KR101819670B1 - Compositon for prevention or treatment of skin disease - Google Patents

Abstract

The present invention relates to a composition for preventing or treating skin diseases, and more particularly, to a composition for preventing or treating an allergic or inflammatory skin disease comprising N-acetyl-LTE ₄ or a pharmaceutically acceptable salt thereof as an active ingredient ≪ / RTI > The N- acetyl sikimyeo _{LTE 4 (N-acetyl Leukotriene E4} ) is to inhibit the water-soluble cytokines, in particular the generation of IL-2 and IL-17 relieve the itching symptoms caused by various causes, at the same time has a control immune effects. Accordingly, the present invention can be applied variously to a composition for relieving itching caused by histamine, chloroquine or contact dermatitis, and a preventive or therapeutic agent for allergic or inflammatory skin diseases including atopic dermatitis.

Description

[0001] The present invention relates to a composition for preventing or treating skin diseases,

The present invention relates to a composition for preventing or treating skin diseases, and more particularly, to a composition for preventing or treating an allergic or inflammatory skin disease comprising N-acetyl-LTE ₄ or a pharmaceutically acceptable salt thereof as an active ingredient ≪ / RTI >

Human skin consists of dermis and epidermis. The dermis is composed mainly of fibroblasts that synthesize collagen and other proteins and produce a small amount of lipids. In contrast, the epidermis is predominantly composed of keratinocytes that mainly produce lipids and do not substantially synthesize collagen. In particular, the epidermis, located at the outermost part of the skin, protects against various physiological stimuli such as chemicals, air pollutants, dry environment, ultraviolet rays, and excessive exudation of body water through the skin The blocking function is performed. This protective function is possible by normally forming and maintaining the stratum corneum composed of keratinocytes.

The stratum corneum (horney layer), which is the outermost part of the epidermis, is formed from keratinocytes and consists of keratinocytes with complete differentiation and a surrounding lipid layer (J. Invest. Dermatol. 1983; 80: 49). The keratinocyte is a cell formed by progressive morphological and functional changes while the basal cell which continuously grows in the stratum basale moves to the stratum corneum. After a certain period of time, old keratinocytes are removed from the skin and new keratinocytes from the epidermis are replaced by their function. This repetitive sequence of changes is called epidermis differentiation or keratinization keratinization. In this keratinization process, the keratinocytes form a stratum corneum while generating natural moisturizing factor (NMF) and intercellular lipids (ceramide, cholesterol and fatty acid), so that the stratum corneum has firmness and flexibility, And functions as a skin barrier acting as a layer.

Such a stratum corneum can easily lose its function due to environmental factors such as excessive washing, lifestyle factors such as bathing, dry air pollution, and endogenous diseases such as atopic skin and senile skin. In fact, the risk factors for skin are increasing in modern times, and as the dietary pattern changes, the rate of generation and elimination of stratum corneum is slowed, and the amount of moisturizing factor and lipid in the stratum corneum is decreased due to the deterioration of keratinocyte function, There is an increasing trend for people with skin whose stratum corneum is unable to exhibit normal skin barrier function.

In particular, atopy dermatitis is a type of hypersensitivity that has recently become increasingly prevalent as its prevalence increases rapidly. Immunoglobulin E (IgE) and allergen (immunoglobulin E) Is defined as a group of allergic diseases that have a strong genetic tendency, accompanied by major symptoms due to immune response. In particular, atopic dermatitis, a representative symptom of atopy, accounts for 0.5 to 1% of the total population and 5 to 10% of the children have serious symptoms, but it is presumed to be related to genetic factors and immune system deficiency The exact cause of the disease has yet to be clarified, and it is expected that it will be alleviated somewhat through the improvement of environment and diet, but there is no fundamental treatment method.

Symptoms of atopic dermatitis include severe itching, skin dryness, rash, dirt, scabs, scaly skin (rabbits), and other symptoms such as emotional anxiety, stress, tension, frustration, Allergic diseases such as urticaria, metal allergy, asthma and allergic rhinitis are often accompanied. Recently, atopic dermatitis has been increasing in adults, and it has been suggested that adults, at risk for atopic dermatitis, have facial and neck lesions, animals, pollen and nickel allergy.

