WO2020138833A1

WO2020138833A1 - Composition comprising corylus heterophylla extract for prevention or treatment of skin disease

Info

Publication number: WO2020138833A1
Application number: PCT/KR2019/018038
Authority: WO
Inventors: 이용준; 김영한
Original assignee: 재단법인 홍천메디칼허브연구소
Priority date: 2018-12-27
Filing date: 2019-12-18
Publication date: 2020-07-02
Also published as: KR20200080555A; KR102162843B1

Abstract

The present invention relates to a pharmaceutical composition comprising a Corylus heterophylla extract or a fraction thereof for prevention or treatment of skin disease, a method for treatment of skin disease, comprising a step of administering the pharmaceutical composition to a subject, a food composition a Corylus heterophylla extract or a fraction thereof for alleviation of skin disease, a quasi-drug composition comprising a Corylus heterophylla extract or a fraction thereof for prevention or alleviation of skin disease, and a cosmetic composition comprising a Corylus heterophylla extract or a fraction thereof for prevention or alleviation of skin disease. The Corylus heterophylla extract of the present invention was found to effectively treat skin diseases such as psoriasis and as such, can find wide applications in the prevention or treatment of various skin diseases.

Description

Composition for preventing or treating skin diseases comprising hazel extract

The present invention relates to a composition for preventing or treating skin diseases comprising the hazel extract, and more specifically, the present invention administers a pharmaceutical composition or pharmaceutical composition for preventing or treating skin disease comprising the hazel extract or a fraction thereof Skin disease treatment method comprising the step of, hazel tree extract or a food composition for improving skin disease comprising a fraction thereof, hazel tree extract or a quasi-drug composition for preventing or improving skin disease comprising a fraction thereof and hazel tree extract or It relates to a cosmetic composition for the prevention or improvement of skin diseases comprising a fraction thereof.

Skin is increasingly damaged by intrinsic factors such as aging or external factors such as ultraviolet rays, external pollutants and stress, and the ability to protect the skin from these factors is weakened, thereby reducing cell protection and proliferation capacity. In this case, the individual starts the regeneration process for the damaged skin, which is a reaction of the skin tissue to the damage and lasts from 2 days to 3 weeks after the skin is damaged.

On the other hand, in recent years, the problem of skin damage due to external environmental factors is gradually increasing, and a representative environmental factor causing the skin damage is a recent rapid increase in fine dust. The dust is divided into total dust (TSP, TotalSuspended Particles) of less than 50㎛ depending on the size of the particles and fine dust (PM, Particulate Matter) with very small particle size, and the fine dust is again fine dust (PM10) with a diameter less than 10㎛. ) And ultra-fine dust (PM2.5) whose diameter is less than 2.5㎛. When fine dust enters the body, immune cells act to remove and discharge dust, and side effects can be accompanied by inflammation. Fine dust penetrates into the pores of the skin as well as the respiratory system such as the airways, bronchi, and lungs, and irritates the skin, causing trouble and inflammation, further exacerbating the skin condition, and further adversely affecting chronic skin diseases such as atopic dermatitis. Can give.

Steroids, which are the most widely used treatments for traditional skin diseases, cause side effects such as diabetes and hypertension, and have become a social problem due to abuse, and recently developed immunomodulators act only on specific targeted areas to safely immunize Antibody preparations that were expected to cause inhibitory effects have been reported to cause unexpected and serious side effects, such as immune disorders, infections, and cancer, and using natural substances to develop a drug that can effectively treat skin diseases without side effects Research is actively underway. For example, Korean Patent Publication No. 2010-0080972 discloses Sanyacho extract using fermented tree sap that can be used to alleviate and improve skin diseases, and Korean Patent Publication No. 2011-0086894 discloses shell or waste coral. Disclosed is a composition for external application for skin comprising a calcined powder of alkali and an alkaline calcium solution extracted from a small ingredient powder of waste shell or coral. However, such a formulation developed using natural products does not exhibit side effects, but has a disadvantage in that the therapeutic effect is insufficient, and thus there is an urgent need for supplementation.

Under these backgrounds, the present inventors have confirmed that the hazel extract shows the effect of effectively treating skin diseases without showing side effects, as a result of earnest research efforts to develop a formulation that can effectively treat skin diseases without side effects, The present invention was completed.

