KR20200095266A - Composition comprising Avenanthramide C for Preventing, Treating or Improving Allergic Disease - Google Patents

Abstract

The present invention relates to a composition for preventing, treating or alleviating allergic diseases containing abenanthamide C as an active ingredient, which inhibits histamine release from mast cells related to an allergic reaction, and also excellently suppresses activities of NF-κB which is a transcription factor, and gene expression of TNF-α and IL-4 which are inflammation-inducing cytokines, thereby being usefully used in the prevention and treatment of allergic diseases, particularly allergic diseases mediated by mast cells.

Description

Composition comprising Avenanthramide C for Preventing, Treating or Improving Allergic Disease, comprising avenanthramide C as an active ingredient

The present invention relates to a composition for preventing, treating or improving allergic diseases comprising abenanthramid C as an active ingredient.

Allergy is a generalization of extensive and complex pathological phenomena and is a systemic or local disorder of a living body based on an immune response. Allergies appearing in the human body are classified into types Ⅰ, Ⅱ, Ⅲ and Ⅳ according to the immune mechanism. Among them, type I allergy, which is an immediate type hypersensitivity reaction, occupies an important part in clinical practice. These include bronchial asthma, hay fever and hay fever.

Around 1953, type I allergy is caused by the activation of mast cells, and it has been reported that the granules of these mast cells contain a large amount of histamine, a mediator of the inflammatory reaction. While elucidating the mechanism of histamine release from mast cells when an allergic reaction occurs, Ishizaka's discovery of immunoglobulin E (IgE) revealed that mast cells are involved in immediate allergic reactions. It was an important opportunity. In other words, there is a high affinity receptor for IgE on the surface of the mast cell, and when the antigen binds again to form a crosslink after IgE binds to this receptor, a degranulation reaction is triggered, and the granule contents such as histamine, serotonin, bradykinin, etc. The same synthesized and stored mediator, protease, and proteoglycan are simultaneously released (Ishizaka, Hosp Pract.; 12(1):57-67, 1977).

Since the 1970s, research results on new lipid-like inflammatory mediators have begun to be reported, and these are prostaglandins, leukotriens, thromboxane, and glycero produced from cell membrane phospholipids accompanied by cell activation. It has been found to be involved in various inflammatory reactions, including allergic reactions of type I, as new mediators exhibiting various physiological activities such as platelet activating factor (PAF), a derivative of glycerophospholipids. These lipid mediators have been reported to be produced and released in parallel with the degranulation reaction in mast cells when the IgE receptor forms a bridge.

Type Ⅰ allergy is known as active amines and proteases, which are granular contents released by the activation of mast cells, lipid mediators produced from cell membrane phospholipids, and IL-3 (mast cell proliferation factor), which is well known as a modulator of immune response. , IL-4, IL-5, such as cytokines (cytokine) is a natural phenomenon in the living body that is involved (Galli SJ et al., Curr Opin Immunol.; 3(6):865-872, 1991; Bradding P et al., J Immunol.; 151(7):3853-3865, 1993). In addition, when the skin is exposed to the allergen, antigen-specific IgE binds to IgE receptors on the surface of Langerhans cells, and is delivered to T cells on the surface of the antigen to activate T cells. In this case, unlike normal, in allergic skin diseases, in particular, atopic dermatitis, Th2 is activated to secrete cytokines such as IL-4, IL-5, IL-6, IL-8, IL-10, and IL-13. As a result, the production of IgE in B cells is promoted, and cytokines and histamine are released by promoting the degranulation of activated mast cells (Baruah CC et al., Pharmacol Res.; 38(6):487-92, 1998; Karadag CH et al., Braz J Med Biol Res.; 33(3):327-330, 2000; Paolini R. et al., Nature.; 353(6347):855-858, 1991).

Allergy treatment drugs currently used clinically include degranulation inhibitors, chemical transporter inhibitors, and chemical transporter synthesis inhibitors, depending on their mechanism of action. Among these drugs, chemical transporter inhibitors and chemical transporter synthesis inhibitors have a relatively certain drug action point, but the mechanism of action of degranulation inhibitors is unclear, and these drugs have various side effects due to long-term administration. Can result. Accordingly, it can be said that for the treatment of type I allergy, it is very important to develop a substance capable of clarifying the mechanism of action on the production and release of these physiologically active substances in mast cells and minimizing side effects from long-term use.

Accordingly, the present inventors completed the present invention by conducting a study to discover substances for improving allergic diseases.