Accordingly, the inventors of the present invention have been studying a method for remarkably improving an allergic or inflammatory skin disease including itching and atopy, confirming that N-acetyl-LTE ₄ has an excellent therapeutic effect, Respectively.

Korean Patent Publication No. 10-2010-0046301 Korean Patent Publication No. 10-2013-0009161

An object of the present invention is to provide an N- _{acetyl-LTE 4 (N-acetyl Leukotriene E4} ) or, allergic, or for the prevention and treatment of inflammatory skin diseases, the composition including a pharmaceutically acceptable salt thereof as an active ingredient.

It is still another object of the present invention to provide a composition for alleviating or suppressing itching comprising the active ingredient.

In order to achieve the above object, the present invention is N- _{acetyl-LTE 4 (N-acetyl Leukotriene E4} ) or pharmaceutical for pharmaceutically the prophylaxis or treatment of allergic or inflammatory skin diseases including acceptable salt thereof as an active ingredient Lt; / RTI >

The present invention also provides an external preparation for skin for the prevention or treatment of an allergic or inflammatory skin disease comprising the active ingredient.

The present invention also provides a health food composition for preventing or ameliorating an allergic or inflammatory skin disease containing the active ingredient.

The present invention also provides a cosmetic composition for alleviating or inhibiting itching comprising the active ingredient.

N- _{acetyl-LTE 4 (N-acetyl Leukotriene E4} ) of the present invention mitigates the symptoms of itching caused by various causes to inhibit the water-soluble cytokines, in particular the generation of IL-2 and IL-17, while the control immune effect I have. Accordingly, the present invention can be applied variously to a composition for relieving itching caused by histamine, chloroquine or contact dermatitis, and a preventive or therapeutic agent for allergic or inflammatory skin diseases including atopic dermatitis.

FIG. 1 shows the effect of histamine-induced itching improvement upon administration of N-acetyl-LTE ₄ .
FIG. 2 shows the effect of improving the itching caused by chloroquine upon administration of N-acetyl-LTE _4. FIG.
FIG. 3 shows the effect of improving the itching caused by contact dermatitis upon administration of N-acetyl-LTE ₄ .
FIG. 4 is a graph showing the results of confirming the improvement effect of atopic dermatitis upon administration of N-acetyl-LTE ₄ through H & E staining.
FIG. 5 is a graph showing the results of bio-analyzing the changes of in vivo factors upon administration of N-acetyl-LTE ₄ .

The present invention provides a N- _{acetyl-LTE 4 (N-acetyl Leukotriene E4} ) or, allergic, or for the prevention and treatment of inflammatory skin diseases, the composition including a pharmaceutically acceptable salt thereof as an active ingredient.

Hereinafter, the present invention will be described in more detail.

As one aspect, the invention provides a N- _{acetyl-LTE 4 (N-acetyl Leukotriene E4} ) or, allergic or pharmaceutical composition for the prevention or treatment of inflammatory skin diseases comprising a pharmaceutically acceptable salt thereof as an active ingredient do.

In the present invention, the active ingredient, the N- acetyl-LTE ₄ is also referred to as, and N-acetyl Leukotriene E4, a material which is mainly metabolised in the bile, spleen, kidneys, and also found in the skin and lungs, LTE ₄ which is produced by metabolism It is a substance that is distinguished from LTE ₄ as a material. This material can be specified by the following formula (1).

[Chemical Formula 1]

In the present invention, the pharmaceutically acceptable salt of N-acetyl LTE ₄ represented by the general formula (1) includes salts of an acidic or basic group which may exist in the compound of the general formula (1) unless otherwise indicated. For example, pharmaceutically acceptable salts include the sodium, calcium and potassium salts of hydroxy groups or carboxy groups, and other pharmaceutically acceptable salts of amino groups include hydrobromide, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, There may be mentioned the hydrogen phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p-toluenesulfonate (tosylate) salts, Or through a manufacturing process.

In the present invention, the term "prevention" means any action that prevents or alleviates the disease by eliminating or early detection of the pathogenesis of an allergic or inflammatory skin disease.

In the present invention, the term "treatment" means any action that alleviates or alleviates symptoms caused by allergic or inflammatory skin diseases by the composition of the present invention.

