KR20200101561A - Composition for preventing, ameliorating or treating inflammatory bowel disease comprising extract of Ircinia species - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of inflammatory bowel diseases, comprising an Ircinia sp. extract or a fraction thereof as an active ingredient. The fraction of the Ircinia sp. extract of the present invention relieves weight loss and clinical symptoms such as diarrhea and bloody stool in mice induced with chronic enteritis by dextran sodium sulfate (DSS), and reduces the activity of infiltrating macrophages in intestinal tissues, and thus can be usefully employed in treating inflammatory bowel diseases.

Description

Composition for preventing, ameliorating or treating inflammatory bowel disease comprising extract of Ircinia species, comprising extract or fractions thereof as an active ingredient {Composition for preventing, ameliorating or treating inflammatory bowel disease)

The present invention relates to a pharmaceutical composition for the prevention, improvement or treatment of inflammatory bowel disease comprising the extract or fractions thereof as an active ingredient.

Inflammatory bowel disease is a disease that causes inflammation in the digestive tract chronically for more than 6 months, repeating improvement and deterioration, and the exact cause or mechanism of the onset has not yet been identified. It belongs to the category, and in a broad sense, it can include infectious enteritis such as bacterial, viral, amoebic, and tuberculous enteritis, and all inflammatory diseases that occur in the intestine such as ischemic enteritis, radiation enteritis, and chronic enteritis.

It is thought that one of the promising causes of inflammatory bowel disease identified so far is that intestinal homeostasis is destroyed due to excessive production of proinflammatory mediators (Non-Patent Document 1). Inflammatory mediators play an important role in the defense and reconstruction of foreign substances in innate immunity, but excessively secreted inflammatory mediators destroy intestinal immune homeostasis, and also induces the invasion of immune cells into the intestinal tissue, which is persistent inflammation. It expresses mediators and in severe cases develops into cancer.

Recently, the incidence of outbreaks is increasing in Korea due to the influence of lifestyle habits that are becoming westernized. In particular, patients with inflammatory bowel disease are more than 6 times more likely to develop into the large intestine than normal people, so timely treatment is essential, but a therapeutic agent that can fundamentally treat inflammatory bowel disease has not yet been developed, and prostaglandins. ), which blocks the production of 5-aminosalicylic acid (5-ASA), such as sulfasalazine, or an immunosuppressant such as steroids, but the treatment effect is insignificant, abdominal weakness, headache , Liver disease, peptic ulcer, diabetes, steroidal kidney disease, and other side effects are problematic.

Therefore, development of effective therapeutic agents for treating inflammatory bowel disease without side effects using natural products has been made. Korean Patent Laid-Open Patent No. 10-2014-0004935 discloses an extract of Obaeja as a therapeutic agent for inflammatory bowel disease caused by tobacco smoke, and Korean Patent Publication No. 10-2007-0117889 discloses an extract of Kanghwa Yakushi as a therapeutic agent for inflammatory bowel disease. Are doing.

On the other hand, sponges are metazoans with a simple structure, and about 1,000 species are currently known as inferior animals that do not have differentiation in muscle, nervous system, digestive system, and embryonic design. There is an identification cell, and it accepts the food that came through the ingestion hole, and the identification cell at this time has one flagella, which is considered similar to the flagellate of protozoa. In distinction from other metazoan animals, it is also referred to as a'surface animal'. Sponge animals' lives are mostly attached to other objects, except for very few living on the bottom of the sea. In particular, beach sponges, which are common on the beach with strong waves, live in close contact with rocks. Sponge animals are divided into four classes: calcarea, hexactinellida, demospongiae, and Homoscleromorpha, depending on the characteristics of the bone fragment. Mushroom Sponge, Utenapal Sponge, White Bugs Sponge, Concave Sponge, etc. belong to the Heukbang Sponge River, the sea scrubber and the tip of Sangmo belong, and the Ordinary Sponge River includes bathing sponges, purple sponges, freshwater sponges, and volcanic sponges.

