WO2012002777A2 - Composition containing plant extracts for inhibiting the granule secretion of mast cells - Google Patents

Abstract

The present invention relates to an inhibitor for the granule secretion of mast cells, comprising, as active ingredients, one or more extracts selected from a group consisting of Larix leptolepis extracts, Maackia amurensis extracts, Corydalis speciosa extracts, Agrimonia pilosa extracts, Codonopsis pilosula extracts, Salix koreensis extracts, Viburnum wrightii extracts, Platycarya strobilacea extracts, Amorpha fruticosa extracts, Pyrus pyrifolia extracts, Lespedeza cuneata extracts, Potentilla cryptotaeniae extracts and Salix graciliglans extracts. The present invention also relates to a composition comprising said extracts as active ingredients for preventing or treating allergic diseases, and to a food composition, a quasi-drug composition, and a cosmetic composition comprising said extracts as active ingredients for alleviating or preventing allergic diseases. The present invention also relates to a method for preventing or treating allergic diseases, and to the uses of said extracts.

Description

Composition for inhibiting granule secretion of mast cells containing plant extracts

The present invention is a Japanese extract of Larix leptolepis , Maackia amurensis extract, Coraldalis speciosa extract, Agrimonia pilosa extract, Codonopsis pilosula extract, Willow ( Salix koreensis ) extract , Viburnum wrightii extract, Platycarya strobilacea extract, Weasel ( Amorpha fruticosa ) extract, Pyrus pyrifolia extract, Lespedeza cuneata extract, Potentilla cryptotaeniae extract, and Granules secretion inhibitor of mast cells comprising at least one extract selected from the group consisting of eyelids ( Salix graciliglans ) extract, a composition for preventing or treating allergic diseases comprising the extract as an active ingredient, allergic diseases Food composition, quasi-drug composition, and cosmetic composition for improvement or prevention, Method for preventing or treating allergic diseases, and relates to the use of the extract.

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 the occurrence of allergies, whereas anti-allergic drugs are used as a preventive agent to suppress the occurrence of allergic reactions, so should be used for 2-4 weeks or more until the effect is shown. 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 form that is more accessible and safer to consumers in the final product development, and is easier to industrial application than synthetic pharmaceutical materials. The inventors of the present invention include Larix leptolepis , Maackia amurensis , Coraldalis speciosa, Agrimonia pilosa , Codonopsis pilosula , Salix koreensis , and Viburnum wrightii. ), Extracts of Platycarya strobilacea , Weasel ( Amorpha fruticosa ), Pear ( Pyrus pyrifolia ), Lespedeza cuneata , Potentilla cryptotaeniae , and Salix graciliglans extracts from mast cells It has a secretory inhibitory effect, which has been shown to improve allergic diseases associated with the secretion of granules from mast cells. By the present invention it has been completed.

An object of the present invention is a Japanese leaf ( Larix leptolepis ) extract, Maackia amurensis extract, Coraldalis speciosa extract, Agrimonia pilosa extract, Codonopsis pilosula extract, Willow ( Salix koreensis) ) Extract, Viburnum wrightii Extract, Platycarya strobilacea Extract, Weasel ( Amorpha fruticosa ) Extract, Pyrus pyrifolia Extract, Lespedeza cuneata Extract, Potentilla cryptotaeniae Extract To provide a granule secretion inhibitor of mast cells comprising, as an active ingredient, at least one extract selected from the group consisting of Salix graciliglans extract.

Another object of the present invention is the Japanese leaf larch extract, elm extract, mountain fire bag extract, hay fever extract, ginseng extract, willow extract, mountain viburnum extract, oyster tree extract, weasel extract, nectarine extract, non-repair extract, It is to provide a pharmaceutical composition for the prevention or treatment of allergic diseases, including one or more extracts selected from the group consisting of water palm flower extract, and eyeball willow extract as an active ingredient.

Still another object of the present invention is to provide a food composition for improving or preventing allergic diseases including the extract as an active ingredient.

Another object of the present invention to provide an quasi-drug composition for improving or preventing allergic diseases comprising the extract as an active ingredient.

Still another object of the present invention is to provide a cosmetic composition for improving or preventing allergic diseases including the extract as an active ingredient.

