one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients, and may be preferably formulated into a pharmaceutical composition.

The formulation form of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, or injectable solutions. For example, for formulation in the form of 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. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders are, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, lacquercanth 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.

As acceptable pharmaceutical carriers for compositions formulated as liquid solutions, sterilization and biocompatible, saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

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

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, and the like, preferably oral administration.

A suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, mode of administration, age, body weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity of the patient, and is usually As such, a skilled physician can easily determine and prescribe an effective dosage for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g/kg.

The pharmaceutical composition of the present invention may be prepared in a unit dosage form by formulation using a pharmaceutically acceptable carrier and/or excipient, or may be prepared by placing it in a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or a stabilizer.

In addition, the present invention provides a method for improving, preventing or treating inflammatory bowel disease or leaky intestinal syndrome, comprising administering an effective amount of red pepper leaf extract to a mammal.

The term "mammal" as used herein refers to a mammal that is the subject of treatment, observation or experimentation, and preferably refers to a human.

The term "effective amount" as used herein refers to an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, which is considered by a researcher, veterinarian, doctor or other clinician, which Contains an amount that induces relief of the. The effective amount and frequency of administration for the active ingredient of the present invention may be changed according to the desired effect. Therefore, the optimal dosage to be administered can be easily determined by a person skilled in the art, and the degree of symptoms of inflammatory bowel disease, the degree of intestinal permeability to leaky intestinal syndrome, the content of active ingredients and other ingredients contained in the composition, the type of formulation, and It can be adjusted according to various factors, including the age, weight, general health status, sex and diet of the patient, the time of administration, the route of administration and the secretion rate of the composition, the duration of treatment, and drugs used simultaneously. In the prophylaxis, treatment or improvement method of the present invention, in the case of an adult, it is preferable to administer the red pepper leaf extract in a dose of 0.001 g/kg to 10 g/kg when administered once to several times a day.

In the treatment method of the present invention, the composition comprising the red pepper leaf extract as an active ingredient is in a conventional manner through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered, and can be preferably administered orally or intraperitoneally.

Hereinafter, a preferred embodiment is presented to aid the understanding of the present invention, but the following examples are only illustrative of the present invention, and it is obvious to those skilled in the art that various changes and modifications are possible within the scope and spirit of the present invention, It is natural that such modifications and modifications fall within the appended claims.

Example 1. Hot water extract of red pepper leaves

Dried domestic red pepper leaves produced by Yecheon Nonghyup in Gyeongbuk were purchased and used. Red pepper leaves were dried in a drying oven at 50° C. for 24 hours and then pulverized to a size of 1.0 mm or less with a grinder to prepare red pepper leaf powder.

Distilled water equivalent to 20 times the volume was added to 5 g of red pepper leaf powder, followed by reflux extraction at 80° C. for 3 hours to obtain a hot water extract. After filtering the hot water extract, it was concentrated under reduced pressure using a rotary vacuum evaporator, and lyophilized for 72 hours. The hot water extract of pepper leaves obtained by the above method was described as WEPL (Water-soluble extract from pepper leaves).

Example 2. Red pepper leaf polymer fraction

Dried domestic red pepper leaves produced by Yecheon Nonghyup in Gyeongbuk were purchased and used. Red pepper leaves were dried in a drying oven at 50° C. for 24 hours and then pulverized to a size of 1.0 mm or less with a grinder to prepare red pepper leaf powder.

Distilled water corresponding to 20 times the volume was added to 100 g of red pepper leaf powder, followed by reflux extraction at 80° C. for 3 hours to obtain a hot water extract. After filtering the hot water extract, it is concentrated under reduced pressure to be 1/10 of the extract using a rotary vacuum evaporator, and 80% alcohol corresponding to 4 times the volume (v/v) of the hot water extract concentrated under reduced pressure Was added to the hot water extract and precipitated for 24 hours. This was centrifuged (6000 rpm, 635 g, 30 minutes) to obtain a precipitate, and a small amount of distilled water was added thereto to redissolve the precipitate and a dialysis membrane (MW cut off 12,000, Sigma-Aldrich Co., St. Louis, MO, USA) was performed for 2 to 3 days to remove low molecular weight substances, and freeze-dried for 72 hours to obtain a polymer fraction of red pepper leaves. The red pepper leaf polymer fraction obtained by the above method was named HFPL (High-molecular fraction from pepper leaves).

