KR20210058781A - Composition for regulating skin hypersensitivity comprising hesperetin - Google Patents

Abstract

The present invention relates to a composition comprising hesperetin for controlling skin hypersensitivity reactions. Hesperetin according to an embodiment exhibits low cytotoxicity and has a desirable effect in inhibiting enlargement of epithelial tissues in skin lesions of atopic dermatitis and inhibiting infiltration of mast cells into skin lesions, thereby being able to be safely used in cosmetics, pharmaceuticals, and food for the alleviation or treatment of allergic skin diseases including atopic dermatitis.

Description

Composition for regulating skin hypersensitivity including hesperetin {Composition for regulating skin hypersensitivity comprising hesperetin}

It relates to a composition for controlling skin hypersensitivity reactions comprising hesperetin.

The immune response is an important defense mechanism that occurs in our body. However, an abnormal immune response leads to an overactive immune response that causes a variety of diseases. There are four types of irritable immune responses. The allergic reaction is a typical type I hypersensitivity immune response and is induced by histamine and cytokines secreted by mast cells. Atopic dermatitis is a typical chronic inflammatory skin disease and is caused by a persistent allergic reaction.

Atopic dermatitis is accompanied by chronic skin inflammation and severe itching (pruritus). Itching, which arouses the feeling of wanting to scratch, is induced by itching mediating substances such as histamine.In the case of normal people, the symptoms are easily relieved when scratching, but in the case of atopic patients, the more scratched, the more the feeling of wanting to scratch is formed, causing severe scratching. Due to this scratching behavior of atopic patients, the skin barrier collapses and secondary infection is caused, which further exacerbates the inflammation. Atopic dermatitis can be temporarily relieved, but it recurs and worsens due to irritants such as the environment and food, and it is a phenomenon that deterioration and remission are repeated. Therefore, in order to improve skin diseases such as atopic dermatitis, there is a need for a material capable of restoring weakened skin barrier function by effectively inhibiting the mechanism of hypersensitivity immune reaction in the skin.

Hespretin is one of the flavonones and is abundant in the peels of citrus fruits such as oranges and lemons. According to previously reported, hesperetin is known to have protective and antioxidant effects against irradiation of radiation (Korean Patent Laid-Open Patent 10-2015-0104950), but studies on the efficacy of hesperetin on atopic dermatitis have been insignificant. Actually.

One aspect is to provide a pharmaceutical composition for preventing or treating skin allergic diseases comprising hesperetin, or an enantiomer or racemic mixture thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. .

Another aspect is to provide a composition for external application for skin for preventing or treating skin allergic diseases comprising hesperetin, or an enantiomer or racemic mixture thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. .

Another aspect is to provide a cosmetic composition for preventing or improving skin allergic diseases comprising hesperetin, or an enantiomer or racemic mixture thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. .

Another aspect provides a health functional food composition for preventing or improving skin allergic diseases comprising hesperetin, or an enantiomer or racemic mixture thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. It is to do.

One aspect provides a pharmaceutical composition for preventing or treating skin allergic diseases comprising hesperetin, or an enantiomer or racemic mixture thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The hesperetin is a compound having a molecular formula of C ₁₆ H ₁₄ O ₆ and a molecular weight of 302.27 represented by the following Formula 1, and a substance in which hesperidin is hydrolyzed in the form of aglycone by a glycolytic enzyme in the body. Can mean The hesperidin is a flavanone glycoside combined with a disaccharide rutinose, and may be present in the pericarp of citrus plants such as oranges and lemons. Hesperetin may be absorbed into the stomach and intestines in the body, and then exhibit physiological activity in vivo, and may be detected in the form of hesperetin in tissues such as blood and liver.

[Formula 1]

The “enantiomers” are compounds that have the same molecular formula as a specific compound and the same order of bonding of atoms, but differ in the three-dimensional configuration of atoms in the space, and are related to the relationship between a specific compound and its mirror image. It may mean a compound that is present.