In the present invention, allergies and inflammation are one of defensive reactions of biological tissues. When the organism is brought into contact with a certain adventitious substance, the allergic reaction causes a sudden change in the reaction ability in vivo, Is a sensitive response. In addition, inflammation refers to complex lesions involving tissue degeneration, circulatory disturbances and exudates, and tissue proliferation.

An allergic or inflammatory skin disease referred to in the present invention refers to atopic dermatitis, allergic dermatitis, contact dermatitis, acne, acne, , Seborrheic dermatitis, rash, urticaria or psoriasis, more preferably atopic dermatitis, contact dermatitis or itching, but the present invention is not limited thereto.

According to the present invention, N-acetyl LTE ₄ inhibits the production of water-soluble cytokines, particularly IL-2 and IL-17, thereby relieving the itching symptoms caused by various causes, and at the same time, Medicines for treating inflammatory skin diseases, health functional foods, and the like.

In the present invention, the composition may further comprise at least one known active ingredient having the effect of preventing or treating allergic or inflammatory skin diseases, together with the N-acetyl LTE ₄ or a pharmaceutically acceptable salt thereof.

In the present invention, the composition may further comprise a pharmaceutically acceptable additive. Examples of the pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, hydrogen phosphate Calcium carbonate, lactose, mannitol, sugar, arabic gum, pregelatinized starch, corn starch, powdered cellulose, hydroxypropylcellulose, opaques, starch glycolate sodium, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, Calcium stearate, white sugar, etc. may be used. The pharmaceutically acceptable additives according to the present invention are preferably included in the composition in an amount of 0.1 to 90 parts by weight, but not limited thereto.

In the present invention, the composition may be administered orally or parenterally in various clinical formulations. In the case of formulation, a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, , And those suitable for use in the art are disclosed in Remington ' s Pharmaceutical Sciences (Mack Publishing Company, Easton PA). Examples of carriers, excipients and diluents that can be contained in the composition include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil and the like.

The solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions and syrups. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like May be included.

The preparation for parenteral administration includes sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like. The parenteral administration can be carried out using an external or intraperitoneal injection, intramuscular injection, subcutaneous injection, intravenous injection, intramuscular injection or intra-thoracic injection.

For the purposes of the present invention, the term "administering" is meant to provide any desired composition of the invention to a subject in any suitable manner.

In the present invention, the preferable dosage of the composition varies depending on the condition and the weight of the individual, the degree of disease, the type of the drug, the administration route and the period, but can be appropriately selected by those skilled in the art. For example, for a desired effect, the N-acetyl LTE ₄ of the present invention or a pharmaceutically acceptable salt thereof may be administered in an amount of 0.1 to 10,000 mg / kg (body weight) per day. The composition may be administered once a day or divided into several doses.

In the present invention, the composition may be administered to a subject by various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular or subcutaneous injection, and may be administered by any route, such as by way of being tabulated on the skin.

In the present invention, the composition may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers for the prevention and treatment of allergic or inflammatory skin diseases.

Further, the present invention provides a N- _{acetyl-LTE 4 (N-acetyl Leukotriene E4} ) or a pharmaceutically, or preventing allergic or skin external preparations for the treatment of inflammatory skin diseases including acceptable salt thereof as an active ingredient.

In the present invention, the composition may further contain one or more known active ingredients having the effect of preventing or treating allergic or inflammatory skin diseases together with the N-acetyl LTE ₄ or a pharmaceutically acceptable salt thereof .

In the present invention, the composition is not particularly limited depending on the desired body part, and may be prepared into any formulation conventionally produced by referring to known techniques in the art. For example, it can be used in the form of liquid, ointment, cream, lotion, spray, patch, oil, wax, emulsion, suspension, gel or aerosol.

In the present invention, the composition may contain conventional additives such as preserving agents, solvents to aid drug penetration, emollients in the case of ointments and creams, and may contain conventional carriers such as ethanol or oleyl alcohol have.

In the present invention, the composition is not limited to the above-described components, and may include other components incorporated in a usual cosmetic composition or pharmaceutical composition, if necessary. For example, it is possible to use a water-soluble organic solvent such as a preservative, a humectant, an emollient, a surfactant, an organic or inorganic pigment, an organic powder, an ultraviolet absorbent, an antiseptic, a bactericide, an antioxidant, a plant extract, Cold agents, antiperspirants, purified water, and the like.

