KR101986008B1 - Pharmaceutical composition for preventing or treating atopic dermatitis comprising extract of resveratrol-enriched rice or resveratrol enriched rice callus as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating atopic dermatitis, a cosmetic composition for improving atopic dermatitis, a composition for preventing or treating atopic dermatitis, a composition for preventing or treating atopic dermatitis, which contains, as an active ingredient, an extract of resveratrol biosynthesis rice or callus thereof deposited under accession number KCTC12529BP Wherein the extract of the resveratrol biosynthesis rice or the callus thereof does not exhibit cytotoxicity and improves the symptoms of atopic dermatitis in mice that have induced atopic dermatitis and the inflammatory cytokine IL-6 or IL-1β It was confirmed that the expression level was decreased.

Description

[0001] The present invention relates to a pharmaceutical composition for preventing or treating atopic dermatitis comprising resveratrol rice or a resveratrol callus extract. [0002] The present invention relates to a pharmaceutical composition for preventing or treating atopic dermatitis comprising resveratrol rice or resveratrol callus extract,

The present invention relates to a pharmaceutical composition for preventing or treating atopic dermatitis including resveratrol rice and resveratrol callus extract, and a cosmetic composition for improving atopic dermatitis, an external preparation for skin or a health food.

Atopic dermatitis is a refractory immune disease, and environmental influences are on the rise. The prevalence of atopic dermatitis was 22.6% in elementary school students and 12.9% in middle school students. The incidence of atopic dermatitis was higher in Korea than in 2000 The incidence of disease was significantly higher than that of 13.4% and 6.7%. Between 1990 and 2010, Africa grew from 9.9% to 20.9%, from Europe from 18.9% to 19.6% and from Asia to Japan from 10.1% to 13.6%. It is highly prevalent in western society and immigrant group due to urbanization and is more frequent in developed countries than Western Europe and developing countries compared to Eastern Europe. It can also be attributed to differences in atmospheric substances, climate, food, and microbiological environment that cause allergies. The high prevalence of atopic dermatitis in western society is due to the clean environment, It is explained that the system becomes more sensitive to the allergen of the environment and the autoimmune reaction easily occurs.

Although corticosteroids, immunosuppressants, antihistamines, skin moisturizers, and softening agents are all used to treat dermatitis, topical steroids and antihistamines are among the drugs used for pharmacological treatment, and most of the drugs currently in use have serious side effects Because of the emergence of phototherapy and drug therapy in combination with the treatment effect is to increase the side effect is to reduce.

In terms of traditional knowledge theory, atopic dermatitis is expressed as ~, and the major cause of it is fever, hemorrhage, hemorrhage, etc. In this case, it is known that the condition is inharmonious as the environmental factor, which is an environmental factor invading from the outside, and causes problems such as skin and muscles.

In general, atopic dermatitis, lymphocytes, macrophages, and granular mast cells are infiltrated into lesions, and eosinophils are elevated in the blood and IgE (Immunoglobulin E) is elevated in the serum. Atopic dermatitis has a complicated pathogenesis due to various factors such as genetic pollution, environmental factor, pharmacological action, abnormal skin barrier and immunological factors. In particular, inflammatory factors such as nitric oxide, IL-6, IL- 1β play an important role in the invention of atopic dermatitis.

The most appropriate treatment for atopic dermatitis is an immunosuppressive agent, and steroids are frequently used as the first drug for the treatment of itodic dermatitis. However, since steroid drugs exhibit various side effects including skin atrophy during long-term use, development of safer and more effective drugs is required. In recent years, there has been an increasing trend of researches related to atopic treatments using natural extracts.

The basic research on various physiological activities was developed for the first time in the world by applying the concept of 'resveratrol', which is already reported by researchers all over the world, and 'rice', which is the main grain, The effects of metabolic diseases such as anti-cholesterol, anti-diabetic, anti-obesity, and resveratrol biosynthesized rice seeds and callus materials have been reported through preclinical experiments. In addition, studies on the skin protection effect of resveratrol have been conducted in Korea, and the effect of UVB-induced killing of keratinocyte cell lines has been reported (Park et al., 2008), and application of resveratrol biosynthesized skin to melanin- The effect of UVB irradiation on the suppression of hypercholesterolemia has been reported by controlling expression, but there has been no research on atopy of resveratrol rice.

Thus, the present inventors confirmed that the resveratrol biosynthesis rice extract and the resveratrol biosynthesis rice callus extract did not show toxicity in the keratinocyte cell line, which is a keratinocyte cell line, and decreased inflammation-related factors. As a result of treatment with the atopic animal model, The present inventors completed the present invention by confirming the efficacy of improving and treating symptoms of atopic dermatitis and itching in physical evaluation and immunological tests.

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating atopic dermatitis including resveratrol rice and resveratrol callus extract, and a cosmetic composition for improving atopic dermatitis, an external preparation for skin or a health food.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating atopic dermatitis, which contains, as an active ingredient, an extract of seed or callus of resveratrol biosynthesis rice deposited with Accession No. KCTC12529BP do.

Further, the present invention provides a cosmetic composition for improving atopic dermatitis, which contains, as an active ingredient, an extract of seed or callus of resveratrol biosynthesis rice deposited with a deposit number KCTC12529BP.

In addition, the present invention provides an external preparation for skin for improving atopic dermatitis, which contains, as an active ingredient, an extract of seed or callus of resveratrol biosynthesized rice deposited with accession number KCTC12529BP.

In addition, the present invention provides a health functional food for improving atopic dermatitis, which contains, as an active ingredient, an extract of seeds or callus of resveratrol biosynthesized rice deposited with accession number KCTC12529BP.

