KR20170122327A - Compositions for Improving, Preventing or Treating for Th2-immune mediated Disease comprising Extracts from Costaria costata - Google Patents

Abstract

Provided is a pharmaceutical composition for ameliorating, preventing, or treating Th2-mediated immune diseases containing an effective amount of a Costaria costata extract and a carrier. Moreover, provided is a food composition. According to the present invention, by containing the Costaria costata extract, which is a natural ingredient contained as the active ingredient and is capable of ameliorating, preventing, or treating allergic diseases, the pharmaceutical composition ensures cost-effective production costs and user convenience.

Description

TECHNICAL FIELD The present invention relates to a composition for improving, preventing or treating Th2-immunity mediated diseases,

The present invention relates to a composition for the improvement, prevention or treatment of Th2-immunity mediated diseases,

 Atopic dermatitis (skin hypersensitivity reaction) is a disease with typical skin lesions. The cause of the disease has not yet been clarified. However, it is known that genetic factors, immune function imbalance and environmental factors are related. In particular, atopic dermatitis may be transferred to bone marrow transplant patients with atopic dermatitis, atopic dermatitis appears to be due to imbalance of immune cells derived from bone marrow.

The immunologic mechanism of atopic dermatitis has been suggested as an imbalance between Th1 / Th2 cells and abnormal cytokine system. In particular, atopic dermatitis is an immune response to the Th2 response among Th1 and Th2, and IL-4 and IL-10, which are cytokines expressed in Th2, are increased. Thus, IL-2 and INF- as a result of a decrease in the production of [gamma]. Th2 cells are known to induce atopic dermatitis progression by producing factors such as chemokines and cytokines. The resulting Th2 cytokines induce the production and differentiation of B lymphocytes and induce hypersensitive responses of eosinophils. The increase in IgE expression is induced by isotype switching of B lymphocytes by Th2 cytokine. IgE is an immunoglobulin expressed in most atopic dermatitis patients and induces degranulation of eosinophils and neutrophils, Lt; RTI ID = 0.0 > inflammatory < / RTI > Based on these immunological mechanisms, currently known methods of treatment include application of external moisturizing agents, antihistamines, and immunosuppressive agents, but they show very effective results in symptom relief, but there is a risk of side effects in long-term use and expecting fundamental treatment Is difficult. Therefore, there is a need for safe and effective new atopic treatments, and studies are being actively conducted to search for natural substances having ameliorating effects against atopic dermatitis.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors have sought to find an atopic dermatitis immunomodulating material derived from natural products. As a result, the extract of Costaria costata inhibited the increase of Th2 type cytokine and inhibited the proliferation of B cells, confirming the inhibitory effect of mast cell activation and allergen induction of histamine production by IgE production Thus completing the present invention.

Accordingly, it is an object of the present invention to provide a pharmaceutical composition for improving, preventing or treating a Th2-mediated immune disorder.

It is another object of the present invention to provide a food composition for improving or preventing a Th2-mediated immune disorder.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, the present invention provides a pharmaceutical composition comprising (a) a therapeutically effective amount of an extract of Costaria costata ; And (b) a pharmaceutically acceptable carrier. The present invention also provides a pharmaceutical composition for improving, preventing or treating a Th2-mediated immune disorder.

According to another aspect of the present invention there is provided a pharmaceutical composition comprising: (a) a pharmacologically effective amount of an extract of Alaska pollack; And (b) a food-acceptable carrier.

The present inventors have sought to find an atopic dermatitis immunomodulating material derived from natural products. As a result, it was confirmed that Costaria costata inhibited the increase of Th2 type cytokine and inhibited the proliferation of B cells, thereby suppressing the activation of mast cells and the allergic induction of histamine production by IgE production .

The marxa extract used in the composition of the present invention is a desalinized (or desalted) marine marine extract.

When the extract of Sukheum cane used in the composition of the present invention is obtained by treating an extractive solvent in Sukhumi, various extraction solvents may be used. Preferably, a polar solvent or a non-polar solvent can be used. Suitable polar solvents are (i) water, (ii) alcohols (preferably methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, Or ethylene glycol), (iii) acetic acid, (iv) dimethyl-formamide (DMFO) and (v) dimethyl sulfoxide (DMSO). Suitable nonpolar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1- But are not limited to, pentane, 1-chlorobutane, 1-chloropentane, o -xylene, diisopropyl ether, 2- chloropropane, toluene, 1- chloropropane, chlorobenzene, benzene, diethyl ether, diethylsulfide, Methane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride, and THF.

