KR20120105403A - Composition for inhibiting the degranulation of mast cell comprising plant extracts - Google Patents

Abstract

PURPOSE: A composition containing plant extract for suppressing granule secretion of mast cells is provided to effectively prevent and treat allergic diseases and to be used in a food composition and cosmetic composition. CONSTITUTION: A composition for preventing or treating allergic diseases contains Salix koreensis extract as an active ingredient. The extract is isolated from Salix koreensis stems using methanol. A method for preparing Corydalis speciosa Maxim. extract comprises: a step of pulverizing Salix koreensis; a step of extracting pulverized Salix koreensis with water, organic solvent, or a mixture solvent thereof and drying; and a step of dissolving the dried material with water, organic solvent, or a mixture thereof.

Description

Composition for inhibiting the degranulation of mast cell comprising plant extracts

The present invention is Japanese larch ( Larix) leptolepis ), elm ( Maackia) amurensis ), massif bag (Corydalis speciosa), straw bud ( Agrimonia) pilosa ), Ginseng ( Codonopsis pilosula ), Willow ( Salix) koreensis ), viburnum ( Viburnum wrightii ), Platycarya strobilacea ), Weasel ( Amorpha) fruticosa ), Pyrus pyrifolia , and non-vulture ( Lespedeza) cuneata ), potentilla cryptotaeniae ), and eyelids ( Salix graciliglans ) relates to a composition for inhibiting granule secretion of mast cells comprising at least one extract selected from the group consisting of active ingredients.

To treat allergic diseases, antihistamines, sympathetic stimulants, antiallergic drugs, theophylline (Theophylline), and steroids are used a lot.

Antihistamines are relatively safe drugs and are most commonly used as a popular remedy for allergic diseases. Antihistamines have a similar effect of blocking histamine action, but the side effects are different from each other. The most common side effects are tremor and hypnosis, and other factors such as excitement, irritability, insomnia, anxiety, dizziness, nightmares and hallucinations Excitatory effects can also occur, and there have been reports of unusual excitatory effects, particularly in children.

While antihistamines are fast-acting by directly antagonizing histamine action, antiallergic agents (Cromolyn Sodium (DSCG)) inhibit the production and maintenance of allergens such as histamine and leukotriene from cells involved in allergic reactions. Therefore, antihistamines are mainly used to suppress the symptoms after an allergy, whereas anti-allergic drugs are used as a prophylactic agent to suppress the occurrence of allergic reactions. Therefore, it cannot be used as a drug to suppress acute symptoms.

In allergic diseases, steroid preparations have anti-inflammatory and immunosuppressive effects, making them very useful in lethal acute allergic reactions such as anaphylaxis, asthma and exfoliative dermatitis. However, long-term administration requires attention because there are side effects that patients may be aware of, such as mental disorders such as nervousness, insomnia and anxiety, obesity and skin abnormalities, and risks such as glaucoma, cataracts, gastritis and osteoporosis.

As described above, the synthetic medicines currently used as treatments for allergic diseases have the disadvantage of improving the disease while causing serious side effects.

Therefore, the development of products that suppress the secretion of mast cell granules using natural plant extracts is a natural product, so it is safe for consumers to approach the final product development. , The inventors of the Japanese larch (Larix leptolepis), Elm (Maackia amurensis), Massif firebags (Corydalis speciosa), straw sprouts (Agrimonia pilosa), Ginseng (Codonopsis pilosula), Bud tree(Salix koreensis), Mountain viburnum (Viburnum wrightii), Oyster tree (Platycarya strobilacea), WeaselAmorpha fruticosa), Pear tree (Pyrus pyrifolia), Non-repair (Lespedeza cuneata), Hydrangea (Potentilla cryptotaeniae), And snowballs (Salix graciliglansThe extract of)) has an effect of inhibiting the secretion of granules of mast cells, and in particular, the Japanese larch extract has completed the present invention by confirming through an animal experiment that the effect of improving allergic diseases such as atopic dermatitis.

The purpose of the present invention is Japanese larch ( Larix) leptolepis ), elm ( Maackia) amurensis), massif bag (Corydalis speciosa), straw bud ( Agrimonia) pilosa ), Codonopsis pilosula ) Willow ( Salix) koreensis ), viburnum wrightii , oyster tree ( Platycarya) strobilacea ), Weasel ( Amorpha) fruticosa ), Pyrus pyrifolia ), vulgaris ( Lespedeza cuneata), to provide a water Potentilla Fragarioides (Potentilla cryptotaeniae), and eye pussy willow granule secretion composition for inhibiting mast cell comprising one or more extracts selected from the group consisting of (Salix graciliglans) as an active ingredient.

An object of the present invention to provide a pharmaceutical composition for the prevention or treatment of allergic diseases comprising the composition.

It is an object of the present invention to provide a food composition for improving or preventing allergic diseases comprising the composition.

An object of the present invention is to provide a quasi-drug composition for improving or preventing allergic diseases comprising the composition.

An object of the present invention is to provide a cosmetic composition for improving or preventing allergic diseases comprising the composition.

