KR102047074B1 - Anti-allergic Composition Comprising neolicuroside as an Active Ingredient - Google Patents

Abstract

The present invention relates to a composition for the prevention or treatment of allergic diseases comprising neolicuroside as an active ingredient, specifically, atopic dermatitis, asthma, allergic rhinitis or allergy including neorikuroside as an active ingredient. A composition for preventing or treating sex conjunctivitis, and a method for preventing or treating allergic disease, comprising administering the composition to a subject.
Since the composition using the neorikuroside of the present invention as an active ingredient can inhibit inflammatory mediators and degranulation and effectively suppress allergic reactions at the level of signaling mechanisms, it can be usefully used as a composition for preventing or treating allergic diseases. Can be.

Description

Anti-allergic Composition Comprising neolicuroside as an Active Ingredient

The present invention relates to a composition for the prevention or treatment of allergic diseases comprising neolicuroside as an active ingredient, specifically, atopic dermatitis, asthma, allergic rhinitis or allergy including neorikuroside as an active ingredient. A composition for preventing or treating sex conjunctivitis, and a method for preventing or treating allergic disease, comprising administering the composition to a subject.

Recently, allergic diseases are on the rise, and research and development of therapeutic agents are also progressing. An allergic disease refers to a disease caused by immune hypersensitivity reactions. An allergic reaction is a complex reaction involving various cells and mediators and also causes various diseases. Therefore, the treatment of various diseases is expected to be possible through the control of allergic reactions, and research on this continues. Allergic diseases include atopic dermatitis, asthma, allergic rhinitis, allergic conjunctivitis, allergic dermatitis, and the like.

Atopic dermatitis is a worldwide disease that can occur at any age, with 70% to 95% occurring in infants under 5 years of age. Even in Korea, the problem of atopic dermatitis has reached a serious social and medical level. According to various reports, about 15% of Koreans are atopic dermatitis patients, and 18 to 22 of 100 infants aged 1 to 4 years are known as atopic patients. Recently, the increase in atopic dermatitis in children as well as in adults is becoming serious, and in severe cases, it causes serious social problems such as employment and marriage. In such cases, severe mental stress may cause extreme avoidance and depression. Although atopic dermatitis has become a serious social and medical problem, there is no effective treatment to date.

Allergic rhinitis is the most common allergic disease, and its incidence is gradually increasing, affecting the quality of life, and becoming a socioeconomic problem due to decreased production capacity and increased burden of treatment costs, but there is no clear treatment yet. It is true. Currently used drug therapy is a basic treatment for allergic rhinitis, but recurrences are common and serious side effects when the drug is discontinued. Immunotherapeutic may be the only treatment that can restore the only modified immune system, but there are cost problems. .

Allergic conjunctivitis is an allergic disease that refers to an acute inflammatory disease of the conjunctiva (white egg) caused by certain allergens.Allergens such as pollen in the spring, dust in the air, and dandruff in animals come into contact with the conjunctiva of the eye. When triggered allergic reactions through immune cells, such as mast cells, eosinophils or basophils, various inflammatory substances, such as histamine, are secreted to cause an inflammatory response in the conjunctiva. Such allergic conjunctivitis can be treated with glucocorticoid steroid eye drops, but such anti-inflammatory drugs may cause sudden systemic weakness, fever, increased intraocular pressure, such as acute adrenal insufficiency, if the medication is suddenly discontinued or prolonged overuse after long-term use. Side effects such as glaucoma and cataract may occur, and there is a need for an antiallergic agent without such problems.

Under these backgrounds, the present inventors have diligently researched to develop a drug that can effectively treat allergic diseases. As a result, neocururoside exhibits excellent antiallergic effect with little cytotoxicity, thus preventing or treating allergic diseases. Confirmed that the present invention was completed.

One object of the present invention is to provide a pharmaceutical composition for preventing or treating allergic diseases, including Neolicuroside and its pharmaceutically acceptable salts.

Another object of the present invention is to provide a dietary supplement for preventing or ameliorating allergic diseases, including neorikuroside and its food acceptable salt.

Still another object of the present invention is to provide a quasi-drug composition for preventing or ameliorating allergic diseases, including neorikuroside and pharmaceutically acceptable salts thereof.

Still another object of the present invention is to provide a cosmetic composition for preventing or ameliorating allergies including neorikuroside and a cosmetically acceptable salt thereof.

Yet another object of the present invention is to provide a method for preventing or treating allergic diseases, comprising administering to a subject, except humans, a composition comprising neorikuroside and a pharmaceutically acceptable salt thereof.