Another object of the present invention is to provide a method for preventing or treating an allergic disease, comprising administering to a subject a therapeutically effective amount of the pharmaceutical composition.

Another object of the present invention in the manufacture of a medicament for the prevention or treatment of allergic diseases, Japanese leaf larch extract, elm tree extract, fire bulge extract, hay fever extract, ginseng extract, willow extract, mountain viburnum extract, To provide the use of one or more extracts selected from the group consisting of oyster bark extract, weasel extract, nectarine extract, non-repair extract, water flower extract, and snow willow extract.

In order to achieve the above object, the present invention in one aspect, the present invention is Japanese larch ( Larix leptolepis ) extract, Elm ( Maackia amurensis ) extract, Coraldalis speciosa extract, Agrimonia pilosa extract, Ginseng ( Codonopsis pilosula) ), Willow ( Salix koreensis ) extract, Viburnum wrightii extract, Platycarya strobilacea extract, Amorpha fruticosa extract, Pyrus pyrifolia extract, Lespedeza cuneata extract and it provides a granule secretion inhibitors of the mast cell that contains one or more extracts selected from the group consisting of water Potentilla Fragarioides (Potentilla cryptotaeniae) extract, and eye pussy willow (Salix graciliglans) extract as an active ingredient.

The inventors of the present invention are Larix leptolepis , Maackia amurensis , Coraldalis speciosa, Agrimonia pilosa , Codonopsis pilosula , Salix koreensis , Viburnum viburnum granules of mast cells in extracts of wrightii ), Platycarya strobilacea , Amorpha fruticosa , Pyrus pyrifolia , Lespedeza cuneata , Potentilla cryptotaeniae , and Salix graciliglans extract It was confirmed that there was an effect of suppressing secretion. 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, Japanese larch extract, which showed the best inhibitory effect of β-hexosaminidase release among 13 plant extracts, was applied to mice that induced atopic dermatitis, and the amount of IgE in serum was significantly increased. The decrease in the number of mast cells is significantly suppressed (FIG. 3) (FIG. 4), and it was confirmed that the Japanese larch extract has an excellent effect of 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.

As used herein, the term 'granular secretion inhibitor of mast cells' refers to a substance capable of blocking or reducing the efficiency of secretion of inflammatory mediators collected in the granules 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, 'larix leptolepis ' has a height of about 30m, leaves are stripe-shaped, gathered on short branches of about 40-50 pieces, and the 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 leafy tree with leaves and a height of about 10-20 meters. 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, 'corydalis speciosa ' is a corydalis color family distributed in Korea, Japan, northeastern China, Heilong River, 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 alternated and split into two pieces of feathers. The length is about 10-15cm, and the butterfly is about 4-6cm. Branches are egg-shaped, split like feathers, and the last branch is a long oval, oval, with a sharp tip. Flowers bloom in yellow in April to 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, ' Agrimonia pilosa ' is a perennial herb of a dicotyledonous plant Rosaceae, and is distributed in Korea, Japan, China, India, Himalayas, Mongolia, Amur, Siberia, Europe, Sakhalin and the like. 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 to 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. Flowers bloom yellow in June-August and run in the form of gunshot inflorescences on the stems and branches. 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 upside down or round shape, 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, the 'Ginseng ( Codonopsis pilosula )' is a vine perennial plant of the dicotyledon plant Campanula Campanula grows in deep 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, ' Salix koreensis ' is a deciduous broad-leaved tree belonging to the genus Salicaceae, blooms in April, and the fruit tree is fruiting in May. Flowers are female and female. A male flower is about 1 ~ 2cm long, honey glands and two stamens, and hairs under the operating table. 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 capsules, ripe in May, with hairy seeds. It is distributed in Korea, Japan, and northeastern China. The willow extract may be extracted from various organs of natural, hybrid, and variegated plants, and may be extracted from plant tissue culture, and extracted from stems according to a specific embodiment of the present invention.

In the present invention, ' Viburnum wrightii ' has leaves facing each other, oval ellipsoidal, and pointed apex. 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, ' Pulpy carya ( Platycarya strobilacea )' is a deciduous tree that grows up to about 10m inhabiting sunny places such as lowland riversides, flowers ripen around May to June, fruit ripening around 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, and according to a specific embodiment of the present invention, it is extracted from flowers, particularly inflorescences.