The purpose of the present invention is Japanese larch ( Larix) leptolepis ), elm ( Maackia) amurensis), massif bag (Corydalis speciosa), straw bud ( Agrimonia) pilosa ), Codonopsis pilosula ) Willow ( Salix) koreensis ), viburnum wrightii , oyster tree ( Platycarya) strobilacea ), Weasel ( Amorpha) fruticosa ), Pyrus pyrifolia ), vulgaris ( Lespedeza pulverizing at least one selected from the group consisting of cuneata ), Potentilla cryptotaeniae and Salix graciliglans , extracting the ground matter with a solvent selected from water, an organic solvent or a mixed solvent thereof and drying the same. It provides a method for producing an extract of the 13 kinds of plants comprising the step of, and adding a solvent selected from water, an organic solvent or a mixed solvent thereof to the dried material.

In order to achieve the above object, in one aspect the present invention is Japanese larch ( Larix) leptolepis), elm ( Maackia) amurensis ), massif bag (Corydalis speciosa), straw bud ( Agrimonia) pilosa ), Codonopsis pilosula ), willow ( Salix koreensis ), viburnum ( Viburnum wrightii ), oyster bark ( Platycarya strobilacea ), weasel ( Amorpha) fruticosa ), Pyrus pyrifolia ), vulgaris ( Lespedeza cuneata ), potentilla cryptotaeniae ), and eyelids ( Salix graciliglans ) provides a composition for inhibiting granule secretion of mast cells comprising at least one extract selected from the group consisting of.

The present inventors used Japanese leaves ( Larix) leptolepis ), elm ( Maackia) amurensis), massif bag (Corydalis speciosa), straw bud ( Agrimonia) pilosa ), Codonopsis pilosula ) Willow ( Salix) koreensis ), viburnum wrightii , oyster tree ( Platycarya) strobilacea ), Weasel ( Amorpha) fruticosa ), Pyrus pyrifolia ), vulgaris ( Lespedeza cuneata), water Potentilla Fragarioides (Potentilla cryptotaeniae), and eyes pussy willow (Salix graciliglans ) extracts have an effect of inhibiting the secretion of granules of mast cells. More specifically, the present inventors have tested the inhibitory effect of beta-hexosaminidase release according to the treatment of the plant extracts, the plant extracts have the effect of inhibiting mast cell granule secretion of 50% or more It was confirmed (Table 2 and FIG. 2).

In addition, the amount of IgE in the serum was significantly reduced as a result of applying the Japanese larch extract, which showed the best inhibitory effect of β-hexosaminidase release among the 13 plant extracts, to mice inducing atopic dermatitis. And (Fig. 3) the increase in the number of mast cells is significantly inhibited (Fig. 4), it was confirmed that the Japanese larch extract has an excellent effect of relieving allergic diseases including atopic dermatitis.

As a result, according to a specific embodiment of the present invention, it was confirmed that the 13 kinds of plant extracts have an inhibitory effect on the secretion of mast cell granules and thereby alleviate allergic diseases.

In the present invention, a 'mast cell' is a cell involved in an inflammatory reaction, when an IgE antibody is activated and crosslinked by an external antigen or allergen, various various signaling mechanisms (signalling) Pathway releases histamine, beta-hexosaminidase, and pro-inflammatory agents such as leukotriene to plasma membranes. Inflammatory substances released in this way act to intensify the inflammation by activating the receptors and cytokines that cause inflammation. In particular, the release of these substances in mast cells is closely associated with allergic diseases. Allergic diseases are commonly genetically / immunologically related to IgE antibodies and are known to be caused by sensitization by contact with or exposure to antigenic factors, ie environmental and mental factor factors. Among them, immunological factors are currently targeted for treatment, and in immunological terms, they can be applied as therapeutic agents for allergic diseases by controlling the release of these allergens into the extracellular membrane.

In the present invention, the term 'granular secretion of mast cells' refers to the secretion of pro-inflammatory substances trapped in the granules of the mast cells, and the pro-inflammatory substances include histamine, beta-hexosaminidase (β-hexosaminidase), Leukotriene and the like.

As used herein, the term 'inhibition of granule secretion of mast cells' refers to blocking or reducing the efficiency of secretion of inflammatory mediators collected in the granules of mast cells. As a result, it is possible to ameliorate, prevent, and treat various allergic diseases induced by inflammation-causing substance secretion through inhibiting granule secretion of mast cells.

In the present invention, 'beta-hexosaminidase' is trapped in granules of mast cells and then released into the outer membrane when the cells are activated and used as a degranulation marker of mast cells. (Cheong H, Choi EJ, Yoo GS, Kim KM, Ryu SY, Ho C. Planta Med 1998; 64; 577). According to a specific embodiment of the present invention, the beta-hexosaminidase (β-hexosaminidase) inhibition experiment according to the treatment of the 13 kinds of plant extracts in order to determine the inhibitory effect of the granule secretion of mast cells of the plant extract using this property As a result, it was confirmed that the release of beta-hexosaminidase (β-hexosaminidase) is significantly inhibited.

In the present invention, the Japanese leaves ( Larch) leptolepis ) 'is about 30m high, leaves are row-shaped, gathered on short branches of about 40 ~ 50, and its length is about 2 ~ 3cm. The flower is a male flower, the male ear is long oval, and the female ear is wide egg-shaped and blooms around May. Fruits are coniferous, broad egg-shaped, about 2 ~ 3.5cm long, ripened yellowish brown in September ~ October. The one-year-old branch is slightly thin and reddish brown, and the convex scale fragment is reversed at the end. The Japanese leaf larch extract can be extracted from various organs of natural, hybrid, and variegated plants, and can be extracted from plant tissue culture, according to a specific embodiment of the present invention. Hereinafter, unless the problem in particular in the present specification, larch also refers to the Japanese larch.