As one embodiment for solving the above problems, the present invention provides a pharmaceutical composition for the prevention or treatment of allergic diseases, including Neolicuroside and its pharmaceutically acceptable salts.

Hereinafter, the anti-allergic composition which uses neorikuroside as an active ingredient in this invention is demonstrated concretely.

The term "Neolicuroside" of the present invention is a compound having a structure of Formula 1, and is also named Isoliquiritin apioside. However, the function of neorikuroside is not well known.

[Formula 1]

The neorikuroside may be obtained through extraction and purification from natural products, or may be purchased commercially, but is not limited thereto.

The neorikuroside may be degranulation inhibition, inflammatory mediator production inhibition, arachidonic acid chain reaction inhibition or FcεRI signaling mechanism.

In one embodiment of the present invention, as a result of treating the neorikuroside in mast cells induced allergic reaction, it was confirmed that effectively suppress the degranulation reaction that is an indicator of allergic reaction (Fig. 1).

In one embodiment of the present invention, as a result of analyzing the supernatant of the mast cells treated with the compound, it was confirmed that the inhibitory effect of the production of inflammatory mediators compared to the control (Fig. 3).

In one embodiment of the present invention, after inducing an allergic reaction, the resultant protein was lysed by lysing mast cells treated with the compound to quantify allergic reaction mechanism-related proteins. And 5).

As used herein, the term "allergic disease" refers to a disease caused by an immune hypersensitivity reaction, and an allergic reaction is a complex reaction involving various cells and mediators and at the same time causes various diseases. Specifically, the allergic disease may be any one selected from the group consisting of allergic rhinitis, asthma, atopic dermatitis and allergic conjunctivitis, but is not limited thereto.

As used herein, the term "prevention" may mean any action of inhibiting or delaying the development of allergic diseases by administering neorikuroside according to the present invention to a subject.

As used herein, the term "treatment" may refer to any action of administering the composition of the present invention to a subject suspected of developing an allergic disease so as to improve or benefit from an allergic disease.

The pharmaceutical composition of the present invention may also be used as a single agent, and may be prepared and used as a complex preparation, further including a drug known to have a recognized allergic disease treatment effect, and formulated using a pharmaceutically acceptable carrier or excipient. It may be prepared in unit dose form or incorporated into a multi-dose container.

As used herein, the term "pharmaceutically acceptable carrier" may refer to a carrier or diluent that does not interfere with the biological activity and properties of the compound to be injected without stimulating the organism. The kind of the carrier usable in the present invention is not particularly limited, and any carrier can be used as long as it is a conventionally used and pharmaceutically acceptable carrier in the art. Non-limiting examples of the carrier include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol and the like. These may be used alone or in combination of two or more thereof. The carrier may include a non-naturally occuring carrier.

In addition, if necessary, other conventional additives such as antioxidants, buffers and / or bacteriostatic agents may be added and used, and diluents, dispersants, surfactants, binders, lubricants, and the like may be additionally added to provide a solution such as an aqueous solution, a suspension, an emulsion, or the like. It may be formulated into a use formulation, pills, capsules, granules or tablets.

In addition, the pharmaceutical composition of the present invention may include a pharmaceutically effective amount of neorikuroside. As used herein, the term “pharmaceutically effective amount” means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, generally in an amount of 0.001 to 1000 mg / kg, preferably 0.05 The amount of to 200 mg / kg, more preferably 0.1 to 100 mg / kg may be administered once to several times daily. However, for the purposes of the present invention, the specific therapeutically effective amount for a particular patient is determined by the specific composition, including the type and extent of the reaction to be achieved, whether or not other agents are used in some cases, the age, weight, general health of the patient, It is desirable to apply differently depending on various factors and similar factors well known in the medical field, including sex and diet, time of administration, route of administration and rate of composition, duration of treatment, drugs used with or co-specific with the specific composition.

The pharmaceutical compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents and may be administered sequentially or simultaneously with conventional therapeutic agents. And single or multiple administrations. In consideration of all the above factors, it is important to administer an amount that can achieve the maximum effect in a minimum amount without causing side effects, and can be easily determined by those skilled in the art.

As used herein, the term "administration" means introducing a pharmaceutical composition of the present invention to a patient in any suitable manner, and the route of administration of the composition of the present invention may be various oral or parenteral routes as long as the target tissue can be reached. It can be administered through.

