KR20150106331A - Composition comprising Maillard reaction product for preventing or treating inflammatory bowel disease - Google Patents

Abstract

The present invention relates to a composition comprising a maillard reaction product as an active ingredient for preventing or treating inflammatory bowel diseases. A maillard reaction product according to the present invention inhibits inflammatory reactions in human intestinal cells, has excellent effects on preventing and treating inflammatory bowel diseases like recovering weight loss, a shortening colon, rectal bleeding, increase in the number of diarrheal evacuation and historrhexis in intestines at normal level in a colitis animal model and thus can be useful in a medicine and a health functional food, etc. for preventing, alleviating or treating inflammatory bowel diseases.

Description

[0001] The present invention relates to a composition for preventing or treating inflammatory bowel disease comprising a Myial reaction product as an active ingredient,

The present invention relates to a composition for preventing or treating an inflammatory bowel disease comprising a Myial reaction product as an active ingredient.

Inflammatory Bowel Disease (IBD) is a disease caused by chronic or acute inflammation of the unknown origin that occurs in the intestines. The causes are not known precisely yet, but these diseases are genetically and environmentally affected and are related to immunity It is known that inflammation is mediated (Functional food, functional food, mathematics, 2011, p. 126-127).

Inflammatory bowel disease is largely divided into ulcerative colitis and Crohn's disease. One of the causes of the disease is increased expression of proinflammatory cytokines. The expression of cytokines and maintenance of intestinal homeostasis in the intestine are known to be important factors for the development of inflammatory bowel disease. (PAN, Min-Hsiung; LAI, Ching-Shu; HO, Chi-Tang.) Anti-inflammatory activity of natural dietary flavonoids. Food & function, 2010, 1 (1): p.

Inflammatory bowel disease is a common disease in western countries, but it has been recognized as a rare disease in Asia. Recently, however, the incidence of inflammatory bowel disease has been increasing rapidly in East Asia due to increasingly Westernized lifestyle. According to a study by Seoul National University Hospital, the number of inflammatory bowel disease patients in Korea has increased rapidly since the 1980s, and the incidence is expected to increase rapidly in the future (Yang Suk Gyun, chronic inflammation is a source of panacea. . Recently, many studies related to the improvement of inflammatory bowel disease have been attempted, and efforts to find the therapeutic or preventive material from nature have been continued.

Amino acids, on the other hand, form muscles to generate energy, stimulate vitality, and activate various metabolisms in the body. Each amino acid has a variety of physiological functions in the body, and many diseases such as obesity, high cholesterol, diabetes and insomnia are caused by disorders of metabolism.

In recent years, along with the accumulation of scientific data on the physiological activity of individual amino acids, various physiological activities and functions of amino acids have been discussed beyond the conventional category of nutrient supply as a basic unit or a nitrogen source constituting a protein.

One way to impart functionality to amino acids or peptides is to use a Maillard reaction (or glycation reaction), which begins with the condensation of the amino group of amino acids, peptides, and proteins with the carbonyl group of the reducing sugar . This reaction produces a number of reactive substances called MRPs (Maillard Reaction Products).