In the present invention, ' Amorpha fruticosa ' is a deciduous shrub belonging to the legume 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 mutant plants and can be extracted from plant tissue culture, according to a specific embodiment of the present invention.

In the present invention, ' Pyrus pyrifolia ' is a deciduous arborescent growing in the southern central regions, grows up to about 5m in height, and leaves rejuvenate. 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, 'Lespedeza cuneata ' is a perennial herb. 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 rosacea rosaceae, distributed in Korea, northeastern China, Japan, the Usuri River basin, Amur, etc., and inhabits deep streams in the mountains. Flowers bloom yellow in July-August and run in the form of pickled flowers 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, ' 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.

Inhibitor 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 period of time because it is safe and has no toxicity or side effects. In addition, the inhibitor may be used not only in humans, but also in animals such as cows and dogs, in which allergic diseases may develop.

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

Therefore, in another embodiment, the present invention is the mast cell granule secretion inhibitor, that is, Japanese larch extract, elm tree extract, fire bulge extract, hay fever extract, ginseng extract, willow extract, mountain viburnum extract, oyster tree Pharmaceutical composition for the prevention or treatment of allergic diseases and treatment of the composition comprising one or more extracts selected from the group consisting of extracts, weasel extract, nectarine extract, non-repair extract, water flower extract, and snow willow extract Provided is a method for preventing or treating an allergic disease, comprising administering a pharmaceutically effective amount to an individual.

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, hey fever, vasomotor neuritis, hypertrophic rhinitis, allergic bronchitis, and Rheoplung syndrome (transient lung infiltration). In the digestive system, dietary allergic gastritis, allergic diarrhea, allergic stomatitis, enteropathic disease, etc. in the circulatory system, nodular periarteritis, obstructive arteritis, angina, endocarditis, etc. Skin relations include hives, quinque edema (vascular neuroedema), nodular erythema, purpura, atopic dermatitis, allergic dermatitis, etc. In the eye relationship, flictene, sympathetic ophthalmitis, allergic conjunctivitis, allergic keratitis And others include diffuse glomerulonephritis and migraine headaches. In addition, allergic diseases can be classified into immediate type (anawheelax type) and late type (tuberculin type). Immediate forms include high fever, bronchial asthma, urticaria, quinque edema, digestive tract allergies, insect allergies, food allergies, and drug allergies.

As used herein, the term "individual" means all animals, including humans, and allergic diseases including primates and livestock, breeding, pet or sport animals such as dogs, wheat, cats, sheep, pigs, cattle, and the like. This means any animal in need of treatment and requiring treatment, and by administering the pharmaceutical composition of the present invention to an individual, allergic disease can be effectively prevented or treated.

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. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are 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, ie a therapeutically 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 embodiment, the present invention is Japanese leaf larch extract, elm tree extract, mountain bag fire extract, hay fever extract, dumplings extract, willow extract, mountain viburnum extract, oyster bark extract, weasel extract, stone extract, It provides a food composition for the improvement or prevention of allergic diseases, including as an active ingredient at least one extract selected from the group consisting of non-repair extract, birch flower extract, and snow willow extract.

The allergic diseases include allergic symptoms, and allergic symptoms include symptoms appearing due to allergic diseases or symptoms appearing before allergic diseases occur.

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.

Examples of the food composition to which the extract according to the present invention may be added include, for example, various foods, for example, beverages, gums, teas, vitamin complexes, health supplements, and the like. 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 embodiment, the present invention is Japanese leaf larch extract, elm tree extract, mountain bag fire extract, hay fever extract, dumplings extract, willow extract, mountain viburnum extract, oyster bark extract, weasel extract, stone extract, It provides a quasi-drug composition for improvement or prevention of allergic diseases, including as an active ingredient at least one extract selected from the group consisting of non-repair extract, birch flower extract, and snow willow extract.

The allergic diseases include allergic symptoms, and allergic symptoms include symptoms appearing due to allergic diseases or symptoms appearing before allergic diseases occur.