Recently, as various secondary metabolites have been found to exist in the sea surface, research on the discovery of physiologically active substances using the sea surface is active, and aurantoside and spongistatin 1 found in the sea surface. Compounds have been found to have antifungal activity and are used commercially. However, there is still a bar confirming the effect of the bioactive going into the cavernous chamber (Ircinia sp.) Up.

Accordingly, the present inventors prepared an extract and a fraction thereof and administered it to an animal model in which chronic enteritis was induced. As a result, the weight loss of mice, diarrhea and bloody stool symptoms were alleviated, the length of the colon was restored, and the infiltration of the intestinal tissue. By confirming that it reduces the activity of one macrophage, the present invention was completed.

Korean Patent Publication No. 10-2014-0004935 Korean Patent Publication No. 10-2007-0117889

Singh UP et al., Chemokine and cytokine levels in inflammatory bowel disease patients, Cytokine. 2016 Jan;77:44-9.

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating inflammatory bowel disease.

Another object of the present invention is to provide a health functional food for preventing or improving inflammatory bowel disease.

In order to achieve the above object, the present invention provides a fine thread surface of the sea in (Ircinia sp.) Extract or pharmaceutical composition for preventing or treating inflammatory bowel disease comprising a fraction thereof as an active ingredient.

The present invention also provides a fine thread surface of the sea in (Ircinia sp.) Extracts or dietary supplement for the prevention and improvement of inflammatory bowel diseases, including fractions thereof as an active ingredient.

Fractions of the extract of fine spongiform spongiformis of the present invention relieve weight loss, diarrhea and bloody stool symptoms in chronic enteritis-induced mice induced by DSS (dextran sodium sulfate), restore colon length, and activate macrophages infiltrating in the intestinal tissue. By confirming that it reduces, the present invention was completed.

1 is a diagram showing the body weight of each mouse group during an experiment in which mice were fed with DSS to induce chronic enteritis.
(Normal: normal control;
VC: negative control (chronic enteritis induction group);
PC: positive control (chronic enteritis induction group + 5-aminosalicylic acid administration);
Ircinia species extract: Fine spongy spongiform administration group (mixture of fine spongy MR3 and MR4 fractions)).
Figure 2 is a diagram showing the disease activity (disease activity index, DAI) calculated based on the weight and symptoms of enteritis of each mouse group during the experiment period in which chronic enteritis is induced by feeding DSS to mice.
(Normal: normal control;
VC: negative control (chronic enteritis induction group);
PC: positive control (chronic enteritis induction group + 5-aminosalicylic acid administration);
Ircinia species extract: fine spongy spongiform administration group (mixture of fine spongy spongi MR3 and MR4 fractions)).
3 is a diagram showing the results of a one-way batch variance analysis performed for each mouse group by obtaining an area under the curve (AUC) value of disease activity of each mouse group.
(Normal: normal control;
VC: negative control (chronic enteritis induction group);
PC: positive control (chronic enteritis induction group + 5-aminosalicylic acid administration);
Ircinia species extract: fine spongy spongiform administration group (mixture of fine spongy spongi MR3 and MR4 fractions)).
Figure 4 is a diagram showing the length of the large intestine and photographs taken of the large intestine extracted from each mouse group after the end of the experiment inducing chronic enteritis by feeding DSS to the mouse.
(Normal: normal control;
VC: negative control (chronic enteritis induction group);
PC: positive control (chronic enteritis induction group + 5-aminosalicylic acid administration);
Ircinia species extract: fine spongy spongiform administration group (mixture of fine spongy spongi MR3 and MR4 fractions)).
5 is a result of measuring myeloperoxidase activity of colon tissues extracted from each mouse group after the end of the experiment inducing chronic enteritis by feeding DSS to mice.
(Normal: normal control;
VC: negative control (chronic enteritis induction group);
PC: positive control (chronic enteritis induction group + 5-aminosalicylic acid administration);
Ircinia species extract: fine spongy spongiform administration group (mixture of fine spongy MR3 and MR4 fractions)).
Figure 6 is a photograph of the state of the fine spongy ( Ircinia sp.).

Hereinafter, the present invention will be described in detail.