MRPs are known to exhibit antioxidant activity in both chemical models and food systems. For example, a heated casein-glucose mixture reduces lipid peroxidation rates in an emulsified linoleic acid model. In addition, the casein-sugar-derived myial reaction products showed antioxidative activities against hydroxyl free radicals induced by Fenton, but no activity against hydrophobic radicals (Gu et al., Food Chemistry (2009) 117 (1): 48-54). In addition, the Maillard reaction has been reported to enhance the thermal stability, emulsification properties and antioxidant activity of whey proteins (Chevalier et al., J Agric Food Chem. (2001), 49 (10): 5031-5038). The antioxidative effects of DPPH radical scavenging activity on plasma proteins and MRPs produced by glucose, galactose and fructose were confirmed by Soottawat Benjakul, Wittayachai Lertittikul, Friedrich Bauer, and Maillard reaction products from a porcine plasma protein-sugar model system , Food Chemistry, 93 (2), 2005, p.189-196), studies have shown that MRPs of fructose and arginine inhibit the formation of lipid peroxides and antioxidative effects of erasing hydrogen peroxide (Nagatoshi Ide, Benjamin HS Lau, Kenjiro Ryu, Hiromichi Matsuura, Yoichi Itakura, Antioxidant effects of fructosyl arginine, a Maillard reaction product in aged garlic extract, The Journal of Nutritional Biochemistry, 10 (6), 1999, p.372-376). In addition, it has been reported that furrosine is extracted from MRPs produced during rye bread production and has antioxidant activities such as hydrogen peroxide radicals and ABTS radical scavenging (Anna Michalska, Miriam Amigo-Benavent, Maria Dolores del Castillo, (2004), pp. 123-132). In this study, we investigated the effects of the antioxidant activity of rye bread on the growth of the rats.

However, the prophylactic or curative effect of the Myial reaction products on inflammatory bowel disease is not known.

Accordingly, the inventors of the present invention have conducted studies to find a novel agent for the prophylaxis or treatment of inflammatory bowel disease. As a result, the present inventors have found that the Myial reaction product inhibits the inflammatory response in human intestinal cells and has excellent therapeutic activity against inflammatory bowel disease in an animal model of colitis The present invention has been completed.

Accordingly, an object of the present invention is to provide a composition for the prevention or treatment of inflammatory bowel disease comprising an Myial reaction product as an active ingredient.

In order to accomplish the above object, the present invention provides a pharmaceutical composition for preventing or treating inflammatory bowel disease comprising an Myial reaction product as an active ingredient.

The present invention also provides a food composition for preventing or ameliorating an inflammatory bowel disease comprising a Myial reaction product as an active ingredient.

The Myial reaction product according to the present invention suppresses the inflammatory reaction in human intestinal cells and reduces weight loss, shortening of colon, rectal bleeding, increase in the number of diarrhea defecation and restoration of intestinal tissue collapse to normal level in an animal model of colitis And has excellent effects of preventing and treating inflammatory bowel disease, and thus can be usefully used for medicines and health functional foods for prevention, improvement, or treatment of inflammatory bowel disease.

1 shows the effect of the myial reaction product according to the present invention on cytotoxicity (A) induced by H ₂ O ₂ and ROS production (B) induced by PMA in human intestinal cells (A 0.0 &gt; p < 0.01, < / RTI &gt;
FIG. 2 is a graph showing the effect of the myial reaction product according to the present invention on the expression of inflammatory cytokines (TNF-α, IL-1β, MCP-1 and IL-8) induced by LPS in a mutual culture system ( p < 0.05).
Figure 3 shows the effect of administration of the Maillal reaction product according to the present invention on body weight (A) and disease activity index (B) in an animal model of colitis ( p <0.01 in A, p <0.001 in B) .
FIG. 4 shows the effect of administration of the Myial reaction product according to the present invention on the length (A and B) and the length / weight (C) of the intestine in an animal model of colitis ( p <0.001 for C, p &Lt; 0.05).
FIG. 5 shows the effect of administration of the Maillal reaction product according to the present invention on enterocolitis lesions in an animal model of colitis by H & E staining.
FIG. 6 shows the effect of administration of the Myial reaction product according to the present invention on the expression of inflammatory cytokines (TNF-.alpha., IL-1.beta., IL-6, IL-10 and NF-.kappa.B) ( P < 0.05).
FIG. 7 is a graph showing the effect of the Maillal reaction product according to the present invention on the MPO activity in colon tissues in an animal model of colitis ( p <0.05).
FIG. 8 is a graph showing the effect of administration of the Myial reaction product according to the present invention on serum NO production (A) and lysozyme activity (B) in an animal model of colitis ( p <0.05).

Hereinafter, the present invention will be described in more detail.