The extract of the present invention may be added to the quasi-drug composition for the purpose of preventing or improving allergic diseases. When the extract 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 embodiment, the present invention is Japanese leaf larch extract, elm tree extract, mountain bag fire extract, hay fever extract, dumplings extract, willow extract, mountain viburnum extract, oyster bark extract, weasel extract, stone extract, It provides a cosmetic composition for the improvement or prevention of allergic diseases, including as an active ingredient at least one extract selected from the group consisting of non-repair extract, hydrangea extract, and snow willow extract.

The allergic diseases include allergic symptoms, and allergic symptoms include symptoms appearing due to allergic diseases or symptoms appearing before allergic diseases occur.

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 embodiment, the present invention is Japanese larch ( Larix leptolepis ), Elm ( Maackia amurensis ), Massif (Corydalis speciosa), Agrimonia pilosa , Ginseng ( Codonopsis pilosula ), Willow ( Salix koreensis ) , Viburnum wrightii , Platycarya strobilacea , Weasel ( Amorpha fruticosa ), Pyrus pyrifolia , Lespedeza cuneata , Potentilla cryptotaeniae , and Salix graciliglans Grinding one or more selected from the group consisting of: extracting and drying the ground product with a solvent selected from water, an organic solvent or a mixed solvent thereof, and drying the solvent with water, an organic solvent, or a mixed solvent thereof. Japanese leaf larch, elm tree, mountain fire bag, straw shinnyum, ginseng, willow, mountain viburnum, gulpina Provided is a method for preparing at least one plant extract selected from the group consisting of radish, weasel, pear tree, non-prey, birch, 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.

In another aspect, the present invention, in the manufacture of a medicament for the prevention or treatment of allergic diseases, Japanese leaf larch extract, elm extract, mountain bag fire extract, straw shinnyum extract, dumplings extract, willow extract, mountain viburnum Provided is the use of one or more extracts selected from the group consisting of extracts, oyster tree extracts, weasel extracts, nectarine extracts, non-repair extracts, birch extracts, and snow willow extracts.

Since the granule secretion inhibitor of mast cells according to the present invention, the composition comprising the same is effective for the prevention, improvement, treatment of allergic diseases, pharmaceutical compositions, food compositions, quasi-drug compositions for improving, preventing or treating allergic diseases, and It can be used as a cosmetic composition and the like. In addition, natural plant extracts can be used to treat allergic diseases and to prevent or ameliorate allergic diseases without side effects.

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 FCεRI and crosses allergens in mast cells.

Figure 2 shows the beta-hexosaminidase inhibition rate analysis to determine the inhibitory effect of the granule secretion formation of the plant extract, showing the natural plant extracts showing an inhibitory effect of more than 50%. The numbers at the bottom of Figure 2 show the serial numbers in Table 1.

3 is After 7 weeks of age, Nc / Nga female mice were treated with DNCB solution to induce atopic dermatitis. 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 Extracts 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.

Table 1

Experimental Example 1 Inhibitory Effect of Granule Secretion on RBL-2H3 Cells

1.1. RBL-2H3 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 measurement

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.

TABLE 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 β-hexosaminidase inhibitory effect. Seven species (PL65, PL18, PL55, PL73, PL39, PL35, PL46, PL54) including leafy tree extract (stem, PL65) showed an inhibitory effect on granule secretion of mast cells of 80% or more.

Experimental Example 2 Amelioration effect of atopic dermatitis symptoms

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 Measurement

To measure serum IgE, blood was collected from mouse eyes using microtubules at the beginning and end of treatment of the test substance. Serum was separated by centrifugation at 1000 g, 4 ° C. and then stored at room temperature for 1 hour after blood collection, and stored at -70 ° C. freezer until use. Serum IgE levels were measured according to the methods presented using the BD OptEIA ^™ ELISA set provided by BD Bioscience. As a result of measuring the amount of IgE in serum 4 weeks after the application of the sample, the amount of IgE was significantly decreased in the positive control group and the Japanese larch plant extract 0.1% treated group compared to the negative control group (FIG. 3).

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.

Since 13 kinds of plant extracts according to the present invention can inhibit degranulation of mast cells, they can be used as a prophylactic or therapeutic agent for allergic diseases, and can be used as cosmetic preparations, quasi-drug preparations, and food preparations containing the extracts. .

