KR20200104996A - A composition for preventing or treating inflammation comprising an extract of Ilex cornuta and uses thereof - Google Patents

Abstract

The present invention relates to a composition for use in preventing or treating inflammation containing an extract of Ilex cornuta; and a method for preventing, alleviating or treating inflammation of an individual. According to the extract of Ilex cornuta according to an aspect, since the extract of Ilex cornuta has excellent effects of alleviating skin inflammation or skin irritation, the extract of Ilex cornuta can be applied to various fields such as cosmetics, foods, pharmaceutical compositions and the like.

Description

A composition for preventing or treating inflammation comprising an extract of Ilex cornuta and uses thereof

It relates to a composition for use in preventing or treating inflammation comprising a holly tree extract and a method of preventing, ameliorating or treating inflammation in a subject.

Inflammation causes many types of polykaryotic leukocytes (PMNs) and immune substances to increase, and these increased cells secrete various types of proteolytic enzymes and cytokines, which are inflammatory cell products, so that they can be treated and defended. Allows. However, this action may cause detrimental damage to adjacent tissue cells and non-cellular components. Therefore, under appropriate conditions, normal function is restored after the initial state of inflammation has passed, but if the irritant that stimulates inflammation does not disappear or is made continuously, as a result, chronic inflammation occurs, leading to more serious tissue damage.

Representative types of skin troubles related to inflammation include atopy and acne, and atopy is described as follows. Skin disease called atopic dermatitis or atopic eczema is a chronic and recurrent disease that generally occurs in infants and children. Symptoms such as severe itching (pruritus) and skin eczema are worsened, relieved, and recurred due to various causes. The vicious cycle continues. The direct cause of the clinical symptoms of atopic dermatitis is that the skin defense and protective functions against allergens are weakened, or the skin immune response is hypersensitive. The first reason is that the keratinocytes are dysfunctional and the moisture-retaining function and the skin barrier function are deteriorated due to structural defects in the stratum corneum. In normal skin, bacteria and foreign substances that are difficult to penetrate penetrate the skin, causing irritation and allergic reactions easily. . The second cause is skin immunological hypersensitivity reaction, and various immunological abnormalities that occur when atopic dermatitis occurs are increased synthesis of immunoglobulin (IgE), increased immunoglobulin against various antigens, and B lymphocytes and monocytes. Increased expression of IgE receptors, increased histamine secretion by mast cells/basophils, increased delayed hypersensitivity reactions, increased eosinophils, and interleukin IL-4, IL-5 by type 2 auxiliary T lymphocytes (Th2 lymphocytes), Increased secretion of IL-13, decreased secretion of interferon INF-γ by type 1 auxiliary T lymphocytes (Th1 lymphocytes), and the like. And some of the various inflammatory cytokines that appear due to immunological hypersensitivity reactions cause dysfunction of keratinocytes, which is caused by dysfunction of the skin barrier, which induces the inflammatory reaction, and the induced inflammatory reaction is again caused by dysfunction of the skin barrier. Atopic dermatitis is sometimes exacerbated by the circulatory process that induces. On the other hand, anti-inflammatory action is a mechanism to defend against inflammation, and inflammation is a local physiological reaction that protects a living body from factors that are harmful or potentially harmful to tissues. When such an inflammatory phenomenon is caused excessively, cells and connective tissues are damaged by proteolytic enzymes, which are inflammatory cell products, and damage to the connective tissue decreases skin elasticity and promotes skin aging, so that healthy skin cannot be maintained.

On the other hand, the holly tree (Ilex cornuta) is a dicotyledonous plant belonging to the Aquifoliaceae family, and grows up to 4 ~ 5m in height. The leaves are thick elliptical hexagonal, the ends of the corners are thorns, and the red fruits ripen in September to October. It grows naturally in the south of Korea's Byeonsan Peninsula, Wando, and Jeju Island. In oriental medicine, leaves and fruits are used as medicine. The leaves have the effect of geopung and tonic, and the fruit has the effect of gangjeong, and it helps blood circulation (「Sanjacho, which is good for the body」, 2009. 11. 15., Nexus Co., Ltd.). However, no effect on skin anti-inflammatory and sensitivity improvement against holly has been reported.