Comparative Example 1. Garcinia cambogia extract

Garcinia cambogia extract was purchased and used from Unicon Natural Products PVT, India. At this time, the garcinia cambogia extract was used that contains 600 mg/g or more of HCA.

<Test Example>

Test Example 1. Analysis of yield, sugar and protein content of hot water extract of red pepper leaf of Example 1 and polymer fraction of red pepper leaf of Example 2

For the analysis of neutral sugars, the method of Albersheim et al. was partially modified to derivatize each constituent sugar with alditol acetate and aldonolactone after hydrolysis, and GC (Gas Chromatography ACME-6100, Young-Lin Co. Ltd. Anyang, Korea) ) Was used. The sample of Example 1 or 2 was hydrolyzed by reacting for 1.5 hr at 121° C. in 2 M TFA (trifluroacetic acid), and then dissolved in 1 ml of 1 M NH ₄ OH (ammonia solution) and 10 mg of NaBH ₄ It was reduced by 4 hr. After adding an appropriate amount of acetic acid to remove the remaining NaBH ₄ , methanol was added thereto and repeated drying to remove excess acetic acid and converted to alditol. 1 ml of acetic anhydride was added to the alditol and reacted at 121° C. for 30 min to convert to alditol acetate, which was separated and extracted with a chloroform/H ₂ O two-phase solvent system. And, the extract was dried and dissolved in a small amount of acetone to be used as a sample for GC analysis.

Protein quantification was measured according to the Bradford method, and BSA (Bovine Serum Albumin, sigma aldrich) was used as a standard and diluted at a constant multiple with 1 mg/ml as the highest concentration. After adding 20 µl of each standard and sample to 980 µl of Bradford reagent, the absorbance was measured at 595 nm using a microplate reader. Protein quantification was converted to the amount for BSA.

Total sugar was measured according to the phenol-sulfuric acid method, and after stirring the same amount of 5% phenol solution to 0.5 mL of the sample solution, 2.5 mL of concentrated sulfuric acid (98%, v/v) was added and reacted at room temperature for 20 minutes to make a microplate. Absorbance was measured at 470 nm using a reader. For the determination of total sugar, a calibration curve was prepared using a glucose standard (sigma aldrich) and converted into the amount.

Total polyphenol content was analyzed according to the Folin Denis method using gallic acid (sigma aldrich).

In the case of constituent sugar, the oligosaccharides of the sample were hydrolyzed to monosaccharides, and then HPAEC-PAD (high-performance anion-exchange chromatography-pulsed amperometric detector) was used, and the data were analyzed using Chromelon 6.80 software (Dionex). Monosaccharides (fucose, rhamnose, arabinose, galactose, glucose, xylose) were used as standard solutions.

As a result, as a result of analyzing the physicochemical properties such as the content of main components of the hot water extract and the polymer fraction from red pepper leaves, it is shown in Table 1 below.

division Example 1 (WEPL) Example 2 (HFPL) Extraction yield (%) 12.66 2.68 Chemical composition (%) Protein 4.5 ± 0.2 4.3 ± 0.4 Neutral sugar 70.7 ± 2.2 52.4 ± 0.5 Uronic acid 16.9 ± 0.1 36.7 ± 1.1 KDO-liked material 0.1 ± 0.1 2.5 ± 0.5 polyphenol 7.8 ± 0.4 4.0 ± 0.7 Component sugar (PMP) (Mole %) Rhamnose (Rhamnose) 9.3 ± 0.1 3.6 ± 0.1 Fucose (fucose) - 0.2 ± 0.0 Arabinose (Arabinose) 2.5 ± 0.2 4.9 ± 0.2 Xylose 0.6 ± 0.1 0.6 ± 0.1 Mannose 0.8 ± 0.0 0.8 ± 0.0 Galactose 8.5 ± 0.1 7.1 ± 0.1 Glucose 54.8 ± 0.5 34.6 ± 0.6 Glucuronic acid - 1.5 ± 0.0 Galacturonic acid 11.1 ± 0.8 36.0 ± 1.1

* The content ratio (%) of the chemical component represents the weight% in the dry sample.