Korean Patent Registration No. 10-1733085

One object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of allergic diseases comprising abenanthramid C represented by the following Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a cosmetic composition for preventing or improving allergic dermatitis comprising abenantramid C represented by the following formula (1) as an active ingredient.

[Formula 1]

One aspect of the present invention provides a pharmaceutical composition for preventing or treating allergic diseases comprising abenanthramid C represented by the following Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

The term "allergy" used in the present invention refers to a phenomenon in which a living body in contact with a foreign substance exhibits a reaction different from normal to the substance. Allergy is a generalization of extensive and complex pathological phenomena and is a systemic or local disorder of a living body based on an immune response. Allergies appearing in the human body are classified into types Ⅰ, Ⅱ, Ⅲ and Ⅳ according to the immune mechanism. Among them, type Ⅰ allergy, which is an immediate type hypersensitivity reaction, occupies an important part in clinical practice. Atopic dermatitis, allergic rhinitis, These include bronchial asthma, hay fever and hay fever. The allergic disease may preferably be an inflammatory allergic disease.

The term "prevention" used in the present invention refers to any action that suppresses or delays progression of an allergic disease by administration of the composition of the present invention.

As used herein, the term "treatment" refers to any action in which symptoms of an allergic disease are improved or advantageously changed by administration of the composition of the present invention.

The pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are those commonly used in the manufacture of drugs, lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin. , Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, etc. It is not limited. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (22th ed., 2013).

The pharmaceutical composition according to an embodiment of the present invention may be administered together with a substance exhibiting activity in the prevention or treatment of one or more allergic diseases.

In addition, the pharmaceutical composition according to an embodiment of the present invention may be used alone or in combination with procedures, hormone therapy, drug therapy, and/or methods of using a biological response modifier for the treatment of allergic diseases.

The pharmaceutical composition of the present invention may contain various bases and/or additives necessary and appropriate for the formulation of the formulation, and within a range that does not deteriorate the effect, nonionic surfactants, silicone polymers, extender pigments, fragrances, preservatives , Disinfectant, oxidation stabilizer, organic solvent, ionic or nonionic thickener, softening agent, antioxidant, free radical destroying agent, opacifying agent, stabilizer, emollient, silicone, α-hydroxy acid, antifoaming agent, moisturizing agent , Vitamins, insect repellents, flavors, preservatives, surfactants, anti-inflammatory agents, substance P antagonists, fillers, polymers, propellants, basification or acidifying agents, or coloring agents, and other known compounds.

A suitable dosage of the pharmaceutical composition of the present invention is formulated in various ways depending on factors such as formulation method, mode of administration, age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity. Can be. The dosage of the pharmaceutical composition of the present invention may be 0.001 to 1000 mg/kg on an adult basis.

The pharmaceutical composition of the present invention may be administered parenterally, and may be administered transdermally by, for example, applying, dropping or spraying to the affected area, but is not limited thereto.

The pharmaceutical composition of the present invention can be administered in a variety of dosage forms when administered parenterally. Solid preparations include tablets, pills, powders, granules, capsules, and the like, and liquid preparations include suspensions, liquid solutions, emulsions, and syrups. In addition to water, liquid, and paraffin, which are commonly used simple diluents, various excipients, for example, wetting agents, sweeteners, fragrances, and preservatives may be included. Specifically, formulations for parenteral administration may include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions and freeze-dried formulations. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. In addition, calcium or vitamin D3 may be added to enhance the efficacy of the therapeutic agent.

Such compositions may be presented in unit-dose (single) or multi-dose (several batches) containers, e.g., sealed ampoules and vials, and immediately prior to use, in a sterile liquid carrier, e.g., water for injection. It can be stored under freeze-drying conditions requiring only the addition of. Ready-to-use preparations and suspensions can be prepared from sterile powders, granules and tablets.

In the present invention, the allergic disease may preferably be an allergic disease caused by mast cells.

According to one embodiment of the present invention, the allergic disease is contact dermatitis, atopic dermatitis, asthma, allergic conjunctivitis, periodontitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout , Ankylosing spondylitis, rheumatoid fever, lupus, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, peri-scapular joint infection, tendinitis, tendonitis, peritonitis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, anaphylaxis It may be any one disease selected from the group consisting of anaphylactic shock, multiple sclerosis, acute inflammation, chronic inflammatory disease, hives, hay fever, digestive tract allergy, drug allergy, food allergy and hay fever.