The present invention provides a fine thread surface of the sea in (Ircinia sp.) Extract or pharmaceutical composition for preventing or treating inflammatory bowel disease comprising a fraction thereof as an active ingredient.

The fine spongy spongy extract may be prepared by a manufacturing method comprising the following steps:

1) extracting by adding an extraction solvent to the fine sea sponge;

2) filtering the extract of step 1); And

3) A step of drying after concentrating the filtered extract of step 2) under reduced pressure.

In the above method, if the spongy fine spongy in step 1) belongs to the sp. of fine spongy, it can be used regardless of species.

In addition, the extraction solvent of step 1) may be water, a lower alcohol of C ₁ to C ₂ , or a mixed solvent thereof. The lower alcohol of C ₁ to C ₂ may be methanol or ethanol.

The extraction method of step 1) may be shaking extraction, Soxhlet extraction, or reflux extraction. In this case, the extraction temperature may be 15 to 35 °C, specifically 17 to 33 °C, more specifically 20 to 30 °C, more specifically 22 to 28 °C, but is not limited thereto. In addition, the extraction solvent may be added in an amount of 0.1 to 1 ml per 1 g of the wet weight of the thin sponge used for extraction, and specifically, may be added in an amount of 0.1 to 0.8 ml, more specifically, It may be added in an amount of 0.2 to 0.6 ml, but is not limited thereto. In addition, the extraction time may be 24 to 72 hours, specifically 30 to 60 hours, more specifically 35 to 55 hours, but is not limited thereto. In addition, the number of extractions may be 1 to 5 times, specifically 1 to 3 times, but is not limited thereto. In a range outside the conditions of the extraction temperature, extraction time, or number of extractions, extraction may not be sufficiently performed, so that the content of the active ingredient in the fine spongy extract may be insignificant. Alternatively, the extraction yield may no longer increase, so that the efficiency of the extraction operation may decrease.

The vacuum concentration in step 3) may be performed using a vacuum vacuum concentrator or a vacuum rotary evaporator. In addition, the drying may be vacuum drying, vacuum drying, boiling drying, spray drying, or freeze drying.

The fraction may be prepared by a manufacturing method comprising the following steps:

1) distributing the extract of fine spongy spongy spongiformis in water and methylene chloride;

2) redistributing the distributed extract of step 1) with a hexane and methanol aqueous solution solvent; And

3) obtaining a fraction by fractionating the methanol layer obtained by redistribution in step 2) with a solvent in which water and methanol are mixed.

The fine spongy spongy extract of step 1) can be obtained by performing the above-described method.

The solvent in which water and methanol are mixed in step 3) may be a mixture of water and methanol in a ratio of 1: 1 to 9. Specifically, the solvent in which water and methanol are mixed may be a mixture of water and methanol in a ratio of 1: 1 to 8, more specifically 1:1 to 6, and more specifically 1: 2 to 5 have.

The inflammatory bowel disease may be ulcerative colitis, Crohn's disease, Behcet's disease, or chronic enteritis.

In a specific embodiment of the present invention, the present inventors distribute the fine spongy extract into water and methylene chloride, redistribute the distributed extract with a hexane and methanol aqueous solution solvent, and then redistribute the methanol separation layer obtained by water and methanol. MR1 (water 50: methanol 50), MR2 (water 40: methanol 60) by sequentially fractionating with a mixed solvent of 50:50, 40:60, 30:70, 20:80, 10:90 and 0:100, Fractions of MR3 (water 30: methanol 70), MR4 (water 20: methanol 80), MR5 (water 10: methanol 90) and MR6 (water 0: methanol 100) were obtained.

In addition, the present inventors orally administered a mixture of MR3 and MR4 fractions having excellent inhibitory activity against inflammatory cytokines among the obtained MR1 to MR6 fractions to an animal model in which chronic enteritis was induced, resulting in significant weight loss compared to the negative control group. It was confirmed that the recovery was (FIGS. 1 to 3 ).

In addition, the present inventors confirmed that diarrhea and bloody stool symptoms were alleviated as a result of oral administration of a mixture of MR3 and MR4 fractions to an animal model in which chronic enteritis was induced (FIGS. 2 and 3).