As shown in Table 1, the hot-water extract of red pepper leaves prepared from Example 1 was found to have a yield of 12.66%, a protein content of 4.5% by weight, and a neutral sugar content of 70.7% by weight, indicating a high sugar content. In addition, as a result of analyzing the constituent sugars of the hot water extract of red pepper leaves prepared in Example 1, the main constituent sugar was glucose (54.8 Mole %), and galacturonic acid, rhamnose, and galactose were contained in 11.1, 9.3, and 8.5 Mole%, respectively. , arabionose, mannose, and xylose were found to be present in trace amounts of 2.5, 0.8 and 0.6 Mole%, respectively.

On the other hand, the yield of the red pepper leaf polymer fraction prepared in Example 2 was found to be as low as 2.68%, but it was confirmed that the protein content was 4.3% by weight, the neutral sugar content was 52.4% by weight, and the uronic acid was 36.7% by weight. In addition, in the red pepper leaf polymer fraction prepared from Example 2, the constituent sugar content of galacturonic acid and glucose was detected as 36.0 and 34.6 Mole%, respectively, and galactose, arabinose, rhamnose, glucuronic acid, mannose, xylose, and fucose were 7.1, 4.9, respectively. It was confirmed that there were traces of 3.6, 1.5, 0.8, 0.6, and 0.2 Mole %. It was confirmed that the sugar content of the red pepper leaf polymer fraction (HEPL) of Example 2 was reduced overall compared to the hot water extract of red pepper leaf of Example 1, but the contents of Arabinose, Fucose, and Glucuronic acid slightly increased. That is, it can be seen that there is a clear difference between the specific components present in the two extracts.

Test Example 2. Analysis of Factors Related to Inflammation of Hot Water Extract of Pepper Leaves of Example

1) Experimental animals

First, 45 6-week-old male C57BL/6J mice were purchased through a central laboratory animal. The experimental animals were divided into five groups as follows. Nine experimental animals were used for each group, and after 8 weeks of diet, the intestinal immune modulating efficacy was evaluated.

-Normal feed group (NC): An experimental group that fed normal feed (AIN-93G) and administered only vehicle orally

-High fat diet group (HFD): An experimental group that fed high fat (60% kcal fat) feed and administered only vehicle orally

-Positive control group (GC): An experimental group fed with a high fat feed and orally administered 50 mg/kg of Garcinia cambogia extract

-Experimental group 1 (WEPL50): an experimental group in which 50 mg/kg of hot water extract of red pepper leaves prepared in Example 1 was orally administered after feeding high fat feed

-Experimental group (WEPL100): A high-fat feed was fed, and 100 mg/kg of hot water extract of red pepper leaves prepared in Example 1 was administered orally.

At this time, a modified AIN 76A purified rodent diet (60 cal% fat) was used as a high-fat diet, which increased calorie by 60 cal% by adding Lard.

2) analysis

After the 8-week diet period was over, the experimental animals of each group were fasted from 6 hours before analysis. Next, the experimental animals were anesthetized and the abdomen was opened. After collecting blood from the inferior vena cava using a sterile syringe, it was stored in a sterile tube treated with heparin. The obtained blood was centrifuged (12,000 × g, 10 minutes, 4° C.), and the supernatant was taken and used for analysis. The concentrations of immunoglobulin A, IL-6, and leukotriene B4 in blood were determined using Mouse IgA ELISA kit (Abcam, ab157717), Mouse IL-6 Quantikine ELISA kit (R&D systems, M6000B), LTB4 Parameter Assay Kit (R&D systems, KGE006B), respectively. Quantified.

3) Analysis result

Intestinal inflammation related indicators for the hot water extract of red pepper leaves prepared from Example 1 were confirmed, and the results are shown in FIGS. 1 to 3.

1 is a graph showing the measurement of the concentration of IgA in serum for the normal feed group (NC), the positive control group (GC), the first experimental group (WEPL50), and the second experimental group (WEPL100). As shown in FIG. 1, in the normal feed group (NC), the blood IgA concentration was 16.25 µg/ml, but in the positive control group (GC), it was confirmed that the intestinal immunity was enhanced to 26.17 µg/ml. In the first experimental group in which the hot water extract of red pepper leaves prepared in Example 1 was orally administered at a concentration of 50 mg/kg, 25.52 ㎍/㎖ of IgA was confirmed. In addition, 30.91 µg/ml IgA was detected in the second experimental group, in which the hot water extract of red pepper leaves prepared in Example 1 was orally administered at a concentration of 100 mg/kg.