The present invention relates to a pharmaceutical composition for preventing or treating atopic dermatitis, a cosmetic composition for improving atopic dermatitis, a composition for preventing or treating atopic dermatitis, a composition for preventing or treating atopic dermatitis, which contains, as an active ingredient, an extract of resveratrol biosynthesis rice or callus thereof deposited under accession number KCTC12529BP Wherein the extract of the resveratrol biosynthesis rice or the callus thereof does not exhibit cytotoxicity and improves the symptoms of atopic dermatitis in mice that have induced atopic dermatitis and the inflammatory cytokine IL-6 or IL-1β It was confirmed that the expression level was decreased. In addition, the resveratrol biosynthesis rice or its callus extract of the present invention showed a superior atopy treatment effect than the resveratrol compound.

1 is a schematic diagram showing an experimental schedule using an atopic mouse model.
FIG. 2 is a graph showing cell viability after treatment of resveratrol biosynthesis rice, general rice, resveratrol biosynthesis rice callus or extract of normal rice callus, or resveratrol compound on keratinocytic cell line HaCaT cell.
FIG. 3 is a graph showing changes in secretion of IL-6, which is a major inflammatory factor, after treatment of resveratrol biosynthesis rice, general rice, resveratrol biosynthetic rice callus or extract of normal rice callus or resveratrol compound to keratinocyte HaCaT cell .
FIG. 4 is a graph showing changes in the amount of IL-β secretion, which is a major inflammatory factor, after treatment of resveratrol biosynthesis rice, general rice, resveratrol biosynthesis rice callus or extract of normal rice callus, or resveratrol compound on the keratinocyte HaCaT cell.
FIG. 5 is a graph showing the mitigation effect of atopic dermatitis through the treatment of the extract of the present invention with NC / Nga mice in which atopy is induced by 1-chloro-2,4-dinitrobenzene (DNCB)
Figure 5a: Dexamethasone, resveratrol biosynthetic rice callus extract or resveratrol compound to confirm clinical changes in skin such as mice;
Figure 5b: Clinical changes in skin such as mice after treatment with dexamethasone, resveratrol biosynthesis rice, ordinary rice or common rice callus extract;
Figure 5c: Changes in the dermatitis index of skin after treatment with resveratrol biosynthesis rice, ordinary rice, resveratrol biosynthesis rice callus or extracts of common rice callus, or resveratrol compound;
Normal: DNCB untreated group;
Control: DNCB 0.4% treated group;
Positive: treated with 0.4% DNCB, treated with 0.1% dexamethasone;
R1: treated with 0.4% of DNCB and 1% of normal rice extract;
R1 ': treated with 0.4% DNCB and 2.5% of normal rice extract;
R2: treated with 0.4% DNCB and 1% resveratrol biosynthesis rice extract;
R2 ': treated with DNCB 0.4%, treated with 2.5% resveratrol biosynthesis rice extract;
R3: treated with 0.4% of DNCB and 1% of normal rice callus extract;
R3 ': treated with 0.4% DNCB, 2.5% treated with normal rice callus extract;
R4: treated with 0.4% DNCB and treated with 1% resveratrol biosynthesis rice callus extract;
R4 ': treated with 0.4% DNCB and treated with 2.5% resveratrol biosynthesis rice callus extract;
R5: group treated with 1% resveratrol compound after treatment with 0.4% DNCB; And
R5 ': treated with 0.4% DNCB and 2.5% resveratrol compound.
FIG. 6 shows changes in transepidermal water loss (TEWL) of mice in NC / Nga mice treated with DNCB for 5 weeks:
Normal: DNCB untreated group; And
Control, Positive, r1 to r5 and r1 'to r5': DNCB processing group.
FIG. 7 shows changes in the skin hydration of the mouse for 5 weeks in which NC / Nga mice were treated with DNCB;
Normal: DNCB untreated group; And
Control, Positive, r1 to r5 and r1 'to r5': DNCB processing group.
Figure 8 shows the change in pH of the mice for 5 weeks in NC / Nga mice treated with DNCB:
Normal: DNCB untreated group; And
Control, Positive, r1 to r5 and r1 'to r5': DNCB processing group.
Figure 9 shows the change in erythema of the mouse for 5 weeks in NC / Nga mice treated with DNCB:
Normal: DNCB untreated group; And
Control, Positive, r1 to r5 and r1 'to r5': DNCB processing group.
FIG. 10 shows the effect of inhibiting transepidermal water loss (TEWL) through treatment of the extract of the present invention with NC / Nga mice in which atopy was induced by treatment with DNCB for 5 weeks:
FIG. 11 is a graph showing the effect of increasing the skin hydration by treatment of the extract of the present invention with NC / Nga mice in which atopy was induced by treatment with DNCB for 5 weeks:
FIG. 12 is a graph showing changes in pH of NC / Nga mice treated with DNCB for 5 weeks by treatment with the extract of the present invention.
FIG. 13 shows changes in epidermal erythema through treatment of the extract of the present invention with NC / Nga mice in which atopy was induced by treatment with DNCB for 5 weeks:
FIG. 14 is a graph showing changes in scratching time through treatment of the extract of the present invention with NC / Nga mice in which atopy has been induced by treatment with DNCB for 5 weeks.
FIG. 15 is a view showing immunohistochemical staining of the expression level of the inflammatory cytokine factor IL-6 through the treatment of the extract of the present invention with NC / Nga mice in which atopy has been induced by treatment with DNCB.
FIG. 16 is a graph showing changes in blood IgE content by treatment with the extract of the present invention in NC / Nga mice in which atopy has been induced by treatment with DNCB.
FIG. 17 shows the expression of IL-6 through the treatment of the extract of the present invention in an artificial skin model:
Figure 17a: The expression level of the inflammatory cytokine factor IL-6 is confirmed by immunohistochemical staining;
Figure 17b: Confirmation of secretion of inflammatory cytokine factor IL-6 to control;
Normal: DNCB untreated group;
Control: DNCB 0.4% treated group;
Positive 0.1%: treated with DNCB 0.4%, treated with 0.1% dexamethasone;
Positive 1%: treated with DNCB 0.4%, treated with 1% dexamethasone;
R1: treated with 0.4% of DNCB and 1% of normal rice extract;
R1 ': treated with 0.4% DNCB and 2.5% of normal rice extract;
R2: treated with 0.4% DNCB and 1% resveratrol biosynthesis rice extract;
R2 ': treated with DNCB 0.4%, treated with 2.5% resveratrol biosynthesis rice extract;
R5: group treated with 1% resveratrol compound after treatment with 0.4% DNCB; And
R5 ': treated with 0.4% DNCB and 2.5% resveratrol compound.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for preventing or treating atopic dermatitis, which comprises, as an active ingredient, an extract of seed or callus of resveratrol biosynthesis rice deposited with Accession No. KCTC12529BP.