More preferably, the extraction solvent used in the present invention is (a) water, (b) anhydrous or hydrated lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.) (E) ethyl acetate, (f) chloroform, (g) butyl acetate, (h) 1,3-butylene glycol, (i) hexane and (j) diethyl ether. . Most preferably, the Alnus maritima extract of the present invention is obtained by treating water, ethanol, or a combination thereof, on a soya bean.

According to one embodiment of the present invention, the extract of Safflower marigold is a solvent extract of water of Safflower marigold, anhydrous or lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate, butyl acetate or 1,3-butylene glycol.

As used herein, the term " extract " means that it is used in the art as a crude extract as described above, but broadly includes fractions obtained by further fractionating the extract. That is to say, the extract of Dappermint is not only obtained by using the above-mentioned extraction solvent but also by additionally applying a purification process thereto. For example, a fraction obtained by passing the above extract through an ultrafiltration membrane having a constant molecular weight cut-off value, and a separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity) The fractions obtained by the purification method are also included in the extract of Sadomai extract of the present invention.

The extract of Alnus maritima used in the present invention can be prepared in powder form by an additional process such as vacuum distillation and freeze drying or spray drying.

As used herein, the term " comprising as an active ingredient " is meant to include an amount sufficient to achieve the efficacy or activity of the following extract. The present invention is a composition extracted from a natural plant material, dandelion. There is no adverse effect on the human body even when administered in an excessive amount. The quantitative upper limit of the composition of the present invention can be selected by a person skilled in the art within a suitable range.

The compositions of the present invention can be used to ameliorate, prevent or treat various Th2-mediated immune diseases, diseases or conditions.

As used herein, the term " Th2-mediated immune disease " refers to diseases in which IgE and mast cells are involved by the production and activity of allergen-specific Th2 cells.

As used herein, the term " Th2 cell " refers to a subset of helper T cell lymphocytes that are specified in terms of gene expression, protein secretion, and functional activity. For example, Th2 cells exhibit IL-4, IL-5, IL-10, and IL-13 cytokine expression patterns. Th2 cells are involved in the humoral immune response.

The Th2-mediated immune diseases to which the composition of the present invention is applied are not particularly limited, and preferably apply to allergic diseases.

According to one embodiment of the present invention, the allergic disease is selected from the group consisting of atopy dermatitis, allergic rhinitis, bronchial asthma, contact dermatitis, allergic conjunctivitis, ), Urticaria, vascular edema, food allergies, physical allergies, irritable pulmonary enteritis, occupational allergic diseases or drug allergies.

The composition of the present invention has an effect of reducing a Th2-mediated immune response.

As demonstrated in the following examples, the compositions of the present invention are (i) inhibiting the proliferation of B cells; (Ii) improvement of Th1 / Th2 cytokine imbalance; (Iii) decreased IgE production; And (iv) inhibiting histamine production.

The compositions of the present invention may be prepared with pharmaceutical compositions.

According to a preferred embodiment of the present invention, the composition of the present invention comprises: (a) a pharmaceutically effective amount of the above-mentioned Sadducein extract of the present invention; And (b) a pharmaceutically acceptable carrier. As used herein, the term " pharmaceutically effective amount " means an amount sufficient to achieve the efficacy or activity of the above-described marigold extract.

When the composition of the present invention is manufactured from a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. The pharmaceutically acceptable carriers to be contained in the pharmaceutical composition of the present invention are those conventionally used in the present invention and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. It is not. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention may be administered orally or parenterally, preferably by oral non-administration.

The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, . Typical dosages of the pharmaceutical compositions of this invention are in the range of 0.001-100 mg / kg on an adult basis.