The present invention provides a composition for the prophylaxis or treatment of inflammatory bowel disease comprising a Myial reaction product as an active ingredient.

The composition comprises a pharmaceutical composition or a food composition.

In the present invention, &quot; Maillard reaction &quot; means that a carbonyl group of an amino acid and a sugar (reducing sugar) are condensed to form a new substance.

The amino acid includes glycine, alanine, valine, leucine, isoleucine, threonine, serine, cysteine, methionine, aspartic acid, asparagine, glutamic acid, glutamine, lysine, arginine, histidine, phenylalanine, tyrosine, tryptophan, proline Is not limited to lysine. The amino acid includes peptides or proteins in which amino acids are bonded, and food by-products or other by-products containing them, but the present invention is not limited thereto.

The sugar includes glucose, fructose, galactose, sucrose, maltose, lactose, xylose and the like, preferably glucose but not limited thereto. In addition, the sugar includes, but is not limited to, a polysaccharide having a sugar-bound form and a disaccharide-containing polysaccharide and a food by-product or other by-product containing them.

In one embodiment of the present invention, Maillard reaction products (MRPs) were obtained using lysine as an amino acid and glucose as a sugar.

The Myial reaction product according to the present invention suppresses the inflammatory reaction in human intestinal cells and reduces weight loss, shortening of colon, rectal bleeding, increase in the number of diarrhea defecation and restoration of intestinal tissue collapse to normal level in an animal model of colitis And has an excellent effect of preventing and treating inflammatory bowel disease.

Therefore, the Myial reaction product according to the present invention can be usefully used for medicines and health functional foods for the prevention or treatment of inflammatory bowel disease.

The term &quot; inflammatory bowel disease &quot; in the present invention includes ulcerative colitis, Crohn's disease, Behcet's disease, intestinal tuberculosis, enteritis (amoebic enteritis, radiitis enteritis, drug-induced enteritis and the like) as a generic term for inflammation in the intestinal tract.

Ulcerative colitis is a disease in which erosions (erosions) or ulcers are continuously formed in the mucous membrane of the large intestine. Blood, dyspepsia, diarrhea, and abdominal pain occur in the ulcerative colitis, and systemic symptoms such as fever, weight loss, . In addition, Crohn's disease is a disease in which lesions such as ulcers are generated continuously in any part of the digestive tract from the mouth to the anus, and in addition to abdominal pain, diarrhea and stool, fever, hemorrhage, weight loss, Anemia, and the like. Both ulcerative colitis and Crohn's disease are chronic refractory diseases in which recurrence is repeated after symptoms are temporarily improved, and it is highly likely to develop into colon cancer.

The pharmaceutical compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. In addition, it can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, oral preparations such as syrups and aerosols, external preparations, suppositories and sterilized injection solutions according to a conventional method. Suitable formulations known in the art are preferably those as disclosed in Remington ' s Pharmaceutical Science, recently, Mack Publishing Company, Easton PA.

Examples of carriers, excipients and diluents that can be contained in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When the composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which are prepared by mixing at least one excipient such as starch, calcium carbonate, sucrose, lactose, It is prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of suppository bases include withexol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The term "administering" as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.

The preferred dosage of the composition of the present invention varies depending on the condition and the weight of the individual, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. For example, for a desired effect, the composition of the present invention may be administered in an amount of 0.1 to 10,000 mg / kg (body weight) per day. The composition may be administered once a day, or divided into several doses.

The compositions of the present invention may be administered to a subject in a variety of routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

The composition of the present invention may contain one or more known active ingredients having an effect of preventing or treating inflammatory bowel disease as well as a myial reaction product.

The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the prevention or treatment of inflammatory bowel disease.

The Myial reaction product of the present invention may be added to a health functional food for the purpose of preventing or improving inflammatory bowel disease.

In the present invention, a health functional food refers to a food having a biological control function such as prevention and treatment of disease, bio-defense, immunity, recovery after disease, and aging inhibition.

