KR101634737B1 - Pharmaceutical composition for Atopic dermatitis - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for prevention or treatment of atopy, which comprises a monocotyledonous tree-derived material, and more particularly to a pharmaceutical composition for preventing or treating atopy comprising interleukin-6, interleukin-1b, interleukin-4, interleukin-13, tumor necrosis factor which inhibits the expression of TNF-alpha, beta-hexosaminidase and inhibitor of kappa B alpha, and is effective for preventing or treating atopic dermatitis .

Description

Pharmaceutical composition for atopic dermatitis < RTI ID = 0.0 >

The present invention relates to a pharmaceutical composition for preventing or treating atopic dermatitis, and more particularly, to a pharmaceutical composition for prevention or treatment of atopic dermatitis, , Β-hexosaminidase and an inhibitor of kappa B alpha, which are effective for the prevention or treatment of atopic dermatitis.

Atopic dermatitis is a chronic inflammatory skin disease that occurs most often during infancy and childhood, and is repeated and aggravated, accompanied by rash and severe itching by external stimulants. Recently, industrialization, urbanization and environmental pollution have rapidly increased the incidence of atopic dermatitis, which is becoming a social problem. Although the pathogenesis of atopic dermatitis has not yet been elucidated, pathological factors such as hyperactivity due to an increase in IgE antibody or functional deficiency of T lymphocytes due to deterioration of cell-mediated immune function are revealed And is thought to be a disease in which immunological abnormalities are now involved. In addition, it is reported that extrinsic atopic dermatitis, which accounts for most of atopic dermatitis, is caused by IgE-related immune mechanism, and delayed immune response due to T-cell abnormality is involved rather than immediate type immune response to specific allergens. Recently, Th2-related cytokines such as Il-4, which induce IgE production from B cells, have been reported to be the cause of atopy (JS Kang et al., Inhibition of atopic dermatitis by topical application of silymarin in NC / Nga mice Intl Immunopharm. (2008) 8 1475-1480).

On the other hand, for the treatment and management, it is common to prescribe a moisturizing agent that maintains moisture on the skin surface and a steroid hormone, that is, a topical corticosteroid preparation, to improve the inflammatory reaction.

However, local adrenocortical hormone is troublesome for long-term use because it causes various skin side effects such as skin atrophy, vasodilatation, pigment desalination, and swelling ancestry. Therefore, the development of atopic treatments or adjuvants using natural drug candidates having anti-atopic efficacy and very low side effects is proceeding in the field of pharmaceuticals and cosmetics.

In the related art disclosed in Japanese Patent Application Laid-Open No. 10-2011-0113028 (published on Oct. 14, 2011), there is disclosed a composition for preventing or treating atopy containing a leaf extract as an active ingredient and a method for producing the composition, This is because it uses an extract using an organic solvent which is not suitable for use in atopic skin, which is a sensitive skin, and is not suitable for use in the human body directly.

Therefore, it is urgent to develop a pharmaceutical composition containing a natural extract having a low side effect suitable for use in skin of a sensitive infant and having high activity for the prevention and treatment of atopic dermatitis.

The present invention has been made in order to solve the above-mentioned problems, and it is an object of the present invention to provide a pharmaceutical composition for preventing or treating inflammation, which comprises interleukin-6, interleukin-1b, interleukin-4, interleukin-13, tumor necrosis factor- It is effective for the prevention or treatment of atopic dermatitis by inhibiting the expression of β-hexosaminidase and inhibitor of kappa B alpha, and is suitable for use as an agent because it does not use organic solvent and has no side effects. To provide an effective composition.

The present invention provides a pharmaceutical composition for prevention or treatment of atopy comprising a compound represented by the following formula (1): < EMI ID =

(Wherein R ₁ is an olefin group of C ₂ to C ₄ , R ₂ is an olefin group of C ₂ to C ₄ , and R ₃ is a C ₁ to C ₃ alkyl group).

According to a preferred embodiment of the present invention, the pharmaceutical composition may inhibit the expression of at least one of the group consisting of interleukin-6, interleukin-1b, interleukin-4 and interleukin-13.

According to another preferred embodiment of the present invention, the compound represented by the above formula (1) may be derived from a plain tree.

According to another preferred embodiment of the present invention, the pharmaceutical composition may be a powder, a gel, an ointment, a cream, a liquid or an aerosol formulation.

According to another preferred embodiment of the present invention, 1 to 20% by weight of the total weight of the pharmaceutical composition may contain the compound represented by the formula (1).

The present invention also provides a cosmetic composition for prevention or improvement of atopy comprising a compound represented by the following Formula 1:

[Chemical Formula 1]

(Wherein R ₁ is an olefin group of C ₂ to C ₄ , R ₂ is an olefin group of C ₂ to C ₄ , and R ₃ is a C ₁ to C ₃ alkyl group).

According to a preferred embodiment of the present invention, the compound represented by the formula (1) may be derived from a plain tree.

According to another preferred embodiment of the present invention, the cosmetic composition according to the present invention is used for the treatment and / or prevention of tumor necrosis factor-alpha (TNF-alpha), beta-hexosaminidase and inhibitors of kappa beta of kappa B alpha). < / RTI >

According to another preferred embodiment of the present invention, the cosmetic composition is one of the group consisting of a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant- Lt; / RTI >

Further, the present invention provides a health functional food for prevention or improvement of atopy comprising a compound represented by the following formula (1): < EMI ID =

[Chemical Formula 1]

(Wherein R ₁ is an olefin group of C ₂ to C ₄ , R ₂ is an olefin group of C ₂ to C ₄ , and R ₃ is a C ₁ to C ₃ alkyl group).

In addition, the present invention separates and separates the essential oil from the essential oil using a mobile phase developing solvent 1 containing nucleic acid and ethyl acetate in a volume ratio of 7 to 9: 1 and thin-layer chromatography (TLC) Obtaining water; And the mobile phase developing solvent 2 comprises fractionating the separated product using open column chromatography using a nucleic acid and ethyl acetate to obtain a fraction, wherein the mobile phase developing solvent 2 is 48 to 52: 1 1 to 5: 1, and 1 to 3: 1 by volume in the range of from 1: 1 to 3: 1, .