One aspect provides a composition for use in preventing, improving, or treating skin inflammation or skin irritation comprising a hollywood extract as an active ingredient.

Another aspect provides a method of preventing, ameliorating or treating skin inflammation or skin irritation in an individual comprising administering to the individual an amount effective to prevent, ameliorate, or treat skin irritation or skin irritation. .

One aspect provides a composition for preventing or improving skin inflammation or skin irritation comprising an extract of Ilex cornuta as an active ingredient.

In one aspect, the hollywood extract is excellent in inhibiting the expression of inflammatory cytokines, inflammatory chemokines, and TSLP (Thymic Stromal Lymphopoietin) caused by irritation and skin irritation. Or it can be useful for mitigation.

The term “preventing, improving or treating skin inflammation” may be used interchangeably with “anti-inflammatory” and may mean an action of suppressing NO production, inflammatory cytokines, inflammatory chemokines, or TSLP expression by mitigating an immune response.

The composition is excellent in preventing, improving or treating atopic dermatitis, itching, redness, erythema, eczema, seborrheic dermatitis, psoriasis, or acne due to its anti-inflammatory effect.

The term "included as an active ingredient" means that the hollywood extract is added to the composition of the present invention to the extent that it can exhibit the effect of improving skin inflammation and alleviating irritation, and various ingredients are used as auxiliary ingredients for drug delivery and stabilization. It is meant to include being formulated in various forms by adding.

The term "improvement" can mean any action that at least reduces the severity of a parameter, such as a symptom, associated with remission or treatment of the condition.

The term "prevention" refers to any action that becomes close to a normal control group by suppressing or delaying skin inflammation or skin irritation symptoms by administration of a composition comprising the hollywood extract.

The term "treatment" refers to any action in which the skin inflammation or skin irritation symptom is improved or beneficially altered to the same or similar to that of a normal control group by administration of a composition comprising the hollywood extract.

The holly tree extract, which has the activity of preventing and improving skin inflammation or skin irritation, can be extracted from various organs of natural, hybrid, and variety plants, such as roots, stems, leaves, flowers, trunks of fruits, and peels of fruits. In addition, it can be extracted from plant tissue culture, and it is most preferable to extract it from the leaves of holly.

The term "holly tree" is a dicotyledonous plant belonging to the Aquifoliaceae family, and grows up to 4 to 5 m in height. The leaves are thick elliptical hexagonal, the ends of the corners are thorns, and the red fruits ripen in September to October. It grows naturally in the south of Korea's Byeonsan Peninsula, Wando, and Jeju Island. In oriental medicine, leaves and fruits are used as medicine. It is known that the leaves have the effect of geopung and tonic, and the fruit has the effect of gangjeong and help blood circulation. The holly tree according to one aspect may be a holly tree (Ilex x wandoensis) grown in Wando.

The hollywood extract may be a solvent extract extracted with an extraction solvent, a fraction obtained by adding a fractionation solvent to an extract prepared by extraction with an extraction solvent, or a purified product obtained through chromatography on the fraction.

The extraction solvent may be water, an organic solvent, or a mixed solvent thereof that can be used for extracting natural products. The extraction solvent may be water, an alcohol having 1 to 6 carbon atoms or a mixture thereof, such as water or ethanol.

The holly tree extract may be prepared according to a conventional method for producing a plant extract. More specifically, the preparation of the hollywood extract may be performed by adding an extraction solvent to the pulverized pulverized product of the hollywood from which impurities have been removed, followed by extraction. The extraction method using the solvent may be a cold needle extraction method, a warm needle extraction method, a reflux extraction method, or an ultrasonic grinding extraction method. The extract may be obtained by incubating holly in a solvent to extract the active ingredient from the solvent. The incubation is 4 ℃ to reflux temperature, 4 ℃ to 100 ℃, 4 ℃? To 80°C, 4°C? To 60°C, 4°C? To 50°C, 4°C? To 40°C, 4°C? To 35°C, 4°C? To 30°C, 4°C? To 25 ℃, or may be carried out at room temperature. The incubation can be carried out for a time sufficient for the active ingredient to be extracted into the solvent. The time is 1 hour to 2 weeks, 1 hour to 1 week, 1 hour to 5 days, 1 hour to 3 days, 12 hours to 5 days, 12 hours to 3 days, 12 hours to 2 days, 24 hours to 5 days , Or 24 hours to 3 days.