The mode of administration of the pharmaceutical composition according to the present invention is not particularly limited, and may be in accordance with methods commonly used in the art. As a non-limiting example of the mode of administration, the composition may be administered by oral or parenteral administration. The pharmaceutical compositions according to the invention may be prepared in various formulations depending on the desired mode of administration.

The frequency of administration of the composition of the present invention is not particularly limited, but may be administered once a day or several times in divided doses.

As used herein, the term `` pharmaceutically acceptable salt '' refers to a formulation that does not impair the biological activity and properties of the neorikuroside administered. The pharmaceutically acceptable salts are acids that form non-toxic acid addition salts containing pharmaceutically acceptable anions, such as inorganic acids, such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydroiodic acid, and tartaric acid. Organic carboxylic acids such as formic acid, citric acid, acetic acid, trichloroacetic acid, trichloroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, salicylic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfate Acid addition salts formed by sulfonic acids and the like such as phonic acid and the like. For example, pharmaceutically acceptable carboxylic acid salts include metal salts or alkaline earth metal salts formed by lithium, sodium, potassium, calcium, magnesium, amino acid salts such as lysine, arginine, guanidine, dicyclohexylamine, N Organic salts such as -methyl-D-glucamine, tris (hydroxymethyl) methylamine, diethanolamine, choline and triethylamine and the like.

In another aspect, the present invention provides a dietary supplement for preventing or ameliorating allergic diseases, including neorikuroside and its food acceptable salt.

Functional food is the same term as Food for special health use (FOSHU), and means foods with high medical effects and processed foods to efficiently display bioregulatory functions in addition to nutrition. In addition, the food may be prepared in various forms such as tablets, capsules, powders, granules, liquids, pills, etc. in order to obtain useful effects in preventing or ameliorating diseases.

The food composition of the present invention may further comprise a food acceptable carrier.

There is no restriction | limiting in particular in the kind of food which can add the composition containing neorikuroside of this invention, For example, there are various drinks, gum, tea, a vitamin complex, a health supplement food, etc. The food composition may be added to other ingredients that do not interfere with the prevention or improvement effect of allergic diseases, the kind is not particularly limited. For example, various herbal extracts, food acceptable additives, natural carbohydrates, and the like may be contained as additional ingredients, such as conventional foods.

The food supplement additive is added to prepare the health functional food of each formulation can be used by those skilled in the art as appropriate. For example, various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic and natural flavors, colorants and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters , Stabilizers, preservatives, glycerin, alcohols, carbonation agents used in the carbonated beverage, and the like, but the type is not limited by the above examples.

At this time, the content of the extract included in the food is not particularly limited, but may be included in 0.01 to 100% by weight, more preferably 1 to 80% by weight relative to the total weight of the food composition.

When food is a beverage, it may be included in a ratio of 1 to 30 g, preferably 3 to 20 g based on 100 ml. In addition, the composition may include additional ingredients that are commonly used in food compositions to improve the smell, taste, time and the like. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid, and the like. Also, minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn) and copper (Cu) may be included. It may also contain amino acids such as lysine, tryptophan, cysteine, valine and the like. In addition, preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetic acid, etc.), fungicides (bleaching powder and highly bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisol (BHA), butylhydroxytoluene ( BHT), etc.), colorants (such as tar pigments), colorants (such as sodium nitrite, sodium nitrite), bleach (sodium sulfite), seasonings (sodium glutamate (MSG), etc.), sweeteners (ducin, cyclate, saccharin, sodium Food additives such as fragrances (vanillin, lactones, etc.), swelling agents (alum, potassium D-tartrate, etc.), reinforcing agents, emulsifiers, thickeners (pigments), coating agents, gum herbicides, antifoaming agents, solvents, and improving agents ( Food additives may be added. The additive is selected according to the type of food and used in an appropriate amount.

The health functional food of the present invention can be prepared by a method commonly used in the art, and the preparation can be prepared by adding raw materials and ingredients commonly added in the art. In addition, unlike the general medicine has the advantage that there is no side effect that can occur when taking a long-term use of the drug with food as a raw material, can be excellent in portability.

As another aspect, the present invention provides a quasi-drug composition for preventing or ameliorating allergic diseases, including neorikuroside and pharmaceutically acceptable salts thereof.

As used herein, the term "quasi drug" refers to articles that have a lesser action than drugs among articles used for the purpose of diagnosing, treating, ameliorating, alleviating, treating or preventing a disease of a human or animal. For example, quasi-drug products under the Pharmaceutical Affairs Law exclude products used for the purpose of medicines, and include products used for the treatment or prevention of diseases of humans / animals, and products with slight or no direct action on the human body.