In the present invention, ' Maackia amurensis ) 'is a broad-leaved tall tree with leaves, about 10 to 20 meters high. The inside of the log is black in color. Leaves are alternately attached, odd feather-shaped double leaves of about 5-11 leaflets. The leaf leaves are oval-shaped chicken eggs, fringes and hairless on both sides of the leaves. At the end of the branches in August, a single flower is formed, and many white butterfly-shaped flowers bloom. Fruits are pods, flat, long oval, about 5 cm long. Several seeds are contained in it, and it starts around October. The elm extract may be extracted from various organs of natural, hybrid, and variegated plants, and can be extracted from plant tissue culture, and extracted from leaves or stems according to a specific embodiment of the present invention.

In the present invention, the 'fire bulge ( Corydalis speciosa ) is a prefecture color family distributed in Korea, Japan, northeastern China, Heilong River and Ussuri River, and grows in wet mountains. The height is about 40cm. The main stem is hollow, grows straight, branches are split, and the whole is white. The leaves are shifted and split into two pieces of feathers. The length is about 10-15cm and the butterfly is about 4-6cm. Forked pieces are egg-shaped, split into feathers, and the last forked is a long oval, strip-shaped, with a sharp tip. Flowers are yellow in April-June, about 2-2.3cm long, hang on inflorescence. Pho is oval lanceolate and sometimes cracked. Corolla is split into lip shape at the end and honeybag at the bottom. There are about 6 surgeries, and then they are divided again. Fruits are capsules, rows, seeds are round, black, and thin, concave dots are scattered on the outside. The massif bag extract can be extracted from a variety of organs of natural, hybrid, mutant plants and can be extracted from plant tissue culture, according to a specific embodiment of the present invention was extracted from the outpost.

In the present invention, the ' sadweed ( Agrimonia pilosa ) is a perennial plant of the dicotyledonous rosacea rosaceae and is distributed in Korea, Japan, China, India, Himalayas, Mongolia, Amur, Siberia, Europe and Sakhalin. It lives in the grass or on the roadside. The stem is about 30-100cm in height and has hairs all over. The leaf is alternately pinnate leaf composed of about 5-7 small leaves. The small leaves are not uniform in size, but the three on the end are similar in size and become smaller toward the bottom, and have a long oval shape or an upside down egg with narrow ends and serrated edges. There are about 1 pair of chin leaves at the bottom of petiole, chin leaves are half-moon-shaped, pointed at the end, and there are several large jagged teeth at the bottom edge. Flower is yellow in June-August and runs in the form of gunshot inflorescence at the end of stem and branch. The calyx is about 3mm long, has a vertical line, and the upper part is divided into about 5 pieces and has a hook-like hair on the outside. Petals are about 5, the eggs are upside down or round, and the surgery is about 5-10. Fruits are achene, about 3mm long, wrapped in calyx, which adheres well to the object because of hairs like hooks in calyx. The straw herb extract may be extracted from various organs of natural, hybrid, and variegated plants, and may also be extracted from plant tissue culture, according to a specific embodiment of the present invention.

In the present invention, ' codonopsis pilosula ) is a dicotyledonous perennial plant that grows deep in the mountains. It is distributed in Korea, China, and Usu River. The leaves are regenerated, but they are viviparous in the short branches, oval or ovate oval, blunt-headed, about 1-5cm long, about 1-3.5cm long, with fine hairs on both sides, especially on the veins, green on the surface, and white on the back. The edges are usually flat and the leaves are 2-3cm long and hairy. A capsule is a circular cone, and the calyx is fully matured and ripened around September-October. Seeds are small and many, long, round, brown without wings, glossy. Flowers bloom in July-August, about 1 at the end of the geodetic branch, and continue to bloom in the mesenchymal below. Calyx is divided into 5, hairless, about 15mm long, and about 5mm long, lanceolate. Corollas are bell-shaped, purple, about 2.5cm long, about 1.5cm in diameter, divided into about 5 ends, lobes are triangular, and about 5mm long. The vines are wound around other objects. Hairs all over, and when cut, latex comes out. Root stocks are club-shaped. Roots are columnar, about 20-30cm long, about 0.5-2 (10) cm in diameter, and yellowish brown on the outside. The ginseng extract can be extracted from a variety of organs of natural, hybrid, mutant plants and can be extracted from plant tissue culture, according to a specific embodiment of the present invention was extracted from the ground.

In the present invention, 'willow ( Salix koreensis ) 'is a deciduous broad-leaved tree belonging to the Salicaceae family, which blooms in April, and the male and female fruits bear fruit in May. Flowers are female and female. A male flower is about 1 ~ 2cm long and has two honey glands and two stamens. The female flower is about 1 ~ 2cm long and has about 1 ~ 2 honey glands. Hairs on stalk, poins are green egg-shaped, hairy. The ovary is egg-shaped, without a sack, hairy, with a slightly long pistil and 4 pistils. Fruits are ripe in May and contain hairy seeds. Distributed in Korea, Japan, northeastern China, etc. The willow extract can be extracted from various organs of natural, hybrid, and varieties of plants and cultures of plant tissues. Extractable in water, according to a specific embodiment of the present invention was extracted from the stem.