In addition, the present invention provides a N- acetyl LTE ₄ dietary supplement for, allergic or prevention or amelioration of inflammatory skin diseases, including (N-acetyl Leukotriene E4) or a pharmaceutically acceptable salt thereof as an active ingredient.

In the present invention, the composition may further contain at least one known active ingredient having N-acetyl LTE ₄ or a pharmaceutically acceptable salt thereof together with a preventive or ameliorative effect of an allergic or inflammatory skin disease.

In the present invention, the term "health functional food" refers to a food having a biological control function such as prevention and improvement of disease, bio-defense, immunity, recovery after disease and aging suppression and should be harmless to human body .

In the present invention, when the above substance is used as a food additive, the N-acetyl LTE ₄ or a pharmaceutically acceptable salt thereof may be directly added or used together with other food or food ingredients, Can be used. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). Generally, N-acetyl LTE ₄ or a pharmaceutically acceptable salt thereof is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material, in the production of food or beverage. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

In the present invention, the kind of the food is not particularly limited. Examples of the foods to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

In the present invention, the composition may contain various flavors or natural carbohydrates as an additional ingredient, such as ordinary beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharine and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may comprise flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention also provides a cosmetic composition, itch relief or suppression, including N- _{acetyl-LTE 4 (N-acetyl Leukotriene E4} ) or a pharmaceutically acceptable salt thereof as an active ingredient.

In the present invention, the composition may further contain one or more known active ingredients having N-acetyl LTE ₄ or a pharmaceutically acceptable salt thereof and having an itching relieving or inhibiting effect.

In the present invention, itching is also known as pruritus and is an uncomfortable sensation that causes a desire to scratch or rub the skin, which is a symptom commonly observed in skin diseases. In particular, the itching referred to in the present invention is not limited to atopic dermatitis, allergic dermatitis, contact dermatitis, dermatitis due to roughness of the skin, acne, seborrheic dermatitis, And allergic reactions caused by one or more of the group consisting of bronchitis, urticaria, urticaria, and psoriasis.

In the present invention, the composition can improve the skin barrier by enhancing skin moisturization or preventing skin and keratinization, and is characterized by having a skin condition improving effect.

In the present invention, in addition to N-acetyl LTE ₄ or a pharmaceutically acceptable salt thereof, the composition may further contain a fatty substance, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, A preservative, a stabilizer, a foaming agent, a perfume, a surfactant, water, an ionic or nonionic emulsifier, a filler, a sequestering and chelating agent, a preservative, a vitamin, a blocking agent, a wetting agent, , Pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredients commonly used in cosmetics, as well as adjuvants commonly used in the cosmetics or dermatological sciences. In addition, the above ingredients may be introduced in amounts commonly used in the field of dermatology.

In the present invention, the composition may be prepared in the form of a general emulsified formulation and a solubilized formulation. Cosmetics of emulsified form include nutrition lotion, cream, essence, etc., and cosmetics of solubilized form have softening longevity. More specifically, the cosmetic composition of the present invention may be in the form of a solution, a gel, a solid or a paste anhydrous product, an emulsion obtained by dispersing an oil phase in water, a suspension, an emulsion, a microemulsion, a microcapsule, In the form of an ionic follicle dispersing agent, in the form of a cream, a skin, a lotion, a powder, a spray, a paste, a pack, a cleanser, a soap, a surfactant-containing cleansing oil, a powder foundation, an emulsion foundation, a wax, Can be provided. It can also be prepared in the form of a foam or in the form of an aerosol which further contains a compressed propellant.

In the present invention, when the composition of the composition is a paste, a cream or a gel, the carrier component may be an animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, Zinc, and the like.

In the present invention, when the formulation of the composition is a powder or a spray, the carrier component may include lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder and the like. Propane / butane, dimethyl ether, and the like.

In the present invention, when the formulation of the composition is a solution or an emulsion, the carrier component may include a solvent, a solubilizing agent, an emulsifying agent and the like. Specific examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, Benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid esters of sorbitan, and the like.

In the present invention, when the formulation of the composition is a suspension, a liquid diluent such as water, ethanol, propylene glycol or the like as a carrier component; Suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester; Sintered cellulose, aluminum metahydroxide, bentonite, agar, trachant, and the like.

In the present invention, when the composition of the composition is a surfactant-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, Fatty acid amide ether sulfate, alkylamido betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, ethoxylated glycerol fatty acid ester, and the like.