The composition of the present invention contains abenanthramid C, which inhibits the release of histamine in mast cells and inhibits the expression of inflammatory cytokines, so that it can exhibit activity in the prevention or treatment of various allergic diseases. .

According to one embodiment of the present invention, the pharmaceutical composition may be transdermally administered in a total dose of 0.1 mg/kg to 10.0 mg/kg.

When the pharmaceutical composition of the present invention is transdermally administered, it is possible to prevent or treat excellent allergic skin diseases by being administered at a total dose of 0.1 mg/kg to 10.0 mg/kg, and in particular, when administered at 10.0 mg/kg, As a mechanism of transcriptional inhibition, it can exhibit remarkably excellent preventive or therapeutic effects of allergic skin diseases even at the same dose compared to dexamethasone, which has strong anti-inflammatory action.

Another aspect of the present invention provides a cosmetic composition for preventing or improving allergic dermatitis comprising abenantramid C represented by the following Chemical Formula 1 as an active ingredient.

[Formula 1]

The cosmetic composition of the present invention further comprises fatty substances, organic solvents, solubilizing agents, thickening and gelling agents, softening agents, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or non- Ionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any other ingredients commonly used in cosmetics, and It may further include adjuvants commonly used in the same cosmetic field.

The cosmetic composition of the present invention may contain various bases and/or additives necessary and appropriate for the formulation of the formulation, and within a range that does not impair the effect, nonionic surfactants, silicone polymers, extender pigments, fragrances, preservatives, Fungicides, oxidation stabilizers, organic solvents, ionic or nonionic thickeners, softening agents, antioxidants, free radical destroying agents, opacifying agents, stabilizers, emollients, silicones, α-hydroxy acids, antifoaming agents, moisturizing agents, Vitamins, insect repellents, flavors, preservatives, surfactants, anti-inflammatory agents, substance P antagonists, fillers, polymers, propellants, basification or acidifying agents, or colorants, and other known compounds may be further included.

According to one embodiment of the present invention, the allergic dermatitis may be contact dermatitis or atopic dermatitis.

According to one embodiment of the present invention, the cosmetic composition may be any one formulation selected from the group consisting of lotion, gel, lotion, ointment, cream, pack, essence, patch, or spray.

According to the composition for the prevention, treatment or improvement of allergic diseases comprising abenanthamide C as an active ingredient, it inhibits histamine release from mast cells related to allergic reactions, and inflammation-inducing cytokines TNF-α and IL-4 It has an excellent effect of inhibiting gene expression and NF-ĸB activity, which is a transcription factor, so it can be usefully used in the prevention and treatment of allergic diseases, particularly allergic diseases mediated by mast cells.

1A is a graph showing the histamine release inhibitory activity of abenanthramide C (Avn C) in RBL-2H3 cells (Dexa: dexamethasone).
1B is a graph showing the histamine release inhibitory activity of abenanthramide C in rat peritoneal mast cells (RPMCs).
2 is a graph showing the intracellular calcium reducing activity of abenanthramid C in RBL-2H3 cells.
3 is a graph showing the inhibitory activity of abenanthamide C on the TNF-α and IL-4 cytokines in RBL-2H3 cells.
Figure 4 is a graph showing the activity of inhibiting IκBα decomposition and NF-ĸB activity of abenanthramid C.
5A is a photograph showing the effect of abenanthramid C on improving skin allergy in mice in which a passive local skin allergic reaction (passive cutaneous anaphylaxis, PCA) was induced.
5B is a graph showing a change in the color of an ear for skin allergies according to concentrations of avenantramid C in mice in which a passive local skin allergic reaction (passive cutaneous anaphylaxis, PCA) was induced.
5C is a graph showing changes in ear thickness for skin allergies according to concentrations of abenanthramid C in mice in which a passive local skin allergic reaction (passive cutaneous anaphylaxis, PCA) was induced.
6 is a graph showing the cytotoxicity of abenanthramid C in RBL-2H3 cells.

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

Example 1. Confirmation of the inhibitory effect of abenanthramid C on histamine release

1-1. Reagents and cell culture

Anti-dinitrophenyl(DNP)-IgE (Cat No. D8406) and DNP-human serum albumin (HSA) (Cat No. A6661) were purchased from Sigma (USA). Human mast cell line RBL-2H3 cells (purchased from the Korea Cell Line Bank) were cultured in DMEM (Dulbecco's modified eagle medium, Cat No. 12800-017) of Gibco (USA) to which 10% FBS inactivated by heat was added. .