One object of the present invention is to provide a pharmaceutical composition for preventing or treating skin diseases comprising a hazel extract or a fraction thereof.

Another object of the present invention is to provide a method for treating skin diseases comprising the step of administering the pharmaceutical composition to a suspected skin disease subject.

Another object of the present invention is to provide a food composition for improving skin diseases comprising a hazel extract or a fraction thereof.

Another object of the present invention is to provide a quasi-drug composition for the prevention or improvement of skin diseases comprising a hazel extract or a fraction thereof.

Another object of the present invention is to provide a cosmetic composition for the prevention or improvement of skin diseases comprising a hazel extract or a fraction thereof.

Since the hazel extract of the present invention has no side effects, it has been confirmed that it can effectively treat skin diseases such as psoriasis, so the hazel extract of the present invention may be widely used for prevention or treatment of various skin diseases.

1 is a graph showing the results of analyzing the cytotoxicity of hazel extract.

Figure 2 is a graph showing the results of comparing the change in the amount of IL-6 production according to the concentration of hazelnut extract.

Figure 3 is a photograph showing the results of Western blot analysis showing the result of comparing the change in the phosphorylation level of the Stat3 protein according to the treatment concentration of the hazel extract.

4 is a schematic diagram schematically illustrating an experimental process for verifying the effect of hazel extract using an animal model of skin disease.

5 is a graph showing the change in body weight of Balb/c mice treated with imiquimod (IMQ), hazel extract (CH), or dermobait (CLO).

Figure 6 is a photograph showing the change in appearance of the ear over the breeding time of Balb/c mice treated with imiquimod (IMQ), hazel extract (CH) or dermobait (CLO).

Figure 7 is a imiquimod (IMQ), hazel extract (CH) or dermobait (CLO) treated Balb / c mice showing the change in the thickness of the ear skin over the breeding time

It is a rap.

8 is a graph showing changes in the erythema level of the ear skin according to the lapse of breeding time in Balb/c mice treated with imiquimod (IMQ), hazel extract (CH), or dermobait (CLO).

Figure 9 is an optical micrograph showing the results of staining the skin tissue of Balb/c mice treated with imiquimod (IMQ), hazel extract (CH) or dermobait (CLO) with hematoxylin and eosin.

10 is a graph showing the results of quantitative analysis of the epidermal thickness of skin tissue of Balb/c mice treated with imiquimod (IMQ), hazel extract (CH) or dermobait (CLO).

One embodiment of the present invention for achieving the above object provides a pharmaceutical composition for the prevention or treatment of skin diseases comprising a hazel extract or a fraction thereof.

Another embodiment of the present invention provides a preventive or therapeutic use for skin diseases comprising a hazel extract or a fraction thereof.

The term "cobryl (Corylus heterophylla)" of the present invention means a kind of deciduous tree growing on the slope of the forest edge, sunny roadside. It reaches 1~2m in height, the fruit is a round nut, runs 1~3, and ripens in October. Use the fruit as edible or medicinal. Unlike other hazel varieties, this species is distinguished by a flattened feature, with the tip of a leaf being cut with a knife, without the lengthening of the flower wrappers. It is distributed all over Korea and overseas, and is grown all over Japan, China, North America, and Russia.

In the present invention, the hazel tree may be used by purchasing a commercially available one, or using one harvested or cultivated in nature, but is not limited thereto.

The term "extract" of the present invention means a resultant product such as a liquid component obtained by immersing a desired substance in various solvents and then extracting it for a period of time at a normal temperature or a warm temperature, and a solid component obtained by removing a solvent from the liquid component. Means In addition, in addition to the above results, it can be comprehensively interpreted to include all of the dilutions of the above results, their concentrates, their modulators, and tablets. Accordingly, the hazel extract provided in the present invention is an extract obtained by extracting a hazel flower, leaf, fruit, stem, root, bark, sap, etc., a diluent or concentrate of the extract, and a dried product obtained by drying the extract , It may be interpreted to include extracts of all formulations that can be formed using the extracts themselves and the extracts, such as a crude product or a purified product of the extract, or a mixture thereof.

In the hazel extract of the present invention, the method for extracting the mixture is not particularly limited, and can be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method may include a hot water extraction method, an ultrasonic extraction method, a filtration method, a reflux extraction method, and the like, which may be performed alone or in combination of two or more methods.