The “racemic mixture” may mean that two enantiomers are mixed in 1:1, that is, in the same ratio.

The "pharmaceutically acceptable" is suitable for use in contact with a subject, for example, human tissue, with a reasonable benefit/risk ratio without excessive toxicity, irritation, allergic reaction or other problems or complications, and is within the scope of sound medical judgment. It may mean a phosphorus compound or composition.

The "salt" may be an acid addition salt formed by a pharmaceutically acceptable free acid. The free acid may be an inorganic acid or an organic acid, and the inorganic acid may be hydrochloric acid, bromic acid, nitric acid, sulfuric acid, perchloric acid, phosphoric acid, etc., and the organic acid may be citric acid, acetic acid, lactic acid, maleic acid, fumaric acid, gluconic acid, methanesulfuric acid. Fonic acid, glycolic acid, succinic acid, tartaric acid, galacturonic acid, embonic acid, glutamic acid, aspartic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, 4-toluenesulfonic acid, Salicylic acid, citric acid, benzoic acid or malonic acid, and the like. In addition, the salt may include an alkali metal salt (sodium salt, potassium salt, etc.) and an alkaline earth metal salt (calcium salt, magnesium salt, etc.). For example, the acid addition salt is acetate, aspartate, benzate, besylate, bicarbonate/carbonate, bisulfate/sulfate, borate, camsylate, citrate, edisylate, esylate, formate, fumarate, Gluceptate, Gluconate, Glucuronate, Hexafluorophosphate, Hybenzate, Hydrochloride/Chloride, Hydrobromide/Bromide, Hydroiodide/Iodide, Isethionate, Lactate, Malate, Male Eate, malonate, mesylate, methyl sulfate, naphthylate, 2-naphsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, saccha Rate, stearate, succinate, tartrate, tosylate, trifluoroacetate, aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, Potassium, sodium, tromethamine, zinc salts, and the like may be included.

The term “skin allergic disease” may mean that symptoms appear on the skin among allergic diseases. The symptoms may be acute or chronic, and may be local or systemic symptoms in the skin.

In one embodiment, the skin allergic disease may be any one selected from the group consisting of atopic dermatitis, allergic dermatitis, acute and chronic skin inflammatory diseases, contact dermatitis, eczema, and urticaria, resulting from It may include a secondary disease. For example, the skin allergic disease may be atopic dermatitis and may include a secondary disease resulting from atopic dermatitis. Specifically, the secretion of histamine and cytokines of mast cells is excessively increased due to an abnormal hypersensitivity immune reaction, causing skin itching or skin inflammatory reaction, and repetitive exacerbation and remission causes the skin barrier to collapse and cause secondary infection. I can.

In one embodiment, the composition may restore a weakened skin barrier function. For example, it may be to restore a skin barrier function weakened by a skin allergic disease such as atopic dermatitis.

In one embodiment, the composition may be to inhibit epithelial tissue hypertrophy. For example, the hypersensitivity immune response mechanism, such as atopic dermatitis, may suppress the enlargement of epithelial tissue.

In one embodiment, the composition may inhibit mast cell infiltration into skin lesions. For example, it may be to suppress the invasion of mast cells into skin lesions by a mechanism of an overactive immune response such as atopic dermatitis.