The composition may be one to suppress the inflammatory response by reducing the expression of TSLP, inflammatory cytokines, and inflammatory chemokines.

The composition may be one to alleviate skin irritation by reducing the expression of TSLP, inflammatory cytokines, and inflammatory chemokines increased by stimulation.

The composition according to an embodiment may contain the extract in an effective amount or as an active ingredient. The effective amount can be appropriately selected according to the individual. The severity of the disease or condition, the age, weight, health, sex of the subject, the sensitivity to the extract of the subject, the time of administration, the route of administration and the rate of excretion, the period of administration, factors including other compositions used in combination or concurrent with the composition, and others It can be determined according to factors well known in the field of physiology or medicine.

The composition for preventing, improving, or treating skin inflammation or skin irritation according to an embodiment may further include a cosmetically, food, or pharmaceutically acceptable excipient or carrier. The carrier may be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be sodium starch glycolate, anhydrous calcium monohydrogen phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

The composition may be formulated as a parenteral dosage form. The parenteral dosage form may be an injection or an external preparation for skin. The external preparation for skin may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal delivery patch, drug-containing bandage, lotion, or a combination thereof.

The above skin external preparations are ingredients commonly used in external preparations for skin such as cosmetics and pharmaceuticals, such as aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances. , Colorants, various skin nutrients, etc. can be appropriately formulated as needed.

The external preparations for the skin include metal sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid, caffeine, tannin, bellapamil, licorice extract, glavidine, and caline. Hot water extract of fruit, various herbal medicines, drugs such as tocopherol acetate, glytilithic acid, tranexamic acid and derivatives or salts thereof, vitamin C, ascorbic acid magnesium phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, Sugars such as trehalose can also be appropriately blended.

The composition according to an embodiment may be a cosmetic composition. In this case, the extract is a lotion (skin lotion), skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, essence, nutrition essence, pack , Soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, suspension, gel, powder, paste, mask pack, or sheet or aerosol composition.

In addition, it can be applied in various properties such as liquid, cream, paste, and solid. Compositions of these formulations can be prepared according to methods conventional in the art. The cosmetic composition may further contain preservatives, stabilizers, surfactants, solubilizers, moisturizers, emollients, ultraviolet absorbers, preservatives, disinfectants, antioxidants, pH adjusters, organic and inorganic pigments, fragrances, cooling agents or limiting agents. have. The blending amount of additional ingredients such as the moisturizing agent can be easily selected by a person skilled in the art within the range that does not impair the object and effect of the present invention, and the blending amount is 0.001 to 5% by weight, specifically 0.01 to 5, based on the total weight of the composition. It may be weight percent.

The composition according to one embodiment may be a food composition. In this case, it may be formulated into a conventional health functional food formulation known in the art. The food composition may be prepared in general formulations such as, for example, powders, granules, tablets, pills, capsules, suspensions, emulsions, syrups, needles, liquids, and extracts, and meat, sausages, bread, chocolate, candy, snacks , Confectionery, pizza, ramen, other noodles, gums, jelly, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and may be prepared in any health food form such as vitamin complex. For the formulation of the health food, a food pharmaceutically acceptable carrier or additive may be used, and any carrier or additive known to be usable in the art may be used to prepare a formulation to be prepared. As the additive, used in various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonated beverages It may contain a carbonation agent and the like. In addition, it may contain flesh for the production of natural fruit juice, fruit juice beverage and vegetable beverage.

These additive components may be used independently or in combination, and the proportion of the additive may be 0.001 to 5% by weight, specifically 0.01 to 5% by weight, based on the total weight of the composition.

The content of the extract in the food composition may be appropriately determined according to the purpose of use (prevention or improvement). In general, it may be contained in an amount of 0.01 to 15% by weight of the total food weight, and when prepared as a beverage, it may be contained in an amount of 0.02 to 10 g, specifically 0.3 to 1 g, based on 100 mL.