The type of the solvent used for the extraction in the present invention is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the extraction solvent include water, alcohol or a mixed solvent thereof, and these may be used alone or in combination of one or more, and specifically water may be used. In the case of using alcohol as a solvent, alcohol having 1 to 4 carbon atoms may be specifically used.

The term "fraction" of the present invention means a result obtained by performing a fraction to separate a specific component or a specific component group from a mixture comprising various various constituents.

The fractionation method for obtaining the fraction in the present invention is not particularly limited, and may be performed according to a method commonly used in the art. Non-limiting examples of the fractionation method include a solvent fractionation method performed by treating various solvents, an ultrafiltration fractionation method performed by passing through an ultrafiltration membrane having a constant molecular weight cut-off value, and various chromatography (size, charge, hydrophobicity) Or chromatographic fractionation for performing separation according to affinity), and combinations thereof. Specifically, a method of obtaining a fraction from the extract by treating a predetermined solvent to the extract obtained by extracting the hazel of the present invention.

In the present invention, the type of fractional solvent used to obtain the fraction is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the fractional solvent include polar solvents such as water and alcohols having 1 to 4 carbon atoms; Non-polar solvents such as hexane, ethyl acetate, chloroform, and dichloromethane; Or mixed solvents thereof. These may be used alone or in combination of one or more, but are not limited thereto.

In addition, the extract or fraction may be prepared and used in a dry powder form after extraction, but is not limited.

The term "skin disease" of the present invention means a disease in which abnormal symptoms such as inflammation, erythema, thickening, fibrosis, and keratin appear on the skin due to genetic factors, physiological factors, environmental factors, and the like.

In the present invention, the skin disease is not particularly limited as long as symptoms can be prevented, improved, alleviated, or treated by treatment with a hazel extract or a fraction thereof, but as an example, excessive secretion of inflammatory cytokine It may be an inflammatory skin disease induced by, and as another example, atopic dermatitis, allergic dermatitis, psoriasis, seborrheic dermatitis, contact dermatitis, erythema, which is a type of inflammatory skin disease induced by excessive secretion of IL-6 It can be reflective lupus, acne, and papule hives.

The term "prevention" of the present invention means all actions to suppress or delay skin diseases by administration of a composition containing the extract.

The term "treatment" of the present invention means any action in which symptoms of skin disease are improved or beneficially changed by administration of a composition containing the extract.

The pharmaceutical composition of the present invention may include the extract in an amount of 0.001 to 80, specifically 0.001 to 70, and more specifically 0.001 to 60% by weight based on the total weight of the composition, but is not limited thereto.

In addition, the pharmaceutical composition may further include a pharmaceutically acceptable carrier, excipient, or diluent commonly used in the manufacture of a pharmaceutical composition, and the carrier may include a non-naturally occurring carrier. have. The carrier, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In addition, the pharmaceutical compositions are tablets, pills, powders, granules, capsules, suspensions, intravenous solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, transdermal formulations according to a conventional method, respectively. It can be formulated in the form of absorbents, gels, lotions, ointments, creams, patches, cataplasmas, pastes, sprays, skin emulsions, skin suspensions, transdermal delivery patches, drug-containing bandages or suppositories. Specifically, when formulated, it may be prepared using diluents or excipients such as fillers, weights, binders, wetting agents, disintegrating agents, surfactants, etc., which are commonly used. Solid preparations for oral administration include, but are not limited to, tablets, pills, powders, granules, capsules, and the like. These solid preparations may be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose, lactose, gelatin, and the like. In addition, lubricants such as magnesium stearate and talc may be used in addition to simple excipients. In addition to liquids for oral administration and liquid paraffin, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, may be added to prepare. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

Another embodiment of the present invention provides a method for treating skin diseases, comprising administering the pharmaceutical composition to a suspected skin disease individual.

The term "administration" of the present invention means the act of introducing a composition comprising said extract to an individual in an appropriate manner.