The pharmaceutical composition of the present invention may be formulated into a unit dose form by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by a person having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container. The formulations may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of excipients, powders, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

The composition of the present invention can be provided as a food composition. When a composition for preventing, ameliorating or treating a Th2-mediated immune disease comprising the active ingredient of the present invention is used as a food composition, not only an extract of Safflower extract as an active ingredient but also a compound And includes, for example, proteins, carbohydrates, fats, nutrients, flavoring agents, and flavoring agents. Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings. For example, when the food composition of the present invention is prepared as a drink, it may further include citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, mulberry extract, jujube extract, licorice extract, .

The composition for preventing, ameliorating or treating Th2-mediated immune diseases comprising the extract of the present invention as an active ingredient can be produced as a health functional food. The health functional food may be any type of food such as a health functional food, a nutritional supplement, a nutrient, a pharmafood, a health supplement, a nutraceutical, a designer food, a food additive, etc. Preferably dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums and ice cream, various soups, drinks, tea, drinks, alcoholic beverages and vitamins And the like.

The health functional food of the present invention includes components that are ordinarily added at the time of food production, and includes, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavors. Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings. In addition to the above-mentioned food, the food of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), dietary components, synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate etc.) , Alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks and the like.

The features and advantages of the present invention are summarized as follows:

(a) The present invention provides a pharmaceutical composition and a food composition for improving, preventing or treating a Th2-mediated immune disease.

(b) The present invention provides an economical production cost and ease of use by including an extract of Sadomai extract which is a natural material capable of improving, preventing or treating an allergic disease as an active ingredient.

Figure 1 shows the effect of the extract of Costaria costata extract on B cell and T cell proliferation in splenocytes.
Figure 2 shows the effect on the production of Th1 cytokines IL-2 and IFN-y, which are the extracts of Safflower extract from spleen cells.
FIG. 3 shows the effect on the production of Th2 cytokines IL-10 and IL-4, which are the extracts of Safflower extract from spleen cells.
Figure 4 shows the effect on the IgE production of a marine extract from spleen cells.
FIG. 5 shows the effect on the histamine production of a marine extract from mast cells.
FIG. 6 shows the effect of chemokine (EOTAXIN and TARC) on the mRNA expression of the extract of Safflower extract in keratinocyte.
Fig. 7 shows the effect of cytokines (TNF-a and IL-6) on the expression of mRNAs of the extract of Allium cepa L. in the kelin-producing cells.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example 1: Preparation of sample

The red sea stew used in the preparation of Costaria costata extract was purchased from Wando and was manufactured by decontamination. The extracts were filtered with Whatman plc (Kent, UK) after refluxing (extraction temperature 90 ° C and time 4 hours) of 10% 24 g powder form (hereinafter referred to as CCW, CCE 10%) obtained by removing the extraction solvent using a concentrator (100 kg, Daemyung ENG Co., Ltd.) was dissolved in the third distilled water again sterilized and used for the experiment.

Experimental Example 1: Measurement of T cell and B cell proliferative capacity in spleen cells

Splenocytes isolated from animals (NC / Nga mouse, 6 weeks old, male) were washed with PBS, and a single cell suspension was prepared using 0.45 μm cell strainer. L-glutamine (Hyclone Laboratories, Logan, Utah, USA), 100 mg / L penicillin-streptomycin (Hyclone Laboratories, Logan, Utah, USA), 10% fetal bovine serum After washing with RPMI-1640 (Hyclone Laboratories, Logan, Utah, USA) supplemented with Hyclone Laboratories, Logan, Utah, USA, red blood cells were treated with erythrocyte lysis buffer (Sigma-Aldrich Co., The spleen cell suspension was prepared by hemolysis, and 200 쨉 l of 1 x 10 cells / well was dispensed into each well of a 96-well plate. 5 ㎍ / ㎖ of concanavalin A (Con A, Sigma-Aldrich Co., St. Louis, Mo., USA) was treated for T cell proliferative activity and LPS (lipopolysaccharide, Sigma -Aldrich Co., St. Louis, Mo., USA), and then the samples were treated together and incubated at 37 ° C for 48 hours. After 48 hours, 20 쨉 l of EZ-Cytox (Deilab INC, Korea) was dispensed and cultured at 37 째 C for 4 hours, and the absorbance was measured at 450 ㎚.

