KR20150083328A - Composition comprising an extract or a fraction of Daphne kamtschatica for preventing or treating inflammatory diseases - Google Patents

Abstract

The present invention relates to a pharmaceutical composition and a food composition comprising a Daphne kamtschatica extract and a fraction thereof as active ingredients for preventing, treating, and alleviating inflammatory disease. A Daphne kamtschatica extract and a fraction thereof of the present invention have no side effects, which are derived from natural substances used as a natural medicine for a long period of time, and inhibit generation of a nitrogen oxide (NO) and TNF-α related to an inflammatory factor. Therefore, a composition comprising the Daphne kamtschatica extract and the fraction thereof of the present invention can be effectively used for preventing or treating inflammatory disease.

Description

[0001] The present invention relates to a composition for preventing or treating an inflammatory disease,

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

Inflammation refers to the tissue response to an injury when external antigens, such as bacteria or viruses, enter the body from outside or when cells or tissue are damaged by some cause. Inflammatory cells release various inflammatory mediators such as active nitrogen and oxygen species that cause DNA damage in the initiation, enhancement of tumor. The inflammation is divided into acute inflammatory and chronic inflammatory. When chronic inflammatory reaction occurs, pre-inflammatory cytokine and chemokine are excessively released, causing damage to surrounding tissues and invasion of tumor cells, And promotes angiogenesis to help tumor growth.

Recently, with the development of molecular biology, attempts have been made to understand inflammatory diseases at the molecular level such as cytokines, and factors affecting these diseases are also being clarified one by one. Among these factors, nitric oxide (NO) is an inflammatory mediator that plays a role in maintaining homeostasis by damaging pathogenic DNA (Kou and Schroder, Ann Sur 221, 220-235, 1995). NO is generated from L-arginine by the three major NOSs, nNOS (neuronal NOS), eNOS (endothelial NOS) and iNOS (inducible NOS), which are isomers of NOS. Of these, NO produced by iNOS excessively reacts with superoxide to form peroxynitrite, which acts as a strong oxidizing agent and damages the cells, thereby activating macrophages by inflammatory stimuli . Macrophages activated by various stimuli and inflammatory mediators produce other inflammatory factors such as tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β) and prostaglandin E2 (PGE2) And NF-κB are known to be involved in chronic diseases such as inflammation, cancer, and arteriosclerosis (Lawrence et al., Nat Med., 7, 1291-1297, 2001).

Studies have been actively conducted to treat inflammatory diseases by modulating intracellular levels of factors involved in the inflammatory reaction. In the early days, attempts were made to control intracellular levels of the factors using chemical substances, but the disadvantage of severe side effects Recently, studies have been actively conducted to regulate intracellular levels of the above factors using natural-substance-derived substances having no or very low level of side effects.

On the other hand, Daphne kamtschatica is a plant that grows mainly in the slightly cooler places of the high mountains such as Mt. Halla, Mt. Jiri, Mt. Taebaek and Mt. The husk is used as a raw material of Korean paper and the toxic fruit is used as a medicine. Regarding the activity of extracts of Dumai mackerel, it is known about skin whitening activity (Korean Patent Laid-Open No. 10-2012-0003267), but the anti-inflammatory activity is not yet known.

Under these circumstances, the inventors of the present invention have made intensive efforts to find natural substances useful for the treatment of inflammatory diseases without causing adverse effects on the human body. As a result, it has been found that the extracts of Dumai nucifera or its fractions are useful for the prevention or treatment of inflammatory diseases And the present invention has been completed.

An object of the present invention is dume mulberry (Daphne The present invention also provides a pharmaceutical composition for preventing or treating an inflammatory disease comprising, as an active ingredient, an extract or a fraction thereof.

Another object of the present invention is to provide a food composition for preventing or ameliorating an inflammatory disease comprising an extract of Dumai nodule or a fraction thereof as an active ingredient.

In one aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating an inflammatory disease comprising Daphne kamtschatica extract or a fraction thereof as an active ingredient.

Specifically, the pharmaceutical composition for preventing or treating an inflammatory disease of the present invention may comprise a Dhammadoe extract or a fraction thereof.