The term "individual" of the present invention refers to all animals, such as mice, mice, livestock, including humans, who may or may have a skin disease. As a specific example, it may be a mammal, including a human.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The term, “pharmaceutically effective amount” means an amount sufficient to treat the disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the individual type and severity, age, sex, activity of the drug, Sensitivity to the drug, time of administration, route of administration and rate of excretion, duration of treatment, factors including co-drugs and other factors well known in the medical arts. For example, the hazel extract may be administered at a dose of 0.01 to 500 mg/kg per day, specifically 10 to 100 mg/kg, and the administration may be administered once or several times a day.

The pharmaceutical composition may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And it can be administered single or multiple. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects, and can be easily determined by those skilled in the art.

In addition, the pharmaceutical composition may be administered orally or parenterally (eg, applied intravenously, subcutaneously, intraperitoneally, or topically) according to the desired method, and the dosage is the patient's condition, weight, and degree of disease. , Depending on the drug form, the route of administration and time, it can be appropriately selected by those skilled in the art.

Another embodiment of the present invention provides a food composition for improving skin disease comprising a hazel extract or a fraction thereof.

At this time, the definition of the hazel, extract, fraction and skin diseases are as described above.

The term "improvement" of the present invention means any action that at least reduces the parameters associated with the condition being treated by administration of a composition comprising said extract, e.g., the severity of symptoms.

The term, "food" of the present invention is meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages , Vitamin complexes, health functional foods and health foods, and includes all foods in the ordinary sense.

Since the food composition of the present invention is derived from hazel that can be ingested on a daily basis, a high skin disease improvement effect can be expected, and thus can be very useful for health promotion purposes.

The functional food (functional) food (food for special health use) (food for special health use, FoSHU) in the same terms, in addition to nutrition, the biomedical control function is processed to appear efficiently, medical and medical effects are high Means food. Here, the term'function (sex)' means obtaining a useful effect for health use such as adjusting nutrients or physiological effects on the structure and function of the human body. The food of the present invention can be prepared by a method commonly used in the art, and the above-described preparation can be made by adding raw materials and ingredients commonly added in the art. In addition, the formulation of the food can be prepared without limitation as long as it is a formulation recognized as food.

The food composition of the present invention can be prepared in various types of formulations, and unlike general medicines, it has the advantage of not having side effects that can occur when taking medicines for a long time using natural substances as raw materials, and is excellent in portability. The food of the invention can be consumed as an adjuvant to enhance the improvement effect of skin diseases.

The health food refers to food having an active health maintenance or enhancement effect compared to general food, and health supplement food means food for health supplement purposes. In some cases, the terms health functional food, health food, and dietary supplement are used interchangeably.

Specifically, the health functional food is a food prepared by adding the hazel extract or fractions thereof of the present invention to food materials such as beverages, teas, spices, gums, confectionery, or by encapsulation, powdering, suspension, etc. If it does, it means that it has a certain effect on health, but unlike general medicines, it has the advantage of not having side effects that can occur when taking medicines for a long time using food as a raw material.

The food composition may further include a physiologically acceptable carrier, and the type of the carrier is not particularly limited, and any carrier commonly used in the art may be used.

In addition, the food composition may include additional ingredients that are commonly used in food compositions to improve odor, taste, and vision. For example, vitamin A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantotenic acid, and the like. In addition, minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr); And amino acids such as lysine, tryptophan, cysteine, and valine.

In addition, the food composition is a preservative (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), fungicides (bleached and highly bleached, sodium hypochlorite, etc.), antioxidants (butyl hydroxyanisole (BHA), butyl hydride) Roxytoluene (BHT, etc.), colorants (such as tar pigment), colorants (sodium nitrite, sodium nitrite, etc.), bleach (sodium sulfite), seasonings (such as MSG sodium glutamate), sweeteners (dulcin, cyclate, saccharin , Sodium, etc.), fragrances (vanillin, lactones, etc.), swelling agents (alum, D-potassium hydrogen sulphate, etc.), strengthening agents, emulsifiers, thickeners (foams), coating agents, gum herbicides, foam inhibitors, solvents, improvers, etc. Food additives. The additive may be selected according to the type of food and used in an appropriate amount.

As an example of the food composition of the present invention, it can be used as a health drink composition, and in this case, it may contain various flavoring agents or natural carbohydrates, etc., as additional components, such as a conventional beverage. The above-described natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Polysaccharides such as dextrin and cyclodextrin; Sugar alcohols such as xylitol, sorbitol, and erythritol. Sweeteners include natural sweeteners such as taumatin and stevia extract; Synthetic sweeteners such as saccharin and aspartame. The ratio of the natural carbohydrate may be generally about 0.01 to 0.04 g, specifically about 0.02 to 0.03 g per 100 ml of the health drink composition of the present invention.