[Chemical Formula 1]

(Wherein R ₁ is an olefin group of C ₂ to C ₄ , R ₂ is an olefin group of C ₂ to C ₄ , and R ₃ is a C ₁ to C ₃ alkyl group).

Hereinafter, terms of the present invention will be described.

The term "atopy" of the present invention means chronic and recurrent inflammatory skin disease, including pruritus (itching), dry skin, and eczema. The composition comprising the compound of Formula 1 according to the present invention may be useful for preventing or treating such diseases.

The term "prevention" used in the present invention of the present invention means all actions that inhibit or delay the onset of atopy by the administration of the composition comprising the compound represented by the formula (1).

Furthermore, the term "treatment" used in the present invention means all the actions of improving or alleviating symptoms of atopy by administration of a composition containing the compound represented by the above-mentioned formula (1).

The present invention also relates to a pharmaceutical composition comprising interleukin-6, interleukin-lb, interleukin-4, interleukin-13, tumor necrosis factor-alpha (TNF-?),? -Hexosaminidase, (Inhibitor of kappa B alpha), which is effective for the prevention or treatment of atopic dermatitis. It is also effective to provide a pharmaceutical composition suitable for use as a medicine because of not using an organic solvent and having few side effects.

Fig. 1 shows the result of fractionation of unshale oil essential oil with an organic solvent in Production Example 1. Fig.
FIG. 2 shows the results of measuring mRNA expression inhibitory effects of beta-hexosaminidase according to the treatment concentrations of fractions 1 to 7 of Preparation Example 1 in Experimental Example 1. FIG.
FIG. 3 shows the results of measuring the inhibitory effect of IL-4 on mRNA expression according to the treatment concentrations of the fractions 1 to 7 of Preparation Example 1 in Experimental Example 1. FIG.
FIG. 4 shows the results of measuring the inhibitory effect of IL-13 on mRNA expression according to the treatment concentrations of the fractions 1 to 7 of Preparation Example 1 in Experimental Example 1. FIG.
5 shows the results of GC-MS analysis of the fraction 5 in Example 1. Fig.
6 shows the results of GC-MS analysis of the fraction 5-1 in Example 1. Fig.
7 shows the results of GC-MS analysis of fraction 5-2 in Example 1. Fig.
8 shows the results of GC-MS analysis of fraction 5-3 in Example 1. Fig.
9 shows the results of GC-MS analysis of fraction 5-4 in Example 1. Fig.
FIG. 10 shows the results of measuring mRNA expression inhibitory effects of beta-hexosaminidase according to the treatment concentrations of fractions 5-1 to 5-4 of Example 1 in Experimental Example 2. FIG.
FIG. 11 shows the results of measuring the inhibitory effect of IL-4 on mRNA expression according to the treatment concentrations of the fractions 5-1 to 5-4 of Example 1 in Experimental Example 2. FIG.
12 shows the results of measuring the inhibitory effect of IL-13 on mRNA expression according to the treatment concentrations of the fractions 5-1 to 5-4 of Example 1 in Experimental Example 2. FIG.
13 shows GC-MS results of the isolated product in Example 2. Fig.
14 shows the results of measurement of beta-hexosaminidase inhibitory activity of the Elemol, Beta-Udesmol, and Gamma-Udesmole obtained in Example 2 in Experimental Example 5. FIG.
15 shows the results of measurement of the IL-4 inhibitory activity of the ELEMOL, beta-UDESMOL and gamma-UDES mol obtained in Example 2 in Experimental Example 5. FIG.
16 shows the results of measurement of IL-13 inhibitory activity of the ELEMOL, beta-yildesmol and gamma-yudes mol obtained in Example 2 in Experimental Example 5. Fig.
17 shows the result of measurement of anti-atopic activity through a mouse model in Experimental Example 7. FIG.
18 is a graph showing the change in the concentration of serum IgE in the blood sample of the mouse model in Experimental Example 7. FIG.
Fig. 19 shows the result of staining a tissue specimen of a mouse model with heptadoxylin and eosin in Experimental Example 7. Fig.
20 is a result of staining a tissue sample of a mouse model with toluidine blue in Experimental Example 7;
FIG. 21 shows the results of measuring the number of mast cells stained with toluidine blue in Experimental Example 7 under a light microscope (400.times.).
22 shows the anti-atopic activity (TNF-α, IL-1b, IL-6 and IκB-α inhibitory activity) of the eleolol measured in Experimental Example 8.

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

As described above, in order to treat and manage atopy, conventionally, it is common to prescribe a moisturizing agent that maintains moisture on the skin surface and a steroid hormone, that is, a topical corticosteroid preparation, in order to improve the inflammatory reaction.

However, local adrenocortical hormone is troublesome for long-term use because it causes various skin side effects such as skin atrophy, vasodilatation, pigment desalination, and swelling ancestry. Therefore, the development of atopic treatments or adjuvants using natural drug candidates having anti-atopic efficacy and very low side effects is proceeding in the field of pharmaceuticals and cosmetics. However, it is urgently required to develop a pharmaceutical composition which does not use an organic solvent which is not suitable for use in skin of sensitive infants, has a low side effect, contains natural extracts, and has high activity of preventing and treating atopic dermatitis.

Accordingly, the present invention has been made to solve the above-mentioned problems by providing a pharmaceutical composition for prevention or treatment of atopy comprising a compound represented by the following formula (1). (IL-6), interleukin-1b, interleukin-4, interleukin-13, tumor necrosis factor-alpha (TNF-alpha), beta-hexosaminidase, It is effective to provide a pharmaceutical composition which is effective for prevention or treatment of atopic dermatitis by inhibiting the expression of an inhibitor of kappa B alpha and is suitable for use as a medicine because of not using an organic solvent and having few side effects.

[Chemical Formula 1]

The present invention provides a pharmaceutical composition for prevention or treatment of atopy represented by the above formula (1).

In this case, R ₁ may be an olefin group of C ₂ to C ₄ , more preferably an olefin group of C ₃ , and most preferably prop-1-en-2-yl.