Claims (12)

1. Larix leptolepis extract, Maackia amurensis extract, Corydalis speciosa extract, Agrimonia pilosa extract, Codonopsis pilosula extract, Salix koreensis extract, Mountain viburnum extract Viburnum wrightii extract, Platycarya strobilacea extract, Amorpha fruticosa extract, Pyrus pyrifolia extract, Lespedeza cuneata extract, Potentilla cryptotaeniae extract, and snow willow Salix graciliglans ) Granule secretion inhibitor of mast cells comprising at least one extract selected from the group consisting of extracts as an active ingredient.
2. The method according to claim 1, wherein the Japanese larch extract is in the stem, the elm extract is in the leaf or stem, the massif fire bag extract in the outpost, hay fever extract in the outpost, the ginseng extract in the ground, willow extract in the stem , Hawthorn extract in leaves, stems, or flowers, oyster bark extract in inflorescences, weasel fronds in leaves or stems, pear tree in leaves or stems, non-fibrous extracts in outposts, hydrangea extracts in outposts, And snow willow extract inhibitor, characterized in that extracted from the inflorescence.
3. The inhibitor of claim 1, wherein the extract is a methanol extract.
4. The inhibitor of claim 1, wherein the composition inhibits secretion of histamine, beta-hexosaminidase, or leukotriene in the granules of mast cells.
5. Japanese Leaf Larch Extract, Elm Extract, Mountain Bulgar Extract, Straw Sprout Extract, Manchurian Extract, Willow Extract, Mountain Viburnum Extract, Oyster Extract, Weasel Extract, Rhododendron Extract, Non-Iris Extract, Water Lily Extract, and Snow Pharmaceutical composition for the prevention or treatment of allergic diseases comprising at least one extract selected from the group consisting of pussy willow extract as an active ingredient.
6. Japanese Leaf Larch Extract, Elm Extract, Mountain Bulgar Extract, Straw Sprout Extract, Manchurian Extract, Willow Extract, Mountain Viburnum Extract, Oyster Extract, Weasel Extract, Rhododendron Extract, Non-Iris Extract, Water Lily Extract, and Snow Food composition for improvement or prevention of allergic diseases comprising at least one extract selected from the group consisting of pussy willow extract as an active ingredient.
7. Japanese Leaf Larch Extract, Elm Extract, Mountain Bulgar Extract, Straw Sprout Extract, Manchurian Extract, Willow Extract, Mountain Viburnum Extract, Oyster Extract, Weasel Extract, Rhododendron Extract, Non-Iris Extract, Water Lily Extract, and Snow A quasi-drug composition for improvement or prevention of allergic diseases comprising at least one extract selected from the group consisting of pussy willow extract.
8. Japanese Leaf Larch Extract, Elm Extract, Mountain Bulgar Extract, Straw Sprout Extract, Manchurian Extract, Willow Extract, Mountain Viburnum Extract, Oyster Extract, Weasel Extract, Rhododendron Extract, Non-Iris Extract, Water Lily Extract, and Snow Cosmetic composition for improvement or prevention of allergic diseases comprising at least one extract selected from the group consisting of pussy willow extract.
9. The method of claim 5, wherein the allergic disease is anaphylaxis, allergic bronchitis, allergic rhinitis, asthma, urticaria, Atopic dermatitis, contact dermatitis, allergic dermatitis, hey fever, insect allergies, food allergies, drug allergies, allergic gastritis, allergic diarrhea ), Allergic stomatitis, allergic conjunctivitis and allergic keratitis.
10. A method of preventing or treating an allergic disease, comprising administering to a subject a therapeutically effective amount of the pharmaceutical composition of claim 5.
11. The method of claim 10, wherein the allergic disease is anaphylaxis, allergic bronchitis, allergic rhinitis, asthma, urticaria, atopic dermatitis, Contact dermatitis, allergic dermatitis, hey fever, insect allergy, food allergy, drug allergy, allergic gastritis, allergic diarrhea, allergic stomatitis , Allergic conjunctivitis and allergic keratitis.
12. In the manufacture of a medicament for the prophylaxis or treatment of allergic diseases, the Japanese Larch extract, the elm tree extract, the firefly bag extract, the straw bud extract, the ginseng extract, the willow extract, the mountain viburnum extract, the oyster bark extract, the weasel extract The use of at least one extract selected from the group consisting of, pear tree extract, non-repair extract, water flower extract, and snow willow extract.

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