The term " Daphne " kamtschatica "is a deciduous broad-leaved shrub of a red-bellied azalea bloom, distributed mainly in Korea (Jeonnam, Gangwon, Pyongbuk, Hambuk), Sakhalin, Kamchatka, Amur, It is 30-40 cm high and has no hairs on the branches. Leaves are alternate phyllotaxis and long eggs are inverted. The ends are sharp or thick. The bark is used as a substitute for paper raw materials and ropes, and has been mainly used for ornamental purposes.

In the present invention, the term " Dumai mackerel extract " means an extract obtained by extracting Dumai mackerel. The Dumai mushroom extract is prepared by dissolving the crushed Dumai mulberry leaf in water having a volume of about 2 to 20 times, preferably about 3 to 5 times the dry weight, of ₁ to 4 carbon atoms (C ₁ to 4) such as methanol, ethanol, ₄ ) or a mixed solvent having a mixing ratio of about 1: 0.1 to 1:10 of them is used as an elution solvent, and the extraction temperature is 20 to 100 ° C, preferably room temperature, and the extraction period is about Extraction can be carried out using an extraction method such as hot water extraction, cold extraction, reflux cooling extraction or ultrasonic extraction for 12 hours to 4 days, preferably 3 days, but a method of extracting a substance having an activity for preventing or treating inflammatory diseases And can be used without limitation. Preferably, the extract is continuously extracted one to five times by cold extraction, filtered under reduced pressure, and the filtrate is concentrated under reduced pressure at 20 to 100 ° C, preferably at room temperature, in a vacuum rotary condenser to be dissolved in water, a lower alcohol or a mixed solvent thereof However, the present invention is not limited thereto, and it may be a dried product obtained by drying an extract, a diluted solution of the extract, a concentrate, or an extract of the extract, as long as it can exhibit the activity of preventing or treating the inflammatory disease of the present invention , Or both of these controlled products or tablets. The extract can be extracted from various organs of natural, hybrid, and variant plants, and can be extracted from, for example, root, top, stem, leaf, fruit or plant tissue culture, It can be extracted from leaves and stems.

The term " fraction " in the present invention means a product obtained by a fractionation method for separating a specific component or a specific group from a mixture containing various constituents. The dumaiwa extract of the present invention can be obtained by suspending Dumai michiensis extract and then fractionating it using a polar solvent such as water, methanol or ethanol or a nonpolar solvent such as hexane or ethyl acetate to obtain a polar solvent fraction and a nonpolar solvent fraction, respectively have. Specifically, after suspending the dume mulberry crude extract or the like of distilled water, from about 1 to 100 times of the suspension, preferably from about 1 to a volume of five times with water, alcohol or chloroform with a carbon number of ₁ (C 1) to ₄ (C 4) , A polar or nonpolar solvent such as ethyl acetate, hexane, butanol, or a mixed solvent thereof is added, and the polar or non-polar solvent soluble layer is extracted and separated from 1 to 10 times, preferably 2 to 5 times have. Further, a conventional fractionation process may be performed (Harborne JB Plant Pathology, 1998, 3rd Ed. P6-7). More specifically, the Dumeye mackerel extract was suspended in water and then extracted continuously with water, methanol or a mixed solvent thereof (10: 90 → 100: 0 (v / v)) as a solvent. (V / v) methanol as a solvent, but the present invention is not limited thereto (Production Example).

The term " inflammatory disease " in the present invention means a disease in which inflammation is the main lesion, and includes, but is not limited to, edema, allergy, asthma, conjunctivitis, periodontitis, rhinitis, otitis, , Ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, acne spondylitis, rheumatic fever, lupus, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, shoulder circumference, tendonitis, mildew, myositis, hepatitis, cystitis, nephritis , Sjogren's syndrome, and multiple sclerosis, but the present invention is not limited thereto.

In the present invention, the term "prevention" means any action that inhibits or delays the onset of an inflammatory disease upon administration of the composition, and "treatment" .

The pharmaceutical composition comprising the Dhammoeae extract of the present invention or a fraction thereof may further comprise an appropriate carrier, adduct or diluent conventionally used in the production of a pharmaceutical composition. At this time, the content of the Dumai mateo extract or its fraction contained in the composition is not particularly limited, but it may include 0.001% by weight to 99% by weight, preferably 0.01% by weight to 50% by weight based on the total weight of the composition have.