When the myial reaction product of the present invention is used as a food additive, the myial reaction product may be directly added or used together with other food or food ingredients, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). Generally, the myial reaction product of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material, in the production of food or beverage. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, it may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the type of food that can contain the Myial reaction product of the present invention. Examples of foods to which the myial reaction product of the present invention can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, , Tea, a drink, an alcoholic beverage, and a vitamin complex, all of which include health foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharine and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may comprise flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, preferred embodiments and experimental examples are provided to facilitate understanding of the present invention. However, the following examples and experimental examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples and experimental examples.

Example  1. Using amino acids and sugars My Yal  Preparation of reaction product

Lysine (1 mol / L) was used as an amino acid and glucose (1 mol / L) was used as a sugar. The product was reacted at a molar ratio of 1: MRPs). More specifically, lysine and glucose were mixed in a shaking water bath at 105-110 ° C for 30 minutes at 120 rpm to carry out the Myial reaction. After completion of the reaction, the reaction product was rapidly cooled and then freeze-dried to remove moisture, and a Meal reaction product was obtained and stored at -70 ° C until use.

Experimental Example  One. in vitro  On My Yal  Verification of improvement of intestinal inflammation of reaction products

In order to confirm the effect of the Maillal reaction product prepared in Example 1 on the intestinal inflammation reaction in vitro , the following experiment was conducted.

First, mononuclear cells were cultured in RPMI-1640 medium containing 5 mM mercaptoethanol and 10% FBS (37 C, &lt; RTI ID = 0.0 &gt; 5% CO ₂ condition). Caco-2 cells, human intestinal cells, were cultured in DMEM medium containing 10% FBS (37 ° C, 5% CO ₂ ).

When Caco-2 cells grew into a single layer state in a flask, the Myial reaction product prepared in Example 1 was treated with 2 mM H ₂ O ₂ , which is an oxidative stress-inducing substance, and cell viability Were measured. In order to measure ROS production inhibitory activity, Caco-2 cells were treated with 0.1 μM phorbol myristate acetate (PMA), which is an oxidative stress inducing substance, and the Myel reaction product prepared in Example 1, Was measured. Control did not do anything. In all experiments, significance was evaluated through one-way analysis of variance. The results are shown in Fig.

As shown in FIG. 1, the Myial reaction product (Glc-Lys MRPs) of the present invention alleviates cytotoxicity induced by H ₂ O ₂ in human intestinal cells in a concentration-dependent manner, and ROS generation induced by PMA Respectively.

Then, THP-1 cells were cultured in the lower layer of the trans-well, and Caco-2 cells were cultured in the upper layer of the trans-well as a single layer in order to examine the anti-inflammatory response using the co-culture system I made it the same as my model. Intracellular inflammatory response was induced using 2 μg / mL LPS (lopopolysaccharide), an inflammation inducer. In the case of the control (control), nothing was treated. In the case of the experimental group, the Myial reaction product prepared in Example 1 was treated with LPS. After 12 hours, the medium was removed and RNA was isolated from the cells using trizol. Then, RT-PCR was performed to detect the expression of TNF-α (Tumor necrosis factor-alpha), IL-1β (Intereukin-1 beta), MCP-1 (monocyte chemoattractant protein-1) and IL-8 were measured. The primer sequences used in the experiments are shown in Table 1, and the results are shown in FIG.