Taken together, it can be seen that the hot water extract of red pepper leaves of Example 1 also enhances intestinal immune activity like the positive control (GC). In particular, it was confirmed that the intestinal immunity activity was also increased as the dose concentration of the hot water extract of red pepper leaves (Example 1) was increased, and it was found that the hot water extract of red pepper leaves according to the present invention has the effect of reducing inflammation by enhancing intestinal immunity. I can.

2 is a measurement of the expression level of cytokine IL-6 in serum for the normal feed group (NC), the high fat diet group (HF), the positive control group (GC), the first experimental group (WEPL50), and the second experimental group (WEPL100). This is the graph shown.

IL-6 is a representative cytokine known to be increased in inflammatory bowel disease. In the present invention, while inducing inflammatory bowel disease through a long-term high-fat diet for 8 weeks, the hot water extract of red pepper leaves of Example 1 was mixed and administered to investigate the effect of preventing or treating inflammatory bowel disease. As shown in FIG. 2, it can be seen that the high-fat diet group (HF) in which inflammatory bowel disease was induced through a high-fat diet for 8 weeks increased the concentration of IL-6 significantly to 12.37 pg/ml. In the case of the first and second experimental groups in which the high-fat diet and hot water extract of red pepper leaf of Example 1 were co-administered at various concentrations, 1.78 pg/ml and 1.96 pg/ml of IL-6 were detected, respectively. This is lower than the concentration of IL-6 (2.89 pg/ml) in the positive control group (GC), which was co-administered with Garcinia cambogia extract, which is known to have therapeutic effect on inflammatory bowel disease, and lower than that of the normal feed group (NC) (2.78 pg/ml) It's a shame. That is, it can be seen that the hot water extract of red pepper leaves prepared in Example 1 significantly inhibits IL-6 production than the normal feed group or garcinia cambogia extract. Through this, it can be seen that the hot water extract of red pepper leaf according to the present invention has an improvement, prevention, or treatment effect for inflammatory bowel disease by suppressing the inflammatory reaction in the intestine. Garcinia cambogia is known to accompany various side effects such as dry mouth, dizziness, headache, or diarrhea when ingested. It has a great advantage in that it is a stable substance with no toxicity detected.

Figure 3 is a normal feed group (NC), high fat diet group (HF), positive control group (GC), the first experimental group (WEPL50) and the second experimental group (WEPL100) in serum cytokine leukotriene B4 (LTB ₄ ) expression amount It is a graph showing by measuring.

leukotriene B4 (LTB ₄ ) is known to be involved in the activity of inflammatory cells by inducing an inflammatory response in immune cells. Therefore, the level of inflammation was analyzed by measuring the LTB ₄ concentration from experimental animals in which inflammatory bowel disease was induced by a long-term high fat diet. As shown in FIG. 3, when inflammatory bowel disease was induced through a high-fat diet, it was found that the concentration of LTB ₄ was significantly increased to 1357 pg/ml compared to the normal feed group (NC). In contrast, it was confirmed that the concentration of LTB ₄ decreased to 976 pg/ml in the positive control group (GC) administered with garcinia cambogia extract in combination with a high fat diet.

It was also confirmed that the concentrations of LTB ₄ decreased to 1177 pg/ml and 1026 pg/ml, respectively, in the first and second experimental groups, in which the hot water extract of red pepper leaf prepared in Example 1 was administered in parallel with a high fat diet.

In addition, the hot water extract of red pepper leaves according to the present invention showed a decrease in the concentration of LTB ₄ similar to that of the positive control group and the normal feed group, through which it can be seen that it has an effect of inhibiting LTB ₄ production equivalent to that of the garcinia cambogia extract.

Although the 1st and 2nd experiment groups to which hot water extract of red pepper leaves were administered had less decrease in LTB ₄ concentration than the positive control group, there is a problem that various side effects may occur in the case of the extract of Garcinia cambogia, whereas the hot water extract of red pepper leaves of the present invention is very As a stable substance, it can be seen that it has a remarkably excellent effect in that it exhibits a remarkable anti-inflammatory effect without showing any cytotoxicity.

It was confirmed that the hot water extract of red pepper leaves according to the present invention can exhibit an effect in improving, preventing or treating inflammatory bowel disease by inhibiting the activity of inflammatory cells. Furthermore, in order to obtain a remarkable activity of inhibiting LTB ₄ production at the level of a normal feed group, the hot water extract of red pepper leaves is preferably administered at a concentration of 100 mg/ml.