The term " rice " of the present invention is the main food resource of more than half of the world's population, especially crops grown in Asia as edible crops. It is said to be cultivated as grains among the plants in the forests. There are 20 to 30 kinds of wild species in rice, but only two species are known to be Oryza sativa and Oryza glabemima. In the present invention, rice may be a natural type rice or a transformed rice, and preferably a transformed rice of Dongjinbyeon or Dongjinbyeon which is developed and supplied in Korea.

The seed may be brown rice or white rice. The term " rice " of the present invention means seeds of rice. This includes all states, whether or not they are seeds, whether or not they are untreated, semi-fixed, or frozen

The "Rice callus" mentioned above is obtained by separating the tissue of rice at a distance of about 2 to 3 mm and culturing it in a nutrient-rich test tube to continue cell division to form a cell mass (callus) A cell mass, originally an oily tissue that appears when a plant is injured, can be obtained by culturing plant tissue or cells in a medium containing plant hormones. In particular, the cells cultured in a fixed medium such as agar are called calli, and do not include cells in suspension culture.

The extract of resveratrol biosynthesis rice or callus thereof deposited with the accession number KCTC12529BP is preferably, but not limited to, produced by a method comprising the following steps:

1) extracting the resveratrol biosynthesis rice deposited with the deposit number KCTC12529BP by adding an extraction solvent;

2) filtering the extract of step 1); And

3) Concentrating the filtrate obtained in step 2) under reduced pressure and drying to prepare a resveratrol biosynthesis rice extract.

In the above method, the resveratrol biosynthesis rice of step 1) can be used without limitation such as cultivated or commercially available rice.

In the above method, it is preferable to use water, an alcohol or a mixture thereof in the extraction solvent of the step 1). As the alcohol, a C ₁ to C ₄ lower alcohol is preferably used, and as the lower alcohol, ethanol, methanol or propyl alcohol is preferably used. As the extraction method, it is preferable to use shaking extraction, Soxhlet extraction or reflux extraction, but it is not limited thereto. The extraction solvent is preferably added by 1 to 10 times the amount of the dried rice or callus, more preferably by 2 to 3 times the extraction. The extraction temperature is preferably 20 占 폚 to 100 占 폚, more preferably 20 占 폚 to 40 占 폚, and most preferably room temperature, but is not limited thereto. The extraction time is preferably 10 to 48 hours, more preferably 15 to 30 hours, most preferably 24 hours, but is not limited thereto. In addition, the extraction number is preferably 1 to 5 times, more preferably 3 to 4 times, and most preferably, 3 times, but not limited thereto.

An ultrasonic extraction step may be added to the extract of step 2). The ultrasonic extraction time may be 10 minutes to 5 hours and may be 30 minutes to 2 hours.

In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in step 3), but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.

The composition reduces trans epidermal water loss (TEWL), inhibits protein expression of IL-6 or IL-1β, and decreases the amount of IgE (Immunoglobulin E) in the serum.

In a specific embodiment of the present invention, we obtained resveratrol biosynthesis rice (Accession No. KCTC12529BP) and resveratrol biosynthetic rice callus from RDA, pulverized and then extracted twice with 80% methanol for 4 hours. Ultrasonic extraction was performed for 1 hour and concentrated filtration was used to make freeze-dried powder. The resveratrol content of the resveratrol biosynthesis rice extract, resveratrol biosynthesis rice callus extract, general rice extract, and general rice callus extract was found to be 0.433 μg resveratrol in 2.5% of resveratrol biosynthesis rice extract, 2.5% resveratrol biosynthesis rice callus extract, 0.008 [mu] g of resveratrol was present (see Table 1).

The present inventors confirmed that there was no cytotoxicity due to the extract of 100 μg / ml of the extract (see FIG. 2), and confirmed the decrease in the secretion amount of IL-6 and IL-1β depending on the extract treatment concentration (see FIGS. 3 and 4) .