The pharmaceutical composition may be any one selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, solutions, emulsions, syrups, sterilized aqueous solutions, nonaqueous solvents, suspensions, emulsions, And may be oral or parenteral formulations of various forms. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may contain one or more excipients such as starch, calcium carbonate, sucrose or lactose lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like may also be 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 non-aqueous solvent and the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

In one embodiment of the present invention, the extract of Dumai michiensis or its fractions inhibits the production of NO (Example 3) without cytotoxicity (Example 2), and inhibits the production of TNF-a, an inflammation-related factor (Example 4). In particular, it was confirmed that the inhibitory activity of NO and TNF-α production was significantly higher in the treatment groups of fractions 9 and 10.

Accordingly, the extract of Dumai mukuba or fractions thereof of the present invention can be usefully used for the prevention or treatment of inflammatory diseases.

In another aspect, the present invention provides a method of preventing or treating an inflammatory disease, comprising the step of administering the pharmaceutical composition to a suspected individual of an inflammatory disease.

In the present invention, the suspected individual of the above-mentioned inflammatory disease means all the animals including humans who have developed or can develop the disease. By administering the pharmaceutical composition of the present invention to suspected individuals having an inflammatory disease, have. The pharmaceutical composition and inflammatory diseases are as described above.

The term "administering" as used herein means introducing the pharmaceutical composition of the present invention into a suspected individual of an inflammatory disease by any suitable method, and the administration route includes various routes of oral or parenteral administration ≪ / RTI >

The pharmaceutical composition of the present invention may be administered in a pharmaceutically effective amount.

The term "pharmaceutically effective amount" as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will vary depending on the species and severity, age, sex, The type of drug, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of the treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and can be easily determined by those skilled in the art.

The pharmaceutical composition of the present invention is not particularly limited as long as it is an object for an inflammatory disease, and any of them can be applied. For example, any non-human animal such as a monkey, a dog, a cat, a rabbit, a guinea pig, a rat, a mouse, a cattle, a sheep, a pig or a goat can be used. Subcutaneous, intraperitoneal, intrapulmonary, and intranasal, and may be administered by a suitable method, including localized administration, if necessary, for localized treatment. The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and body 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, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

Suitable total daily doses may be determined by the treatment within the scope of sound medical judgment and are generally in the range of 0.001 to 1000 mg / kg, preferably 0.05 to 200 mg / kg, more preferably 0.1 to 100 mg / kg can be administered once or several times a day.

In another aspect, the present invention provides a food composition for preventing or ameliorating an inflammatory disease, which comprises an extract of Dhammayuka or a fraction thereof as an active ingredient.

The above-mentioned Dumai mackerel, its extracts, fractions and inflammatory diseases are as described above.

The term "improvement" in the present invention is intended to mean all kinds of diseases, such as inflammatory diseases, which are prevented or treated by using a composition comprising Dumaiabea extract or a fraction thereof as an active ingredient, .

Specifically, the extract of the present invention or a fraction thereof may be added to a food composition for the purpose of preventing or improving an inflammatory disease.

The food composition of the present invention may be in the form of pills, powders, granules, infusions, tablets, capsules or liquid preparations, and there is no particular limitation on the kind of food to which the Dumasi mica extract of the present invention or its fractions can be added , For example, various drinks, gum, tea, vitamin complex, and health supplement foods.

 The food composition may be supplemented with other ingredients in addition to the Dumai mateobokus extract or the fraction thereof, and the kind thereof is not particularly limited. For example, it may contain various herbal medicine extracts, food-acceptable food-aid additives or natural carbohydrates such as ordinary food, but is not limited thereto.

 The term "food supplementary additive " in the present invention means a component which can be added to foods in a supplementary manner, and is appropriately selected and used by those skilled in the art as added to produce health functional foods of each formulation. Examples of food-aid additives include flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, , a pH adjusting agent, a stabilizer, a preservative, a glycerin, an alcohol, and a carbonating agent used in a carbonated beverage. However, the types of the food auxiliary additives of the present invention are not limited by the above examples.

Examples of the natural carbohydrate include monosaccharides such as glucose and fructose; Disaccharides such as maltose, sucrose and the like; And polysaccharides such as dextrin and cyclodextrin and sugar alcohols such as xylitol, sorbitol and erythritol. In addition to the above, natural flavorings (such as tau mart), stevia extracts (rebaudioside A, Etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used.