Further, R ₂ is an olefin group of C ₂ to C ₄ , more preferably an ethenyl group which is an olefin group of C ₂ .

Furthermore, R ₃ is a C ₁ -C ₃ alkyl group, and more preferably a methyl group.

As shown in Experimental Example 5, Experimental Example 8, Figures 16 to 18 and 24, the pharmaceutical composition inhibits the expression of at least one of the group consisting of interleukin-6, interleukin-1b, interleukin-4 and interleukin- Which is effective in prevention and treatment of atopy.

The compound represented by the above formula (1) is a natural material-derived substance, and may be preferably one derived from a whitish tree.

The pharmaceutical composition of the present invention may further comprise a carrier. The pharmaceutically acceptable carrier to be contained in the pharmaceutical composition of the present invention is one commonly used in the present invention. Examples of the pharmaceutically acceptable carrier include lactose, dextrose, sucrose, sorbitol, But are not limited to, mannitol, starch, acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, Talc, magnesium stearate, mineral oil, and the like.

The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition is not particularly limited as long as it is a formulation suitable for use for prevention or treatment of atopy, but may be in the form of an external preparation for skin, more preferably a powder, gel, ointment, cream, liquid or aerosol formulation Lt; / RTI >

In the present invention, the external preparation for skin type gel base includes Carbopol, Carbomer, Polyethyleneglycol, Polypropylene Glycol, Polyacrylic acid, Carboxymethyl cellulose Hydroxymethylcellulose, polyvinylpyrrolidone, gelatine, alginate salt, chitin or chitosan derivatives, hyaluronic acid, hyaluronic acid, Collagen may be used, but the present invention is not limited thereto.

The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as the formulation method, the administration method, the age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate and responsiveness of the patient, Usually, a skilled physician can readily determine and prescribe dosages effective for the desired treatment or prophylaxis.

In addition, the content of the compound of formula (1) is not particularly limited as far as it contains the compound of formula (1), but it is preferably 1 to 20% by weight of the total weight of the pharmaceutical composition By weight, and more preferably 2 to 18% by weight.

When the compound of Formula 1 is contained in an amount of less than 1% by weight, interleukin-6, interleukin-1b, interleukin-4, interleukin-13, tumor necrosis factor- The inhibitory activity of inhibitors of kappa B alpha is low and thus it is not suitable for the prevention or treatment of atopy, and when it contains more than 20% by weight , The inhibitory activity against the addition amount of the compound of formula (1) may be low, which is not economical.

In addition, the present invention provides a cosmetic composition for preventing or ameliorating atopy comprising the compound represented by the above formula (1).

In this case, R ₁ may be an olefin group of C ₂ to C ₄ , more preferably an olefin group of C ₃ , and most preferably prop-1-en-2-yl.

Further, R ₂ is an olefin group of C ₂ to C ₄ , more preferably an ethenyl group which is an olefin group of C ₂ .

Furthermore, R ₃ is a C ₁ -C ₃ alkyl group, and more preferably a methyl group.

As can be seen in Experimental Example 5, Experimental Example 8, Figures 16 to 18 and 24, the pharmaceutical composition can be used for tumor necrosis factor-alpha (TNF-alpha), beta-hexosaminidase (beta- hexosaminidase) and inhibitor of kappa B alpha (inhibitor of kappa B alpha), which is effective for prevention or improvement of atopy.

The compound represented by the above formula (1) is a natural material-derived substance, and may be preferably one derived from a whitish tree.

The cosmetic composition is not particularly limited so long as it is a formulation suitable for use for prevention or treatment of atopy. For example, it may be formulated into solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays, But is not limited thereto. More specifically, it can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

When the formulation of the present invention is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters.

The ingredients contained in the cosmetic composition of the present invention include, in addition to the active ingredient and the carrier ingredient, the ingredients conventionally used in cosmetic compositions and include conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, .

When the composition of the present invention is identified as a soap composition, the soap composition of the present invention can be prepared by incorporating an active ingredient into a soap base, and can be prepared as an additive including a skin moisturizer, an emulsifier, a water softener and the like.

Examples of the soap base material include vegetable oils such as palm oil, palm oil, soybean oil, perm free oil, olive oil and palm kernel oil or animal fats such as tallow, lard, sunshine and fish oil. Examples of the skin moisturizers include glycerin, erythritol, polyethylene glycol , Propylene glycol, butylene glycol, pentylene glycol, hexyl glycol, isopropyl myristate, silicone derivatives, aloe vera, sorbitol and the like. Examples of the emulsifier include natural oils, wax fatty alcohols, hydrocarbons, May be used. As the water softening agent, tetrasodium ethylenediate and the like may be used.

The soap composition of the present invention may further contain, as an additive, an antibacterial agent, a dispersant, a foam inhibitor, a solvent, a scouring inhibitor, a corrosion inhibitor, a fragrance, a dye, a sequestering agent, an antioxidant and an antiseptic.

In the soap composition of the present invention, the soap base or additive may be included in a content commonly used in the art, wherein the soap base generally comprises from 99.999% to 50% by weight, based on the total weight of the soap composition And the additive may be added in an amount of 1 wt% to 50 wt%.

In addition, the present invention provides a health functional food for prevention or improvement of atopy comprising the compound represented by the general formula (1).

In this case, R ₁ may be an olefin group of C ₂ to C ₄ , more preferably an olefin group of C ₃ , and most preferably prop-1-en-2-yl.

Further, R ₂ is an olefin group of C ₂ to C ₄ , and more preferably an ethenyl group which is an olefin group of C ₂ .

R ₃ is a C ₁ -C ₃ alkyl group, and more preferably a methyl group.

The kind of health functional food is not particularly limited as long as it is usually produced and / or sold. For example, dairy products such as meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums and ice cream, soups, drinks, tea, drinks, alcoholic beverages, And can be used in the form of pills, powders, granules, infusions, tablets, capsules or beverages, all of which include health functional foods in the conventional sense.

The health beverage composition of the present invention is not particularly limited as long as it contains the compound represented by the formula (1), and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages.