In the present invention, 'Viburnum ( Viburnum wrightii ) 's leaves are opposite, oval-shaped, and acute. There is a preoccupation on the back, and the vein is clearly extended to the edge. There is tooth tooth on the edge. There are many thin branches and brown. Flowers bloom white in late spring. Fruits are elliptical ovate with a nucleus, ripening red in autumn. Although it is amniotic, it grows well in the shade, and does not cover soil but grows well in fertile sandy loam. It has strong pollution resistance and flame resistance. The mountain viburnum extract may be extracted from various organs of natural, hybrid, and variegated plants and may be extracted from plant tissue culture, and according to a specific embodiment of the present invention, the extract is extracted from leaves, stems, and flowers.

In the present invention, the ' hulled tree ( Platycarya strobilacea ) 'is a deciduous tree that grows up to about 10m and lives in sunny places such as lowland riversides. Flowers ripen in May to June and fruits ripen in September. The leaves are about 15 ~ 30cm long. Lobules about 7 ~ 19, narrow oblong, about 5 ~ 11cm long, basal, cut, left and right, large on top, heavy, with hairs on both sides. Unghwasu is about 5 ~ 8cm long, Jahwa water is about 2 ~ 4cm long, and the fruit tree is long oval, about 3 ~ 5cm long and has no dark brown hairs. The fragments are lanceolate without falling off. The oyster tree extract may be extracted from various organs of natural, hybrid, and variegated plants, and may be extracted from plant tissue culture, according to a specific embodiment of the present invention.

In the present invention, 'weasel ( Amorpha) fruticosa ) is a deciduous shrub belonging to the legume and grows about 3m tall. The leaves of about 11 ~ 25 sheets of leaf are shifted. The leaf is ovate and the edge is flat. The purple flowers bloom in clusters in a gunshot inflorescence at the end of the stem from May to June. The fruit, containing about one seed, ripens around September. The weasel extract can be extracted from various organs of natural, hybrid, and variegated plants, and can be extracted from plant tissue cultures, and extracted from leaves and stems according to specific embodiments of the present invention.

In the present invention, ' Pyrus ( Pyrus) pyrifolia ) 'is a deciduous arborescent growing in the southern central region, growing up to about 5m in height, and its leaves are regenerated. It is similar to the heart type and is about 7-12 cm long. Petioles and peduncles have no hairs or downy hairs. Small fruits about 3cm in diameter run, and unlike mountain dorsum, calyx has pointed tip, and when it bears fruit, it falls off early. Flowers bloom before leaves in April-May, fruits are dark brown, and fruit is produced in October. The pear tree extract may be extracted from various organs of natural, hybrid, and variegated plants, and may be extracted from plant tissue culture, and extracted from leaves and stems according to a specific embodiment of the present invention.

In the present invention, ' Lyspedeza cuneata ) 'is a perennial plant. Stems stand straight, branched a lot, hairy, and have vertical ridges. The leaves are alternately attached, and the leaf stalk is a short leaf of about 3 leaflets. The leafs are strip-shaped wedges with dull or concave ends, with silk on the edges. In July-August, several small butterfly-shaped flowers with purple streaks bloom. The calyx is divided into about 5 branches, and the fragment is willow-shaped and hairy. There are about 10 hydrosurgeries, two hydrosurgeries. Fruits are round pods, with net-like lines on the outside, with fine hairs. Contains about 1 seed. The non-repair extract can be extracted from various organs of natural, hybrid, and mutant plants and can be extracted from plant tissue culture, according to a specific embodiment of the present invention.

In the present invention, ' potentilla cryptotaeniae ) is a perennial plant of the dicotyledonous rosaceae, distributed in Korea, northeastern China, Japan, the Usuri River basin, and Amur, and lives in deep mountain streams. Flowers bloom in yellow in July and August and run in a pickled inflorescence at the end of branches. The diameter of the flower is about 1cm, the calyx is egg-shaped basso, and the jaw is upside down. Petals are about 5, round in the shape of an upside down egg, about 4mm long, roughly the same as calyxes, about 20 stamens, and many pistils. The stem is hairy throughout and the leaves from the roots disappear when the flowers bloom, the leaves from the stems are alternate and the leaves come out with 3 small leaves, the petioles are long at the bottom of the stem but short at the top, and the chin leaves are More than half attach to petioles. The birch flower extract can be extracted from various organs of natural, hybrid, mutant plants and can be extracted from plant tissue culture, according to a specific embodiment of the present invention was extracted from the outpost.

In the present invention, 'snowballs ( Salix graciliglans ) 'is a deciduous broad-leaved shrub belonging to the willow tree Willow family. It is native to Korea and is distributed in Asia. The size of the tree is about 1 ~ 2m, and the leaves are narrow lanceolate, about 10cm long. The snow willow extract can be extracted from various organs of natural, hybrid, and variegated plants, and can be extracted from plant tissue culture, according to a specific embodiment of the present invention.

The composition of the present invention is based on natural extracts, such as the 13 kinds of plant extracts, there is an advantage that it can be taken for a long time because there is little safety and no toxicity or side effects. In addition, the composition may be used not only in humans, but also in animals such as cows and dogs, in which allergic diseases may develop.