The food composition of the present invention may include a health functional food. The term " health functional food " as used in the present invention refers to foods prepared and processed in the form of tablets, capsules, powders, granules, liquids, and rings using raw materials and ingredients having useful functions in the human body. Here, the term "functionality" means that the structure and function of the human body are controlled to obtain nutritional effects or effects useful for health use such as physiological actions. The health functional food of the present invention can be manufactured by a method commonly used in the art and can be prepared by adding raw materials and ingredients which are conventionally added in the art. Also, unlike general medicine, there is an advantage that there is no side effect that can occur when a medicine is used for a long time by using food as a raw material, and it is excellent in portability.

The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). In general, the dumpling extract of the present invention or a fraction thereof may be added in an amount of 1 to 50% by weight, preferably 5 to 10% by weight, of the raw material composition, but is not limited thereto. However, in the case of long-term ingestion intended for health and hygiene purposes or for the purpose of controlling health, the amount can also be used in the above-mentioned range.

There is no particular limitation on the kind of the food. Examples of the food to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include health functional foods in a conventional sense.

The extract of Dumai mateobee or a fraction thereof of the present invention is derived from a natural product which has been used for a long time as a natural medicinal substance and has no side effects and inhibits the production of nitrogen oxides (NO) and TNF-a which are inflammation-related factors. Therefore, The compositions of the present invention comprising fractions can be usefully used in the prevention or treatment of inflammatory diseases.

FIG. 1 is a graph showing the results of MPME (Medium Pressure Liquid chromatography) analysis of the extracts of Daphnia amurensis and stem methanol.
FIG. 2 is a graph showing the results of analysis of PDA (Photodiode-Array detection) and MS (Mass Spectrometry) using UPLC (Ultra Performance Liquid Chromatography) fractions of fractions of the leaves of Daphnia amurensis and stem methanol (fractions 1 to 13).
FIG. 3 is a graph showing the results of analysis of fraction (fractions 1 to 13) of the extracts of Daphnia amurensis and stem methanol by UV and CAD (Charged Aerosol Detector).
4 is a graph showing the cell survival rate of fractions of cuttlefish leaves and stem methanol extract (fractions 1 to 13).
FIG. 5 is a graph showing the inhibition rate of nitric oxide (NO) formation in the fractions of cuttlefish leaves and stem methanol extract (fractions 1 to 13).

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

Manufacturing example : Dummawood  Leaves, stem extracts and Fraction  Produce

The present inventors dried the shrub leaves and 30 g of the stem, which had been collected and dried in Jecheon, Chungcheongbuk-do, by natural drying (shade) using a dryer (50-55 ° C), and pulverized to a size of about 1 cm. Based on the dry weight of the pulverized powder sample, 1 L of methanol was added and extracted at room temperature. Then, after filtration and concentration under reduced pressure, 4.58 g of a methanol extract of Daphne leaf and stem was obtained. In the subsequent experiments, the obtained extracts of Dumadilla leaves and stem methanol were named as total extracts.

Further, a column (20 mm × 250 mm; Resin: Zeoprep C18, 10 μm) was attached to a medium pressure liquid chromatography (MPLC) apparatus (sepbox 2D 5000) Was loaded in an amount of 2 g. At this time, water / methanol [10: 90 -> 100: 0 (v / v)] was used as a solvent and the elution rate was 40 ml / min and detected at a wavelength of UV 200-400 nm. The analysis time was 150 minutes, and fractions 1 to 13 were obtained as shown in Table 1 (Table 1, Fig. 1).

Dummawood MeOH (%) Analysis time (min) Weight (mg) Fraction 1 0-14 0-18 796.1 Fraction 2 14-22 18-27 105.2 Fraction 3 22-27 27-34 87.4 Fraction 4 27-32 34-40 162.2 Fraction 5 32-42 40-51 178.8 Fraction 6 42-48 51-58 112.2 Fraction 7 48-51 58-62 150.9 Fraction 8 51-60 62-72 121.8 Fraction 9 60-64 72-78 68.1 Fraction 10 64-70 78-84 115 Fraction 11 70-83 84-100 69.1 Fraction 12 83-89 100-107 60.8 Fraction 13 89-100 108-134 60.4

Example  One: Dummawood  Leaves, stem extracts and Fraction  analysis

In order to examine the active fractions of the methanol extract and the active fractions of Dumai mulberry obtained in the above Preparation Examples, they were analyzed by UPLC (ultra performance liquid chromatography) which is liquid chromatography.