Examples of the natural carbohydrate include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; Polysaccharides such as dextrin, cyclodextrins; And sugar alcohols such as xylitol, sorbitol, and erythritol. As natural flavors other than those described above, natural flavors (such as tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin)) and synthetic flavors (saccharin, aspartame, etc.) have.

In addition to the above-mentioned additional components, the health functional food of the present invention may contain flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and aging agents (cheese, chocolate, etc.) Salts of alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like.

In addition, the health functional foods of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination.

Hereinafter, the present invention will be described in detail with reference to examples. However, these examples are intended to further illustrate the present invention, and the scope of the present invention is not limited to these examples.

[ Example ]

Manufacturing example . Manufacture of woodwort-derived materials.

A 500 - gram (500 ~) 1 ㎜ (1 ㎝ - 1 ㎝) leaf and a 4 ℓ distilled water were added to the laboratory - scale steam distillation apparatus to extract the unshiu tree essential oil. The temperature is gradually increased to volatilize the volatile substances contained in the flat leaves at 100 ° C. The volatiles were liquefied and collected in the trap as they passed through the cooler. The volatiles were collected in the form of condensate for 24 hours. The trap contained condensation of water vapor and volatile substances, and water was collected at the bottom and volatile substances in the form of oil at the top, thereby removing water and obtaining volatile substances in the form of oil. These volatile substances were called unrefined wood oil, and the yield of unrefined wood oil was 3.6%.

Then, the eluate was purified by thin layer chromatography (TLC). The eluted fractions were eluted with a mixture of nucleic acid and ethyl acetate (v / v) at a ratio of 20: 1, 16: 1, 12: 1, 10: 1, 8: 1 and 4: 1, respectively. For this purpose, isolates of 8: 1 nucleic acid and ethyl acetate (v / v) were used for subsequent fractions.

Then, open column chromatography was carried out using the separated product. Specifically, 2 kg of silica was added to a column having a diameter of 10 cm and a length of 80 cm as a stationary phase and packed with nucleic acid for 24 hours.

After the packing, 100 grams of unrefined wood essential oil was loaded and fractionation was started. The mobile phase was eluted sequentially with 50: 1, 16: 1, 8: 1, 4: 1 and 2: 1 ratio of nucleic acid and ethyl acetate to obtain 350 fractions.

The 350 fractions were spotted using a thin layer chromatography (hexane: ethyl acetate, 8: 1) as described above. The fractions located on the same spot were separated into a total of 7 fractions (1 to 7).

Preparation Example  1. Cell culture

RBL-2H3 cells were purchased from Korean Cell Line Bank (Seoul, Korea) and incubated with 10% heat-inactivated fetal bovine serum, 2 mM L-glutamine, 100 units / ml penicillin, The cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) containing 100 쨉 g / ml streptomycin while being filled with 80% of 6-well culture dishes at 37 ° C under 5% CO ₂ conditions.

Experimental Example  One. Of fractions Anti-atopic  Active measurement

The RBL-2H3 cell line cultured in Preparation Example 1 was divided into 0.25 × 10 ⁵ cells in 96-wells, and 25 μg / ml concanavalin A (Con A) was added to induce an immune response for 1 hour. Then, the cells were washed twice with PBS, and 1 占 퐂 / ml of cyclosporin A (CsA) and one of the fractions 1 to 7 prepared in Preparation Example 1 were added at 10 ^-7 , 10 ^-6 or 10 ^-5 % (v / v ) And 1 hour later, the RNA was isolated according to the instructions using TriZol (Invitrogen Life Technologies, USA), and cytokine interleukin-4 or interleukin-13 was reacted with real-time polymerase chain reaction -time PCR).

Specifically, in the real-time PCR, 300 ng of oligo (dT) and reverse transcriptase were added to 1 μg of total RNA by measuring the concentration of the separated RNA, and reacted at 42 ° C. for 1 hour to obtain a reaction solution. Then, 200 nM of oligonucleotide, 10 mM Tris-HCl, pH 8.3, 50 mM KCl, 3.0 mM MgCl ₂ and 0.125 units of Taq polymerase (iNtRON Biotechnology, Korea) were added to the reaction solution, PCR was carried out with 25 cycles using a DNA gene amplifier (Thermal cycler, Applied Biosystems) under conditions of 55 ° C for 30 seconds and 72 ° C for 30 seconds.

After electrophoresis on a 1.5% agarose gel, images were analyzed with phorimager analysis (Molecular Dynamics, USA) under UV.

The oligonucleotide used at this time is shown in Table 1 below, and the obtained agarose gel electrophoresis results were numerically analyzed.

target
Cytokine RNA (5'-3 ') division SEQ ID NO: TNF-a CCT GTA GCC CAC GTC GTA G sense RNA SEQ ID NO: 1 GGG AGT AGA CAA GGT ACA ACC C antisense RNA SEQ ID NO: 2 IL-1? GAA ATG CCA CCT TTTG ACA GTG sense RNA SEQ ID NO: 3 CTG GAT GCT CTC ATC AGG ACA antisense RNA SEQ ID NO: 4 IL-6 CTG CAA GAG ACT TCC ATC CAG sense RNA SEQ ID NO: 5 AGT GGT ATA GAC AGG TCT GTT GG antisense RNA SEQ ID NO: 6 Iκ-β GCA ATC ATC CAC GAA GAG AAG CC sense RNA SEQ ID NO: 7 CAG GAT CAC AGC CAG CTT TCA GAA G antisense RNA SEQ ID NO: 8 β-actin TTC TAC AAT GAG CTG CGT GTG sense RNA SEQ ID NO: 9 ACC AGA GGC ATA CAG GGACA antisense RNA SEQ ID NO: 10 IL-4 CCA GGG TTT GGA ATT ATT TC sense RNA SEQ ID NO: 11 GAA GAT AAA CCC TAA GGC TC antisense RNA SEQ ID NO: 12 IL-13 TGA TGT ACC TCC GTG CTT GA sense RNA SEQ ID NO: 13 AGG ACA TGG AAG TGC AGG AC antisense RNA SEQ ID NO: 14

The β-hexosaminidase activity was determined by dividing the RBL-2H3 cell line cultured in Preparatory Example 1 by 2 × 10 ⁵ cells in a 6-well plate and incubating with DNP-specific IgE (800 ng / ml, Sigma, USA ) For 16 hours and the culture medium was diluted with Tyrodes' assay buffer (119 mM NaCl, 4.74 mM KCl, 2.5 mM CaCl ₂ , 1.19 mM MgSO ₄ , 10 mM HEPES, 5 mM glucose 0.1% (w / 7.3).