The composition for inhibiting mast cell granule secretion according to the present invention has an excellent effect on inhibiting the release of beta-hexosaminidase, reducing the amount of IgE in the serum of mice causing atopic dermatitis, and inhibiting the increase in the number of mast cells. It is effective in improving, preventing, and treating allergic diseases.

Therefore, in another embodiment, the present invention is to prevent or treat an allergic disease comprising the composition for inhibiting the secretion of mast cell granules and to prevent the allergic disease by administering the composition for inhibiting secretion of mast cell granules or Provide a method of treatment.

As used herein, the term 'prevention' refers to all actions of inhibiting or delaying the onset of the allergic disease by administration of a composition for inhibiting mast cell granule secretion according to the present invention.

As used herein, the term 'treatment' refers to all actions of improving or beneficially modifying the symptoms of the allergic disease by administration of a composition for inhibiting mast cell granule secretion according to the present invention.

In the present invention, the term 'allergic disease' includes all diseases that can be caused by allergies, and is very similar to the disease in which the antigen-antibody reaction is demonstrated, as well as the pathological phenomenon caused by the antigen-antibody reaction. It also includes things that are not proved, or that the involvement of antigen-antibody reactions has not been proved but is likely. Examples of allergic diseases include organs such as bronchial asthma, hyperthermia, vasomotor neuritis, hypertrophic rhinitis, allergic bronchitis, and allergic bronchitis (transient lung infiltration) in the respiratory system. In the circulatory system, such as sexual diarrhea, allergic stomatitis, enteropolitan purpura, nodular periarteritis, obstructive arteritis, angina pectoris, endocarditis, urticaria, quinque edema (vascular neuroedema), nodular erythema, purpura, atopic dermatitis, eye, etc. Relationships include flicten, sympathetic ophthalmitis, allergic conjunctivitis, allergic keratitis, and other examples include diffuse glomerulonephritis and migraine headaches. In addition, allergic diseases can be classified into immediate type (anawheelax type) and late type (tuberculin type). Immediate type includes high fever, bronchial asthma, urticaria, quinque edema, digestive tract allergy, and delayed type includes contact dermatitis and drug allergy.

Compositions according to the invention can be prepared in pharmaceutical formulations using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. In the preparation of the dosage forms, it is preferred to mix or dilute the compositions according to the invention with the carrier or to enclose them in a carrier in the form of a container.

Accordingly, the pharmaceutical composition of the present invention is a tablet, pills, powders, granules, capsules, suspensions, contents, emulsions, syrups, aerosols, injectable solutions, etc. according to conventional methods for preventing and treating allergic diseases. It can be formulated into a form and used. For example, carriers, excipients and diluents which may be included in the pharmaceutical compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, Calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may include at least one excipient in the composite composition, for example, starch, calcium carbonate, sucrose, lactose, gelatin, etc. Can be prepared by mixing. In addition to the simple excipients, lubricants such as magnesium stearate, talc can also be used. Liquid preparations for oral administration include suspensions, solvents, emulsions, and syrups, and various excipients, for example, wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. Can be used.

Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, suppositories, and the like. As the non-aqueous solvent and the suspension solvent, vegetable oils such as propylene glycol, polyethylene glycol, olive oil, etc., injectable esters such as ethyl oleate, and the like can be used.

In addition, the pharmaceutical compositions of the present invention may further comprise a carrier, excipient or diluent. Carriers, excipients or diluents include lactose, textose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, hydroxy Minerals such as propyl methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxy benzoate, fluoro hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate, silicon dioxide and the like can be used. have.

The dosage of the pharmaceutical composition according to the invention should be a pharmaceutically effective amount. As used herein, the term 'pharmaceutically effective amount' means an amount sufficient to prevent or treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level refers to the method of formulation, the condition and weight of the patient. It may be variously selected by those skilled in the art according to factors such as sex, age, extent of disease, drug form, route and duration of administration, rate of excretion, response sensitivity, and the like. Effective amounts may vary depending on the route of treatment, the use of excipients, and the possibility of use with other agents, as will be appreciated by those skilled in the art.

The pharmaceutical composition of the present invention can be administered to mammals such as mice, livestock, humans, etc. through various routes. All modes of administration can be expected and can be administered, for example, by oral, parenteral, intravenous, intradermal, and subcutaneous injection.

In another aspect, the present invention provides a food composition for preventing or improving allergic diseases, including the composition for inhibiting mast cell granule secretion.

The food composition may be a functional food according to the purpose of the present invention, wherein the 'functional food' is a food that emphasizes the bioregulatory function of the food, and acts and expresses a specific purpose using physical, biochemical and biotechnological methods. It is food added with added value. The components of the functional food are designed and processed to fully exercise the body control functions related to biological defense and control of body rhythm, prevention and recovery of diseases to the living body, and are acceptable food supplement additives, sweeteners or functional ingredients It may contain.

Food compositions to which the composition for inhibiting secretion of mast cells granules according to the present invention can be added include, for example, various foods, for example, beverages, gums, teas, vitamin complexes, health supplements, and the like. Natural fruit juices, fruit juice drinks, vegetable drinks, and the like.

The food composition of the present invention may be prepared in a formulation such as tableting, tablets, granules, powders, capsules, liquid solutions, pills and the like according to known production methods.