In addition, the present inventors confirmed that as a result of oral administration of a mixture of MR3 and MR4 fractions to an animal model in which chronic enteritis was induced, the length of the colon was significantly recovered compared to the negative control group (FIG. 4 ).

In addition, as a result of oral administration of a mixture of MR3 and MR4 fractions to an animal model in which chronic enteritis was induced, the myeloperoxidase (MPO) activity of the colon tissue was significantly lowered compared to the negative control group, resulting in infiltration of macrophages. It was confirmed that the activity of one macrophage was decreased (FIG. 5).

Therefore, the fraction of the extract of the spongy spongiformis of the present invention alleviates the weight loss of chronic enteritis-induced mice, and clinical symptoms such as diarrhea and bloody stool, and reduces the activity of infiltrating macrophages in the intestinal tissue, thus treating inflammatory bowel disease. It can be usefully used for

The pharmaceutical composition according to the present invention may include 10 to 95% by weight of the extract or fractions thereof, which is an active ingredient, based on the total weight of the composition. In addition, the pharmaceutical composition of the present invention may further include one or more active ingredients exhibiting the same or similar functions in addition to the active ingredients.

The pharmaceutical composition of the present invention may contain a carrier, a diluent, an excipient, or a mixture thereof commonly used in biological preparations. Any pharmaceutically acceptable carrier can be used as long as it is suitable for delivering the composition in vivo. Specifically, the carrier is Merck Index, 13th ed., Merck & Co. Inc., saline, sterile water, Ringer's solution, dextrose solution, maltodextrin solution, glycerol, ethanol, or a mixture thereof. In addition, conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as needed.

When formulating the composition, diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants may be added.

The composition of the present invention may be formulated as an oral or parenteral formulation. Oral formulations may include solid formulations and liquid formulations. The solid preparation may be a tablet, a pill, a powder, a granule, a capsule, or a troche, and the solid preparation may be prepared by adding at least one excipient to the composition. The excipient may be starch, calcium carbonate, sucrose, lactose, gelatin, or a mixture thereof. In addition, the solid preparation may contain a lubricant, examples of which include magnesium stearate and talc. On the other hand, the liquid formulation may be a suspension, a liquid formulation, an emulsion or a syrup. At this time, the liquid formulation may contain excipients such as wetting agents, sweetening agents, fragrances, preservatives, and the like.

The parenteral preparations may include injections, suppositories, powders for respiratory inhalation, aerosols for sprays, powders and creams. The injection may include a sterilized aqueous solution, a non-aqueous solvent, a suspension solvent, an emulsion, and the like. At this time, as the non-aqueous solvent or suspension solvent, vegetable oils such as propylene glycol, polyethylene glycol, and olive oil, or injectable esters such as ethyl oleate may be used.

The composition of the present invention may be administered orally or parenterally according to a desired method. Parenteral administration may include intraperitoneal, rectal, subcutaneous, intravenous, intramuscular or intrathoracic injection.

The composition may be administered in a pharmaceutically effective amount. This may vary depending on the type of disease, the severity, the activity of the drug, the patient's sensitivity to the drug, the administration time, the administration route, the treatment period, and the drugs used at the same time. However, for a desirable effect, the amount of the active ingredient contained in the pharmaceutical composition according to the present invention may be 0.0001 to 100 mg/kg, specifically 0.001 to 70 mg/kg. The administration may be once or several times a day.

The composition of the present invention may be administered alone or in combination with other therapeutic agents. When administered in combination, administration may be sequential or simultaneous.

The present invention also provides a fine thread surface of the sea in (Ircinia sp.) Extracts or dietary supplement for the prevention and improvement of inflammatory bowel diseases, including fractions thereof as an active ingredient.

The extract or fraction thereof according to the present invention is as described above.

The inflammatory bowel disease may be ulcerative colitis, Crohn's disease, Behcet's disease, or chronic enteritis.

The composition of the present invention may be added to food as it is, or may be used together with other foods or food ingredients. At this time, the content of the active ingredient to be added may be determined according to the purpose, and generally may be 0.01 to 90 parts by weight of the total food weight.