Then, after 15 minutes of cultivation, one of the fractions 1 to 7 prepared in Preparation Example 1 was treated at a concentration of 10 ^-7 , 10 ^-6 or 10 ^-5 % (v / v), and again with 2,4-nitrofluoro And stimulated with benzene-fetal serum (2,4-Dinitrophenol-bovine-serum-albumin; DNP-BSA, 100 ng / mL, Sigma) for 3 hours. Add the same amount of substrate solution (1 mM p-nitrophenyl N-acetyl-beta- D-glucosamine in 0.05 M citrate buffer, pH 4.5) and incubate at 37 ° C for 1 hour. Add 0.05 M sodium bicarbonate buffer And the absorbance is measured at 450 nm. The measured absorbance was numerically analyzed.

The numerical analysis was confirmed by one-way ANOVA. Tukey's test was used for the analysis of other data. Significance was determined by comparing the control and experimental groups with P <0.05. All analyzes were performed using GraphPad Prism (v4.0). The results of the analysis are shown in the following figures 2 to 4.

Fig. 2 shows results of beta-hexosaminidase activity, Fig. 3 shows results of measurement of IL-4 activity, and Fig. 4 shows results of measurement of IL-13 activity. CsA in the figure is the result of treatment with 1 ㎍ / ml cyclosporin A (CsA).

As shown in Fig. 2, in the cells treated with 10 ^-7 , 10 ^-6 or 10 ^-5 (%, v / v) of fraction 5 of the fractions 1 to 7 prepared in Production Example 1, beta-hexosaminidase Was decreased in a concentration-dependent manner.

As shown in FIG. 3, in the cells treated with the fraction 1, 2, 3, 5, 6 or 7 prepared in Preparation Example 1, compared to the negative control treated with only 25 μg / ml concanavalin A IL-4 expression was found to be about 2-fold lower. On the other hand, the expression of IL-4 in fraction 4 treated cells was decreased compared with the negative control, but it was not significant.

As shown in FIG. 4, IL-13 mRNA expression in the cells treated with the fractions 1 to 7 prepared in Preparation Example 1 was inhibited about twice as much as that of the negative control. In addition, the inhibitory effect of IL-13 mRNA expression by the fractions 1 to 7 was observed to be lower than that of the cells treated with cyclosporin A (CsA) as a positive control. Particularly, .

Example  One.

Fraction 5 was further fractionated on the basis of the results of the anti-atopic activity measurement of Experimental Example 1. The fraction 5 was analyzed using GC-MS (GC: Agilent 6890, MS: Agilent 5973, column: HP5 (25 m × 0.32 mm × 0.52 μm), Carrier gas: helium)

5, fraction 5 of Preparation Example 1 contained terpinene-4-ol (13.6%), elemol (20.96%), cedrol (6.01 %), Cadinol (8.02%), gamma-eudesmol (8.07%) and beta-eudesmol (18.14).

Further, the additional fraction was fractionated in the same manner as in Production Example 1, and after loading 700 g of silica into the column, 5 g of fraction 5 was loaded. The solvent was a mixture of nucleic acid and ethyl acetate in a ratio of 16: 1 (v / v).

As a result of this additional fraction, fraction 5 was further fractionated into four fractions. The fractions 5-1 to 5-4 were analyzed using GC-MS as described above, and the results are shown in FIGS. 6 to 9.

As can be seen in FIG. 6, it was confirmed that fraction 5-1 contained 68.97% terpinene-4-ol and 16.45% cadinol, and as shown in FIG. 7, fraction 5-2 contained terpinene-4-ol 68.97%, cedrol 33.64% and cadinol 35.95%.

In FIG. 8, it was confirmed that fraction 5-3 contained cedrol 7.90%, r-eudesmol 14.46% and cadinol 36.65%. In FIG. 9, fraction 5-4 contained elemol 19.67%, gamma -eudesmol 25.28% and beta -eudesmol 33.01%.

Experimental Example  2.

The fractions 5-1 to 5-4 prepared in Example 1 were treated and the anti-atopic activity was measured in the same manner as in Experimental Example 1 and shown in Figs. 10 to 12. FIG. 10 shows the result of measurement of IL-4 activity, FIG. 12 shows the result of measurement of IL-13 activity, and CsA in the figure shows 1 μg / ml of cyclosporin A (cyclosporin A; CsA).

As shown in Fig. 10, the cells treated with 1 占^10-7 , 1 占^10-6 or 1 占^0-5 (%, v / v) of the fractions 5-1 to 5-4 prepared in Example 1 , The activity of beta-hexosaminidase was significantly decreased and it was found that the concentration of beta-hexosaminidase decreased in a concentration-dependent manner, but there was no significant difference between them.

11, the fractions 5-1 to 5-4 prepared in Example 1 were treated with 1 × 10 ^-7 , 1 × 10 ^-6 or 1 × 10 ^-5 (%, v / v) Cells showed significantly lower expression of IL-4 than the negative control treated with concanavalin A alone. In addition, although the expression of IL-4 tended to decrease depending on the fraction concentration treated, it was confirmed that these were not significant.

As shown in FIG. 12, it was confirmed that the expression of IL-13 mRNA in the cells treated with the fractions 5-1 to 5-4 prepared in Example 1 was suppressed about twice as much as that of the negative control. In particular, it was found that the cells treated with the fractions 5-3 to 5-4 inhibited the expression of IL-13 mRNA in a fraction-dependent manner, compared with cells treated with the cyclosporin A (CsA) In particular, it was confirmed that the cells treated with the fraction 5-4 had a similar immunosuppressive effect as compared with the fraction 5 of the upper fraction.