1-2. Isolation and culture of rat peritoneal mast cells

In order to isolate rat peritoneal mast cells (RPMCs), rats were euthanized with carbon dioxide, and then 20 ml of Tyrod's buffer (137 mM NaCl, 5.6 mM glucose, 12 mM NaHCO ₃ , 2.7 mM KCl, 0.3 mM) NaH ₂ PO ₄ , 0.1% gelatin) was injected into the abdominal cavity, and the left and right sides of the abdomen were gently massaged for about 90 seconds. After dissecting the abdominal cavity of the rat, the buffer solution in the abdominal cavity was recovered using a Pasteur pipette, and centrifuged at room temperature (150 g, 10 minutes) to precipitate. The precipitate was again mixed with 1 ml of a buffer solution, then carefully floated on 2 ml of Histodenz solution (0.235 g/ml) and centrifuged at room temperature (400 g, 10 minutes). After removing the supernatant, 1 ml of a buffer solution was added to the remaining precipitate, washed, and mast cells were identified through toluidine blue staining, and the survival of cells was confirmed by trypan blue staining. . RPMCs were cultured in Alpha minimum essential media (Cat No. 11900-024) of Gibco (USA) to which 10% FBS inactivated by heat was added.

1-3. Confirmation of histamine release inhibitory activity of abenanthamide C

To confirm whether Avenanthramide C can inhibit the release of histamine from mast cells, RBL-2H3 cells cultured in Example 1-1 and rat mast cells isolated in Example 1-2 It was analyzed whether the release of histamine was inhibited by treatment with abenanthamide C, respectively.

Specifically, RBL-2H3 cells and RPMCs were sensitized with anti-DNP-IgE, and then treated with DNP-HSA to activate, thereby liberating histamine, which is a major component of intracellular granules and an allergen inducer. Prior to activation, each mast cell was pretreated with avenantramid C at various concentrations. The culture medium of the activated mast cells was recovered and used in the experiment. That is, 200 µl of a sample was taken in an Eppendorf tube, 80 µl of 0.1N hydrochloric acid and 20 µl of a 60% perchloric acid solution were mixed, and then centrifuged (13,000 g, 20 minutes, 5415R, Eppendorf). After centrifugation, the supernatant was taken 200㎕ 5N NaOH solution 100㎕, n - butanol (n -butanol) 800㎕, placed in a test tube, mixed with 5M NaCl 200㎕, after shaking it centrifugation (13,000g, 20 min.) I did. After centrifugation, 500 µl of a butanol layer was taken, 200 µl of 0.1N hydrochloric acid, 500 µl of n-heptane was added, shaken, and centrifuged again (13,000 g, 20 min) to 150 µl of the obtained aqueous layer. , 1N NaOH 40㎕, 1% o - mixture was added to program taldi aldehyde (o -phthaldialdehyde) solution (Sigma Co., Cat No. P1378) 10㎕, and allowed to stand for 5 minutes. Then, 20 µl of 3N hydrochloric acid was added, and the fluorescence intensity was measured using a fluorescence analyzer (GEMINIEM, Molecular Devices) at an emission wavelength of 440 nm and an excitation wavelength of 360 nm.

As a result, a large amount of histamine was released in activated RBL-2H3 cells (Fig. 1A) and RPMCs cells (Fig. 1B), whereas in mast cells treated with abenanthramid C, the concentration of abenanthamide C was dependently It was confirmed that histamine release was inhibited. In particular, abenanthramid C was found to have a similar level of histamine release inhibitory activity even at a low concentration compared to the positive control dexamethasone.

Through these results, it was confirmed that the improvement effect of abenanthramid C on allergic diseases.

1-4. Confirmation of the mechanism of action of abenanthramide C to inhibit histamine release by decreasing calcium in mast cells

Calcium, as a secondary transducer in cells, plays an important role in the regulation of physiological activity, and is known as a signaling material that regulates degranulation of mast cells. That is, when the amount of calcium in the cell increases, the mast cells are degranulated, and when the amount of calcium decreases, the degranulation of the mast cells is inhibited. Therefore, by analyzing the change of calcium in mast cells, the mechanism of action of inhibiting histamine release of abenanthramid C in mast cells was confirmed.

Specifically, after activating mast cells by treating RBL-2H3 cells with anti-DNP-IgE and DNP-HSA, respectively, the amount of calcium in the activated mast cells and the amount of calcium in the mast cells treated with abenanthamide C The amount was fluorescently stained using Fluo-3, AM (Invitrogen, Cat No. F1242), and then measured and compared with a fluorescence analyzer.