The form and type of the health functional food is not particularly limited. Specifically, the health functional food may be in the form of tablets, capsules, powders, granules, liquids, and pills. The health functional food may contain various flavoring agents, sweetening agents, or natural carbohydrates as additional ingredients. The sweetener may be a natural or synthetic sweetener, and examples of the natural sweetener include taumatin and stevia extract. Meanwhile, examples of synthetic sweeteners include saccharin and aspartame. In addition, the natural carbohydrates may be monosaccharides, disaccharides, polysaccharides, oligosaccharides and sugar alcohols.

In addition to the above-described additional ingredients, the health functional food of the present invention includes nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pexane and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives , Glycerin, alcohol, and the like may be further included. These ingredients can be used independently or in combination. The proportion of the additive may be 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, the present invention will be described in detail by the following examples.

However, the following examples are for illustrative purposes only, and the present invention is not limited thereto.

<Example 1> Fine thread spongy ( Ircinia sp.) Preparation of extracts and fractions thereof

<1-1> extract

First, the sponge used in the experiment is the genus Ircinia sp., belonging to the Dictyoceratida of the common sponge (Demospongiae), has a colonial shape in appearance and has numerous knitting threads, and the individual is very tough. It shows a characteristic that is not easily cut (Fig. 6). Wet weight 4.5 kg of fine thread spongy ( Ircinia sp., Vietnam, No. 0014028) was cut at intervals of 5 cm in length, and water was removed through a freeze-drying process for 5 days. The dried fine sponge was put into a 5 L Erlenmeyer flask, and extracted with 2 L methanol solvent at room temperature for 2 days at room temperature to prepare a methanol extract. The methanol extract prepared by using a cotton filter was separated, and the extraction solvent was removed using a rotary evaporator to obtain a crude extract of the fine spongy spongy. Since the obtained crude extract contains a large amount of salt, 1 L of water and methylene chloride were added to distribute the crude extract. Subsequently, the methylene chloride layer (7 g) was redistributed sequentially with hexane and an 85% methanol aqueous solution to obtain a hexane fraction (2.5 g) and an 85% methanol fraction (4 g).

<1-2> Fraction

In a glass tube (10×7 cm) filled with reverse-phase silica gel (YMC-Gel ODS-A, S-75 μm) in the form of a slurry, the 85% methanol fraction obtained in Example <1-1> was mixed with the reverse-phase resin and the upper layer was Put on. After preparing the six solvents in Table 1 below, flash chromatography was performed to develop each solvent by applying a vacuum to a glass tube to obtain six fractions. The amount of each fraction was 1.52 g for MR1, 451 mg for MR2, 189 mg for MR3, 141 mg for MR4, 192 mg for MR5, and 124 mg for MR6.

Sample name Solvent type MR1 50% water: 50% methanol MR2 40% water: 60% methanol MR3 30% water: 70% methanol MR4 20% water: 80% methanol MR5 10% water: 90% methanol MR6 100% methanol

< Experimental example 1> Evaluation of inflammatory cytokine inhibitory activity

In vitro inflammatory cytokine inhibition experiments of the MR1 to MR6 fractions obtained in Example 1 were conducted.

Specifically, the macrophage cell line RAW264.7 cells were sold and used from the Korea Cell Line Bank. Cells were 5%-CO ₂ in DMEM (Dulbecco's modified Eable's medium) medium supplemented with 10% heat-inactivated FBS (fetal bovine serum), penicillin (P/S) and streptomycin, It was maintained at 37 ℃ culture conditions. RAW264.7 cells were seeded in 96-well plates and incubated for 24 hours. Fractions from MR1 to MR6 were dissolved in dimethyl sulfoxide (DMSO), prepared at a concentration of 100 mg/mL, diluted with cell culture solution, and treated to a final concentration of 20 μg/mL. After 30 minutes after treatment of the fraction, 100 ng/ml of LPS (lipopolysaccharide) was treated, and then the amount of NO (nitric oxide) and inflammatory cytokines IL-10 (IL-10) and IL-1β produced in the cell culture solution were determined by ELISA kit. The cell viability was measured through the MTT assay immediately after taking the cell culture solution in order to check whether the cell culture was toxic due to sample treatment.