Gene Oligonucleotide sequence TNF-a sense, 5'-ATGAGCACTGAAAGCATGATCCGG-3 '
antisense, 5'-GAAGACCCCTCCCAGATAGATGG-3 ' IL-1? sense, 5'-AAACAGATGAAGTGCTCCTTCCAGG-3 '
antisense, 5'-TGGAGAACACCACTTGTTGCTCCA-3 ' IL-8 sense, 5'-CTGGCCGTGGCTCTCTTGGCAGCCTTCCTG-3 '
antisense, 5'-GGCAACCCTACAACAGACCCACACAATACA-3 ' MCP-1 sense, 5'-TCGCGAGCTATAGAAGAATCA-3 '
antisense, 5'-TGTTCAAGTCTTCGGAGTTTG-3 '. GAPDH sense, 5'-AGGTCGGAGTCAACGGATTTG-3 '
antisense, 5'-ACAGTCTTCTGGGTGGCAGTG-3 '

As shown in FIG. 2, the Myial reaction product (Glc-Lys MRPs) of the present invention down-regulated the expression of inflammatory cytokines induced by LPS in a mutual culture system, which is the same model as in intestine.

From the above experimental results, it was confirmed that the Myial reaction product (Glc-Lys MRPs) of the present invention has antioxidant and anti-inflammatory effects in the intestine in vitro .

Experimental Example  2. in vivo On My Yal  Verification of therapeutic effect of inflammatory bowel disease of reaction products

The myial reaction product prepared in Example 1 was in vivo The following experiment was conducted to confirm the effect on inflammatory bowel disease. Male Wistar rats aged 6 to 7 weeks were used as colitis animal models and were divided into 5 groups of 6 animals (group 1 was control group, group 2 was DSS (dextran sulfate sodium), colitis induced group 3 (MRPL: 42.1 mg of Glc-Lys MRPs, 421 mg of Glc-Lys MRPs, GL: 421 mg of GL, 200 mg of MRPH), 4 (MRPH) g rats). Colitis was induced by drinking water with DSS (5%) and then by free water. Each sample was orally administered for 7 days from the induction of colitis to the induction period of colitis for a total of 14 days. At the end of the experiment, the rats were sacrificed 18 hours later, the colon was removed, washed twice with PBS, dried between two filter paper, and then weighed. The length of the colon was then measured from the intersection of the colon to the end of the rectum. The end of the colon was used for microscopy and biochemical analysis. The rest of the colon was homogenized in 100 mg / 1 ml PBS and stored at -70 ° C for subsequent experiments.

2-1. Weight and disease activity index (Disease activity index, DAI ) Measure

Body weights were measured during the experiment and the fecal concentration and status were measured for 5 days from the day before DSS administration using the DAI scoring system. The DAI score was calculated by adding the fecal concentration and blood stain of each rat and dividing by 2. Each score conversion is as follows. Fecal concentrations (0 and 1: normal, 2 and 3: dilute stools, 4: diarrhea), stool (0: negative, 1: ±, 2: +, 3: ++, 4: very severe). The results are shown in Fig.

As shown in FIG. 3, when colitis is induced by DSS as compared with the control group, the body weight is significantly reduced and the disease activity index is increased. However, when the Myial reaction product (Glc-Lys MRPs) of the present invention is administered, The index recovered to normal levels.

2-2. Length measurement of field

As a representative marker of inflammatory bowel disease, the length of each intestine was measured and the length / weight ratio of intestine was calculated. The results are shown in Fig.

As shown in FIG. 4, when the colitis was induced by DSS as compared with the control group, the length of the intestine was remarkably reduced, but when the Myial reaction product (Glc-Lys MRPs) of the present invention was administered, the intestinal length was restored to a normal level . In addition, when the colitis induced by DSS was compared with the control group, the length / weight ratio of the intestine was increased, but when the Myial reaction product (Glc-Lys MRPs) of the present invention was administered, the normal level was recovered.

2-3. Histopathological examination

The end of the colon separated from each rat was fixed in 10% formalin solution for 24 hours, washed in PBS, and dehydrated by serial dilution with alcohol. Each sample was fixed with paraffin, reacted at 56 ° C for 24 hours, and then prepared according to a microtome slicing method with a thickness of 4 μm. After deparaffinization, the cells were stained with hematocylin and eosin, and observed with an optical microscope. The lesion score was measured. The results are shown in FIG. 5 and Table 2.