The beverage may further contain ingredients other than the extract, and may further contain various flavoring agents or natural carbohydrates that are commonly used in beverages. The natural carbohydrates include monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, etc.), polysaccharides (e.g., dextrin, cyclodextrin, etc.) and conventional sugars such as xylitol, sorbitol, erythritol, etc. Sugar alcohol of may be contained. In addition, natural flavoring agents (eg, taumatin, stevia extract, etc.) and synthetic flavoring agents (eg, saccharin, aspartame, etc.) may be included as flavoring agents. The ratio of the natural carbohydrate may be generally contained in about 1 to 20 g, specifically about 5 to 12 g per 100 mL of beverage.

The composition according to an embodiment may be a pharmaceutical composition. Specifically, the pharmaceutical composition may be a pharmaceutical composition for preventing or treating skin diseases. The skin disease may be a skin inflammatory disease or a disease caused by skin irritation.

The skin inflammatory disease may mean all diseases caused by skin irritation and immune stimulation. For example, it may include atopic dermatitis, itching, redness, erythema, eczema, seborrheic dermatitis, psoriasis, acne, etc., but is not limited thereto. The pharmaceutical composition according to an embodiment may prevent or treat skin inflammatory diseases through effects such as maintaining skin moisture, preventing skin moisture loss, strengthening skin barrier, and anti-inflammatory.

The damage to the skin barrier function may mean all changes that appear on the skin due to a decrease or damage to the function of the skin barrier. For example, skin wrinkles may increase, dryness, dermatitis, atopic dermatitis, allergic dermatitis, acne, etc., but are not limited thereto. The pharmaceutical composition according to an embodiment may prevent or treat damage to the skin barrier function by increasing the recovery rate of the amount of transdermal moisture loss.

The skin aging may mean all changes in tangible and intangible forms that appear on the skin with age. For example, it may include, but is not limited to, increase in skin wrinkles, dryness, decrease in wound healing ability, decrease in skin immune function, and the like. The pharmaceutical composition according to an embodiment may prevent or treat skin aging through effects such as strengthening the skin barrier, maintaining skin moisture, and preventing skin moisture loss.

Another aspect provides a method of soothing the skin of a subject comprising administering the composition to the subject. The composition is the same as described above.

Specifically, the method may be a method of improving or treating inflammation of an individual, or a method of improving or treating skin irritation of an individual.

The terms “administering”, “introducing” and “transplanting” are used interchangeably and one embodiment into a subject by a method or route that results in at least partial localization of a composition to a desired site according to one embodiment. It may mean the arrangement of the composition according to. The extract of the composition according to one embodiment or at least a portion of the extract component may be administered by any suitable route that delivers to a desired location in a living subject.

Administration may be administered by a method known in the art. Administration can be administered directly to a subject by any means, for example by routes such as intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration. I can. The administration can be administered systemically or locally.

The subject may be a mammal, for example a human, cow, horse, pig, dog, sheep, goat, or cat. The subject may be a subject in need of skin soothing, eg anti-inflammatory, atopic dermatitis treatment.

The administration is 0.1 mg to 1,000 mg per subject per day, for example, 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1,000 mg per individual. , 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg, 5 mg To 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg may be administered. However, the dosage may be variously prescribed according to factors such as formulation method, administration mode, patient's age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate, and response sensitivity. Taking these factors into account, the dosage can be appropriately adjusted. The number of administration can be once a day or two or more times within the range of clinically acceptable side effects, and the administration site can be administered at one or two or more locations, daily or at intervals of 2 to 5 days. The number of days of administration can be administered from 1 to 30 days per treatment. If necessary, the same treatment can be repeated after an appropriate time period. For animals other than humans, the same dose per kg as that of humans or, for example, the volume ratio (e.g., average value) of the target animal and human organs (heart, etc.) The converted amount can be administered.

According to the hollywood extract according to an aspect, since it has an excellent effect of improving skin inflammation or skin irritation, it can be applied to various sales such as cosmetics, foods, and pharmaceutical compositions.