In addition, any of the above components may be compounded within a range that does not impair the objects and effects of the present invention, but is preferably 0.01-10% by weight based on the total weight , More preferably from 0.01% to 5% by weight.

The redundant contents are taken into consideration in the complexity of the present specification, and terms not otherwise defined herein have the meanings commonly used in the art to which the present invention belongs.

EXAMPLES Hereinafter, the present invention will be described in detail with reference to Examples and Preparation Examples. However, the following examples and preparative examples are merely illustrative of the present invention, and the scope of the present invention is not limited to the following examples and preparative examples. The embodiments and manufacturing examples of the present invention are provided to enable those skilled in the art to more fully understand the present invention.

Example  1. N-Acetyl- LTE ₄ (N-acetyl Leukotriene  E4)

N-acetyl-LTE ₄ (N-acetyl Leukotriene E4) used in the following examples was supplied from Santa Cruz biotechnology (Dallas, Texas, USA). The N-acetyl-LTE ₄ is specified by the following formula.

Example  2. N-Acetyl- LTE ₄ on  Confirms the effect of relieving itching by

2-1. Experimental animal

All animal - related experiments were carried out in accordance with the guideline of the experimental animal committee of Chonbuk National University. Balb / c mice were purchased from Orient Biotech (Seongnam, Korea) and weighing 6-8 weeks. Balb / c mice were purchased from Oriental Biotech (Seongnam, South Korea) - specific pathogen free (SPF) laboratory. In an in vivo experiment, mice were randomly divided into groups containing at least 6 mice per group, and then experiments were performed with either a control group or an experimental group.

2-2. Check for itching relief

In order to confirm the effect of N-acetyl-LTE ₄ on the relief of itching, histamine and chloroquine, which are known to cause itching in the intradermal neck, and contact dermatitis induced by DNBF, Lt; RTI ID = 0.0 > N-acetyl-LTE < / RTI > ₄ . Specifically, it is as follows.

2-2-1. Identification of the itching relief effect caused by histamine

Acetyl LTE _{4 at} a concentration of 0 to 1 μM (1 nM, 10 nM, 100 nM, or 1 μM) and 200 μg of histamine in 100 μl of PBS (phosphate buffered saline) . After injection, one mouse was placed per observation chamber (11 cm x 11 cm, 18 cm height). The behavior of the mouse was recorded with a video camera and the number of scratches of the mouse for one hour was counted. The results are shown in Fig.

1, the case where only the N- acetyl -LTE ₄ without histamine in mice injected 86 ± 8.1 (n = 5) were itching of the boat caused, injection itching according to the injection amount of the N- acetyl-LTE ₄ which is considerably . In particular, administration of 1 μM of N-acetyl LTE ₄ showed maximal inhibitory activity against itching (62%).

2-2-2. On chloroquine  Confirm the itching relief effect caused by

Acetyl LTE _{4 at} a concentration of 0 to 1 μM (1 nM, 10 nM, 100 nM, or 1 μM) and 400 μg of chloroquine in 100 μl of PBS (phosphate buffered saline) . After injection, one mouse was placed per observation chamber (11 cm x 11 cm, 18 cm height). The behavior of the mouse was recorded with a video camera and the number of scratches of the mouse for one hour was counted. The results are shown in Fig.

2, the case where the mouse N- acetyl -LTE only chloroquine injection without ₄ to 244 ± 56 (n = 5) were itching of the boat caused, injection itching according to the injection amount of the N- acetyl-LTE ₄ which is considerably . In particular, administration of 1 μM of N-acetyl LTE ₄ showed the maximum inhibitory activity against itching (84%).

2-2-3. Confirm the itching relief effect caused by contact dermatitis

In order to measure the therapeutic effect of itching induced by contact dermatitis caused by administration of 1-fluoro-2,4-dinitrobenzene (DNFB), the back of the mouse ear was epilated with Dorco laser, and 0.3% RTI ID = 0.0 > 1-fluoro-2,4-dinitrobenzene. ≪ / RTI > After 5 days, DNFB diluted to 0.15% was treated at the same site and repeated 3 times, once every 3 days. N-acetyl-LTE ₄ was treated locally 30 minutes after the last DNFB treatment. The results are shown in Fig.