In addition, the food composition of the present invention may further contain conventional various flavors, natural carbohydrates and the like. Flavors include natural flavors such as tautin and stevia extract, and synthetic flavors such as saccharin and aspartame. The natural carbohydrates described above may include glucose, monosaccharides such as fructose, maltose, disaccharides such as sucrose, dextrins, polysaccharides such as cyclodextrins, sugar alcohols such as xylitol, sorbitol, erythritol, and the like. . In addition, the food composition of the present invention is a variety of nutrients, vitamins, minerals (electrolytes), flavors such as synthetic or natural flavors, colorants, neutralizing agents (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and its Salts, organic acids such as citric anhydride, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, natural fruit juices, fruit juice beverages, and pulp for making vegetable drinks It may further contain an acceptable food supplement additive. Such additives may be used independently or in combination.

In another aspect, the present invention provides a quasi-drug composition for preventing or improving allergic diseases, including the composition for inhibiting mast cell granule secretion.

The composition of the present invention may be added to an over-the-counter composition for the purpose of preventing or ameliorating allergic diseases. When the composition for inhibiting mast cell granule secretion according to the present invention is used as an quasi-drug additive, the 13 kinds of plant extracts may be added as they are or may be used together with other quasi-drugs or quasi-drug components, and may be appropriately used according to a conventional method.

The mixed amount of the active ingredient may be suitably determined depending on the purpose of use (prevention, health or therapeutic treatment). Preferably, the quasi-drug composition may be used for the preparation of antiseptic cleaners, shower foams, gagrins, wet tissues, detergent soaps, hand washes, humidifier fillers, masks, ointments, coating agents or filter fillers.

In another aspect, the present invention provides a cosmetic composition for preventing or improving allergic diseases, including the composition for inhibiting mast cell granule secretion. The cosmetic composition according to the present invention may be added with a pH adjusting agent, flavoring agent, emulsifier, preservative, etc. as necessary and containing solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, and surfactants in a conventional cosmetic preparation method. It can be formulated as a cleansing, oil, powder foundation, emulsion foundation, wax foundation, cream, essence and spray, and the like. Compositions include, but are not limited to, conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments, flavors, and the like, and carriers.

If the formulation is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, tracants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components, It is not limited. If the formulation is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray additionally chlorofluorohydrocarbons, propane / butane or Propellants such as dimethyl ether may be included but are not limited thereto. If the formulation is a surfactant-containing cleansing, the carrier component may be aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivatives, methyltaurate, sarcosinate, fatty acid amide ether sulfate, Alkyl amidoventins, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like may be used, but are not limited thereto.

In another aspect, the present invention is a Japanese leaf ( Larix) leptolepis ), elm ( Maackia) amurensis ), massif (Corydalis speciosa), straw bud ( Agrimonia pilosa ), ginseng ( Codonopsis) pilosula ) Willow ( Salix) koreensis ), viburnum wrightii , oyster tree ( Platycarya) strobilacea ), Weasel ( Amorpha fruticosa ), Pyrus ( Pyrus) pyrifolia ), vulgaris ( Lespedeza cuneata ), potentilla crushing at least one selected from the group consisting of cryptotaeniae ), and Salix graciliglans , extracting and grinding the ground matter with a solvent selected from water, an organic solvent or a mixed solvent thereof, water in the dry matter, Japanese leaf larch, elm tree, mountain fire bag, straw shinnasu, mansam, willow, mountain viburnum, oyster tree, weasel, pear tree, non-repair, comprising the steps of adding and dissolving a solvent selected from an organic solvent or a mixed solvent thereof. It provides a method for producing one or more extracts selected from the group consisting of hydrangea, and snow cover.

13 kinds of plant extracts according to the present invention in one step, the leaves of red oak tree, elm tree, mountain fire bag, straw buds, dumplings, willow, mountain viburnum, oyster tree, weasel, pear tree, non-ice, water flower, or snow willow Grinding the process. Preferably, it is dried for a predetermined time and crushed. The ground product may be obtained by extraction in two steps using water, an organic solvent or a mixed solvent thereof, and may be extracted according to various extraction methods such as cold needle extraction, heat extraction, ultrasonic extraction, and cold extraction as known in the art. Can be. The extraction method is not particularly limited and may be extracted at room temperature or warmed under conditions where the active ingredient is not destroyed or minimized. In addition, as known in the art, it may be dried according to various drying methods such as hot air drying, freeze drying, and vacuum drying. In three steps, a solvent selected from water, an organic solvent, or a mixed solvent thereof is added to the dried product to dissolve, thereby obtaining 13 plant extracts.

According to a specific embodiment of the present invention, 1000 ml of anhydrous methanol was added to the natural product (100 g), followed by warm extraction at 80 ° C. for 2 hours in an extractor equipped with a reflux condenser. After filtering the extract, the remaining residue (Residue) was extracted one or more times in the same manner as described above, and the resulting extract was combined and concentrated under reduced pressure and then lyophilized to obtain a dry extract.

Organic solvents that can be used for extraction and dissolution include ethanol, methanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF), dimethyl Sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol or a mixed solvent thereof, preferably C ₁ ~ C ₄ lower alcohol, such as methanol, ethanol can be used, specific embodiments of the present invention Methanol was used.