Experimental Example  3. Cytotoxicity experiment

The RBL-2H3 cell line cultured in Preparation Example 1 was divided into 96-wells at 1 × 10 ⁴ cells and cultured at 37 ° C. for 24 hours at 5% CO ₂ . Embodiment one of the Example 1 was prepared in fractions 5-1 to 5-4 after ^{1 × 10 -12%, 1 ×} 10 -10%, 1 × 10 -8%, 1 × 10 -6%, 1 × 10 - ⁴ %, or 1 × 10 ^-2 % (v / v), and incubated at 37 ° C in a 5% carbon dioxide incubator for 24 hours. After the reaction, Cell Counting Kit-8 (Dojindo Laboratories, Tokyo, Japan) was treated for 1 hour and the absorbance was measured at 450 nm using a microplate reader (Molecular Devices, Sunnyvale, CA, USA) . The absorbance as a measurement result was numerically analyzed in the same manner as in Experimental Example 1. The results of the absorbance numerical analysis are shown in Table 2 below.

Fraction IC ₅₀ Fraction 5-1 7.9 x ^10-5 % (v / v) Fraction 5-2 2.9 x ^10-5 % (v / v) Fraction 5-3 1.1 x 10 ^-4 % (v / v) Fraction 5-4 4.1 * 10 ^-5 % (v / v)

As shown in Table 2, it was confirmed that the cytotoxicity was the lowest in fraction 5-3 and the highest in fraction 5-2. However, fraction 5-3 did not show a high inhibitory effect on mRNA expression of IL-4 and IL-13 among fractions 5-1 to 5-4, and IL-4 and IL-13, similar to fractions 5-1 and 5-3, 13 mRNA expression was inhibited.

On the other hand, fraction 5-2 showed cytotoxicity at least 1.5 times and at most 27 times higher than that of fractions 5-1 to 5-4. On the contrary, fraction 5-4 showed the best effect for pharmacological pharmacological effects on IL-4 and IL-13 mRNA expression inhibition and cytotoxicity compared to fractions 5-1 to 5-3 I could.

Experimental Example  4. Fraction  The detection of the constituents of 5-4 and the Anti-atopic  Active measurement

The components of the fraction 5-4 prepared in Example 1 were fractionated by column chromatography and the components were analyzed by GC-MS. As a result of the component analysis, fractions 5-4 were found to contain bornyl acetate, terpineol, Elemol (including β-eudesmol and γ-eudesmol), borneol, and terpinen-4-ol.

The mRNA expression inhibitory effect of beta-hexosaminidase, IL-4 and IL-13 according to the components of fraction 5-4 as described above was measured.

As a result, the mRNA expression of IL-4 increased 2.3-fold by ConA to about 1.5-fold at the concentration of bornyl acetate (1 × 10 ^-8 %, 1 × 10 ^-6 %, 1 × 10 ^-4 % But no concentration-dependent inhibition of expression was observed. In addition, IL-4 mRNA expression was suppressed to approximately 1.7-fold levels in cells treated with 1 × 10 ^-8 % α-terpineol. In addition, Elemol (including β-eudesmol and γ-eudesmol) significantly decreased IL-4 mRNA expression at concentrations of 1 × 10 ^-8 %, 1 × 10 ^-6 %, and 1 × 10 ^-4 % But there was no significant difference in treatment concentration.

In addition, IL-13 mRNA expression, which was increased approximately 3.6-fold by Con A, was expressed at some concentrations of (-) borneol, (+) terpinen-4-ol, Elemol (including eudesmol [β, γ]) and a-terpineol , And the highest inhibitory effect was observed by inhibiting the expression of IL-13 mRNA at a level of 2.3 times by Con A in the cells treated with 1 × 10 ^-8 (%) of (-) borneol.

Elemol (including eudesmol [β, γ]) showed the highest anti-atopic activity among the corresponding components, based on the above-described reactivity and IL-4 and IL-13 expression inhibitory effects.

Example  2.

Based on the anti-atopic activity and cytotoxicity measurement results of Experimental Examples 2 to 4, it was confirmed that elemol, beta-eudesmol and gamma-yudesmol, which are components of fraction 5-4, eudesmol) was isolated.

Specifically, a 5 cm diameter column was filled with 150 g of silica and 1 g of fraction 5-4 was loaded. The solvent was isolated with nucleic acid and ethyl acetate (16: 1, v / v). The results of the isolation analysis using GC-MS are shown in FIG.

As can be seen from Fig. 13, elemol having a purity of 89.06% was obtained as an isolation product.

Experimental Example  5. Anti-atopic  Active measurement

IL-4 and IL-13 inhibitory activities were measured in the same manner as in Experimental Example 1 using anti-atopic activity of the ELEMOL, beta-yudesh mol and gamma-yudesmol obtained in Example 2, Respectively. Fig. 14 shows the result of beta-hexosaminidase activity, Fig. 15 shows the results of measurement of IL-4 activity, Fig. 16 shows the results of measurement of IL-13 activity and CsA in the figure shows 1 mu g / ml cyclosporin A (cyclosporin A; CsA).

As shown in FIG. 14, the concentration of beta-hexosaminidase induced by DNP-BSA in the mast cells was about 2-fold higher than that of the normal group, which was higher than that of the ELMOR, beta- It was confirmed that the activity of beta-hexosaminidase was significantly reduced in cells treated with 1 × 10 ^-7 , 1 × 10 ^-6 or 1 × 10 ^-5 (%, v / v) In particular, it was observed that the cells treated with 1 × 10 ^-6 % and 1 × 10 ^-5 % of the elemol were inhibited to a level of about 1.5 times.

In addition, the inhibitory effect of beta-hexosaminidase at 1 × 10 ^-5 % concentration of fraction 5-4, the upper fraction of Elemol, was similar to that of 1 × 10 ^-6 % of Elemol, In comparison with the 1 × 10 ^-6 % concentration group of Elemol and the 1 × 10 ^-5 % concentration group of the fraction 5-4 at a 10-fold higher concentration, similar immunosuppressive activity was observed in the effect of lowering the blood concentration of beta-hexosaminidase .