First, for the UPLC analysis, the methanol extract and the active fractions obtained from the above Preparation Example were once filtered with a 0.25 占 퐉 membrane filter for UPLC. A column (Waters BEH C18 column, 2.1 x 100 mm, 1.7 mu m) was loaded on a UPLC instrument (Waters UPLC-QTOF-MS) and each fraction filtered was loaded with 5 uL aliquots. At this time, acetonitrile + 0.1% formic acid / water + 0.1% formic acid [10: 90 → 100: 0 (v / v)] was used as a solvent and the elution rate was 0.4 ml / min. ), Mass spectrometry (MS) and collision-activated decomposition (CAD) (Fig. 2 and Fig. 3).

Example  2: Dummawood  Extract or its The fraction  Analysis of effect on cell survival rate

Raw 264.7 cells were cultured in DMEM supplemented with 5% Fetal Bovine Serum (Dulbecco's Modified Eagle Medium, Gibco) to 1 × 10 5 cells / Ml / ml, and then 100 [mu] l of the solution was inoculated into a 96-well plate. After 4 hours, the samples were each treated at a concentration of 50 μg / ml. After culturing for 24 hours, 10 쨉 l of 5 ㎎ / ml MTT solution was added to each well, followed by further incubation for 4 hours. Then, the supernatant was removed, 100 μl of DMSO was added, and the absorbance was measured at 570 nm. Cell viability was calculated according to the following equation 1 with a negative control value of 0.2% DMSO treated as 100%.

[Equation 1]

As a result, the cell survival rate was 90% or more, except for the fractions 11 (73.06 ± 5.0%) and fraction 13 (73.98 ± 8.63%) in the above production example. Indicating no cytotoxicity (Fig. 4).

Example  3: Dummawood  Extract or its Fraction  Nitrogen oxides ( NO ) Production inhibitory effect

In order to investigate the inflammation inhibitory effect of Dumai mackerel extract or its fractions, the inhibitory rate of lipopolysaccharide (LPS) - induced nitric oxide (NO) production in Raw264.7 cells, a macrophage of mice, was measured.

Specifically, 5% fetal bovine serum (Fetal Bovine Serum) was added to a phenol-Red free DMEM medium (Dulbecco's Modified Eagle Medium, Gibco), and cells were suspended at a concentration of 2.5 x 10 ⁵ cells / , And 200 [mu] L of the solution was inoculated into a 96-well plate. After inducing cell adhesion for 4 hours, the samples were treated with 50 μg / ml of each and incubated for 1 hour, followed by treatment with 0.5 μg / ml of LPS and further incubation for 24 hours. Then, 100 쨉 l of the supernatant was collected and dispensed into a new 96-well plate. The same amount of a grease reagent (Sigma) was added thereto and reacted at room temperature for 10 minutes. Thereafter, a microplate reader (Bio-Rad The absorbance was measured at a wavelength of 540 nm. Calibration curves were prepared using sodium nitrite. The amount of nitrogen oxides in the culture was calculated on the basis of this, and the inhibition rates of each sample were calculated using the amount of nitrogen oxides produced in the LPS-treated group as 100%.

As a result, it was confirmed that the production of NOx was significantly increased in the cells treated with LPS alone, whereas the amount of production of NOx was decreased in the cells treated with LPS after pretreatment of the extract or its fraction (Fig. 5). Particularly, it was confirmed that fractions 9 to 13 in the above-mentioned production examples showed a NO generation inhibiting rate of 40% or more.

Example  4: Dummawood  Extract or its Fraction  Tumor necrosis factor-alpha ( tumor  necrosis factor - alpha , TNF -α) production inhibitory effect

In order to examine the effect of Dumai mackerel extract or its fractions on the production of TNF-α, the amount of TNF-α in the cell supernatant treated as in Example 3 above was confirmed using a TNF-α ELISA kit. The NO production inhibition rate and the TNF-? Production inhibition rate of fractions 9-13 of Dumai mulberry and the extract (total) of Dumai mulberry obtained in the above-mentioned Examples, which were judged to be effective fractions, were compared.