In addition, the present inventors have found that treating atopy-induced mice with resveratrol rice extract (R2 and R2 ') or resveratrol rice callus extract (R4 and R4') resulted in symptom relief in five evaluation items indicating the degree of severity of atopic dermatitis (See Figs. 5A to 5C). In addition, the present inventors confirmed that the skin moisture loss was greatly reduced by treatment with 2.5% resveratrol rice extract (R2 ') in atopic-induced mice (see FIG. 10), and skin moisture increased (see FIG. 11) (See Fig. 13). In addition, it was confirmed that the number of scratches in 1% and 2.5% resveratrol biosynthesis rice callus extract treated groups (R4 and R4 ') was greatly reduced (see FIG. 14). In addition, IL-6 expression was decreased in the group treated with 1% resveratrol rice extract (R2), resveratrol 1% (R5), resveratrol rice callus extract 1% (R4), and normal rice callus extract 2.5% (R3 ' (See FIG. 15), and it was confirmed that the amount of IgE production was significantly decreased in the group treated with 2.5% concentration of resveratrol rice extract (see FIG. 16).

The present inventors also measured the amount of IL-6 expression in the experimental group (R5) treated with a resveratrol compound at a concentration of 1% and the experimental group (R2 and R2 ') treated with a resveratrol biosynthesis rice extract at a concentration of 1% or 2.5% (See Figs. 17A and 17B).

Accordingly, the resveratrol biosynthesis rice extract or resveratrol biosynthesis rice callus extract of the present invention is a natural extract and has few side effects and exhibits an atopy treatment effect superior to the resveratrol compound, and thus can be used as a pharmaceutical composition for preventing or treating atopic dermatitis.

The composition of the present invention may further comprise a pharmaceutically acceptable additive, wherein pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose Starch glycolate, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, calcium stearate, , White sugar, dextrose, sorbitol and talc. 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 are not limited thereto.

That is, the composition of the present invention can be administered in various formulations of oral and parenteral administration at the time of actual clinical administration. In the case of formulation, a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, . ≪ / RTI > Solid formulations 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 may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions and syrups, and various excipients such as wetting agents, sweetening agents, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used . Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the non-aqueous solvent and the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The composition of the present invention may be administered orally or parenterally in accordance with the intended method, and may be administered orally, parenterally or intraperitoneally, rectally, subcutaneously, intravenously, intramuscularly, . The dosage varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity.

The dosage of the composition of the present invention varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate and severity of disease, The dose is 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg, and can be administered 1 to 6 times a day.

The compositions of the present invention may be used alone or in combination with other therapies, such as procedures, radiotherapy, hormone therapy, chemotherapy, and methods using biological response modifiers, for the improvement of atopic dermatitis.

Further, the present invention provides a cosmetic composition for improving atopic dermatitis, which comprises, as an active ingredient, an extract of seed or callus of resveratrol biosynthesis rice deposited with Accession No. KCTC12529BP.

The composition reduces trans epidermal water loss (TEWL), inhibits protein expression of IL-6 or IL-1β, and decreases the amount of IgE (Immunoglobulin E) in the serum.

The cosmetic composition may be in the form of a skin, a lotion, a cream, an essence, an emulsion, a gel, a lipstick, a cleansing foam, a cleansing cream, a cleansing water, a spray, a shampoo, a rinse, a treatment, a body cleanser, , A foundation, a two-way cake, and a makeup base. However, the present invention is not limited thereto.

In a specific example of the present invention, the present inventors confirmed that there was no cytotoxicity due to the extract (see Fig. 2), and confirmed that IL-6 and IL-1? 4).

In addition, the present inventors have found that treating atopy-induced mice with resveratrol rice extract (R2 and R2 ') or resveratrol rice callus extract (R4 and R4') resulted in symptom relief in five evaluation items indicating the degree of severity of atopic dermatitis (See Figs. 5A to 5C). In addition, the present inventors confirmed that the skin moisture loss was greatly reduced by treatment with 2.5% resveratrol rice extract (R2 ') in atopic-induced mice (see FIG. 10), and skin moisture increased (see FIG. 11) (See Fig. 13). In addition, it was confirmed that the number of scratches in 1% and 2.5% resveratrol biosynthesis rice callus extract treated groups (R4 and R4 ') was greatly reduced (see FIG. 14). In addition, IL-6 expression was decreased in the group treated with 1% resveratrol rice extract (R2), resveratrol 1% (R5), resveratrol rice callus extract 1% (R4), and normal rice callus extract 2.5% (R3 ' (See FIG. 15), and it was confirmed that the amount of IgE production was significantly decreased in the group treated with 2.5% concentration of resveratrol rice extract (see FIG. 16).

The present inventors also measured the amount of IL-6 expression in the experimental group (R5) treated with a resveratrol compound at a concentration of 1% and the experimental group (R2 and R2 ') treated with a resveratrol biosynthesis rice extract at a concentration of 1% or 2.5% (See Figs. 17A and 17B).

Therefore, the resveratrol biosynthesis rice extract or the resveratrol biosynthesized rice callus extract of the present invention is a natural extract and has less side effects and exhibits an atopy treatment effect superior to the resveratrol compound, and thus can be used as a cosmetic composition for improving atopic dermatitis.

When the extract of the present invention is used as a cosmetic composition, for example, a solution, a gel, a solid or a paste anhydrous product, an emulsion obtained by dispersing an oil phase in a water phase, a suspension, a microemulsion, a microcapsule, , In the form of a non-ionic follicular dispersion, in the form of creams, skins, lotions, powders, ointments, sprays or conical sticks. It can also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

The cosmetic composition may further contain, in addition to the extract of the present invention, a lipid, an organic solvent, a solubilizer, a thickener and a gelling agent, a softener, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, As used herein means any emulsion or emulsion which is commonly used in emulsifying or emulsifying agents, fillers, sequestering and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics And may contain adjuvants conventionally used in the cosmetics field, such as other ingredients.

In addition, the present invention provides an external preparation for skin for improving atopic dermatitis, which comprises, as an active ingredient, an extract of seed or callus of resveratrol biosynthesized rice deposited with a deposit number KCTC12529BP.