Specifically, the quasi-drug may include external skin preparations and personal hygiene products. More specifically, it may be, but is not limited to, a disinfectant cleaner, a shower foam, a gagreen, a wet tissue, a detergent soap, a hand wash, or an ointment.

When the composition according to the present invention is used as an quasi-drug additive, the composition may be added as it is or used with other quasi-drugs or quasi-drug components, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the intended use.

As another aspect, the present invention provides a cosmetic composition for preventing or ameliorating allergies, including neorikuroside and its cosmetically acceptable salts.

In the present invention, the cosmetic composition may further include one or more cosmetically acceptable carriers formulated in general skin cosmetics, and as a typical component, for example, oil, water, surfactants, moisturizers, lower alcohols , Thickeners, chelating agents, pigments, preservatives, fragrances and the like may be appropriately blended, but is not limited thereto.

The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation of the cosmetic composition.

When the formulation of the present invention is an ointment, paste, cream or gel, the carrier component is animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide and the like. May be used, but is not limited thereto. These may be used alone or in combination of two or more thereof.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder and the like can be used, and especially in the case of a spray, additionally chlorofluorohards. Propellants such as carboxylic, propane / butane or dimethyl ether may be included, but are not limited thereto. These may be used alone or in combination of two or more thereof.

When the formulation of the present invention is a solution or an emulsion, a solvent, a solubilizer or an emulsifier may be used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Propylene glycol, 1,3-butylglycol oil, and the like can be used, and in particular, cottonseed oil, peanut oil, corn seed oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid ester May be used, but is not limited thereto. These may be used alone or in combination of two or more thereof.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, micro Crystalline cellulose, aluminum metahydroxyde, bentonite, agar or tracant may be used, but is not limited thereto. These may be used alone or in combination of two or more thereof.

When the formulation of the present invention is a soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolyzates, isethionates, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugars and the like can be used as carrier components. May be, but is not limited thereto. These may be used alone or in combination of two or more thereof.

As another aspect, the present invention provides a method for preventing or treating an allergic disease, comprising administering to a subject other than a human a composition comprising neorikuroside and a pharmaceutically acceptable salt thereof.

As used herein, the term "individual" may mean any animal, including a human, who has or is likely to develop an allergic disease. The animal may be a mammal such as, but not limited to, a human, a cow, a horse, a sheep, a pig, a goat, a camel, a antelope, a dog, a cat, and the like, which require treatment of similar symptoms.

The method for preventing or treating the present invention may specifically include administering the composition in a pharmaceutically effective amount to an individual with or at risk of developing an allergic disease.

As used herein, the term "administration" refers to introducing a pharmaceutical composition of the present invention to a patient by any suitable method, and the route of administration of the composition of the present invention may be various oral or parenteral routes as long as the target tissue can be reached. It can be administered through.

Since the composition using the neorikuroside of the present invention as an active ingredient can suppress inflammatory mediators and degranulation and effectively suppress allergic reactions at the level of signaling mechanisms, it can be usefully used as a composition for preventing or treating allergic diseases. Can be.

1 is a diagram showing the results of inhibiting mast cell degranulation according to the concentration of neolicuroside (Neolicuroside). ** P <0.01 compared to IgE / DNP-HSA group. A relates to the activity of β-hexosaminidase, and B relates to the amount of histamine released. In this case, DNP-IgE (Dinitrophenyl-immunoglobulin E) and DNP-HSA (Dinitrophenyl-human serum albumin) mean that the mast cells are sensitized (+) or not (-).
Figure 2 shows the results of the cytotoxicity according to the concentration of neorikuroside. At this time, DNP-IgE and DNP-HSA means that (+), or not (-) sensitizing mast cells with the substance.
Figure 3 is a result of the inhibitory effect of neorikuroside on the production of inflammatory mediators. * P <0.05, ** P <0.01 compared to IgE / DNP-HSA group. A is TNF-α, B is IL-4, C is LTC ₄ and D is the expression level of PGD ₂ . At this time, DNP-IgE and DNP-HSA means that (+), or not (-) sensitizing mast cells with the substance.
4 shows the results of plotting the inhibitory effect of neorikuroside on the arachidonic acid chain reaction. It relates to the expression level of p-cPLA ₂ , COX-2, p-5-LO and L-PGDS.
Figure 5 shows the results of the efficacy of neorikuroside in the FcεRI signaling. A is p-Syk, p-Lyn and p-Fyn, B is p-PLCγ1, p-PLCγ2 and p-PKCδ, C is the expression level of p-ERK, p-JNK, p-p38 and p-Akt will be. At this time, β-Actin was used as a loading control.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