Group Acute ulcer Mucosal erosion Inflammatory cell infiltration in the mucosa Submucosal edema Inflammatory cell infiltration in the submucosa Control 0.00 ± 0.00 0.00 ± 0.00 ^a 0.00 ± 0.00 ^a 0.00 ± 0.00 ^a 0.00 ± 0.00 5% DSS 0.60 ± 1.3 1.8 ± 0.84 ^b 2.00 ± 1.00 ^b 0.80 ± 0.45 ^b 0.60 + - 0.55 5% DSS + MRPL 0.00 ± 0.00 0.20 ± 0.45 ^a 1.00 0.71 ^c 0.80 ± 0.45 ^b 0.40 + - 0.55 5% DSS + MRPH 0.00 ± 0.00 0.40 + - 0.55 ^a 0.80 + 0.45 ^ac 0.00 ± 0.00 ^a 0.00 ± 0.00 5% DSS + GL 0.60 ± 1.3 1.6 ± 1.3 ^b 1.60 ± 0.89 ^bc 0.40 ± 0.55 ^ab 0.20 ± 0.45

(Means within a column with different letters are significantly different (p <0.05), according to one-way analysis of variance.)

As shown in FIG. 5 and Table 2, when the colitis was induced by DSS, the score of typical lesions of enteric inflammation (Acute ulcer, Mucosal erosion, Inflammatory cell infiltration in the mucosa, Submucosal edema, Inflammatory cell infiltration in the submucosa) , But the scores of the lesions were restored to normal levels when the Myial reaction product (Glc-Lys MRPs) of the present invention was administered.

2-4. Measurement of cytokine expression

The expression level of cytokines such as TNF-α, IL-1β, IL-6, IL-10 and NF-κB in the colon tissues of each rat was measured by quantitative real-time RT-PCR . The primer sequences used in the above experiment are shown in Table 3, and the results are shown in FIG.

Gene Oligonucleotide sequence TNF-a sense, 5'-CGTGTTCATCCGTTCTCTAC-3 '
antisense, 5'-CAGCGTCTCGTGTGTTTC-3 ' IL-1? sense, 5'-GCACCTTCTTTTCCTTCATC-3 '
antisense, 5'-GCCGTCTTTCATCACACA-3 ' IL-6 sense, 5'-AAGTCGGAGGCTTAATTACATATGTTC-3 '
antisense, 5'-TCATCGCTGTTCATACAATCAGAA-3 ' IL-10 sense, 5'-CGGGGTGACAATAACTGC-3 '
antisense, 5'-CCTGGGGCATCACTTCTAC-3 ' NF-κB sense, 5'-GGCAGCACTCCTTATCAA-3 '
antisense, 5'-GGTGTCGTCCCATCGTAG-3 ' β-Actin sense, 5'-TCAGGAGGAGCAATGATCTTGA-3 '
antisense, 5'-GACAGGATGCAGAAGGAGATCAC-3 '

As shown in FIG. 6, the expression of inflammatory cytokines (TNF-α, IL-1β, IL-6, IL-10 and NF-κB) Administration of the Myial reaction product (Glc-Lys MRPs) of the present invention resulted in a very low level of expression of inflammatory cytokines.

2-5. MPO ( Myeloperoxidase ) Active measurement

MPO enzyme activity was measured in the colonic tissue homogenate of each rat using a fluorimetric detection kit. This is done by measuring absorbance at 450 nm using a reagent that detects and oxidizes nonpolar substances in the presence of hydrogen peroxide and MPO. The activity of MPO was expressed as U / mg protein. The results are shown in Fig.

As shown in FIG. 7, when colitis caused by DSS as compared with the control group, the activity of MPO, which is a marker of inflammatory bowel disease, is remarkably increased. However, when the Myial reaction product (Glc-Lys MRPs) of the present invention is administered MPO activity was reduced to a very low level.