As a result, as shown in the following Table 2, it was confirmed that the total extract of Duma mallow had a production inhibitory rate of NO and TNF-α of 30% or more, and fractions 9 to 13 thereof showed higher NO and TNF- α production inhibition rate. In particular, it was confirmed that fraction 9 and fraction 10 had the highest inhibitory rate of production.

Samples (50 mu g / ml) NO  Inhibition rate
(%, Ave ± SD ) TNF -α inhibition rate
(%, Ave ± SD ) DMSO + LPS treated group 0.00 ± 3.19 0.00 ± 11.29 Dumedak extract (Total) + LPS treated group 30.83 + - 0.43 32.40 ± 1.52 Dumedak fraction (No. 9) + LPS treated group 85.26 ± 1.70 70.83 + - 1.42 Dumedak fraction (No. 10) + LPS treated group 88.12 + - 4.04 80.69 + - 2.66 Dumedak fractions (No. 11) + LPS treated group 59.25 + - 3.19 49.44 + 0.76 Dumedak fraction (No. 12) + LPS treated group 51.43 + - 2.76 38.37 ± 2.18 Dumedak fractions (No. 13) + LPS treated group 50.83 + - 0.21 32.47 ± 3.32

From the above results, it was confirmed that the extract of Dumai mudberry or its fractions according to the present invention is effective for prevention, treatment or improvement of inflammatory diseases.

Examples of formulations for the composition of the present invention are illustrated below, but are not limited thereto.

Formulation example  1: Preparation of pharmaceutical preparations

The pharmaceutical preparations containing the Dhammoe leaf extract or fractions of the present invention were prepared according to a conventional method as follows.

Formulation example  1-1: Sanje  Produce

2 mu m < 2 >

Lactose 1 g

The above components were mixed and packed in airtight bags to prepare powders.

Formulation example  1-2: Preparation of tablets

100 mg < RTI ID = 0.0 >

Corn starch 100 mg

Lactose 100 mg

2 mg of magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

Formulation example  1-3: Preparation of capsules

100 mg < RTI ID = 0.0 >

Corn starch 100 mg

Lactose 100 mg

2 mg of magnesium stearate

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

Formulation example  1-4: Manufacture of rings

Each of the dumpling extracts or fractions prepared in Preparation Example 1 g

Lactose 1.5 g

Glycerin 1 g

0.5 g of xylitol

After mixing the above components, they were prepared so as to be 4 g per one ring according to a conventional method.

Formulation example  1-5: Preparation of granules

150 mg < / RTI > of the Dumai mushroom extract or fraction, respectively,

Soybean extract 50 mg

200 mg of glucose

600 mg of starch

After mixing the above components, 100 mg of 30% ethanol was added and the mixture was dried at 60 캜 to form granules, which were then filled in a capsule.

Formulation example  2: Preparation of Health Functional Food

Healthy functional foods containing the dumai moohwadoe extract or fractions of the present invention were prepared according to the conventional method as follows.

Formulation example  2-1: Manufacture of Flour Food

0.5 to 5.0 weight of Dumai mackerel extract or fraction prepared in the preparation example of the present invention was added to wheat flour, and bread, cake, cookie, cracker and noodle were prepared using this mixture to prepare a food for health promotion.

Formulation example  2-2: Dairy products ( dairy products )

5 to 10% by weight of Dumai mackerel extract or fraction prepared in the Preparation Example of the present invention was added to milk and various dairy products such as butter and ice cream were prepared using the milk.

Formulation example  2-3: Solar  Produce

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh. Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer. The dried Dumai mackerel extract or fractions prepared in the preparation examples of the present invention were concentrated under reduced pressure in a vacuum concentrator, dried by spraying and dried in a hot-air drier, and pulverized to a size of 60 mesh with a pulverizer to obtain a dry powder.

The grains, seeds, and dry powder of the extract or fraction of Dumai mushroom, prepared above, were blended in the following proportions.