As shown in FIG. 3, it was confirmed that the itching caused by contact dermatitis was remarkably inhibited by the dose of N-acetyl LTE ₄ .

Example  3. N-Acetyl- LTE ₄ on  Of atopic dermatitis

3-1. Experimental animal  - Atopic animal model

All animal - related experiments were carried out in accordance with the guideline of the experimental animal committee of Chonbuk National University. NC / Nga mice (male, 10 weeks old) were used. Experimental animals were maintained at a temperature of 20 ° C and a humidity of 50%, and were adapted for one week in a conventional breeding room where the light and shade changes every 12 hours. First, the dorsal part of the NC / Nga mouse was shaved to the upper part of the ear wheel to completely remove the hair. Later, the fat content of skin was removed by spraying 4% SDS aqueous solution on the site to be coated with N-acetyl-LTE _4. After the material was completely dried, 100 mg of Biotir AD (house dust mite ) Was evenly applied to the dorsal part of the auricle. The application of the reagent was repeated 10 times for 2 weeks, 5 weeks, to produce an atopy-induced experimental animal.

3-2. Confirmation of relieving atopic dermatitis by H & E staining

The H & E staining method was used to confirm whether atopic dermatitis was alleviated by N-acetyl-LTE ₄ . To this end, a total of 1 μM solution of N-acetyl-LTE ₄ dissolved in ethanol was prepared, and 100 μl of the solution was applied to the atopy-induced experimental animal of Example 3-1. This was repeated every day for at least three weeks after the onset of atopy. On the other hand, 100 μl of ethanol was added to the negative control group instead of N-acetyl-LTE ₄ . The mice were sacrificed after a total of 5 weeks after the onset of atopic dermatitis. The back skin was removed from the sacrificed mice and the tissue was fixed with an aquintin-formalin fixer to prepare a paraffin block. The paraffin block was cut to a thickness of 5 μm, and hematoxylin / eosin staining was performed. Then, a change in the thickness of the epidermis and dermis was observed using a microscope. The results are shown in FIG. 4A. FIG. 4B shows a graph of the contents observed through FIG. 4A.

As shown in FIG. 4, it was confirmed that dermatitis occurring in the epidermis of an animal induced by atopy by N-acetyl-LTE ₄ was alleviated. That is, N-acetyl-LTE ₄ has a remarkable effect of improving atopic dermatitis.

Example  4. BioMAP  Analysis of N-acetyl- LTE ₄ on  Identification of Biological Factor Changes by

BioMAP assays were performed with the aid of DiscoveRx (Fremont, Calif.) Using primary human cells. This study adheres to the guidelines for the study of human subjects under the US HHS Human Specimen Discipline (45 CFR Part 46).

First, endothelial cells were prepared and cultured for BioMAP analysis. More specifically, human umbilical vein endothelial cells (HUVECs) were obtained from various donors, cultured according to standard methods, and transferred to a microtiter plate in the fourth subculture step. Peripheral blood mononuclear cells (PBMC) were obtained from normal donor phosphorus buffy coats according to standard methods. In addition, CD4 + T-cells (TH2) or human blood-derived CD14 + macrophages (All Cells LLC, Emeryville, Calif.) Were added to HUVEC for 14 days for co-cultivation for l TH2 or l Mphg system, Respectively. The BT system consists of CD19 + B-cells (AllCells LLC, Emeryville, CA) stimulated with anti-IgM co-cultured with PBMC in consideration of low levels of TCR stimulation co-cultured for 84 hours. For the HDF3GF system, foreskin fibroblasts (HDFn) from three newborns were isolated and cultured according to the manual provided by the supplier (Lonza, Inc., Allendale, NJ). HDFn was placed in a low-concentration plasma for 24 hours before stimulation with cytokines. Primary human bronchial epithelial cells (Cell Applications, Inc., San Diego, Calif.), Coronary SMC, keratinocytes and HDFn (Lonza, Inc., Allendale, Calif.) Were used for BE3C, BF4T, CASM3C and KF3CT systems. NJ) were cultured according to the method provided by the manufacturer. Peripheral blood mononuclear cells (PBMC) were obtained from buffy coats of normal human donors according to the general methods in the art.