As a result, MR5 was excluded because cytotoxicity was observed at the dose concentration of 20 ng/ml, and the MR3, MR4, and MR6 fractions showed excellent cytokine production inhibitory activity. In the comparative chromatogram experiment of the extract using HPLC, the MR3 and MR4 fractions showed a very similar pattern, and the in vivo experiment was performed in an amount of 120 mg of the two fractions combined.

% Compared to untreated control NO IL-10 IL- 1β MR1 82.2 5.3 112.3 MR2 83.6 3.2 113.7 MR3 56.1 2.3 104.1 MR4 21.0 2.0 45.6 MR6 38.5 35.6 56.3

< Experimental example 2> Preparation of chronic enteritis animal model and fine spongy Fraction administration

Female C57BL/6 mice purchased from Japan SLC were given a period of acclimatization of 10 days or more to increase their weight sufficiently, and when their weight reached an average of 19±2 g, they were randomly separated into 4 groups of 8 animals and placed in individual wire cages. Put the experiment to proceed. The four groups were the normal control, negative control, positive control, and fine spongiform administration group, respectively. In order to induce human inflammatory bowel disease (IBD) to mice of the rest of the group except for the normal control group, a method of administering dextran sodium sulfate (DSS) through negative water was used. Specifically, 1.5% DSS (colitis grade DSS from P bioscience) was fed negatively for 7 days and then sterilized breeding water was fed three times for 7 days, followed by a total of 42 days (Table 3). For 42 days, the mixture of fine spongiform MR3 and MR4 fractions prepared in Example 1 was administered to mice in the fine spongiform spongiform group at a dose of 15 mg/kg, and 5-aminosalicylic acid in mice of the positive control group. ) Was orally administered once daily at a dose of 50 mg/kg, and none of the mice of the negative control group were administered.

1 cycle
2 cycle 3 cycle Purification period 1.5% DSS
(7 days) water
(7 days) 1.5% DSS
(7 days) water
(7 days) 1.5% DSS
(7 days) water
(7 days) End of experiment Observation of clinical symptoms and weight measurement

< Experimental example 3> Observation of clinical symptoms and weight change

In order to confirm whether the fine spongy fraction affects the clinical symptoms and body weight of the chronic enteritis animal model, chronic enteritis symptoms and body weight changes in each mouse group were measured during the experiment.

Specifically, body weight was measured between 9 am and 10 am every day for the duration of the experiment, and the presence of diarrhea and bloody stool was recorded once a day on white paper under the cage for enteritis symptoms. Based on the measured weight and symptoms of enteritis, disease activity (disease activity index, DAI) was calculated according to Table 4 below. Disease activity (DAI) was modified based on existing references, and disease activity analysis was performed by one-way analysis of variance for each mouse group by obtaining AUC (area under the curve) values.

As a result, it was confirmed that the group in which chronic enteritis was induced, excluding the normal control group, showed a rapid body weight change when DSS was administered, and the width of the weight loss decreased in the 2nd and 3rd cycles and gradually approached the body weight of the normal control group. Among them, it was confirmed that the fluctuation width of body weight in the fine spongiform administration group was smaller than that of the negative control group and the positive control group (FIG. 1). In addition, it was confirmed that the DAI value of the fine spongiform administration group was on average 35.77±21.45, which was significantly reduced from the DAI value of 72.62±31.48 of the negative control group (FIGS. 2 and 3).

Weight loss ① Average weight on the day of the negative control group-Body weight on the day of all chronic enteritis induction groups
(However, negative values are evaluated as 0) diarrhea
(loose stool) ② 0=no symptoms (negative), 1=moderate, 2=severe Bloody stool
(blood in the stool) ③ 0=no symptoms (negative), 2=symptoms (positive)
(Evaluate with weight on blood) DAI ①+②+③

< Experimental example 4> Measure the length of the large intestine

In order to confirm whether the fraction of the thin spongy spongiform affects the length of the colon of the chronic enteritis animal model, after the experiment was completed, each mouse was euthanized, and the colon was removed by autopsy, and the length of the colon was measured by spreading it from the gyrus junction to the anus. I took a picture.