Test Example 3. Effect of hot water extract of pepper leaves on improving, preventing or treating leaky gut syndrome

1) Experimental animals

First, 45 6-week-old male C57BL/6J mice were purchased through a central laboratory animal. The experimental animals were divided into five groups as follows. Nine experimental animals were used for each group, and after 8 weeks of diet, the efficacy of improvement, prevention and treatment for intestinal leak syndrome were evaluated.

-Normal feed group (NC): An experimental group that fed normal feed (AIN-93G) and administered only vehicle orally

-High fat diet group (HFD): An experimental group that fed high fat (60% kcal fat) feed and administered only vehicle orally

-Positive control group (GC): An experimental group fed with a high fat feed and orally administered 50 mg/kg of Garcinia cambogia extract

-Experimental group 1 (WEPL50): an experimental group in which 50 mg/kg of hot water extract of red pepper leaves prepared in Example 1 was orally administered after feeding high fat feed

-2nd experimental group (WEPL100): A high-fat feed was fed, and 100 mg/kg of hot water extract of red pepper leaves prepared in Example 1 was administered orally.

At this time, a modified AIN 76A purified rodent diet (60 cal% fat) was used as a high-fat diet, which increased calorie by 60 cal% by adding Lard.

2) analysis

After the 8-week diet period was over, the experimental animals of each group were fasted from 6 hours before analysis. In order to check the intestinal permeability from each experimental animal, FITC-dextran 400 mg/kg body weight dissolved in sterile saline was administered orally, followed by microvenous blood collection 0, 2, and 4 hours later. The obtained 100 μL of blood was centrifuged (12,000 × g, 10 min, 4 ℃) to take the supernatant, and the excitation wavelength was 485 nm and the emission wavelength was 535 nm using a microplate reader. It was set and the absorbance was measured. The concentration was calculated using the standard curve of FITC-dextran, and the concentration of FITC-dextran by time was expressed as an area under the curve (AUC).

3) Analysis result

Figure 4 shows the detection concentration of FITC-dextran in the normal feed group (NC), the high fat diet group (HFD), the positive control group (GC), the first experimental group (WEPL50) and the second experimental group (WEPL100) over time. It is a graph. 5 is a graph showing the area under the curve of the result measured from FIG. 4.

As shown in Figs. 4 and 5, when the normal feed group (NC) (normal control) was set to 100%, the high fat diet group (HFD) in which intestinal leakage syndrome was caused through a high fat diet was 399.61%, and the positive control group (normal control) was 399.61%. GC) was detected as 271.71%. That is, it can be seen that even if a high-fat diet was provided, intestinal permeability was suppressed when garcinia cambogia extract was administered in combination.

However, in the 1st and 2nd experimental groups to which the hot water extract prepared from Example 1 was co-administered, it was measured as 125.97% and 104.98%, respectively, and it can be seen that the intestinal permeability was suppressed and decreased to a value substantially similar to that of the normal feed group. have. Particularly, it was confirmed that the hot water extract of red pepper leaf prepared in Example 1 was administered at least 100 mg/kg to have the best effect.

Compared to the high fat diet group (HFD), intestinal permeability decreased by 3.17 times and 3.81 times in the first and second experimental groups, respectively. Therefore, it can be seen that the hot water extract of red pepper leaves according to the present invention has an improvement, prevention, or treatment effect on leaky intestinal syndrome.

Hereinafter, examples of the formulation of the composition containing the powder of the present invention are described, but the present invention is not intended to limit it, but is intended to be described in detail.

Formulation Example 1. Preparation of powder

Hot water extract of red pepper leaves obtained in Example 1 or polymer fraction of red pepper leaves obtained in Example 2 500 mg

100 mg lactose

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2. Preparation of tablet

Hot water extract of red pepper leaves obtained in Example 1 or polymer fraction of red pepper leaves obtained in Example 2 300 mg

Corn starch 100 mg

Lactose 100 mg

2 mg of magnesium stearate

After mixing the above ingredients, tablets are prepared by tableting according to a conventional tablet preparation method.

Formulation Example 3. Preparation of Capsule

Hot water extract of red pepper leaves obtained in Example 1 or polymer fraction of red pepper leaves obtained in Example 2 200 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare a capsule.

Formulation Example 4. Preparation of injection

Hot water extract of red pepper leaves obtained in Example 1 or polymer fraction of red pepper leaves obtained in Example 2 600 mg

Mannitol 180 mg

2974 mg of sterile distilled water for injection

Na ₂ HPO _4, 12H ₂ O 26 mg

It is prepared with the above ingredients per ampoule according to a conventional injection preparation method.