1 is a graph showing the result of measuring the change of TSLP mRNA expression in human keratinocytes according to ethanol extract treatment of holly leaves (a), stem (b), and root (c) by real-time PCR (-for control ##: p<0.01; + **: p<0.01, *: p<0.05 for control).
Figure 2 is a graph showing the result of measuring the MDC mRNA expression change in human keratinocytes according to ethanol extract treatment (10 ppm) of holly leaves, stems, and roots by real-time PCR (##: p<0.01; + ** for control group: p<0.01, *: p<0.05).
3 is a graph showing the result of measuring the TARC mRNA expression change in human keratinocytes according to ethanol extract treatment (10 ppm) of holly leaves, stems, and roots by real-time PCR (##: p<0.01; + ** for control group: p<0.01, *: p<0.05).
Figure 4 is a graph showing the result of measuring the TARC protein expression change in human keratinocytes according to ethanol extract treatment (10 ppm) of holly leaves, stems, and roots by ELISA (##: p<0.01 for the control group. ; + ** for the control group: p<0.01).
Figure 5 is a graph showing the result of measuring the change in IL-6 mRNA expression of human keratinocytes according to ethanol extract treatment (10 ppm) of holly leaves, stems, and roots by real-time PCR (## for the control group: p<0.01; + ** for control: p<0.01).
Figure 6 is a graph showing the result of measuring the change in IL-8 mRNA expression of human keratinocytes according to ethanol extract treatment (10 ppm) of holly leaves, stems, and roots by real-time PCR (## for the control group: p<0.01; + ** for control: p<0.01).

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

Example 1: Preparation of holly extract

In this example, holly (Ilex cornuta, Ilex x wandoensis) was purchased at Gyeongdong Market (Seoul, Korea). The leaves, stems, and roots of the purchased holly were dried and powdered, and 100 ml of 70% (v/v) ethanol per 10 g of each powder was added, and then allowed to stand at room temperature for 1 day. The filtrate obtained by filtering the solution with filter paper was concentrated under reduced pressure at 40-60° C. to obtain 3.7 g of the extract (hereinafter referred to as “ethanol extract”).

Experimental example One: TSLP Inhibition evaluation

1.1. Human skin Keratinocyte line HaCaT Cell culture

Human keratinocytes HaCaT were distributed from the Korea Cell Line Bank and cultured in DMEM medium containing 10% fetal bovine serum (FBS) and antibiotics. The culture vessel used a 75T-flask and a 6-well plate, and cultured in a 37°C incubator supplied with 5% CO ₂ . The medium was changed every 3-4 days, and when cells were excessively proliferated, they were subcultured. HaCaT was dispensed at ⁵ ×10 ⁵ /well and cultured for 24 hours, followed by washing with phosphate buffered saline solution (PBS). Washed HaCaT was added to each well in DMEM medium without FBS, Poly I:C was added at 10 μg/ml, IL-4 was added at 10 ng/ml, and ethanol extracts of holly leaves, stems, and roots were added to the final concentration. Diluted to 0.1, 1, and 10 ppm, respectively, added to each well, and cultured.

1.2. TSLP ( Thymic Stromal Lymphopoietin ) Measurement of expression change

TSLP (thymic stromal lymphopoietin) is classified as a top-level cytokine that initiates a type 2 helper T-cell mediated immune bias that is identified as the cause of atopic dermatitis. After incubation with Poly I:C/IL-4 for 4 hours, cDNA was synthesized from HaCaT cells, and real-time polymerase chain using SYBR green master mix and primers using the synthesized cDNA as a template The reaction (real-time PCR) was performed to measure the level of gene expression of TSLP. As a positive control, dexamethasone (Dex) was treated at -concentration. The relative values were compared through corrected quantification of the internal control gene β-actin by the Ct (threshold cycle) method.

The primer sequences used in the examples of the present invention are shown in Table 1 below.

gene order TSLP Forward 5'- GCTATCTGGTGCCCAGGCTAT-3' SEQ ID NO: 1 TSLP Reverse 5'- CGACGCCACAATCCTTGTAAT-3' SEQ ID NO: 2 TARC Forward 5'-CTTCTCTGCAGCACATCC-3' SEQ ID NO: 3 TARC Reverse 5'-AAGACCTCTCAAGGCTTTG-3' SEQ ID NO: 4 MDC Forward 5'- GCATGGCTCGCCTACAGACT-3' SEQ ID NO: 5 MDC Reverse 5'- GCAGGGAGGGAGGCAGAGGA-3' SEQ ID NO: 6 IL-6 Forward 5'-TACCCCCAGGAGAAGATTCC-3' SEQ ID NO: 7 IL-6 Reverse 5'-TTTTCTGCCAGTGCCTCTTT-3' SEQ ID NO: 8 IL-8 Forward 5'-CCAACACAGAAATTATTGTAAAGC-3' SEQ ID NO: 9 IL-8 Reverse 5'-TGAATTCTCAGCCCTCTTCAA-3' SEQ ID NO: 10 β-actin Forward 5'-GGCCATCTCTTGCTCGAAGT-3' SEQ ID NO: 11 β-actin Reverse 5'-GACACCTTCAACACCCCAGC-3' SEQ ID NO: 12