The term “prevention” may comprehensively mean that the composition is administered in a pharmaceutically effective amount to prevent skin allergic diseases such as atopic dermatitis in advance, or to reduce the likelihood or frequency of occurrence. For example, it may be to lower the probability of onset or lower the probability of recurrence in a patient with a possibility of developing a skin allergic disease such as atopic dermatitis or a patient who has had it. The term "treatment" may comprehensively mean improving skin allergic diseases such as atopic dermatitis by administering the composition in a pharmaceutically effective amount, and alleviating symptoms of skin allergic diseases in a shorter time compared to natural healing. Or it may be to provide a cure, and may be to improve one symptom or most of the symptoms caused by skin allergic diseases such as atopic dermatitis. The "pharmaceutically effective amount" refers to the type of disease, the patient's age, weight, health, sex, the patient's sensitivity to the drug, the route of administration, the method of administration, the number of administration, the duration of treatment, the combination, or the drug used simultaneously. It can be easily determined by a person skilled in the art according to factors well known to. When administered to an individual, the "effective amount" may mean an amount that inhibits histamine secretion of mast cells, suppresses epithelial tissue hypertrophy, inhibits mast cell infiltration into a skin lesion, or reduces a skin lesion site. . The "individual" may be a mammal, for example, a human, cow, horse, pig, dog, sheep, goat, or cat, and may be a cell, tissue, organ, etc. derived from a mammal.

The "pharmaceutical composition" may be formulated in an oral or parenteral dosage form. The oral dosage form may be a granule, a powder, a liquid, a tablet, a capsule, a dry syrup, or a combination thereof. The parenteral dosage form may be an injection or an external preparation for skin.

In one embodiment, the pharmaceutical composition may further include a pharmaceutically acceptable carrier or excipient. The "carrier or excipient" may include any physiologically compatible solvent, dispersion medium, coating, antibacterial and antifungal agent, isotonic and absorption delaying agent, and the like. For example, 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, lubricants, wetting agents, sweetening agents, flavoring agents, emulsifying agents, suspending agents, preservatives, and the like.

Another aspect provides a composition for external application for skin for preventing or treating skin allergic diseases comprising hesperetin, or an enantiomer or racemic mixture thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In the composition for external application for skin, hesperetin, enantiomer, racemic mixture, pharmaceutically acceptable, salt, skin allergic disease, prevention or treatment are as described above.

The "external preparation for skin" may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal delivery patch, drug-containing bandage, lotion, or a combination thereof. The above skin external preparations are ingredients commonly used in external preparations for skin such as cosmetics and pharmaceuticals, such as aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances. , Colorants, various skin nutrients, or a combination thereof and may be appropriately formulated according to need. The external preparations for the skin include metal sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid, caffeine, tannin, bellapamil, licorice extract, glavidine, and caline. Hot water extract of fruit, various herbal medicines, drugs such as tocopherol acetate, glytilithic acid, tranexamic acid and derivatives or salts thereof, vitamin C, ascorbic acid magnesium phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, Sugars such as trehalose can also be appropriately blended.

Another aspect provides a cosmetic composition for preventing or improving skin allergic diseases comprising hesperetin, or an enantiomer or racemic mixture thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In the cosmetic composition, hesperetin, enantiomer, racemic mixture, pharmaceutically acceptable, salt, skin allergic disease, and prevention are as described above.

The term "improvement" may comprehensively mean alleviating skin conditions caused by skin allergic diseases such as atopic dermatitis by administering the composition in a pharmaceutically effective amount, and skin allergic diseases in a shorter time compared to natural healing. It may be to provide relief or cure of the symptoms of, and may be to improve one symptom or most of the symptoms caused by skin allergic diseases such as atopic dermatitis. The pharmaceutically effective amount and the effective amount are as described above.

The “cosmetic composition” may be prepared in various forms, and in one embodiment, the composition is an emulsion, lotion, cream (oil-in-water type, water-in-oil type, multi-phase), solution, suspension (anhydrous and water-based), It may be any one formulation selected from the group consisting of emulsions, anhydrous products (oil and glycol), pastes, gels, masks, packs, powders, soaps, powders, and sprays.

The cosmetic composition may further include an acceptable carrier in a cosmetic formulation. The "acceptable carrier in cosmetic formulations" is a compound or composition that is already known and used that can be included in a cosmetic formulation, or is a compound or composition to be developed in the future, and has no toxicity, instability or irritation beyond which the human body can adapt when in contact with the skin. Say that. The carrier may be included in the cosmetic composition of the present specification in about 1% by weight to about 99.99% by weight, preferably about 90% by weight to about 99.99% by weight, based on the total weight of the cosmetic composition. Examples of the carrier include alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, moisturizing agents, viscosity modifiers, emulsions, stabilizers, sunscreen agents, color developing agents, fragrances, and the like.