As a result, it was confirmed that the average length of the colon of the fine spongiform administration group was 6.54±0.35 cm, and that of the negative control group was 5.94±0.09 cm. 4).

< Experimental example 5> Myeloperoxidase ( MPO , myeloperoxidase ) Activity measurement

In order to confirm whether the fine spongy fraction affects the activity of infiltrating macrophages in the colon tissue of the chronic enteritis animal model, each mouse was euthanized after the end of the experiment, and the colon was excised by autopsy, and some colon tissue was collected. Myeloperoxidase (MPO, myeloperoxidase) activity was measured. MPO activity was measured using the Oxiselect ^TM myeloperoxidase peroxidation activity assay kit of Biolabs.

As a result, the average MPO activity of the fine spongiform administration group was 32.81±15.24 μU/mg, and the average MPO activity of the negative control group was 75.00±37.80 μU/mg. Confirmed (Fig. 5).

As a result of administering the fine spongiform fraction to chronic enteritis-inducing mice, clinical symptoms such as weight loss and diarrhea were alleviated, and macrophage activity in intestinal tissue was also reduced. Therefore, the fine spongy fraction can be usefully used in the treatment of chronic enteritis.

Claims (12)

The surface of the sea in fine thread (Ircinia sp.) Extracts or fractions thereof, the inflammatory bowel prevention or treatment a pharmaceutical composition of the disease, including as an active ingredient.
According to claim 1, wherein the extract is extracted with water, C ₁ to C ₂ lower alcohol or a mixed solvent thereof, a pharmaceutical composition for preventing or treating inflammatory bowel disease.
According to claim 2, The lower alcohol is ethanol or methanol, the pharmaceutical composition for preventing or treating inflammatory bowel disease.
The pharmaceutical composition for preventing or treating inflammatory bowel disease according to claim 1, wherein the fraction is prepared by a manufacturing method comprising the following steps:
1) distributing the extract of fine spongy spongy spongiformis in water and methylene chloride;
2) redistributing the distributed extract of step 1) with a hexane and methanol aqueous solution solvent; And
3) obtaining a fraction by fractionating the methanol layer obtained by redistribution in step 2) with a solvent in which water and methanol are mixed.
The pharmaceutical composition for preventing or treating inflammatory bowel disease according to claim 4, wherein the solvent in which water and methanol are mixed in step 3) is a mixture of water and methanol in a ratio of 1: 1 to 9.
The pharmaceutical composition of claim 1, wherein the inflammatory bowel disease is ulcerative colitis, Crohn's disease, Behcet's disease or chronic enteritis.
Health functional food for the prevention or improvement of inflammatory bowel disease, comprising the extract of Ircinia sp. or a fraction thereof as an active ingredient.
The health functional food for preventing or improving inflammatory bowel disease according to claim 7, wherein the extract is extracted with water, C ₁ to C ₂ lower alcohol or a mixed solvent thereof.
According to claim 8, The lower alcohol is ethanol or methanol, health functional food for preventing or improving inflammatory bowel disease.
The health functional food for preventing or improving inflammatory bowel disease according to claim 7, wherein the fraction is prepared by a manufacturing method comprising the following steps:
1) distributing the extract of fine spongy spongy spongiformis in water and methylene chloride;
2) redistributing the distributed extract of step 1) with a hexane and methanol aqueous solution solvent; And
3) obtaining a fraction by fractionating the methanol layer obtained by redistribution in step 2) with a solvent in which water and methanol are mixed.
The health functional food for preventing or improving inflammatory bowel disease according to claim 10, wherein the solvent in which water and methanol are mixed in step 3) is a mixture of water and methanol in a ratio of 1: 1 to 9.
According to claim 7, wherein the inflammatory bowel disease is ulcerative colitis, Crohn's disease, Behcet's disease or chronic enteritis, health functional food for preventing or improving inflammatory bowel disease.

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