1 is a graph showing the result of measuring the change of TSLP mRNA expression in human keratinocytes according to ethanol extract treatment of holly leaves (a), stem (b), and root (c) by real-time PCR (-for control ##: p<0.01; + **: p<0.01, *: p<0.05 for control).

(-): negative control group; (+): PolyI:C and IL-4 treatment group; Dex: positive control group treated with dexamethasone; 0.1: holly extract

As shown in FIG. 1, TSLP expression was significantly increased in HaCaT cells treated with poly I:C and IL-4 (+) compared to HaCaT cells not treated with poly I:C and IL-4 (-). The TSLP mRNA expression was decreased in a concentration dependent manner by treatment with holly extract. In particular, it was confirmed that TSLP expression decreased to a level similar to that of HaCaT cells treated with dexamethasone, a positive control, by treatment of 1 ppm or more of holly extract.

Experimental example 2: inflammatory Chemokines Inhibition evaluation

In order to evaluate the inflammatory chemokine inhibitory ability of hollywood extract, changes in the expression of inflammatory chemokines MDC (macrophage induced chemokine) and TARC (thymus and activation regulated chemokine6) were measured in the same manner as in Experimental Example 1.1. Also known as (CC motif) ligand 17 (CCL17).

Figure 2 is a graph showing the result of measuring the MDC mRNA expression change in human keratinocytes according to ethanol extract treatment (10 ppm) of holly leaves, stems, and roots by real-time PCR (##: p< 0.01; + ** for control group: p<0.01, *: p<0.05).

3 is a graph showing the result of measuring the TARC mRNA expression change in human keratinocytes according to ethanol extract treatment (10 ppm) of holly leaves, stems, and roots by real-time PCR (##: p< 0.01; + ** for control group: p<0.01, *: p<0.05).

2 and 3, it was confirmed that the mRNA expression of MDC and TARC increased by poly I:C and IL-4 treatment was significantly reduced by treatment with ethanol extract of holly leaves, stems, and roots.

(-): negative control group; (+): PolyI:C and IL-4 treatment group; Dex: The positive control group treated with dexamethasone.

Additionally, changes in chemokine expression were measured using an ELISA kit. HaCaT (5x10 ⁵ /well) cells were cultured in FBS DMEM medium for 24 hours, then holly leaves, stems, and root extracts and poly I:C 10 μg/ml and IL-4 10 ng/ml were simultaneously added to DMEM without FBS. Treated and further incubated for 72 hours. Then, the supernatant of each well was quantified using a human CCL17/TARC ELISA kit (R&D).

Figure 4 is a graph showing the result of measuring the TARC protein expression change in human keratinocytes according to ethanol extract treatment (10 ppm) of holly leaves, stems, and roots by ELISA (##: p<0.01 for the control group. ; + ** for the control group: p<0.01).

Experimental example 3: inflammatory cytokines Inhibition evaluation

In order to evaluate the inflammatory cytokine inhibitory ability of hollywood extract, changes in the expression of inflammatory cytokines IL-6 and IL-8 were measured in the same manner as in Experimental Example 1.1.

Figure 5 is a graph showing the result of measuring the change in IL-6 mRNA expression of human keratinocytes according to ethanol extract treatment (10 ppm) of holly leaves, stems, and roots by real-time PCR (## for the control group: p<0.01; + ** for control: p<0.01).

6 is a graph showing the result of measuring the change in IL-8 mRNA expression in human keratinocytes according to ethanol extract treatment (10 ppm) of holly leaves, stems, and roots by real-time PCR (## for the control group: p<0.01; + ** for control: p<0.01).