The following materials were added to the subculture microtiter plates of each of the above systems: cytokine (IL1β, 1 ng / ml; TNFα, 5 ng / ml; IFN-γ, 20 ng / ml; IL4, 5 ng / ml (IL-2, 2 ng / ml), activator (SAg, 20 ng / ml; histamine, 10 μM; or LPS, 2 ng / ml) PDGF-BB, 10 ng / ml), or PBMC (7.5x10 ⁴ cells / well).

The DiscoveRx PathHunter technology (DiscoveRx) was used to confirm whether N-acetyl LTE ₄ activates or inhibits ? -Estestin-2 using BioMAP constructed according to the above procedure. This includes enzymatic complementation of the fusion-tagged receptor according to the arestin regulatory regulatory sequence and the beta-arestin-2 protein.

Specifically, CHO-K1 cells (DiscoveRx) expressing various types of G-protein coupled receptors were plated at a density of 2625 cells / well in a plate composed of 384-well black, transparent underside, (DiscoveRx). After 24 hours of incubation, the cells were treated with various concentrations (1.8 μM, 600 nM, 200 nM and 67 nM) of N-acetyl-LTE ₄ in PBS containing 1% dimethylsulfoxide Are shown in red, orange, yellow and green, respectively, in Fig. 5A) and incubated at 37 [deg.] C for 90 minutes. The cells were then added to the DiscoveRx reagent according to the manufacturer's instructions and incubated at room temperature for 30-60 minutes. Fluorescence was measured on a Hamamatsu FDSS μ Cell reader. Collected data were entered into RLUs and compared to the control (N-acetyl-LTE ₄ < RTI ID = 0.0 > Without addition). The results are shown in Fig. 5A. In addition, the cell activity profile (indicated in blue) of the high concentration 3700 ng / ml human gamma globulin and the 600 nM N-acetyl-LTE ₄ A comparison of the cell activity profile (indicated in orange) derived from the addition is shown in Figure 5b.

The readout information is described according to the X-axis, the Y-axis represents log10 as a relative value to the control solvent (ethanol), and the gray area above and below the dotted line represents 95% significance as a control standard .

As shown in Fig. 5A, it was confirmed that no significant cytotoxicity occurred in the treatment with N-acetyl LTE _{4 at} a concentration of 67 nM to 1.8 μM, and at the same time, it had broad anti-inflammatory activity. More specifically, it was confirmed that low levels of IL-17 and IL-2 were regulated in BT and IL-10 was regulated in lMphg. This indicates that N-acetyl LTE ₄ has an immunomodulating effect. It was also confirmed that the overall level of collagen III did not change significantly, indicating that the formulated substrate / tissue was remodeled.

Further, additionally, as shown in Figure 5b, the result of comparing the cellular activity profile derived according to the N- acetyl -LTE ₄ was added and the cell activity profile of human gamma globulin, the cellular activity profile for the N- _acetyl-4 immune -LTE It is confirmed that the activity profile is similar to that of human gamma globulin used as an inhibitor. This indicates that N-acetyl-LTE ₄ in vivo has an immune control function for immune diseases.

All of the above results were expressed as mean ± SEM (n = 5) (* P <0.05, ** P <0.01).

Overall, N- _{acetyl-LTE 4 (N-acetyl Leukotriene E4} ) of the present invention to inhibit the water-soluble cytokines, in particular the generation of IL-2 and IL-17 mitigates the symptoms of itching caused by various causes, at the same time the immune Control effect. Accordingly, the present invention can be applied to a composition for alleviating the itching induced by histamine, chloroquine or contact dermatitis, and a preventive or therapeutic agent for allergic or inflammatory skin diseases including atopic dermatitis.

Formulation example  1. Preparation of pharmaceutical preparations

One. Sanje  Produce

20 mg of N-acetyl LTE ₄

Lactose 100 mg

Talc 10 mg

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

2. Preparation of tablets

N-acetyl LTE ₄ 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

3. Preparation of capsules

N-acetyl LTE ₄ 10 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

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

4. Preparation of injections

N-acetyl LTE ₄ 10 mg

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na ₂ HPO ₄ 2H ₂ O 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