In the atopic control, the proliferative capacity of T cells was significantly decreased and the proliferative capacity of B cells was significantly increased compared to the normal group. T cell proliferative capacity was significantly increased in all groups treated with Daphnia magna extract compared to atopic control. In addition, the proliferative activity of B cells was significantly different from that of the atopic control in all the groups treated with Sadducein extract (Fig. 1). In this study, it was predicted that the normal function of T cells was lost by confirming the decrease of T cell proliferation by atopy. The increase of IgE due to the overgrowth of immunoglobulin due to the increase of B cells, It is activated and promotes histamine production, causing itching and inflammation. It seems that the extract of Daphniaceae inhibited abnormal proliferative activity of T cells and B cells and attenuated atopic dermatitis.

Experimental Example 2. Measurement of Th1 / Th2 cytokine production change test in spleen cells

IL-4, IL-10, and IFN [gamma]) were treated with 5 [mu] g / ml of Con A together with the sample treatment after the splenocytes were dispensed in a 96-well plate at a rate of 1 x 10 & -γ. IL-2, IL-4, and IL-10 were cultured for 24 hours, IFN-γ was cultured for 72 hours, and supernatants were collected. The amount of cytokine in the supernatant was measured using a DuoSet sandwich ELISA mouse kit (R & D System, McKinley Place NE, MN, USA). The primary antibody, which was characterized for each cytokine assay, was diluted with PBS and dispensed into 100-μl portions of the 96-well plate for ELISA for 1 day. The next day, the cells were washed with 1 ml of washing buffer (PBST, 0.05% Tween 20 in PBS) The kit for detecting IL-4 and IL-10 was washed with 1% BSA in PBS, IL-2 and IFN-γ detection kit 0.1% BSA in TBST) for 2 hours and washed with washing buffer. 100 .mu.l of the solution for the standard curve and the seeded spleen cell was incubated for 2 hours in each well. After completion of the reaction, the cells were washed with the washing buffer, and the secondary antibody was diluted in the assay buffer 100 [mu] L of each well was dispensed and treated for 2 hours. At the end of this process, the plates were washed with wash buffer and reacted with 100 μl of substrate reagent to aid color development. The absorbance was measured at 570 nm and the amount of cytokine produced in the cells was calculated using a standard curve.

CD4 + Th cells include Th1 cells that produce Th1-type cytokines and Th2 cells that produce Th2-type cytokines. Th1-type cytokines mainly stimulate phagocytosis by increasing macrophage activity, and Th2-type cytokines stimulate B cell activity and increase antibody production. It has been reported that the Th1 / Th2 type cytokine maintains the immunity balance by complementary regulation, and the Th1 type cytokine decreases and the Th2 type cytokine increases at the atopic state. Such cytokine imbalance lowers the ability to regulate immunity, resulting in chronic development of atopic symptoms.

As a result of the study, it was confirmed that the treatment of the extract of Daphnia magna extract significantly increased the production of Th1 type cytokines IL-2 and IFN-γ in all groups (FIG. 2). Whereas the production of Th2-type cytokines IL-4 and IL-10 was significantly reduced in all groups (Fig. 3). This efficacy was the most efficacious in the 10% extract of Sado marinade. Therefore, the treatment of Daphnia magna extract inhibited the imbalance of Th1 / Th2 type cytokines and confirmed the immunoregulatory ability.

Experimental Example 3: Immunoglobulin measurement test in spleen cells

Splenocytes were dispensed in a 96-well plate at a rate of 1 × 10 6 cells / well in a volume of 100 μl per well, and then immunoglobulin E (IgE) production with IL-4 (50 ng / ml) and LPS (10 μg / ml) And the sample was treated to cultivate for one week, and the supernatant was collected. The primary antibody, which had been characterized for each cytokine assay, was diluted with PBS and dispensed into 100-μl portions of the 96-well plate for ELISA overnight. After overnight treatment, the cells were washed with 1 ml of washing buffer (PBST, 0.05% Tween 20 in PBS) After washing the secondary antibody, an assay buffer (1% BSA in PBS) was added to the plate for blocking the antibody-unbound plate for 1 hour and then washed three times with washing buffer. 100 μl of the solution for the standard curve and the culture solution of the splenocytes seeded on the top were added to each well for 2 hours. After completion of the reaction, the cells were washed 5 times with the washing buffer, and the secondary antibody and SAv-HRP After dilution, 100 쨉 l of each well was dispensed and treated for 30 minutes. At the end of the procedure, wash the plates 7 times with wash buffer, add 100 μl of substrate reagent to aid color development, measure the absorbance at 650 nm, and calculate the amount of cytokine produced in the cells using a standard curve Respectively.