The external preparation for skin reduces trans epidermal water loss (TEWL), inhibits protein expression of IL-6 or IL-1β, and decreases the amount of IgE (Immunoglobulin E) in serum.

The external preparation for skin may be any one selected from the group consisting of an ointment, a cream, a lotion, a liquid, a dressing, a patch, a foaming agent, a tape agent, an aerosol agent, an external agent and a spraying agent.

In a specific example of the present invention, the present inventors confirmed that there was no cytotoxicity due to the extract (see Fig. 2), and confirmed that IL-6 and IL-1? 4).

In addition, the present inventors have found that treating atopy-induced mice with resveratrol rice extract (R2 and R2 ') or resveratrol rice callus extract (R4 and R4') resulted in symptom relief in five evaluation items indicating the degree of severity of atopic dermatitis (See Figs. 5A to 5C). In addition, the present inventors confirmed that the skin moisture loss was greatly reduced by treatment with 2.5% resveratrol rice extract (R2 ') in atopic-induced mice (see FIG. 10), and skin moisture increased (see FIG. 11) (See Fig. 13). In addition, it was confirmed that the number of scratches in 1% and 2.5% resveratrol biosynthesis rice callus extract treated groups (R4 and R4 ') was greatly reduced (see FIG. 14). In addition, IL-6 expression was decreased in the group treated with 1% resveratrol rice extract (R2), resveratrol 1% (R5), resveratrol rice callus extract 1% (R4), and normal rice callus extract 2.5% (R3 ' (See FIG. 15), and it was confirmed that the amount of IgE production was significantly decreased in the group treated with 2.5% concentration of resveratrol rice extract (see FIG. 16).

The present inventors also measured the amount of IL-6 expression in the experimental group (R5) treated with a resveratrol compound at a concentration of 1% and the experimental group (R2 and R2 ') treated with a resveratrol biosynthesis rice extract at a concentration of 1% or 2.5% (See Figs. 17A and 17B).

Therefore, the resveratrol biosynthesis rice extract or the resveratrol biosynthesized rice callus extract of the present invention is a natural extract and has little side effects and exhibits an atopy treatment effect superior to the resveratrol compound, and thus can be used as an external preparation for skin for improving atopic dermatitis.

When the extract of the present invention is used as an external preparation for skin, it may further contain at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickening agents and gelling agents, softening agents, antioxidants, suspending agents, stabilizers, foaming agents, For example, water, ionic or nonionic emulsifiers, fillers, sequestering agents and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or external preparations for skin And any other ingredients used, such as those commonly used in the field of dermatology. In addition, the components can be introduced in amounts commonly used in the dermatology field.

The dose of the extract to the external preparation for skin is 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg, but is not limited thereto. The dose may vary depending on the weight, age, sex, health condition, administration period, elimination rate, severity of disease, etc. of the particular patient.

In addition, the present invention provides a health functional food for improving atopic dermatitis, which contains, as an active ingredient, an extract of seed or callus of resveratrol biosynthesis rice deposited with accession number KCTC12529BP.

The health functional food reduces trans epidermal water loss (TEWL), inhibits protein expression of IL-6 or IL-1β, and decreases the amount of IgE (Immunoglobulin E) in the serum.

In a specific example of the present invention, the present inventors confirmed that there was no cytotoxicity due to the extract (see Fig. 2), and confirmed that IL-6 and IL-1? 4).

In addition, the present inventors have found that treating atopy-induced mice with resveratrol rice extract (R2 and R2 ') or resveratrol rice callus extract (R4 and R4') resulted in symptom relief in five evaluation items indicating the degree of severity of atopic dermatitis (See Figs. 5A to 5C). In addition, the present inventors confirmed that the skin moisture loss was greatly reduced by treatment with 2.5% resveratrol rice extract (R2 ') in atopic-induced mice (see FIG. 10), and skin moisture increased (see FIG. 11) (See Fig. 13). In addition, it was confirmed that the number of scratches in 1% and 2.5% resveratrol biosynthesis rice callus extract treated groups (R4 and R4 ') was greatly reduced (see FIG. 14). In addition, IL-6 expression was decreased in the group treated with 1% resveratrol rice extract (R2), resveratrol 1% (R5), resveratrol rice callus extract 1% (R4), and normal rice callus extract 2.5% (R3 ' (See FIG. 15), and it was confirmed that the amount of IgE production was significantly decreased in the group treated with 2.5% concentration of resveratrol rice extract (see FIG. 16).

The present inventors also measured the amount of IL-6 expression in the experimental group (R5) treated with a resveratrol compound at a concentration of 1% and the experimental group (R2 and R2 ') treated with a resveratrol biosynthesis rice extract at a concentration of 1% or 2.5% (See Figs. 17A and 17B).

Therefore, the resveratrol biosynthesis rice extract or the resveratrol biosynthesis rice callus extract of the present invention is a natural extract and has few side effects and exhibits an atopic treatment effect superior to the resveratrol compound, so that it can be used as a health functional food for improving atopic dermatitis.

The extract according to the present invention can be directly added to the food or used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed may be appropriately determined depending on the purpose of use. In general, the amount of the extract may be 0.01 to 15 parts by weight of the whole food, and the health drink composition may be added in an amount of 0.02 to 5 g Can be added in a proportion of 0.3 to 1 g. However, in the case of long-term intake intended for health and hygiene purposes 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.

There is no particular limitation on the kind of the food. Examples of the food include functional mixed tea, dairy product, various kinds of soup, beverage, and vitamin complex, and include health functional foods in a conventional sense.