In addition to the above, the health drink composition includes various nutrients, vitamins, electrolytes, flavoring agents, colorants, pectic acid, pectic acid salts, alginic acid, alginic acid salts, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, Alcohol or carbonic acid. In addition, it may contain flesh for the manufacture of natural fruit juice, fruit juice beverage, or vegetable beverage. These ingredients may be used independently or in combination. The proportion of these additives is not particularly important, but is generally selected from 0.01 to 0.1 parts by weight per 100 parts by weight of the health drink composition of the present invention.

Another embodiment of the present invention provides a quasi-drug composition for the prevention or improvement of skin diseases comprising a hazel extract or a fraction thereof.

Another embodiment of the present invention provides a preventive or ameliorative use of skin diseases comprising a hazel extract or a fraction thereof.

At this time, the definition of hazel, extract, fraction, skin disease, prevention and improvement is as described above.

The term "quasi-drug" of the present invention is a fiber, rubber product or the like used for the purpose of treating, alleviating, treating or preventing a disease of a human or animal, has a weak effect on the human body or does not act directly on the human body, Or non-machine and similar products, one of the agents used for sterilization, pesticide and similar purposes to prevent infection, used for the purpose of diagnosing, treating, reducing, treating or preventing human or animal diseases Of the items that are not used as devices, machines, or devices, and those used for the purpose of having a pharmacological effect on the structure and function of a person or animal, excluding items that are not devices, machinery, or devices, and external preparations for skin and personal hygiene Includes supplies.

When the hazel extract of the present invention is added to a quasi-drug composition for the purpose of preventing or improving skin disease, the extract can be added as it is or used with other quasi-drug components, and can be suitably used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment).

The quasi-drug composition is not particularly limited, but includes personal hygiene products, external skin preparations, disinfectant cleaners, shower foams, wipes, detergent soaps, hand washes, masks or ointments. The external preparation for skin is not particularly limited thereto, and may be specifically used in the form of an ointment, lotion, spray, patch, cream, powder, suspension, gel, or gel. The personal hygiene products are not particularly limited, but may be soap, wipes, tissues, shampoo, toothpaste, hair care products, air freshener gels or cleaning gels.

Another embodiment of the present invention provides a cosmetic composition for preventing or improving skin diseases comprising a hazel extract or a fraction thereof.

The hazel extract may contain the extract in a proportion of 0.1% to 20% by weight based on the total weight of the composition, as in the pharmaceutical composition, for the composition. Specifically, it is included in a ratio of 0.1 wt% to 5 wt%, more specifically 0.4 to 0.6 wt%, and even more specifically 0.5 wt% based on the total weight of the composition, but is not limited thereto.

The ingredients included in the cosmetic composition of the present invention include ingredients commonly used in cosmetic compositions in addition to the extract, for example, water, surfactant, moisturizer, lower alcohol, chelating agent, fungicide, antioxidant, preservative, pigment And one or more additives selected from the group consisting of perfumes.

In addition, the cosmetic composition may be prepared in any conventionally prepared formulation, for example, solutions, emulsions, suspensions, pastes, creams, lotions, gels, powders, sprays, surfactant-containing cleansing, oils, soaps , Liquid cleaning agents, bathing agents, foundations, makeup bases, essences, lotions, foams, packs, soft water, sunscreen cream or sun oil, etc., but is not limited thereto.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, ethoxylated isostearyl alcohol, suspensions such as polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystals Sex cellulose, aluminum metahydroxide, bentonite, tracant, or the like can be used.

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

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, and in the case of a spray, additionally chlorofluorohydrocarbon, propane /Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide as a carrier component Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

In addition, the ingredients included in the cosmetic composition may be included in an amount generally used in the field of skin science.

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.

실시예 1: 개암나무 추출물의 세포독성 분석Example 1: Analysis of cytotoxicity of hazel extract

The cytotoxicity of the hazelnut extract was analyzed using a sample obtained by dissolving the hazelnut methyl alcohol extract purchased from the Korea Plant Extract Bank in DMSO.