Formulation Example 5. Preparation of liquid formulation

Hot water extract of red pepper leaves obtained in Example 1 or polymer fraction of red pepper leaves obtained in Example 2 4 g

10 g of isomerized sugar

5 g of mannitol

Purified water appropriate amount

According to the usual preparation method of liquid preparation, add each ingredient to purified water to dissolve it, add lemon zest, mix the above ingredients, add purified water and add purified water to adjust the total to 100g, then fill in a brown bottle for sterilization. To prepare a solution.

Formulation Example 6. Preparation of granules

Hot water extract of red pepper leaves obtained in Example 1 or polymer fraction of red pepper leaves obtained in Example 2 1,000 mg

Vitamin mixture right amount

Vitamin A acetate 70 ㎍

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 ㎍

Vitamin C 10 mg

Biotin 10 ㎍

Nicotinic acid amide 1.7 mg

Folic acid 50 ㎍

0.5 mg of calcium pantothenate

Suitable amount of inorganic mixture

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dicalcium phosphate 55 mg

Potassium citrate 90 mg

100 mg of calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for granules, but it is possible to arbitrarily modify the mixing ratio. After mixing the above ingredients according to a conventional granule preparation method, granules And can be used to prepare a health functional food composition according to a conventional method.

Formulation Example 7. Preparation of functional beverage

Hot water extract of red pepper leaves obtained in Example 1 or polymer fraction of red pepper leaves obtained in Example 2 1,000 mg

Citric acid 1,000 mg

100 g oligosaccharides

2 g of plum concentrate

1 g taurine

Total 900 ml by adding purified water

After mixing the above ingredients according to a normal health drink manufacturing method, stirring and heating the resulting solution at 85°C for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, and stored in a refrigerator. It is used to prepare the functional beverage composition of the invention.

The composition ratio is composed of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences, such as the demand class, the country of demand, and the purpose of use.

Claims (12)

A food composition for improving or preventing inflammatory bowel disease or leaky intestinal syndrome using red pepper leaf extract as an active ingredient. The method of claim 1,
The red pepper leaf extract is a food composition for improving or preventing inflammatory bowel disease or leaky gut syndrome, characterized in that it is extracted with water, alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The method of claim 1,
The composition is a food composition for improving or preventing inflammatory bowel disease or leaky gut syndrome, characterized in that it increases immune activity, suppresses inflammatory reactions, and reduces activation of inflammatory cells. The method of claim 1,
The composition is a food composition for improving or preventing inflammatory bowel disease or leaky gut syndrome, characterized in that it enhances IgA expression and inhibits or reduces the expression of IL-6 and LTB ₄ . The method of claim 1,
The inflammatory bowel disease is a disease selected from the group consisting of Crohn's disease, ulcerative colitis, bowel Behcet's disease, infectious enteritis, ischemic bowel disease, and radiation enteritis. . The method of claim 1,
The leaky gut syndrome is a food composition for improving or preventing inflammatory bowel disease or leaky gut syndrome, characterized in that induced by a high fat diet. A pharmaceutical composition for preventing or treating inflammatory bowel disease or leaky intestinal syndrome, using red pepper leaf extract as an active ingredient. The method of claim 7,
The red pepper leaf extract is a pharmaceutical composition for preventing or treating inflammatory bowel disease or leaky gut syndrome, characterized in that it is extracted with water, alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The method of claim 7,
The composition is a pharmaceutical composition for preventing or treating inflammatory bowel disease or leaky gut syndrome, characterized in that it increases immune activity, suppresses inflammatory reactions, and reduces activation of inflammatory cells. The method of claim 7,
The composition is a pharmaceutical composition for preventing or treating inflammatory bowel disease or leaky intestinal syndrome, characterized in that it enhances IgA expression and inhibits or reduces the expression of IL-6 and LTB ₄ . The method of claim 7,
The inflammatory bowel disease is a disease selected from the group consisting of Crohn's disease, ulcerative colitis, enteric Behcet's disease, infectious enteritis, ischemic bowel disease, and radiation enteritis. . The method of claim 7,
The leaky gut syndrome is a pharmaceutical composition for preventing or treating inflammatory bowel disease or leaky gut syndrome, characterized in that induced by a high fat diet.

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