The health functional beverage composition of the present invention has no particular limitation on the other ingredients other than the above-mentioned extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above .

In addition to the above-mentioned foods, the food of the present invention may contain flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and heavies (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the extract 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. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the extract of the present invention.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples.

It should be noted, however, that the following examples and experimental examples are illustrative of the present invention only, and the content of the present invention is not limited by the examples and the experimental examples.

Resveratrol  Biosynthetic rice extract and Resveratrol  Biosynthetic rice Callus  Preparation of extract

Resveratrol biosynthesis rice (Accession No. KCTC12529BP) and resveratrol biosynthesis rice callus were obtained from the Rural Development Administration. After pulverizing each of the above samples, 500 g of each sample was immersed in 5 L of 80% methanol and extracted for 24 hours. The extraction was repeated twice, followed by sonication for 1 hour. The extract was concentrated and filtered to prepare a lyophilized powder.

< Comparative Example  1>

Common rice extract and ordinary rice Callus  Preparation of extract

Normal rice and ordinary rice callus were obtained from the Rural Development Administration. After pulverizing each of the above samples, 500 g of each sample was immersed in 5 L of 80% methanol and extracted for 24 hours. The extraction was repeated twice, followed by sonication for 1 hour. The extract was concentrated and filtered to prepare a lyophilized powder, which was used as a comparative example.

Resveratrol  Comparison of contents

Resveratrol contents of the resveratrol biosynthesis rice extract, resveratrol biosynthesis rice callus extract, general rice extract and common rice callus extract of Example 1 and Comparative Example 1 were measured, and the results are shown in Table 1 below.

Sample (sample) The treated amount (μl) Resveratrol Content (unit) Resveratrol Biosynthesis Rice Extract 2.5% (200 [mu] L) 0.433 / / 200 ㎕ Resveratrol Biosynthesis Rice Callus Extract 2.5% (200 [mu] L) 0.008 / / 200 ㎕ Common rice extract 2.5% (200 [mu] L) 0 Common rice callus extract 2.5% (200 [mu] L) 0 Resveratrol compound (Sigma / R5010) 2.5% (200 [mu] L) 5 / / 200 ㎕

Preparation of atopic dermatitis model

In order to confirm the inhibitory effect of resveratrol rice and resveratrol rice callus extract on atopic dermatitis, 35 male four week old NC / Nga mice, well known as a model of spontaneous atopic dermatitis induction, were purchased and purified in a laboratory environment for 7 weeks, Were divided into 5 groups. Mice were fed at constant temperature (23 ± 3 ° C), humidity (55 ± 15%) and light (7 - 19 hours) and all feed and water were taken freely. Breeding and storage of animals were based on the guidelines of the Animal Laboratory of Cancer Diabetes Center of Gachon University.

As a test group, 1-chloro-2,4-dinitrobenzene (DNCB, Aldrich, USA) was administered to NC / Nga mice to induce human-like atopic dermatitis as a normal group. For the induction of atopic dermatitis, 5-week-old NC / Nga mice were cleaned from the back area and allowed to stand for 24 hours to heal micro wounds. 200 μl of a 1% DNCB solution (acetone: olive oil = 3: 1) was applied to the back area to sensitize allergen, followed by a second application of 1% DNCB solution 3 days after the first application, 200 μl of 0.4% DNCB solution was applied three times to the back region to induce atopic dermatitis. After the DNCB solution was treated for 5 weeks, if dermatitis was sufficiently induced to peel off all of the back skin and new scratching was formed in the area, if the scratching behavior became worse, DNCB 0.4% solution and resveratrol rice extract of Example 1 Resveratrol rice callus extract, general rice extract of Comparative Example 1 or common rice callus extract or resveratrol single compound (Sigma / R5010) were treated three times a week for 6 weeks, treated with dexamethasone as a positive control, Evaluation, sensory evaluation, and physical evaluation experiment (Fig. 1).

< Experimental Example  1>

Identification of cytotoxicity and inflammatory factors

The cytotoxicity of the extracts of Example 1 and Comparative Example 1 was confirmed and the production of IL-6 and IL-1β, which are anti-allergic inflammatory cytokines according to environmental factor stimulation, was measured in human keratinocyte keratinocytes The following experiment was conducted.

Specifically, in order to confirm cytotoxicity, keratinocyte cell line (HeCaT cell) was divided into 96 wells at ⁵ × 10 ⁵ / ml and stabilized for 24 hours. Resveratrol rice extract or resveratrol rice callus extract of Example 1, Comparative Example 1 Or normal rice callus extract at a concentration of 1, 10, 50 or 100 μg / ml, and a resveratrol compound alone at a concentration of 1, 10, 50 or 100 μM. After incubation for 24 hours in a CO ₂ incubator, the culture medium was discarded, washed with PBS, treated with MTT reagent (0.1 μg / ml) and incubated in a CO ₂ incubator for 4 hours. After that, MTT reagent was discarded and treated with DMSO Incubated in a CO ₂ incubator for 30 min and analyzed by a microspectrophotometer at a wavelength of 540 nm.

In order to analyze the production of inflammatory factors IL-6 and IL-1β, keratinocyte cell line (HeCaT cell) was divided into 48 wells at 2 × 10 ⁵ / ml and stabilized for 24 hours. The resveratrol rice extract of Example 1 Resveratrol rice callus extract, ordinary rice extract of Comparative Example 1 or common rice callus extract was treated at a concentration of 1, 10, 50 or 100 占 퐂 / ml, a resveratrol single compound at a concentration of 1, 10, 50 or 100 占,, One hour later, the cells were activated with TNF-α (10 ng / mL) + IFN-γ (10 ng / mL) and cultured in a CO ₂ incubator for 18 hours. The culture broth was diluted by concentration and then subjected to ELISA. And analyzed with a microspectrophotometer at a wavelength of 450 nm.