As can be seen in Fig. 15, the elemoles, beta-yde desmoles and gamma-yde desmoles of Example 2 were added at a concentration of 1 x 10 ^-7 , 1 x 10 ^-6 or 1 x 10 ^-5 (%, v / v ) Significantly lower IL-4 expression than the control group treated with concanavalin A alone. In particular, the expression of IL-4 in IL-4 was decreased in a dose-dependent manner, but it was not found to be significant.

As shown in FIG. 16, IL-13 mRNA expression in the cells treated with the ELEMOL, beta-yDESMOL and gamma-YDESMOL of Example 2 was inhibited about twice as much as the negative control I could. In particular, it was found that the cells treated with 1 × 10 ^-7 % (v / v) Elimol inhibited the expression of IL-13 mRNA 1.7 times more than the cells treated with the positive control, cyclosporin A (CsA) The higher the treatment concentration, the more inhibition of IL-13 mRNA expression was observed. However, the inhibitory activity was not significantly different depending on treatment concentration.

Experimental Example  6. Elemor  Cytotoxicity measurement

The cytotoxicity of the elemol obtained in Example 2 was measured in the same manner as in Experimental Example 3.

As a result, the IC ₅₀ value of elemol was 2.9 × 10 ^-4 %, which is 7 times higher than 4.1 × 10 ^-5 % of fraction 5-4, which is the upper fraction, and in-vitro cytotoxicity And it was confirmed that it was safer from the viewpoint of evaluation. Therefore, it is considered that Eleamol has higher immunosuppressive effect than fraction 5-4, which is the upper fraction, and secures better intracellular safety, thus being suitable as a target substance for development of anti-atopic agent at the previous clinical level.

Preparation Example  2.

To induce atopic dermatitis on 7 week old BALB / c mice, the back of the mouse is epilated from the neck to the pelvis 2 x 4 cm, and then repeatedly epilated at intervals of 2 days. All experimental animals are divided into 6 groups. All experimental animals were handled with the approval of Chungbuk National University Animal Experimental Ethics Committee.

To induce atopic dermatitis in mice with 2,4-dinitro-chlorobenzene (2,4-dinitro-chlorobenzene), the back of the mouse was epilated 2 x 4 cm from the neck to the pelvis, Additional hair removal was done. Each group was divided into two groups: 1. untreated group (normal group), 2. DNCB treated group (negative control group), 3. DNCB and clobetasol propionate (CP, steroid dermal disease ointment) Control group), 4. Elemol-treated group (experimental group; 2%, 5% or 10%) of Example 2 after DNCB treatment and 10 mice of each group.

At the first DNCB treatment, 2% DNCB was sensitized one time, and after one week, 0.2% DNCB treatment was started from the second week of experiment to induce atopic dermatitis. After the symptoms of atopic dermatitis were confirmed, additional application was made twice a week to keep the symptoms constant. The solvent used in the experiment was a mixture of acetone and olive oil in a ratio of 4: 1 (v / v). DNCB and Elemol were diluted to the corresponding concentrations. The mice used in the experiment were anesthetized at 9 weeks of the start of the experiment to collect blood samples and tissue samples.

Experimental Example  7. In atopic dermatitis mouse model Anti-atopic  Active measurement

Atopic dermatitis mice prepared in Preparative Example 2 were visually evaluated in the mouse skin at 9 weeks as of the start of the experiment. The results of the evaluation are shown in FIG. 17. The control (Ve) was treated with a mixture of acetone and olive oil and the positive control (DNCB + CP) was treated with clobetasol propionate (CP) Week. Elemol of Example 2 was applied to the skin for the same period of time as the positive control with the mixed solution containing the concentration of 2%, 5% or 10%.

As can be seen in FIG. 17, it was confirmed that the gross appearance of the hair follicle induced by DNCB is the same as that of atopic dermatitis due to transdermal color change, appearance of red spot, and formation of a crust.

In addition, symptoms of atopic dermatitis induced during 5 weeks were alleviated by treatment with CP. On the 9th day of the start of the experiment, symptoms such as the degree of red spot, transdermal color change, and scar formation were restored to normal levels Respectively.

Furthermore, the Elemol-treated group (2%, 5% or 10%) of Example 2 also showed atopic dermatitis symptoms such as percutaneous color change, degree of red spot, and scar formation to the positive control level were alleviated, , There was no significant difference according to the concentration.

In addition, the degree of severity calculated on the basis of gross evaluation was 5 stages. Based on 9th week of experiment start, the severity of negative control was 5 levels positive, 1.1 ± 0.3 for positive control, 1.3 ± 0.5 for Elemol 2% , elemol 5% treatment group 1.6 + 0.5 and elemol 10% treatment group 1.9 ± 0.6, respectively. However, concentration-dependent Elemol treatment did not show any concentration-dependent relief of atopic dermatitis.

Blood samples were anesthetized by anesthesia on the 9th week starting from Preparation Example 2. The blood samples were taken from the abdominal atery using a 1 ml syringe and centrifuged at 2,000 rpm for 10 minutes. Respectively. Mouse serum IgE was measured using Mouse IgE Ready-SET-Go! ® as a product of Affymetrix eBioscience. This evaluation focuses on whether the increased concentration of serum IgE in the group treated with DNCB, which is an atopic dermatitis inducing substance, is reduced by Elemol in Example 2. The results of the analysis are shown in FIG.

As can be seen in FIG. 18, it was observed that the IgE concentration increased 3.3-fold by DNCB to about 2.8-fold in the CP treatment group, which is a positive control.

In addition, the serum IgE concentration of the Elemol-treated group (2, 5, 10%) of Example 2 was lowered to about 2.3 times, 2.2 times, and 1.6 times, depending on the concentration, The highest anti - atopic activity was found in the treated group.

Tissue specimens collected by anesthesia on the 9th week of the preparation start date of Preparation Example 2 were cut into 10% neutral formalin and then paraffin sections were cut from the skin tissue of atopic dermatitis mice. After the paraffin was removed by xylen, the sections were treated with hepatoxylin and eosin or toluidine blue, followed by dehydration to obtain canada balsam. And then examined. This review focuses on the epidermal and dermal hyperplasia and the appearance of immune cells. All tissue samples were analyzed by storing images with photographs using an OLYPUS DP controller and manager. The results of staining with helipetoxylin and eosin in the analysis results are shown in Fig. 19, and results of staining with toluidine blue are shown in Fig. 20 Respectively.