Since the composition for inhibiting granule secretion of mast cells according to the present invention is effective in preventing, improving and treating allergic diseases, the pharmaceutical composition, food composition, quasi-drug composition, cosmetic composition, etc. Can be used.

1 is a diagram illustrating the activation mechanism of mast cells in vivo, and illustrates a series of signaling mechanisms in which degranulation occurs in cells after IgE attaches to IFCeRI and crosses allergens in mast cells.
Figure 2 shows the natural plant extracts showing a 50% or more inhibitory effect of beta-hexosaminidase inhibition rate analysis to determine the inhibitory effect of granule secretion formation of the plant extract. The numbers in the lower part of Figure 2 shows the serial numbers of Table 1.
3, Atopic dermatitis was induced by applying DNCB solution to Nc / Nga female mice at 7 weeks of age, and then serum IgE concentrations of the groups coated with the Japanese Larch extract, the negative control (vechicle), and the positive control (dex.) Were measured. It is shown.
Figure 4 shows the biopsies of NC / Nga mice, stained with Toluidin Blue, and then stained mast cells (top) and the number (bottom).

Hereinafter, preferred embodiments of the present invention will be presented to assist in understanding the present invention. However, the following examples are merely provided to more easily understand the present invention, and the contents of the present invention are not limited by the examples. In addition, all documents disclosed in the present invention are incorporated by reference.

Example  1: Preparation of natural product extract from plants

All plants used in this experiment were purchased from Korea Plant Extract Bank. The purchased plant (100 g) was washed with purified water, dried and 1000 ml of anhydrous methanol was added thereto, followed by warm extraction at 80 ° C. for 2 hours in an extractor equipped with a reflux condenser (Model: HB 4 basic, manufacturer: IKA). The extract was filtered with Whatman No. 2 filter paper, and the remaining residue was extracted one or more times in the same manner as above, and the filtered extracts were combined and concentrated under reduced pressure (Model: Rotavapor, Manufacturer: Buchi, Temperature: 40 ℃). After freeze drying (model name: FD5518, manufacturer: Bonjaro, temperature: -45 ℃, pressure: 50 mTorr) to obtain a dry extract. All plant extracts were extracted in this same way. The 87 plant names used in this experiment, the fruit name belonging to this plant and the site of the extracted plant are shown in Table 1 below.

Experimental Example  One: RBL -2 H3  Inhibitory effect of granule secretion on cells

1.1. RBL -2 H3  Cell preparation and processing

RBL-2H3 cells were purchased from ATCC and cultured in plates plated with DMEM medium (100 mm dish) containing inactivated 10% fetal bovine serum, antibiotic 4 mM glutamine, 50 U / ml penicillin, and 50 g / ml streptomycin. . Aspirate the medium for subculture, add 2 ml of PBS and pipette to separate the cells from the dish wall, centrifuge at 1,000 x g for 5 minutes to collect the cells, and then pipet into fresh medium to pellet the cells. Was dispersed and placed in a new culture plate and incubated in an incubator at 37 ° C. and 5% CO ₂ gas supplied. The DNP-IgE antibody and DNP-BSA used in the experiment were purchased from SIGMA. After inhaling the medium from the plate of RBL-2H3 cells and washing with PBS, 3 ml of 0.25% Trysin-EDTA was added and cultured in an incubator supplied with 5% CO ₂ gas for 5 minutes. After the cells were removed from the plate wall, the cell number was measured by a hematocytometer and dispersed and inoculated in fresh medium at a concentration of 2 × 10 ⁵ cells / well. After 24 hours, 10ng / ml of the DNP-IgE antibody was added thereto, followed by incubation for 3 hours in a 5% carbon dioxide gas incubator. After the reaction, the medium was removed and washed three times with PBS, followed by 87 kinds of Tyrodes's buffer (10 mM Hepes, 130 mM NaCl, 5 mM KCl, 1.4 mM CaCl ₂ , 1 mM MgCl ₂ , 5.6 mM glucose, and 0.1% BSA, pH 7.4). After adding each plant extract for 30 minutes and adding 0.2ug / ml DNP-BSA, it was reacted in an incubator for 20 minutes, and the plate was placed on ice for 10 minutes to stop the reaction. It was measured whether it inhibited the release of sosaminidase (β-hexosaminidase).

1.2. beta- Hexosaminidase (β- hexosaminidase ) Measure

Beta-hexosaminidase release was performed with substrate solution containing 1.3 mg / ml of p-nitrophenyl-N-acetyl-β-D-glucosaminide in 50ul of supernatant and Citrate buffer (0.1M citrate, pH 4.5). (substrate solution) After the same amount (50ul) was reacted for 90 minutes in a 37 ° C incubator, the reaction was stopped using 150ul 0.2M glycine (pH 10.7), it was measured at 405nm. The experiment was performed at least three times independently, and the analysis was performed by one-way analysis of variance (ANOVA) of the SPSS 10.0 program, and was determined to be statistically significant when P <0.05. Beta-hexosaminidase inhibition rate was calculated according to the following formula.

[Beta-hexosaminidase Inhibition Rate]

β- hexosaminidase inhibition (%) = (1- (T-B-N / C-N)) * 100

Control (C): DNP-BSA (+), test sample (-)

Test (T): DNP-BSA (+), test sample (+)

Blank (B): DNP-BSA (-), test sample (+)

Normal (N): DNP-BSA (-), test sample (-)

The results are shown in Table 2 below, and the plant extracts having an inhibitory effect on mast cell granule secretion of 50% or more in the results of Table 2 are separately indicated in FIG. 2.