Example 1 Determination of Degranulation Inhibitory Effect of Neolicuroside

In order to confirm the degranulation inhibitory effect of neolicuroside, allergic reactions of mast cells were induced. And the efficacy of neorikuroside through the amount of histamine released. Specifically, RBL-2H3 cells were dispensed in 24-well plates at a concentration of 1 × 10 ⁵ cells / well, 100 units / mL penicillin, 100 μg / mL streptomycin and 5% (v / v) cattle. Incubated overnight at 37 ° C., 5% CO ₂ , humidified conditions in MEM-α medium containing fetal serum. After incubation, cells were washed with 1 × PBS buffer and incubated with 0.1 μg / mL of DNP-IgE (Sigma, Co., Ltd) for 24 hours to sensitize the cells.

After incubating for 1 hour after adding neorikuroside by concentration, 0.1 ㎍ / mL of DNP-HSA (Dinitrophenyl-human serum albumin, Sigma, Co., Ltd.) was added to induce hypersensitivity and incubated for another 4 hours. It was. The supernatant was taken and centrifuged at 17,000 g at 4 ° C. for 10 minutes, and the supernatant was stored at −80 ° C. and thawed and used for analysis.

In order to measure the activity of β-hexosaminidase secreted from the cells into the medium by the above method, 10 mM p-NAG prepared with 25 μl of supernatant and 0.1 M sodium citrate buffer (pH 4.5) 50 μl of 4-Nitropheyl N-acetyl-β-D-glucosaminide) was placed in a 96-well plate and incubated at 37 ° C. After 1 hour of incubation, the reaction was terminated with 0.1 M Na ₂ CO ₃ buffer (pH 10.0) and the activity of β-hexosaminidase was determined by measuring the amount of p-nitrophenol with absorbance at 405 nm. It was. Each result is shown as the average of three experiments.

In addition, in order to measure the amount of histamine secreted from the cells to the medium by the above method, the concentration of histamine in the supernatant of allergic-induced mast cells was measured using an ELISA kit (Enzo Life Sciences, Inc.).

As a result, as shown in Fig. 1A, neorikuroside of the present invention can inhibit the secretion of β-hexosaminidase from mast cells in a concentration-dependent manner, it was confirmed that the IC ₅₀ value is 294.1 μM. In particular, neorikuroside showed more than 90% of β-hexosaminidase secretion inhibitory effect compared to the negative control at the concentration of 800 μM, confirming that neorikuroside can suppress allergic reactions by inhibiting degranulation of mast cells. .

In addition, as shown in B of FIG. 1, the neorikuroside of the present invention can inhibit histamine secretion from mast cells in a concentration-dependent manner, and the IC ₅₀ value was 240 μM. In particular, neorikuroside showed an excellent histamine secretion inhibitory effect even at a low concentration of 25 μM, confirming that neorikuroside can inhibit allergic reactions by inhibiting degranulation of mast cells.

Example 2. Confirmation of Cytotoxicity of Neorikuroside

To confirm the cytotoxicity of neorikuroside, RBL-2H3 cells were dispensed in 96-well plates at a concentration of 1 × 10 ⁴ cells / well, followed by MEM containing 5% (v / v) fetal bovine serum. In -α medium, it was incubated for 12 hours at a temperature of 37 ℃. The cells that were cultured the next day were washed with 1 × PBS buffer and incubated with 0.1 μg / mL of DNP-IgE (Dinitrophenyl-immunoglobulin E; Sigma, Co., Ltd.) for 24 hours to sensitize the cells.

Incubated for 1 hour after adding neorikuroside to the sensitized cells by concentration, 10 μl of WST-1 reagent (Daeil Lab Service. Co., Ltd.) and DNP-HSA (Dinitrophenyl-human serum albumin; Sigma, Co., Ltd.) 0.1 μg / mL were added simultaneously and incubated for another 4 hours. In order to confirm cell viability, the cell viability of mast cells was confirmed by measuring absorbance at 450 nm using a micro-plate reader (Emax, Molecular Devices Inc. Sunnyvale, California, USA).

As a result, as shown in FIG. 2, it was confirmed that neorikuroside does not show cytotoxicity.