Cereals (30% by weight of brown rice, 15% by weight of yulmu, 20% by weight of barley)

Seeds (7% by weight of perilla, 8% by weight of black beans, 7% by weight of black sesame seeds)

Dried powder (3% by weight) of Dumai mushroom extract or fraction,

(0.5% by weight),

(0.5% by weight)

Formulation example  3: Manufacturing of beverages

Formulation example  3-1: Health drink  Produce

1000 mg < RTI ID = 0.0 >

Citric acid 1000 mg

100 g of oligosaccharide

Plum concentrate 2 g

Taurine 1 g

Purified water was added to a total of 900 ml

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 ° C for about 1 hour. The resulting solution was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, It is used in the production of the health beverage composition of the invention.

Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.

Formulation example  3-2: Manufacture of vegetable juice

Healthy vegetable juices were prepared by adding 5 g of the Dumai mudberry extract or fractions prepared in the preparation examples of the present invention to 1,000 ml of tomato or carrot juice.

Formulation example  3-3: Manufacture of fruit juice

The fruit juice for health promotion was prepared by adding 1 g of each of the Dumai matsutake extracts or fractions prepared in the preparation examples of the present invention to 1,000 ml of apple or grape juice.

From the above description, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are illustrative in all aspects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention, rather than the above detailed description, and all changes or modifications derived from the equivalents thereof will be included in the scope of the present invention.

Claims (17)

Daphne The present invention relates to a pharmaceutical composition for preventing or treating an inflammatory disease, which comprises an extract or a fraction thereof as an active ingredient.
The method of claim 1, wherein the extract is water, carbon number ₁ (C 1) to 4 will be extracted by using alcohol or a mixed solvent of (C ₄₎ composition.
The composition of claim 1, wherein the extract is extracted with methanol.
The method of claim 1, wherein the fraction by fractionation using water, carbon number ₁ (C 1) to ₄ (C 4) alcohols, chloroform, ethyl acetate, hexane, butanol, or a mixed solvent of methanol extract of dume mulberry ≪ / RTI >
The composition according to claim 1, wherein the fraction is obtained by fractionating the methanol extract of Dumai mukuba with water, methanol or a mixed solvent thereof.
The composition of claim 1, wherein said fraction is obtained by fractionating 60 to 100% (v / v) methanol with a solvent.
2. The composition of claim 1, wherein the Daphne kamtschatica is a Dhammateae stem, a leaf or a mixture thereof.
The composition of claim 1, further comprising a pharmaceutically acceptable carrier, excipient or diluent in said Dumai mateo extract or fraction thereof.
2. The composition of claim 1, wherein the Dhumitrich extract or fraction thereof reduces the production of NO (nitric oxide) or TNF-alpha (Tumor necrosis factor-ahpha) cytokines.
The method of claim 1, wherein the inflammatory disease is selected from the group consisting of edema, allergy, asthma, conjunctivitis, periodontitis, rhinitis, otitis, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, Wherein the composition is selected from the group consisting of lupus, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, shoulder circumflex, tendinitis, hay fever, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome and multiple sclerosis.
Daphne The present invention relates to a food composition for preventing or ameliorating an inflammatory disease comprising an extract or a fraction thereof as an active ingredient.
12. The method of claim 11, wherein the extract is water, carbon number ₁ (C 1) to 4 will be extracted by using alcohol or a mixed solvent of (C ₄₎ composition.
12. The composition of claim 11, wherein the extract is extracted with methanol.
12. The method of claim 11, wherein the fraction by fractionation using water, carbon number ₁ (C 1) to ₄ (C 4) alcohols, chloroform, ethyl acetate, hexane, butanol, or a mixed solvent of methanol extract of dume mulberry ≪ / RTI >
12. The composition of claim 11, wherein the fraction is obtained by fractionating the methanol extract of Dhammayuk tea with water, methanol or a mixed solvent thereof.
12. The composition of claim 11, wherein said fraction is obtained by fractionating 60 to 100% (v / v) methanol with a solvent.
12. The method of claim 11, wherein the inflammatory disease is selected from the group consisting of edema, allergy, asthma, conjunctivitis, periodontitis, rhinitis, otitis, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, Wherein the composition is selected from the group consisting of lupus, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, shoulder circumflex, tendinitis, hay fever, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome and multiple sclerosis.

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