Another aspect provides a health functional food composition for preventing or improving skin allergic diseases comprising hesperetin, or an enantiomer or racemic mixture thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. do.

In the health functional food composition, hesperetin, enantiomer, racemic mixture, pharmaceutically acceptable, salt, skin allergic disease, prevention or improvement are as described above.

In the "health functional food composition", hesperetin may be used alone or in combination with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment). In general, when preparing food or beverage, the health functional food composition may be added in an amount of 15 parts by weight or less based on the raw material. There is no particular limitation on the kind of the health functional food. Among the types of health functional foods, the beverage composition may contain various flavoring agents or natural carbohydrates as an additional component, like ordinary beverages. The natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used. The health functional food composition may also include nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonated beverages It may contain a carbonation agent used for, or a combination thereof. The health functional food composition may also contain natural fruit juice, fruit juice beverage, flesh for the manufacture of vegetable beverages, or a combination thereof.

Another aspect provides a method of preventing or treating a skin allergic disease in a subject comprising administering to the subject a pharmaceutically effective amount of the pharmaceutical composition.

In the method of preventing or treating, the pharmaceutically effective amount, pharmaceutical composition, individual, skin allergic disease, prevention or treatment are as described above.

The term “administration” can be administered by methods known in the art. Administration can be administered directly to a subject by any means, for example by routes such as intravenous, intramuscular, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration. . The administration can be administered systemically or locally. The administration may be topically administered to a site where an allergic skin disease exists.

The administration is 0.1 mg to 1,000 mg of hesperetin per individual, for example, 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1,000 mg, 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg, 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg may be administered.

Hesperetin according to one aspect exhibits low cytotoxicity, has excellent epithelial tissue hypertrophy inhibitory effect in lesions with atopic dermatitis and mast cell infiltration into skin lesions, thus preventing skin allergic diseases including atopic dermatitis. It can be safely used for improving or treating cosmetics, medicines, and foods.

1 is a result of confirming the cytotoxicity according to the concentration of hesperetin on human mast cells (HMC-1).
2 is a method for producing an atopic animal model.
3 is a result of observing the lesion onset of atopic dermatitis after inducing atopy using DNCB in a mouse, and then treatment with hesperetin by concentration. FIG. 3A is a photograph of a lesion onset of atopic dermatitis in a mouse, and FIG. 3B And FIG. 3C is a photograph observed under a microscope by performing H&E staining and toluidine blue O staining, respectively.

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

Example 1. Cytotoxicity test of Hesperetin

To evaluate the cytotoxicity of hesperetin, the human mast cell line HMC-1 (human mast cells-1) was treated with hesperetin (SigmaAldrich) and then Annexin/PI staining was performed to analyze the cell viability with flow cytometry. I did.

Specifically, Iscove's Modified Dulbecco's medium containing 10% fetal bovine serum (FBS), 200IU/ml penicillin, and 200g/ml streptomycin was added to HMC-1 cells, a human mast cell line. Medium; IMDM) at 37° C. and 5% CO ₂ . Human mast cells (HMC-1) cultured in a 25T flask were treated with hesperetin at different concentrations and cultured for 24 hours. After culturing for 24 hours, the cells were recovered and washed twice with PBS to remove the remaining medium. Washed cells were stained using Annexin V Apoptosis Detection Kit I (BD Biosciences). The stained cells were measured using an FC500 (Beckman coulter) instrument, and then analyzed using FlowJo V10 (FlowJo, LLC).

1 is a result of confirming the cytotoxicity according to the concentration of hesperetin on human mast cells (HMC-1).

As a result, as shown in FIG. 1, it was confirmed that hesperetin did not exhibit cytotoxicity up to 200 μM with respect to human mast cells.