Example 3 Confirmation of Inflammatory Mediator Inhibitory Effect of Neorikuroside

To confirm the inflammatory mediator inhibitory effect of neorikuroside, TNF-α, IL-4 (Interleukin 4), PGD ₂ (Prostaglandin D ₂ ) in the supernatant of mast cells induced by allergic reaction by the method of Example 1 And the concentration of LTC ₄ (Leukotriene C ₄ ) was measured using an ELISA kit (e-Bioscience, Inc.) and an EIA kit (Cayman Chemical, Inc.).

As a result, as shown in A, C, and D of FIG. 3, neorikurosides exhibit IC ₅₀ values of 59.50 μM, 200.1 μM, and 119.5 μM for TNF-α, PGD _2, and LTC ₄ , respectively, and FIG. 3. As shown in B, it was confirmed that the excellent IL-4 production inhibitory effect also at a concentration of 400 μM or more for IL-4. Thus, neorikuroside was confirmed to inhibit the production of the inflammatory mediators in a concentration-dependent manner, it was confirmed that it can suppress the allergic disease induced through the inflammatory response.

Example 4 Effect of Inhibiting Allergic Mechanism of Neorikuroside

Example 4-1. Confirmation of the inhibitory effect of arachidonic acid chain reaction of neorikuroside

Arachidonic acid chain reaction is a reaction that produces a bioactive substance such as prostaglandin or leukotriene through an enzyme reaction mechanism from arachidonic acid, eicosapentaenoic acid, and the like, and is closely related to allergic diseases.

Therefore, in order to confirm the efficacy of inhibiting arachidonic acid chain reaction, after lysing the allergic-induced mast cells of Example 1 and extracting total protein, arachidonic acid chain reaction related proteins (p-cPLA ₂ , COX-2, p -5-LO and L-PGDS) were quantified using Western blot. Antibodies used in Western blot were Cell signaling Technology, Inc. It was purchased from and used.

As a result of the quantification, as shown in FIG. 4, neorikuroside inhibits the expression of arachidonic acid chain reaction related proteins p-cPLA ₂ , COX-2, p-5-LO and L-PGDS in a concentration-dependent manner. It confirmed that.

Example 4-2. Confirmation of Neorikuroside Inhibitory Effect on FcεRI Signaling

FcεRI is a high affinity IgE receptor and is known to cause allergic diseases by mediating IgE dependent allergen expression.

Therefore, in order to confirm the effect of inhibiting FcεRI signaling, FcεRI signaling related proteins (p-Syk, p-Lyn, p-Fyn, p-PLCγ1, p-PLCγ2, p) in the same manner as in Example 4-1 -PKCδ, p-ERK, p-JNK, p-p38 and p-Akt) were quantified.

As a result, as shown in Figs. 5A to 5C, the neorikurosides were p-Syk, p-Lyn, p-PLCγ1, p-PLCγ2, p-PKCδ, p-ERK, p-JNK, p-p38 And it was confirmed that the concentration-dependent inhibition of the expression of p-Akt.

Through Example 4, confirming the suppression of allergy-related mechanisms of neorikuroside confirmed that neorikuroside can inhibit the allergic reaction at the level of cellular signaling.

Taken together, Neo Example can suppress degranulation of mast cells, which are indicative of allergic diseases, inhibit the production of inflammatory mediators, and suppress the mechanisms associated with allergic reactions. It was confirmed that the present invention can be used as an antiallergic composition.

From the above description, those skilled in the art will appreciate that the present invention can be implemented in other specific forms without changing the technical spirit or essential features. In this regard, it should be understood that the embodiments described above are exemplary in all respects and not restrictive. The scope of the present invention should be construed that all changes or modifications derived from the meaning and scope of the following claims and equivalent concepts rather than the detailed description are included in the scope of the present invention.

Claims (7)

Pharmaceutical composition for the prevention or treatment of atopic dermatitis, including Neolicuroside and its pharmaceutically acceptable salts.
delete The pharmaceutical composition of claim 1, wherein the neorikuroside inhibits degranulation, inhibits inflammatory mediator production, inhibits arachidonic acid chain reaction, or inhibits FcεRI signaling.
A dietary supplement for the prevention or amelioration of atopic dermatitis, including neorikuroside and its food acceptable salt.
A quasi-drug composition for preventing or ameliorating atopic dermatitis, including neorikuroside and a pharmaceutically acceptable salt thereof.
A cosmetic composition for preventing or ameliorating atopic dermatitis, including neorikuroside and a cosmetically acceptable salt thereof.
A method for preventing or treating atopic dermatitis, comprising administering to a subject, except a human, a composition comprising neorikuroside and a pharmaceutically acceptable salt thereof.

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