Thereafter, the number of mast cells stained with toluidine blue was measured with a light microscope (400 ×), and the number of stained cells / mm 2 in the skin tissue of the atopic dermatitis mice collected thereafter was shown. The results are shown in Fig.

As shown in Fig. 19, hyperplasia was observed in the epidermis and dermis layer by the DNCB-treated group, and many cells suspected to be immune-related cells appeared. In the CP treatment group, the above symptoms were alleviated to the control level. In the Elemol treatment group of Example 2, atopy relieved depending on the concentration of 2%, 5% or 10%.

As shown in FIG. 20, the appearance of mast cells (positive: purple) was observed in the dermal layer of the dermal tissue by the DNCB treatment, and the appearance of mast cells was observed to be decreased by the positive control CP treatment. In addition, it was confirmed that the meconium-treated group of Example 2 showed a decrease in the meconium cells that appeared in a concentration dependent manner on elemol treatment.

As shown in Fig. 21, the appearance of mast cells was increased by DNCB treatment, about 1.6 times in the CP treatment group, 2.4 times in the Elemol 2% treatment group and 1.3 times in the 5% elemol treatment group Respectively. In particular, the highest mast cell decreasing effect was confirmed in the group treated with 10% of Elemol of Example 2.

Experimental Example  8.

TNF-α, IL-1b, IL-6 and IκB-α inhibitory activities were measured in the same manner as in Experimental Example 1 by measuring the anti-atopic activity of the eleolol obtained in Example 2,

As shown in FIG. 22, the expression of inflammation-related factors (TNF-α, IL-1b, IL-6 and IκB-α) increased in expression compared to the normal group by DNCB treatment was evaluated by Elemol And IL-6 and IκB-α were significantly lower than those in the CP treatment group, which is a positive control group.

As can be seen from the examples and experimental examples, the in-vitro evaluation results of the present invention indicate that elemol is considered to be a representative active substance of the anti-atopic activity of the elderly refined wood oil. Through in-vivo evaluation using elemol, , Which is a biological indicator of inflammation of the atopic dermatitis, by inhibiting blood IgE level, hyperplasia of epidermis and dermis, and infiltration of mast cells. As a result, the compound of formula (I) of the present invention was found to have significantly increased anti-atopic activity.

<110> KOREA FOREST RESEARCH INSTITUTE <120> Pharmaceutical composition for Atopic dermatitis <130> 1040489 <160> 14 <170> Kopatentin 2.0 <210> 1 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> oligonucleotide <400> 1 cctgtagccc acgtcgtag 19 <210> 2 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> oligonucleotide <400> 2 gggagtagac aaggtacaac cc 22 <210> 3 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> oligonucleotide <400> 3 gaaatgccac cttttgacag tg 22 <210> 4 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> oligonucleotide <400> 4 ctggatgctc tcatcaggac a 21 <210> 5 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> oligonucleotide <400> 5 ctgcaagaga cttccatcca g 21 <210> 6 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> oligonucleotide <400> 6 agtggtatag acaggtctgt tgg 23 <210> 7 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> oligonucleotide <400> 7 gcaatcatcc acgaagagaa gcc 23 <210> 8 <211> 25 <212> RNA <213> Artificial Sequence <220> <223> oligonucleotide <400> 8 caggatcaca gccagctttc agaag 25 <210> 9 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> oligonucleotide <400> 9 ttctacaatg agctgcgtgt g 21 <210> 10 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> oligonucleotide <400> 10 accagaggca tacagggaca 20 <210> 11 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> oligonucleotide <400> 11 ccagggtttg gaattatttc 20 <210> 12 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> oligonucleotide <400> 12 gaagataaac cctaaggctc 20 <210> 13 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> oligonucleotide <400> 13 tgatgtacct ccgtgcttga 20 <210> 14 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> oligonucleotide <400> 14 aggacatgga agtgcaggac 20

Claims (11)

1. A pharmaceutical composition for prevention or treatment of atopy comprising 1 to 20% by weight of a compound of the following formula (1)
[Chemical Formula 1]

The pharmaceutical composition for prevention or treatment of atopy according to claim 1, wherein the pharmaceutical composition inhibits mRNA expression of at least one member selected from the group consisting of interleukin-6, interleukin-1b, interleukin-4 and interleukin-13 . delete The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is a powder, a gel, an ointment, a cream, a liquid or an aerosol formulation. delete 1. A cosmetic composition for prevention or improvement of atopy comprising 1 to 20% by weight of a chewy wood-derived compound represented by the following formula (1)
[Chemical Formula 1]

delete 7. The cosmetic composition of claim 6, wherein the cosmetic composition is selected from the group consisting of tumor necrosis factor-alpha (TNF-a), beta-hexosaminidase and inhibitor of kappa B alpha Wherein the at least one inhibitor is at least one of the group consisting of: &lt; RTI ID = 0.0 &gt;
9. The cosmetic composition according to claim 6 or 8, wherein the cosmetic composition is one of the group consisting of a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant- Wherein the cosmetic composition is a cosmetic composition for prevention or improvement of atopy.
A health functional food for prevention or improvement of atopy comprising 1 to 20% by weight of a chewy wood-derived compound represented by the following formula 1:
[Chemical Formula 1]

(a) distilling the unicorn tree leaves to obtain a unicorn tree essential oil;
(b) fractionating the elixir seed oil obtained in step (a) by 1 to 2 fractions by using open-column chromatography using a mobile phase developing solvent 2 containing nucleic acid and ethyl acetate in a volume ratio of 5-10: 1, ;
(c) fractionating the fractions obtained in step (b) by open column chromatography using a developing solvent 3 containing nucleic acid and ethyl acetate in a volume ratio of 12 to 20: 1 to obtain an isolated product Lt; RTI ID = 0.0 &gt; (1) &lt; / RTI &gt;
[Chemical Formula 1]

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