5 and 6, it was confirmed that the mRNA expression of IL-6 and IL-8 increased by poly I:C and IL-4 treatment was significantly reduced by treatment with ethanol extract from holly leaves, stems, and roots. I did. In particular, it was confirmed that the mRNA expression of IL-8 was significantly reduced by treatment with ethanol extract of holly leaves compared to other sites.

Therefore, the ethanol extract of holly leaves, stems, and roots inhibits the expression of TSLP, inflammatory chemokines, and inflammatory cytokines, and has excellent inhibitory effect on inflammation and alleviation of irritation. It is effective in preventing, improving, or treating diseases.

Formulation example 1: Preparation of cream

It was prepared by a conventional method according to the composition and composition ratio shown in Table 2 in order to prepare a cream containing hollywood extract.

ingredient Content (% by weight) Holly Leaf Extract One Glyceryl Stearate/Pig-100 Stearate 2.0 Polysorbate 60 0.5 Sorbitan Stearate 0.5 Phytosqualane 5.0 Jojoba oil 3.0 Caprylic/Capric Triglyceride 10.0 Aloe butter 3.0 Beeswax 1.0 Stearyl alcohol 0.5 Beheryl alcohol 0.5 Stearic acid 0.5 glycerin 7.0 Butylene glycol 5.0 Carboxy vinyl polymer 0.2 Arginine 0.3 Dimethicone 0.5 antiseptic a very small amount Spices a very small amount Purified water Balance

Formulation example 2: Preparation of lotion

It was prepared by a conventional method according to the composition components and composition ratio as shown in Table 3 to prepare a lotion containing the hollywood extract.

ingredient Content (% by weight) Holly Leaf Extract One glycerin 3.0 Butylene glycol 2.0 C14-22 alcohol/C12-20 alkyl glucoside 2.0 Glycerin monostearate 0.5 Caprylic/Capric Triglyceride 10.0 Microcrystalline wax 5.0 Carboxy vinyl polymer 0.2 Triethanolamine 0.2 antiseptic a very small amount Spices a very small amount Purified water Balance

<110> COSMAX, INC. <120> A composition for preventing or treating inflammation comprising an extract of Ilex cornuta and uses thereof <130> PN125582 <160> 12 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TSLP forward primer <400> 1 gctatctggt gcccaggcta t 21 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TSLP reverse primer <400> 2 cgacgccaca atccttgtaa t 21 <210> 3 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> TARC forward primer <400> 3 cttctctgca gcacatcc 18 <210> 4 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> TARC reverse primer <400> 4 aagacctctc aaggctttg 19 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> MDC forward primer <400> 5 gcatggctcg cctacagact 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> MDC reverse primer <400> 6 gcagggaggg aggcagagga 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> IL-6 forward primer <400> 7 tacccccagg agaagattcc 20 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> IL-6 reverse primer <400> 8 ttttctgcca gtgcctcttt 20 <210> 9 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> IL-8 forward primer <400> 9 ccaacacaga aattattgta aagc 24 <210> 10 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> IL-8 reverse primer <400> 10 tgaattctca gccctcttca a 21 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> beta actin forward primer <400> 11 ggccatctct tgctcgaagt 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> beta actin reverse primer <400> 12 gacaccttca acaccccagc 20

Claims (9)

A cosmetic composition for preventing or improving skin inflammation comprising an extract of hollywood (Ilex cornuta) as an active ingredient. The cosmetic composition according to claim 1, wherein the prevention or improvement of skin inflammation is one or more prevention or improvement selected from the group consisting of atopic dermatitis, itching, redness, erythema, eczema, seborrheic dermatitis, psoriasis and acne. The cosmetic composition of claim 1, wherein the extract is a holly leaf extract. The cosmetic composition of claim 1, wherein the extract is an extract of C1-C6 alcohol, water, or a mixture thereof. The cosmetic composition of claim 1, wherein the extract is an ethanol extract. A food composition for preventing or improving skin inflammation comprising an extract of Ilex cornuta as an active ingredient. A pharmaceutical composition for preventing or treating skin inflammation comprising an extract of hollywood (Ilex cornuta) as an active ingredient. A composition for external application for skin for preventing or improving skin inflammation, comprising an extract of hollywood (Ilex cornuta) as an active ingredient. A cosmetic composition for preventing or improving skin irritation, comprising an extract of hollywood (Ilex cornuta) as an active ingredient.

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