Roughly, the hazelnut extract was treated on a cell line of cultured human-derived skin keratinocytes capable of simulating skin psoriasis symptoms, and the change in cell viability was measured through a WST 1 assay.

Specifically, 10% (v/v) fetal bovine serum (FBS, Gibco) and antibiotics (Antibiotic antimycotic) added DMEM (Dulbecco's modified Eagle Medium) medium was used, 37 ℃ and 5% CO ₂ HaCaT cell line, a human-derived skin keratinocyte, was cultured under conditions. When the cells grew to about 80% of a 100 mm culture dish, the cells were lightly washed with PBS for detachment, treated with trypsin-EDTA (Gibco/BRL), left at 37° C. for 8 minutes, and then the cells were collected. The culture was performed once every 2-3 days. The collected cells were dispensed at a concentration of 3×10 ⁴ cells/ml in a 96-well plate, pre-incubated for 24 hours while maintaining 37° C. and 5% CO ₂ in the incubator, and various concentrations (100-800 μg/ ML) Hazelnut extract was treated and cultured for 24 hours. Then, water-soluble tetrazolium salt (WST-1) was treated to incubate 1 hour so that WST-1 is reduced by enzymatic action of viable cells. Then, it was measured at 450 nm using a microplate reader (FIG. 1). At this time, cells not treated with hazel extract were used as a negative control, and cells treated with DMSO only were used as a positive control.

As shown in Figure 1, when treated with various concentrations of hazel extract, it was confirmed that the cell viability of all 10% is reduced.

실시예 2: IL-6 생성에 미치는 개암나무 추출물의 효과 분석Example 2: Analysis of the effect of hazel extract on IL-6 production

IL-6, a kind of cytokine, is known to cause skin diseases through an inflammatory reaction, and it was intended to confirm whether hazelnut extract affects the production of IL-6.

Roughly, HaCaT cells were treated with M5 (IL22, IL1a, TNF-α, Oncostatin M, IL17A; JImmunol 2010, 184:5263-5270) to increase the production of IL-6, followed by various concentrations (50, 100 or 200 µg/ml) of the hazel extract was treated to measure the change in IL-6 production (FIG. 2 ). In this case, cells not treated with hazel extract as a negative control group were used, and cells treated only with M5 were used as a positive control group, and cells treated with only various concentrations of hazel extract were not treated with M5 as a comparative group. Was used.

As shown in FIG. 2, when M5 was treated with HaCaT cells, IL-6 production was significantly increased. When M5 and hazel extract were simultaneously treated, IL-6 production was increased by M5 treatment. It was confirmed that this decreases.

From the above results, it was analyzed that the hazel extract would have an effect of treating skin diseases induced by IL-6.

실시예 3: IL-6 관련 신호전달에 미치는 개암나무 추출물의 효과 분석Example 3: Analysis of the effect of hazel extract on IL-6 related signaling

Through Example 2, it was confirmed that the hazel extract inhibits the production of IL-6, and it is known that one of the main causes of IL-6 production is phosphorylation of the Stat3 protein.

Accordingly, it was intended to confirm whether the hazel extract provided in the present invention can inhibit the level of phosphorylated Stat3 protein (pSTAT3).

Roughly, HaCaT cell lines were treated with M5 and hazelnut extracts of various concentrations (100, 200 or 400 µg/ml) alone, or a combination thereof, incubated for 4 hours, and then proteins were extracted from each of these cell lines. Did. Western blot analysis using protein, anti-pSTAT3 antibody or anti-STAT3 antibody and anti-rabbit IgG HRP linked antibody of each cell line was performed to quantitatively analyze the level of pSTAT3 and STAT3 protein expressed in each cell line ( Fig. 3). At this time, β-actin was used as an internal control.

As shown in FIG. 3, when M5 was treated alone in the HaCaT cell line, the level of phosphorylated Stat3 protein (pSTAT3) increased significantly, but when M5 and hazel extract were simultaneously treated in the cell line, M5 treatment was performed. It was confirmed that the level of pSTAT3 increased by is decreased.

From the above results, it was found that the effect of hazel extract inhibiting the production of IL-6 is by reducing the level of phosphorylated Stat3 protein (pSTAT3) that induces the production of IL-6.