Example 2. Evaluation of anti-atopic efficacy of hesperetin

To confirm the anti-atopic effect of hesperetin, after inducing atopy using DNCB, a skin sample was obtained from an atopic animal model treated with hesperetin (SigmaAldrich) by concentration, and tissue staining was performed to develop atopic dermatitis. The lesion was observed.

Specifically, BALB/c mice were divided into four groups (control group, 2,4-dinitrochlorobenzene (2,4-dinitrochlorobenzene; DNCB), 200 μM hespertin+DNCB, 2 mM hespertin+DNCB). 2 is a method for producing an atopic animal model. As shown in FIG. 2, first, a 1% DNCB solution was applied to the back skin of the mouse for 4 days. After having a rest period of 3 days, the control group applied only the solvent in which DNCB was dissolved. On the 8th day, the control group was applied with a solvent, DNCB was treated with 0.5% DNCB, and the hesperetin-treated group applied each concentration of hesperetin with 0.5% DNCB for 11 days. On the 19th day, the mice were euthanized to obtain a skin sample.

The skin tissue obtained in the mouse experiment inducing atopy using the DNCB was fixed in 4% paraformaldehyde and then embedded with an OCT compound. Each sample was cut to 40 μm using a cryosectioning machine, followed by H&E staining and toluidine blue O staining, and observed under a microscope.

3 is a result of observing the lesion onset of atopic dermatitis after inducing atopy using DNCB in a mouse, and then treatment with hesperetin by concentration. FIG. 3A is a photograph of a lesion onset of atopic dermatitis in a mouse, and FIG. 3B And FIG. 3C is a photograph observed under a microscope by performing H&E staining and 3 toluidine blue O staining, respectively.

As a result, as shown in Figure 3A, compared to the control group, compared to the control group treated with DNCB alone, it was confirmed that severe keratin occurred and the skin was keratinized, resulting in atopic dermatitis, and in the group treated with hesperetin, keratinization of the skin It was confirmed that the amount of keratin and atopic dermatitis decreased. In addition, as shown in FIGS. 3B and 3C, it was confirmed that hypertrophy of epithelial tissue and mast cell infiltration into skin lesions occurred in the group treated with DNCB alone compared to the control group, and the group treated with hesperetin In Figure 2, it was confirmed that the enlargement of epithelial tissue and the infiltration of mast cells into skin lesions were inhibited.

This is because hesperetin can restore the skin barrier function damaged by atopic dermatitis, and has the effect of inhibiting the enlargement of epithelial tissue and the invasion of mast cells into skin lesions.Therefore, various skin allergic diseases including atopic dermatitis It means that it can be used usefully.

Claims (6)

As a pharmaceutical composition for preventing or treating atopic dermatitis comprising hesperetin represented by the following formula (1), or an enantiomer or racemic mixture thereof, or a pharmaceutically acceptable salt thereof as an active ingredient,
The composition is to restore weakened skin barrier function,
The pharmaceutical composition is administered transdermally.
[Formula 1]

The pharmaceutical composition of claim 1, wherein the composition inhibits epithelial tissue hypertrophy. The pharmaceutical composition according to claim 1, wherein the composition inhibits mast cell infiltration into skin lesions. The pharmaceutical composition according to claim 1, wherein the composition further comprises a pharmaceutically acceptable carrier or excipient. As a cosmetic composition for preventing or improving atopic dermatitis comprising hesperetin represented by the following formula (1), or an enantiomer or racemic mixture thereof, or a pharmaceutically acceptable salt thereof as an active ingredient,
The composition is to restore weakened skin barrier function,
The composition is a cosmetic composition that is administered transdermally.
[Formula 1]

The method of claim 5, wherein the composition is any one formulation selected from the group consisting of emulsions, lotions, creams, solutions, suspensions, emulsions, pastes, gels, masks, packs, powders, soaps, powders, and sprays. Cosmetic composition.

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