The production of IgE was significantly increased in the atopic control group as compared with the normal control group, and was significantly decreased in all of the soya extract-treated groups (Fig. 4).

Experimental Example 4: Test for measuring histamine in MC / 9 cells (mast cells)

Histamine is stored in mast cells, and when mast cells are stimulated and damaged, it releases and increases the relaxation and permeability of the blood vessels, causing blood vessels and plasma proteins to rapidly move to the site of invasion of bacteria, thus causing an inflammatory reaction. The excessive secretion of histamine due to excessive immune responses may cause allergic reactions and anaphylaxis. The excess IgE produced in B cells stimulates mast cells to increase histamine release and is an important indicator for evaluating allergic symptoms with IgE.

MC / 9 cells were prepared by dividing 100 μl of each 1 × 10 6 cells / cell into 96-well plates, stimulating histamine production with 1 μM of A23187 and 50 nM of phorbol 12-myristate 13-acetate (PMA) The samples were incubated for 24 hours, and the supernatant was collected and measured using a mouse histamine ELISA kit (LDN, Nordhorn, Germany). The reaction plate was dispensed with 25 μl of standard, control and sample, and treated with 25 μl each of acylation buffer and acylation reagent for 1 hour. 200 占 퐇 of distilled water was treated for 30 minutes, and 20 占 퐇 was collected and transferred to a histamine multi-titer strip. 100 [mu] l histamine antiserum was applied to each well and the foil was attached and incubated at 2-8 [deg.] C for 15-20 hours. After rinsing with washing buffer (PBST, 0.05% Tween 20 in PBS), 100 μl of enzyme conjugate was dispensed per well and treated with foil for 1 hour and washed with washing buffer. The substrate was treated with 100 μl of each well for 20-30 minutes and absorbance was measured at 650 nm. The mean value and standard error of the absorbance were determined by five measurements.

Histamine was released by stimulation with PMA and A23187 in MC / 9 cells, and the histamine concentration was significantly increased in the atopic control as compared with the normal control. It was confirmed that all the experimental groups treated with the extract of Safflower marigold were significantly reduced as compared with the atopic control (Fig. 5). Especially, the 10% alcohol extract of Daphni is the lowest level, which is considered to have the most positive effect on atopic dermatitis.

Experimental Example 5: Measurement of expression of chemokines and proinflammatory cytokines in kelin-producing cells (HaCaT cells)

Ketinogen-producing cells (keratinocyte, HaCaT cells) were dispensed in 2-ml portions at 1 × 10 6 cells / well in a 6-well plate and stimulated with hypersensitive skin with 20 ng / ml of TNF-α and 20 ng / After 24 hours from the treatment of the samples, the cells were collected and subjected to RNA extraction using the RNeasy extraction kit (Qiagen, Gaithersburg, Maryland, USA) according to the manufacturer's protocol. cDNA was synthesized using iScript cDNA synthesis kit (Bio-Rad Laboratories Headquarters, Hercules, Calif., USA). Real-time quantitative PCR was performed using SYBR Green (iQ SYBR Green Supermix, Bio-Rad Bio-Rad Laboratories Inc.) and real-time PCR was performed using Applied Biosystems (Foster City, CA, USA) Respectively. The nucleotide sequences of the PCR primers for each gene are shown in Table 1. In a total of 20 μl of the real-time PCR reaction, 2 μl of cDNA and 10 μl of 2 × SYBR mix were added, and 1 μl of 100 pmol / μl of the forward and reverse primers were added, respectively, and the remainder was filled with H ₂ O. The PCR amplification step was as follows and the amplification cycle was carried out for 40 cycles. Annealing at 95 ° C for 8 min at 95 ° C for hot start, denaturation at 95 ° C for 15 sec at the amplification step, annealing at 55 ° C for TARC (Thymus and Activation-regulated Chemokine) and Iotaxin, TNF- [alpha] and IL-6 were repeated at 56 [deg.] C for 1 minute 30 seconds and elongation at 72 [deg.] C for 1 minute, and values were recorded after each cycle of elongation. Melting curve analysis was performed to confirm the specificity of the primers after all cycles were completed. The analysis of the results was performed with the OneStem System Software v2.1 provided by Applied Biosystems.