실시예 4: 피부질환 동물모델을 이용한 개암나무 추출물의 효과 검증Example 4: Verification of the effect of hazel extract using an animal model of skin disease

An animal model inducing skin disease was produced by applying imiquimod (IMQ), which can simulate skin psoriasis symptoms, to mouse skin. The imiquimod used in the test was Aldara cream (5% imiquimod, 3M Health Care Ltd. UK), and the control group was Dermovate (clobetasol 17-propionate 0.5 mg/1 g, Glaxo Operation, UK). Was used.

Approximately, the Aldara cream was applied to the epilated ear area of a 7-week-old Balb/c mouse for 7 days to induce a skin condition similar to skin disease, and an ointment containing hazel extract was applied together from 2 days, Ear skin thickness and erythema levels were measured at 0, 2, 4, and 6 days, respectively (FIG. 4 ). At this time, the thickness of the ear skin was measured using a Digimatic Thickness Gage (Mitutoyo, Japan), and the erythema level was measured using Dermacatch® (Colorix, Swiss). In addition, ointment containing hazel extract white petrolatum: 4521.6 mg, sorbitan sesquioleate: 23.86 mg, propylene glycol: 204.50 mg, hazel extract (5%): 250 mg and an appropriate amount of ointment was 50°C It was prepared by mixing in a bath. In addition, the ointment to be treated as a control was prepared to include white petrolatum, sorbitan sesquioleate, and propylene glycol. Finally, 62.5 mg of the Aldara cream was applied to the skin of the mouse and after 5 hours, 80 mg of ointment or dermobait containing hazel extract was applied.

First, the change in body weight of animals that induced skin conditions similar to skin diseases was measured (FIG. 5). At this time, Balb/c mice treated with a control ointment were used as a control.

As shown in Figure 5, the control group (vehicle) and mice treated with hazel extract alone (CH) did not show any significant weight change, whereas all mice treated with imiquimod (IMQ) had reduced weight, especially , It was confirmed that the mice treated with imiquimod and dermobait (IMQ+CLO) showed the greatest weight change.

Second, changes in the appearance of the ears of the mouse were observed with the naked eye during the experiment procedure (FIG. 6).

As shown in Fig. 6, at the time of 6 days, the ear of the mouse (IMQ+CLO) treated with imiquimod alone (IMQ+Vehicle) or imiquimod and Dermovate showed significant appearance changes. However, no significant change in appearance was observed in the ears of the mice treated with hazel extract (CH/IMQ+CH).

Third, the thickness of the skin of the ear of the mouse was compared during the experiment procedure (FIG. 7).

7 is a graph showing the change in the thickness of the ear skin over the breeding time of Balb/c mice treated with imiquimod (IMQ), hazel extract (CH), or dermobait (CLO).

As shown in FIG. 7, the thickness of the ear skin of the mouse (IMQ+Vehicle) treated with imiquimod alone is significantly higher than that of the control (vehicle) and the mouse (CH) treated with hazel extract alone. Increased and increased ear skin thickness by treatment with imiquimod in mice treated with imiquimod and dermobait (IMQ+CLO) and mice treated with imiquimod and hazel extract (IMQ+CH). Was confirmed to decrease. It was analyzed that the increase in the thickness of the ear skin was caused by an increase in skin keratin.

Fourth, erythema levels in the ears of the mice were compared during the experiment procedure (FIG. 8).

As shown in FIG. 8, the erythema level of the ear skin of the mouse (IMQ+Vehicle) treated with imiquimod alone, compared to the erythema level of the ear skin of the control group (vehicle) and the hazel tree extract treated alone (CH). This markedly increased, increased ear skin by treatment with imiquimod in mice treated with imiquimod and dermobate (IMQ+CLO) and mice treated with imiquimod and hazel extract (IMQ+CH). It was confirmed that the erythema level was decreased.

When the results of FIGS. 5 to 8 were synthesized, it was found that when the hazel extract was treated, it was possible to effectively suppress skin thickening (increased skin keratin) and erythema formation caused by skin diseases.