As a result, as shown in Fig. 2, it was confirmed that no cytotoxicity was shown even at a concentration of 100 占 퐂 / ml for all the extracts (Fig. 2).

As shown in FIG. 3 and FIG. 4, IL-6 and IL-1β secretion decreased significantly depending on extract treatment concentration. In particular, treatment with resveratrol rice callus extract showed the greatest decrease (Figs. 3 and 4).

< Experimental Example  2>

Clinical symptom relief effect of atopy-induced mice

The following experiment was conducted to confirm clinical symptom relief effect through the treatment of the extract of Example 1 in the back region of atopic-induced mice.

Specifically, resveratrol rice extract or resveratrol rice callus extract of Example 1, general rice extract of Comparative Example 1, or common rice callus extracts were inoculated in a ratio of 1% or more per day for 6 weeks to mice of atopy induced by 5-week DNCB treatment for 6 weeks. Or 2.5%, and clinical visual evaluation was carried out at the last 6 weeks. As a positive control, dexamethasone 0.1% was treated. The degree of severity of atopic dermatitis was evaluated by 5 items; There are no symptoms in erythema, pruritus & dry skin, edema & excoriation, erosion, lichenification, no symptoms, 1 symptom, (2 points), and severity (3 points), and then summed up the score of 5 items, indicating a minimum score of 0 (no symptom) to a maximum of 15 (symptom of all items was severe).

As a result, as shown in FIG. 5, the group treated with the resveratrol rice extract (R2 and R2 ') or the resveratrol rice callus extract (R4 and R4') showed significant symptom relief as compared with the positive control group 5a and 5b). In addition, clinical symptom scores indicating severity of atopic dermatitis showed significant symptom relief effects in the group treated with resveratrol rice callus extract (R4 and R4 ') (FIG. 5C).

< Experimental Example  3>

Identification of physical symptom relief effect of atopy-induced mice

The skin moisture loss, skin moisture content and epidermal erythema degree were measured in order to confirm the effect of alleviating the physical symptom through the treatment of the extract of Example 1 with the atopy-induced mouse.

Specifically, resveratrol rice extract or resveratrol rice callus extract of Example 1, general rice extract of Comparative Example 1, or general rice callus extract of the above-mentioned Example 1 was cultured three times a week for 1 week in a mouse on which atopy was induced by DNCB treatment for 5 weeks, or 2.5% of the epidermal water loss (transepidermal water loss, TEWL), epidermal water content (hydration) and skin redness (erythema) was measured after application at a concentration using a ^{Dermalab ® Combosystem (CortecTechnologyAps, Hudsund,} Denmark) of the device. The establishment of the atopy model due to the 5-week DNCB treatment was confirmed by decreasing the skin moisture loss and the skin moisture content, increasing the epidermal erythema and increasing the skin pH (FIGS. 6 to 9).

As shown in Figs. 10 to 13, it was confirmed that the skin moisture loss was reduced due to the treatment with R2 (2.5%) of resveratrol rice extract (Fig. 10) and the skin moisture was increased 11). In addition, it was confirmed that the increased pH was decreased at the atopic dermatitis lesion site, and the degree of erythema was reduced by treating with 2.5% of resveratrol rice extract (Fig. 13).

< Experimental Example  4>

Confirming the effect of atopy-induced mice on behavioral symptoms

The degree of self-scratching of the mice was measured in order to confirm the effect of mitigating the behavioral symptoms by treating the atopy-induced mouse with the extract of Example 1 above.

Specifically, the resveratrol rice extract or resveratrol rice callus extract of Example 1, general rice extract of Comparative Example 1, or general rice callus extract of the above Example 1 was administered once a week for two weeks on the back of the atopic induced mice induced by the DNCB treatment for 5 weeks % Or 2.5%, and the number of scratching bahavior was measured periodically (20 minutes) after the application.

As a result, as shown in Fig. 14, it was confirmed that the number of scratches was greatly reduced in the groups treated with resveratrol biosynthesis rice callus extract (R4 and R4 ') of 1% and 2.5% (Fig. 14).

< Experimental Example  5>

Reduction of Inflammatory Cytokine Expression in Atopic-induced Mouse Skin Tissue

The following experiment was conducted to analyze the effect of the extract of the present invention on the expression of IL-6, an inflammation-related main protein, in atopic skin mice.

Specifically, the resveratrol rice extract or the resveratrol rice callus extract of Example 1, the common rice extract of Comparative Example 1, or the common rice callus extract of Example 1 at a rate of 1% or 2.5% three times a week for 6 weeks on the atopy- After the application, the skin tissue of the mouse was taken for the last 6 weeks, fixed in 4% formaldehyde for 24 hours, rinsed in water for 6 hours, and dehydrated through ethanol to xylene to prepare a paraffin block . The paraffin block was cut to a thickness of 10 ㎛ using a microtome, and paraffin was removed to perform hematoxylin-eosin staining. The expression level of IL-6, a major protein involved in inflammation, Were measured.

As a result, as shown in Fig. 15, a group treated with 1% (R2) of resveratrol rice extract, 1% (R5) of resveratrol, 1% (R4) of resveratrol rice callus extract, and 2.5% The expression of IL-6 was reduced compared to the control group (DNCB alone) (Fig. 15).

< Experimental Example  6>

Atopy-induced mouse blood IgE  Confirm the effect of reducing the content

The following experiment was conducted to analyze the effect of the extract of the present invention on the blood IgE content of atopy-induced mice.