As shown in Table 2, as a result of analyzing beta-hexosaminidase inhibitory effect on RBL-2H3 cells of plant extracts, 16 of 87 plant extracts (PL65, PL18, PL55, PL73, PL39, PL35, PL46, PL54, PL09, PL19, PL26, PL68, PL27, PL53, PL40, PL17) showed more than 50% of beta-hexosaminidase inhibitory effect. Seven species (PL65, PL18, PL55, PL73, PL39, PL35, PL46, PL54), including tree extract (stem, PL65), showed an inhibitory effect on the secretion of granules from mast cells.

Experimental Example  2: atopic dermatitis symptoms relief effect

2.1. : Experimental Animal

All animal experiments used in this experiment were conducted after approval of the Animal Experiment Ethics Committee, which is operated by Gyeonggi Bio Center. Nc / Nga female mice at 7 weeks of age were purchased from a central laboratory animal (Seoul, Korea). All mice were reared in a 12-hour light-dark cycle in a laboratory maintained at an optimal environmental condition with a temperature of 24 ± 2 ° C. and a humidity of 50 ± 10%, and the food was freely eaten. The experiment was started at 8 weeks after the mouse was quarantined and purified for 1 week before the experiment.

2.2. Induced atopic dermatitis

The hair from the lower part of the ear to the upper part of the tail of the mouse was depilated using a depilatory cream, and then, after 1 day, 200 µl of 2,4-dinitrochlorobenzene (DNCB) solution was applied. Dermatitis was induced by first application of 1% DNCB solution (acetone: olive oil = 4: 1) three times per week and twice with 0.2% DNCB solution three times for three weeks.

2.3. Test Material Manufacturing and Processing

The test substance was dissolved in 0.1% of a solvent mixed with EtOH: H ₂ O: butyleneglycol = 1: 4: 5, respectively, and applied for 100 days at 100 μl once a day. The solvent was used as a negative control, dexamethasone (0.1%) dissolved in a solvent as a positive control was applied in the same manner as the test material.

2.4. serum IgE  Measure.

To measure serum IgE, blood was collected from mouse eyes using microtubules at the beginning and end of treatment of the test substance. After collecting blood for 1 hour and stored at room temperature for 1 hour, the serum was separated by centrifugation at 1000g, 4 ℃ conditions, and stored in -70 ℃ freezer until use. Serum IgE levels were measured using the BD OptEIA ^™ ELISA set provided by BD Bioscienc according to the method presented. As a result of measuring the amount of IgE in serum 4 weeks after application, the amount of IgE was significantly decreased in the positive control group and the 0.1% treatment group of the Japanese Larch plant extract compared to the negative control group.

2.5. Histopathological examination.

In order to observe the skin tissue, the skin tissue of the lower part of the ear of the mouse was extracted at the end of the test material treatment. The extracted tissue was fixed in 10% formalin solution, and then paraffin embedding, tissue sections and staining were commissioned to the central laboratory animals. Tissue sections stained with Hematoxylin & Eosin were visually observed 200 times by optical microscope. In addition, mast cells were stained by toluidine blue staining, and magnified 100 times with an optical microscope, and 5 sites were randomly selected to measure the number of mast cells. Tissues of NC / Nga mice were biopsied, stained with Toluidin Blue, and the number of mast cells was measured. Cells were randomly selected after 5 locations on each slide, and statistically treated as a mean value. As a result, a significant decrease was found in the 0.1% treatment group and the 0.1% treatment group of Japanese Larch extract compared to the negative control group.

Claims (11)

A composition for preventing or improving allergic diseases, including willow (Salix koreensis) extract as an active ingredient.
According to claim 1, The willow extract composition for the prevention or improvement of allergic diseases, characterized in that extracted from the stem.
According to claim 1, wherein the extract is a composition for the prevention or improvement of allergic diseases, characterized in that the methanol extract.
According to claim 1, wherein the composition is a composition for preventing or improving allergic diseases, characterized in that the secretion of histamine, beta-hexosaminidase (β-hexosaminidase), or leukotriene in the granules of mast cells.
A pharmaceutical composition for preventing or treating allergic diseases, comprising the composition of claim 1 as an active ingredient.
Food composition for improving or preventing allergic diseases comprising the composition of claim 1 as an active ingredient.
Claim 1 composition for improving or preventing allergic diseases comprising the composition of claim 1 as an active ingredient.
Cosmetic composition for improving or preventing allergic diseases comprising the composition of claim 1 as an active ingredient.
9. The composition of claim 5, wherein the allergic disease is asthma, urticaria, or atopy.
Grinding the willow tree (Salix koreensis);
Extracting and drying the ground material with a solvent selected from water, an organic solvent or a mixed solvent thereof; And
A method for producing an oxidized bag extract, comprising the steps of adding a solvent selected from water, an organic solvent, or a mixed solvent thereof to the dried product.
The method of claim 10, wherein the organic solvent is ethanol, methanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N- dimethylformamide (DMF), dimethyl sulfoxide Said (DMSO), 1, 3- butylene glycol, or a propylene glycol manufacturing method.

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