2-6. NO (Nitric Oxide) Measurement and Plasma lysozyme  activity assay

Blood samples from each rat were centrifuged at 1000 rpm for 10 minutes and serum was separated.

First, 80 μL of serum and 80 μL of griess reagent (1% Sulfanilamide / 5% phosphoric acid solution) were added to a 96-well plate and incubated at room temperature for 10 minutes. Absorbance was measured at 540 nm. The concentration (mM) of NO was calculated based on the quantitative curve of NaNO ₂ standard solution.

For lysozyme activity, lysates of a suspension of Micrococcus lysodeikticus (75 mg / 100 mL of 0.1 M phosphate / citrate buffer with 0.09% NaCl, pH 5.6) were measured. (PH adjustment with 0.09% NaCl after buffer preparation with P4809). 25 μL of serum or standard reagent was added to 175 μL of the suspension of Micrococcus lysodeikticus, left at room temperature for 30 minutes, and then absorbance was measured at 450 nm. As a standard reagent, hen egg white lysozyme (L6876) was dissolved in 0.1 M phosphate / citrate buffer (with 0.09% NaCl (pH 5.8)) and the protein content of each serum sample was measured and expressed as mg lysozyme / mg protein .

The results of the above experiment are shown in Fig.

As shown in FIG. 8, when the colitis was induced by DSS as compared with the control group, the production of NO, which is an inflammatory marker, was markedly increased and the activity of lysozyme was remarkably decreased, but the Myial reaction product (Glc-Lys MRPs) , The NO production and the lysozyme activity recovered to normal levels.

From the above experimental results, it was confirmed that the Myial reaction product according to the present invention has an excellent therapeutic effect in an animal model of inflammatory bowel disease induced by colitis.

Hereinafter, the pharmaceutical composition of the present invention and the pharmaceutical composition of the food composition will be described, but the present invention is not intended to be limited but is specifically described .

Formulation example  1. Preparation of pharmaceutical preparations

One. Sanje  Produce

Meal reaction product 20 mg

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

2. Preparation of tablets

The myial reaction product 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

3. Preparation of capsules

The myial reaction product 10 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

4. Preparation of injections

The myial reaction product 10 mg

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na ₂ HPO ₄ 2H ₂ O 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

5. Liquid  Produce

The myial reaction product 10 mg

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.

Formulation example  2. Manufacture of food preparation

Meal reaction product 100 mg

Vitamin mixture quantity

70 g of vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

0.2 g of vitamin B12

Vitamin C 10 mg

Biotin 10 g

Nicotinic acid amide 1.7 mg

Folate 50 g

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health foods according to conventional methods.

Claims (6)

A pharmaceutical composition for the prophylaxis or treatment of inflammatory bowel disease comprising a Maillard reaction product as an active ingredient.
The pharmaceutical composition for preventing or treating inflammatory bowel disease according to claim 1, wherein the Myial reaction product is produced by a reaction of an amino acid and a sugar.
3. The composition of claim 2, wherein the amino acid is selected from the group consisting of glycine, alanine, valine, leucine, isoleucine, threonine, serine, cysteine, methionine, aspartic acid, asparagine, glutamic acid, glutamine, lysine, arginine, histidine, phenylalanine, tyrosine, tryptophan and proline Wherein the pharmaceutical composition is at least one selected from the group consisting of:
The pharmaceutical composition for preventing or treating inflammatory bowel disease according to claim 2, wherein the sugar is at least one selected from the group consisting of glucose, fructose, galactose, sucrose, maltose, lactose and xylose .
The method according to any one of claims 1 to 4, wherein the inflammatory bowel disease is one or more selected from the group consisting of Chron's disease, ulcerative colitis, intestinal Behcet's disease, &Lt; / RTI &gt; or a pharmaceutically acceptable salt thereof, for the prophylaxis or treatment of inflammatory bowel disease.
A food composition for preventing or ameliorating an inflammatory bowel disease comprising a Maillard reaction product as an active ingredient.

Images