Specifically, the resveratrol rice extract or the resveratrol rice callus extract of Example 1, the common rice extract of Comparative Example 1, or the common rice callus extract of Example 1 at a rate of 1% or 2.5% three times a week for 6 weeks on the atopy- After the application, 1 ml of eyeball blood was collected from the mouse for the last 6 weeks and the serum was separated. The amount of IgE produced in the serum was measured using an ELISA kit (ebioscience, USA).

As a result, as shown in FIG. 16, it was confirmed that the amount of IgE production was significantly reduced in the group treated with 2.5% resveratrol rice extract (R2 ') compared to the control (control) (FIG. 16).

< Experimental Example  7>

Reduction of Inflammatory Cytokine Expression in Artificial Skin Model

The following experiment was conducted to confirm the effect of the extract of the present invention on the improvement of skin inflammation in the artificial skin model.

Specifically, Keraskin (Biosolution, Korea), a human epidermis artificial skin model, was purchased and induced atopic dermatitis with 1-chloro-2,4-dinitrobenzene (DNCB, Aldrich, USA) Was transfected into 24 wells and stabilized for one day and sensitized with inflammatory cytokines (TNF-α (10 ng / mL) + IFN-γ (10 ng / mL)). 20 μl of the resveratrol rice extract of Example 1 or the general rice extract of Comparative Example 1 and 0.1% or 1% of resveratrol compound were dissolved in a solvent (polyphenol glycol: PBS = 1: 1) to treat the artificial skin surface. As a positive control, dexamethasone 0.1% or 1% was treated.

IL-6 and IL-1β, the major inflammatory cytokines, were measured in conditoned medium after 1 week treatment with artificial skin model (R & D system, USA or Takara, Japan). For tissue staining, artificial skin tissue was fixed in 4% formaldehyde for 24 hours, rinsed in water for 6 hours, and dehydrated through ethanol to xylene to prepare paraffin blocks. The paraffin block was cut to a thickness of 10 탆 using a microtome, and then subjected to a paraffin removal process to perform ematoxylin-eosin staining. The expression of major proteins involved in inflammation was then measured.

As a result, as shown in Fig. 17, the expression level of IL-6 was the lowest in the experimental group (R5) treated with 1% resveratrol compound and the experimental group (R2 and R2 ' ) Of IL-6 was also decreased compared to the control (Fig. 17B). In addition, IL-6 expression levels were measured by tissue staining. The results were as follows: Experimental group (R2 and R2 ') treated with 1% or 2.5% resveratrol biosynthetic rice extract and 1% or 2.5% (R5 and R5') resveratrol compound (Fig. 17 (a)).

Claims (15)

A pharmaceutical composition for preventing or treating atopic dermatitis, which comprises an extract of a resveratrol biosynthetic rice seed or callus deposited with a deposit number KCTC12529BP as an active ingredient, wherein the extract is water, C ₁ to C ₄ A lower alcohol, or a mixture thereof. The pharmaceutical composition for preventing or treating atopic dermatitis according to claim 1,
The pharmaceutical composition for preventing or treating atopic dermatitis according to claim 1, wherein the seed is brown rice or white rice.
delete The pharmaceutical composition according to claim 1, wherein the lower alcohol is ethanol or methanol.
The pharmaceutical composition for preventing or treating atopic dermatitis according to claim 1, wherein the composition reduces trans epidermal water loss (TEWL).
The pharmaceutical composition for preventing or treating atopic dermatitis according to claim 1, wherein the composition inhibits protein expression of IL-6 or IL-1 ?.
The pharmaceutical composition for preventing or treating atopic dermatitis according to claim 1, wherein the composition reduces an amount of IgE (Immunoglobulin E) in serum.
A cosmetic composition for improving atopic dermatitis comprising an extract of a seed or callus of resveratrol biosynthesis rice deposited with a deposit number KCTC12529BP as an active ingredient, wherein the extract is water, a C ₁ to C ₄ lower alcohol or Wherein the composition is extracted with a mixture thereof.
The cosmetic composition for improving atopic dermatitis according to claim 8, wherein the composition reduces transepidermal water loss (TEWL).
The cosmetic composition for improving atopic dermatitis according to claim 8, wherein the composition inhibits protein expression of IL-6 or IL-1 ?.
The cosmetic composition for improving atopic dermatitis according to claim 8, wherein the composition reduces the amount of IgE (Immunoglobulin E) in serum.
The cosmetic composition according to claim 8, wherein the cosmetic composition is at least one selected from the group consisting of skin, lotion, cream, essence, emulsion, gel, lipstick, cleansing foam, cleansing cream, cleansing water, spray, shampoo, Wherein the cosmetic composition is any one selected from the group consisting of a make-up base, a face powder, a compact, a foundation, a two-way cake, and a makeup base.
An extract for the treatment of atopic dermatitis, which comprises an extract of a seed or callus of resveratrol biosynthesis rice deposited with a deposit number KCTC12529BP as an active ingredient, wherein the extract is water, a C ₁ to C ₄ lower alcohol or Wherein the composition is extracted with a mixture thereof.
14. The composition according to claim 13, wherein the external preparation for skin is at least one selected from the group consisting of an ointment, a cream, a lotion, a liquid, a dressing, a patch, a foaming agent, a tape agent, an aerosol agent, Skin external preparation for improving dermatitis.
A health functional food for improving atopic dermatitis comprising an extract of seeds or callus of resveratrol biosynthesis rice deposited with a deposit number KCTC12529BP as an active ingredient, wherein the extract is water, a C ₁ to C ₄ lower alcohol Or a mixture thereof. &Lt; / RTI &gt;

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