As a result, it was confirmed that the intracellular calcium of the activated mast cells increased rapidly, while the intracellular calcium was significantly decreased in the mast cells treated with abenanthramid C. In particular, abenanthramid C is an intracellular calcium inhibitor used as a positive control, BAPTA-AM (1,2-Bis(2-aminophenoxy)ethane- N , N , N',N' -tetraacetic acid tetrakis (acetoxymethyl ester). )), it was found to inhibit the degranulation of mast cells at a similar level even at a lower concentration.

Through these results, it was confirmed that abenanthramid C inhibited histamine secretion by inhibiting degranulation of mast cells by reducing the amount of calcium in mast cells.

Example 2. Confirmation of the inhibitory activity of abenanthamide C to induce inflammation

2-1. Confirmation of the inhibitory activity of avenantramid C on the expression of inflammatory cytokines

In order to confirm the effect of abenanthramid C on the expression of inflammatory cytokines such as TNF-α and IL-4, which are physiologically active substances that cause inflammatory allergies by being released or newly synthesized and secreted from mast cells, mast cells Was sensitized with anti-DNP-IgE, activated with DNP-HSA to promote the secretion of inflammatory cytokines, and then measured the secretion of cytokines using an enzyme-linked immunosorbent assay (ELISA).

Specifically, in order to calculate the cytokine concentration in the culture medium by the enzyme-linked immunosorbent assay, a commercially available kit (San Diego, BD Biosciences) was used. 200 µl of a standard sample diluted by culture medium and concentration was added to the well to which the antigen was attached and left to stand. After 120 minutes, the mixture was washed three times with a washing solution, and 100 µl of the primary antibody was added thereto, mixed well, and left for 60 minutes. Then, it was washed three times with a washing solution, and 100 µl of a mixed reagent in which the secondary antibody and enzyme reagent were mixed was added. After the reaction was over, the mixture was discarded, washed 4 times with a washing solution, and then 100 μl of the substrate solution was added at a time and allowed to stand at room temperature for 20 minutes. After the reaction, 50 µl of a stop reagent (2N H ₂ SO ₄ ) was added to stop the reaction, and the absorbance at 450 nm was measured with an absorbance analyzer (Sunnyvale, Molecular Devices).

As a result, the secretion of IL-4, IL-6 and TNF-α cytokines was increased in RBL-2H3 cells activated with anti-DNP-IgE and DNP-HSA, whereas mast cells treated with abenanthramid C It was confirmed that the amount of secretion of these cytokines was decreased depending on the concentration of abenanthramid C (Fig. 3). In addition, it was found that the secretion of inflammation-inducing cytokines was effectively reduced even at a lower concentration than the positive control dexamethasone.

Through these results, it was confirmed that abenanthramid C inhibited inflammation induction more effectively than dexamethasone.

2-2. Confirmation of the inhibitory activity of abenanthamide C on NF-ĸB activation

In order to elucidate a more precise signaling mechanism involved in the process of inhibiting the expression of TNF-α and IL-4, which are inflammation-inducing cytokines, abenanthramid C is a factor that regulates the transcription of inflammatory cytokines. The activation of NF-ĸB, an important signal transducer that mediates the inflammatory response, was measured. On the other hand, NF-ĸB in an inactive state binds to the endogenous inhibitory protein IĸBα and maintains its inactive state. When activated, IĸBα is phosphorylated and decomposed, released from them, moves to the nucleus, and acts as a transcription factor. Therefore, whether or not IĸBα was decomposed was also measured.

Specifically, mast cells were sensitized with anti-DNP-IgE and then activated with DNP-HSA to activate NF-ĸB, and then the amount of each protein in the nucleus and cytoplasm was measured using Western blot. RBL-2H3 cells were extracted with 0.5 mM phenylmethanesulfonyl fluoride (Sigma, Cat No. P7626) and 5 µg/ml leupeptin (Sigma, Cat No. L2884). After dissolving with (0.5% triton X-100, PBS solution containing 1mM Na ₃ VO _4, etc.), DNA was fragmented by sonication. The amount of protein was measured using a protein assay kit (Cat No. 500-0002) manufactured by Bio-Rad using bovine serum albumin as a standard. After electrophoresis of 10 to 50 µg of total cell protein on an 8 to 12% (w/v) polyacrylamide gel containing 0.1% SDS, the protein present in the gel is electroblotting method After transferring to a nitrocellulose (NC) filter, in order to block non-specific binding, put the nitrocellulose filter in a tris-buffered saline-tween (TBS-tween, Sigma) solution containing 5% skim milk and put it at room temperature. It was reacted for 1 hour. The filter was put in a TBS-tween solution containing an antibody against the target protein, left at 4° C. for 12 hours, and then labeled with a secondary antibody labeled with HRP (horseradish peroxidase, Sigma, Cat No. P0889). The band was measured using ECL (Enhanced chemiluminescence, Thermo scientific, Cat No. 34080).