gene Primer sequence Sequence List GAPDH (H) F 5'-ATGGAAATCCCATCACCATCTT-3 ' First sequence R 5'-CGCCCCACTTGATTTTGG-3 ' Second sequence TARC (H) F 5'-GAAGACGTGGTACCAGACATCTGA-3 ' Third sequence R 5'-CCCTGCACAGTTACAAAAACGA-3 ' Fourth sequence Iot Thaksin (H) F 5'-GCGACTAGAGAGCTACAGGAGAATC-3 ' Fifth sequence R 5'-GGTCTTGAAGATCACAGCTTTCTG-3 ' 6th sequence IL-6 (H) F 5'-AGGGCTCTTCGGCAAATGTA-3 ' Seventh sequence R 5'-GAAGGAATGCCCATTAACAACAA-3 ' Eighth sequence TNF-α (H) F 5'-CCACTTCGAAACCTGGGATTC-3 ' Ninth sequence R 5'-TTAGTGGTTGCCAGCACTTCA-3 ' Tenth sequence

The mRNA production of TARC and iotaxin treated with Sukwoo seaweed was significantly decreased compared with the control group of atopic dermatitis. In particular, in the case of iotaxin, it decreased to the normal group level from CCW 1000 μg / ml, CCE 10% 200 μg / ml and CCE 10% 1000 μg / ml, and TARC decreased to the normal group level from CCE 10% 1000 μg / (Fig. 6).

The cytokine-stimulated keratinocytes increased TNF-α, but were significantly reduced in all experimental groups treated with Dextran extract, especially in the CCE 10% 1000 group. In addition, the increased mRNA expression of IL-6 was significantly reduced in all experimental groups treated with Sukwa seaweed extract, especially in CCW 1000, CCE 10% 1000 and CCE 70% 200 (FIG. 7) .

  In conclusion, the results of this study showed that the extract of Daphniaceae showed the inhibitory effect on atopic dermatitis. Especially, the efficacy was confirmed by inhibiting the secretion of histamine in the 10% alcohol extract. These results suggest that the extract of Sadamushi extract may be used as a functional material having atopic dermatitis inhibitory activity in Korea.

The formulation of the composition containing the extract of the present invention is described above, but the present invention is not intended to be limited thereto but is specifically described.

Formulation Example 1: Preparation of powder

CCE10% ---------------------------------------------- 20 mg

Lactose -------------------------------------------- 100 mg

Talc --------------------------------------------- 10 mg

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

Formulation Example 2: Preparation of tablets

CCE 10% -------------------------------------------- 10 mg

Corn starch -------------------------------------- 100 mg

Lactose -------------------------------------------- 100 mg

Magnesium stearate ------------------------------- 2 mg

After mixing the above components, tablets are prepared by tableting according to a conventional tablet preparation method.