5. Liquid  Produce

20 mg of N-acetyl LTE ₄

10 g per isomer

5 g mannitol

Purified water quantity

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

Formulation example  2. Manufacture of food preparation

1. Manufacture of health food

100 mg of N-acetyl LTE ₄

Vitamin mixture quantity

70 g of vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

0.2 g of vitamin B12

Vitamin C 10 mg

Biotin 10 g

Nicotinic acid amide 1.7 mg

Folate 50 g

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

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

2. Manufacture of health drinks

100 mg of N-acetyl LTE ₄

Vitamin C 15 g

Vitamin E (powder) 100 g

19.75 g of ferrous lactate

3.5 g of zinc oxide

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B1 0.25 g

Vitamin B2 0.3 g

Water quantification

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

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Formulation example  3. Cosmetics  Manufacture of pharmaceutical preparations

1. Flexible longevity manufacturing

N-acetyl LTE ₄ 0.1 wt%

5.2% by weight of 1,3-butylene glycol, 1.5% by weight of oleyl alcohol,

3.2% by weight of ethanol

Polysorbate 20 3.2 wt%

Benzophenone-9 2.0 wt%

1.0% by weight of a carboxyl vinyl polymer, 3.5% by weight of glycerin,

A minor amount of fragrance, a small amount of preservative, and a remaining amount of purified water were mixed to prepare a softening water by a conventional method.

2. Milk lotion  Produce

N-acetyl LTE ₄ 0.1 wt%

Glycerin 5.1 wt%

Propylene glycol 4.2 wt%

3.0% by weight of tocopheryl acetate

Liquid paraffin 4.6 wt%

1.0% by weight triethanolamine

Squalane 3.1 wt%

Macadamia nut oil 2.5 wt%

Polysorbate 60 1.6 wt%

1.6% by weight of sorbitan sesquiterate

Propyl paraben 0.6 wt%

Carboxyl vinyl polymer 1.5 wt%

A minor amount of fragrance, a small amount of preservative, and a remaining amount of purified water were mixed to prepare an milk lotion by a conventional method.

3. Nourishing Cream  Produce

N-acetyl LTE ₄ 0.5 wt%

Glycerin 4.0 wt%

Vaseline 3.5 wt%

2.1% by weight triethanolamine

Liquid paraffin 5.3 wt%

Squalane 3.0 wt%

Wax 2.6 wt%

Tocopheryl acetate 5.4 wt%

Polysorbate 60 3.2 wt%

Carboxyl vinyl polymer 1.0 wt%

3.1 weight percent sorbitan sesquioleate

A minor amount of fragrance, a small amount of preservative, and a remaining amount of purified water were mixed to prepare a nutritional cream by a conventional method.

Claims (8)

N- _{acetyl-LTE 4 (N-acetyl Leukotriene E4} ) or a pharmaceutically acceptable salt, including, as an active ingredient, allergic or inflammatory skin diseases, the prevention or treatment a pharmaceutical composition of.
The method according to claim 1,
Wherein the skin disease is selected from the group consisting of atopic dermatitis, allergic dermatitis, contact dermatitis, acne, seborrheic dermatitis, rashes, urticaria and psoriasis.
N- _{acetyl-LTE 4 (N-acetyl Leukotriene E4} ) or a pharmaceutically which, allergic or prevention or the external preparation for skin for the treatment of inflammatory skin diseases including acceptable salt thereof as an active ingredient.
N- _{acetyl-LTE 4 (N-acetyl Leukotriene E4} ) or a pharmaceutically acceptable salt of the active ingredient, allergic or inflammatory skin diseases health food composition for the prevention and improvement of including the.
N- _{acetyl-LTE 4 (N-acetyl Leukotriene E4} ) or a pharmaceutically itch relief, including acceptable salt as an active ingredient or a cosmetic composition for inhibiting.
6. The method of claim 5,
Characterized in that itching is caused by at least one member selected from the group consisting of atopic dermatitis, allergic dermatitis, contact dermatitis, dermatitis due to roughness of the skin, acne, seborrheic dermatitis, rash, erythema, ascites, urticaria and psoriasis &Lt; / RTI &gt;
6. The method of claim 5,
Wherein the cosmetic composition has a skin moisturizing effect or a skin and keratinization effect.
6. The method of claim 5,
The cosmetic composition may be in the form of a solution, a gel, a solid, a paste, an emulsion, a suspension, an emulsion, a microemulsion, a microcapsule, a microgranule, a liposome, , A pack, a cleanser, a soap, an oil, a wax, a conical stick or an aerosol.

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