As a result, it was confirmed that RBL-2H3 cells were sensitized with anti-DNP-IgE and then activated with DNP-HSA to degrade IĸBα and increase NF-κB in the nucleus. On the other hand, it was confirmed that when abenanthramid C was treated with activated RBL-2H3 cells, the degradation of IκBα by the binding of anti-DNP-IgE and DNP-HSA was inhibited, and NF-ĸB in the nucleus was reduced (Fig. 4).

Through these results, it was confirmed that abenanthramid C inhibited the activation of NF-ĸB, thereby suppressing gene expression of inflammation-inducing cytokines.

Example 3. Confirmation of the improvement effect of abenanthramid C on local skin allergic reactions

The improvement effect of abenanthramid C on skin allergies was measured using a passive cutaneous anaphylaxis (PCA), which is widely used when developing drugs with anti-allergic effects.

Specifically, PCA was an allergic reaction mediated by IgE, and an antibody was injected intradermally into the local skin, and 48 hours later, the antigen was administered to the tail vein of mice to artificially induce a local skin allergic reaction. Then, after the concentration of abenanthamide C was applied to the ear once, the degree of redness and the degree of edema of the ear were observed in mice (FIG. 5A).

As a result, depending on the treatment concentration of avenantramid C, edema decreases (Fig. 5B), and the redness site appears to be small (Fig. 5C), and abenanthamide C suppresses local skin allergic reactions in a concentration-dependent manner. It was confirmed to be.

Example 4. The toxicity of abenanthramid C to mast cells

MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) analysis was performed to confirm the cytotoxicity of abenanthamide C.

Specifically, the MTT solution forms formazan by mitochondrial dehydrogenases in living cells to confirm the survival of cells. To confirm cytotoxicity, RBL-2H3 cells were cultured in a 96-well plate at 3×10 ⁴ cells/well at 37° C., and then avenantramid C was treated with 0.01 to 100 μM, respectively, and cultured for 24 hours. After 24 hours, 20 µl of MTT solution was added to each well and incubated for an additional 2 hours, the culture medium was removed, and 100 µl of dimethylsulfoxide was added, and absorbance was measured at 570 nm.

As a result, it was confirmed that abenanthramid C did not show cytotoxicity up to a concentration of 100 μM, and was harmless to the human body (FIG. 6).

So far, the present invention has been looked at around the examples. Those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered in terms of explanation, not limitation. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the equivalent range should be construed as being included in the present invention.

Claims (6)

A pharmaceutical composition for the prevention or treatment of allergic diseases comprising abenanthramid C represented by the following Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

The method of claim 1, wherein the allergic disease is contact dermatitis, atopic dermatitis, asthma, allergic conjunctivitis, periodontitis, rhinitis, otitis media, pharyngitis, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis , Rheumatoid fever, lupus, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, peri-shoulder joint inflammation, tendonitis, tendonitis, tendonitis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, anaphylactic shock shock), multiple sclerosis, acute inflammation, chronic inflammatory disease, urticaria, hay fever, digestive tract allergy, pharmaceutical allergy, food allergy and hay fever, any one disease selected from the group consisting of allergic disease prevention or treatment pharmaceutical composition .
The pharmaceutical composition for preventing or treating allergic diseases according to claim 1, wherein the pharmaceutical composition is transdermally administered at a total dose of 0.1 mg/kg to 10.0 mg/kg.
A cosmetic composition for preventing or improving allergic dermatitis comprising abenanthramid C represented by the following Formula 1 as an active ingredient.
[Formula 1]

The cosmetic composition for preventing or improving allergic dermatitis according to claim 4, wherein the allergic dermatitis is contact dermatitis or atopic dermatitis.
The cosmetic composition for preventing or improving allergic dermatitis according to claim 4, wherein the cosmetic composition is any one formulation selected from the group consisting of lotion, gel, lotion, ointment, cream, pack, essence, patch, or spray.

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