실시예 5: 조직검사를 통한 개암나무 추출물의 효과 검증Example 5: Verification of the effect of hazel extract through biopsy

In Example 4, the Balb/c mice treated with imiquimod (IMQ), hazel extract (CH) or dermobait (CLO) for 7 days were sacrificed, and the imiquimod, hazel extract or dermobait was applied. Skin tissue was removed. After the exfoliated tissue was fabricated, hematoxylin and eosin (H & E) staining was performed, and then observed with an optical microscope to compare the degree of damage to the skin tissue and quantify the thickness of the skin epidermis. Analysis (Figures 9 and 10).

Figure 9 is an optical micrograph showing the results of staining the skin tissue of Balb/c mice treated with imiquimod (IMQ), hazel extract (CH) or dermobait (CLO) with hematoxylin and eosin. Is a graph showing the results of quantitative analysis of the epidermal thickness of skin tissue of Balb/c mice treated with imiquimod (IMQ), hazel extract (CH) or dermobait (CLO).

9 and 10, when imiquimod was treated alone (IMQ+Vehicle), the skin thickness was significantly thickened compared to the control (Vehicle), but hazel extract (IMQ+CH) or dermobait (IMQ+) It was confirmed that the thickening level of the skin thickness was reduced by treatment with CLO). It was analyzed that this increase in skin thickness was caused by an increase in skin keratin.

Summarizing the above results, as a result of evaluating the efficacy of the hazel extract in an in vitro and animal experimental model for the hyperplasia and inflammatory response of keratinocytes, one of the main symptoms of skin disease, in HaCaT cells, keratin was extracted by the hazel extract. The cell proliferation rate of the cells was decreased, and the effect of treating skin diseases could be predicted through the inhibition of keratinocyte proliferation. Confirmed. In addition, in the animal model of skin disease induced by IMQ, it was confirmed that the degree of erythema, thickness, and keratin of the skin was decreased by the hazel extract, and thus it was directly suppressed the symptoms related to skin disease. Therefore, it is judged that the present invention can significantly improve the improvement and treatment efficiency for skin diseases.

From the above description, those skilled in the art to which the present invention pertains will appreciate that the present invention may be implemented in other specific forms without changing its technical spirit or essential features. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as including all changes or modifications derived from the meaning and scope of the following claims rather than the detailed description and equivalent concepts thereof.

Claims

Pharmaceutical composition for the prevention or treatment of skin diseases, comprising hazel extract or a fraction thereof.
According to claim 1,

The hazel extract inhibits the phosphorylation of STAT3 protein to suppress the secretion of inflammatory cytokines, pharmaceutical composition for the prevention or treatment of skin diseases.
According to claim 1,

The extract is selected from the group consisting of flowers, leaves, berries, stems, roots, bark, sap, and combinations of hazel, water, alcohols having 1 to 4 carbons, and mixed solvents thereof. It is obtained by extraction with one or more solvents, pharmaceutical composition for the prevention or treatment of skin diseases.
According to claim 1,

The fraction is selected from the group consisting of hazel extract water, alcohol having 1 to 4 carbon atoms, nucleic acid (Hexane), ethyl acetate (Ethyl acetate), chloroform (Chloroform), dichloromethane (Dichloromethane) and mixed solvents thereof The pharmaceutical composition for prevention or treatment of skin diseases, obtained by fractionation with a solvent.
According to claim 1,

The skin disease is an inflammatory skin disease induced by excessive secretion of inflammatory cytokine, a pharmaceutical composition for the prevention or treatment of skin disease.
The method of claim 5,

The inflammatory skin disease is from a group consisting of atopic dermatitis, allergic dermatitis, psoriasis, seborrheic dermatitis, contact dermatitis, erythematous lupus, purulent acne, papillary urticaria and combinations thereof induced by excessive secretion of IL-6. The selected disease, a pharmaceutical composition for the prevention or treatment of skin diseases.
According to claim 1,

The composition further comprises a pharmaceutically acceptable carrier, excipient or diluent, pharmaceutical composition for the prevention or treatment of skin diseases.
A method for treating skin diseases, comprising administering the pharmaceutical composition of any one of claims 1 to 7 to a subject suspected of skin disease.
Food composition for improving skin diseases, including hazel extract or a fraction thereof.
A quasi-drug composition for the prevention or improvement of skin diseases, including hazel extract or a fraction thereof.
Cosmetic composition for the prevention or improvement of skin diseases comprising hazel extract or a fraction thereof.