Formulation Example 3: Preparation of capsules

CCE 10% ------------------------------------------- 10 mg

Crystalline cellulose - 3 mg

Lactose --------------------------------------- 14.8 mg

Magnesium stearate - 0.2 mg

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

Formulation Example 4: Preparation of injection

CCE 10% ------------------------------------------- 10 mg

Mannitol ------------------------------------------ 180 mg

Sterile sterilized distilled water for injection - 297 mg

Na ₂ HPO ₄ 12H ₂ O ------------------------------------ 26 mg

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

Formulation Example 5. Preparation of a liquid preparation

CCE 10% ------------------------------------- 20 mg

Isomerization Party ------------------------------------------ 10 g

Mannitol --------------------------------------------- 5 g

Purified water --------------------------------------------

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

Formulation Example 6: Preparation of health food

CCE 10% ------------------------------------- 1000 mg

Vitamin mixture -------------------------------------

Vitamin A Acetate ------------------------------ 70 g

Vitamin E ---------------------------------------- 1.0 mg

Vitamin B ₁ --------------------------------------- 0.13 mg

Vitamin B ₂ --------------------------------------- 0.15 mg

Vitamin B ₆ ---------------------------------------- 0.5 mg

Vitamin B ₁₂ --------------------------------------- 0.2 g

Vitamin C ----------------------------------------- 10 mg

Biotin ------------------------------------------- 10 [mu] g

Nicotinic acid amide 1.7 mg

Folic acid --------------------------------------------- 50 μg

Calcium pantothenate ----------------------------------- 0.5 mg

Inorganic mixture -------------------------------------

Ferrous sulfate -------------------------------------- 1.75 mg

Zinc oxide --------------------------------------- 0.82 mg

Magnesium carbonate - 25.3 mg

Potassium phosphate monohydrate 15 mg

Secondary calcium phosphate -------------------------------------- 55 mg

Potassium citrate --------------------------------------- 90 mg

Calcium carbonate ---------------------------------------- 100 mg

Magnesium chloride ----------------------------------- 24.8 mg

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

Formulation Example 7: Preparation of health drinks

CCE10% ---------------------------------------- 1000 mg

Citric acid ---------------------------------------------- 1000 mg

Oligosaccharide --------------------------------------------- 100 g

Plum concentrate --------------------------------------------- 2 g

Taurine ------------------------------------------------- 1 g

Purified water was added.

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 L container, It is used in the production of the health beverage composition of the invention.

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

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

<110> INDUSTRY FOUNDATION OF CHONNAM NATIONAL UNIVERSITY <120> Comositions for Improving, Preventing or Treating for Th2-immune          mediated Disease comprising Extracts from Ecklonia stolonifera <130> PN160106 <160> 10 <170> Kopatentin 2.0 <210> 1 <211> 22 <212> DNA <213> GAPDH (H) forward primer <400> 1 atggaaatcc catcaccatc tt 22 <210> 2 <211> 18 <212> DNA <213> GAPDH (H) reverse primer <400> 2 cgccccactt gattttgg 18 <210> 3 <211> 24 <212> DNA <213> TARC (H) forward primer <400> 3 gaagacgtgg taccagacat ctga 24 <210> 4 <211> 22 <212> DNA <213> TARC (H) reverse primer <400> 4 ccctgcacag ttacaaaaac ga 22 <210> 5 <211> 25 <212> DNA <213> Eotaxin (H) forward primer <400> 5 gcgactagag agctacagga gaatc 25 <210> 6 <211> 24 <212> DNA <213> Eotaxin (H) reverse primer <400> 6 ggtcttgaag atcacagctt tctg 24 <210> 7 <211> 20 <212> DNA IL-6 (H) forward primer <400> 7 agggctcttc ggcaaatgta 20 <210> 8 <211> 23 <212> DNA <213> IL-6 (H) reverse primer <400> 8 gaaggaatgc ccattaacaa caa 23 <210> 9 <211> 21 <212> DNA <213> TNF-a (H) forward primer <400> 9 ccacttcgaa acctgggatt c 21 <210> 10 <211> 21 <212> DNA <213> TNF-a (H) reverse primer <400> 10 ttagtggttg ccagcacttc a 21

Claims (6)

(a) a therapeutically effective amount of an extract of Costaria costata ; And (b) a pharmaceutically acceptable carrier. A pharmaceutical composition for improving, preventing or treating a Th2-mediated immune disorder, comprising a pharmaceutically acceptable carrier.
3. The composition of claim 1, wherein the extract is a desalinized waxy sea weed extract.
The extract of claim 1, wherein the extract is at least one selected from the group consisting of water-in-cane starch, 1-4 anhydrous or lower alcohol, acetone, ethyl acetate, butyl acetate or 1,3-butylene glycol Composition.
2. The composition of claim 1, wherein the Th2-mediated immune disorder is an allergic disease.
The method of claim 4, wherein the allergic disease is selected from the group consisting of atopy dermatitis, allergic rhinitis, bronchial asthma, contact dermatitis, allergic conjunctivitis, wherein the composition is selected from the group consisting of urticaria, vascular edema, food allergies, physical allergies, irritable pulmonary enteritis, occupational allergic diseases or drug allergies.
(a) a pharmacologically effective amount of an extract of Costaria costata ; And (b) a food-acceptable carrier.

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