Composition for inhibiting IL-8 production and / or collagenase production and method for producing same

Abstract

To provide a novel IL-8 production inhibitory and / or collagenase production inhibitory composition, and application products thereof.SOLUTION: A IL-8 production inhibitory and / or collagenase production inhibitory composition contains strawberry leaf extract. The composition is superior in IL-8 production inhibitory action and collagenase production inhibitory action and is less toxic and irritative to the human body. Thus, it is suitable for applications such as cosmetic compositions, functional foods.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a novel composition for inhibiting IL-8 production and / or collagenase production, and applications thereof. [Background technology]

[0002] Interleukin-8 (IL-8) is an inflammatory cytokine that is known to increase neutrophil migration, promote inflammation, and promote the degradation of elastin. Diseases in which IL-8 is involved include inflammatory diseases accompanied by neutrophil infiltration, rheumatism, gout, asthma attacks, and Crohn's disease.

[0003] Matrix metalloproteinases (MMPs) are a group of proteolytic enzymes with metal ions in the active center, and are involved in the decomposition metabolism of extracellular matrices such as collagen, elastin, and proteoglycans. When collagenase, one of the MMPs, works excessively, it causes deterioration of skin tissue, such as wrinkles and sagging. Diseases in which collagenase is involved include rheumatism, osteoarthritis, osteoporosis, periodontitis, and cancer metastasis.

[0004] As such, IL-8 and collagenase can be the cause of various diseases, and therefore research into the components involved in the production of IL-8 and collagenase is ongoing. For example, as substances that have been reported to have an effect of suppressing the production of interleukin-8 (IL-8), Rosa robur fruit extract (Patent Document 1), zinc or zinc salts (Patent Document 2), etc., have been reported to have an effect of suppressing the production of collagenase, while persimmon petal extract (Patent Document 3), one or more plants selected from plants belonging to the genus Psophocarpus of the family Leguminosae, or extracts thereof (Patent Document 4), etc. have been reported to have an effect of suppressing the production of collagenase. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2018-058806 A [Patent Document 2] JP 2003-146892 A [Patent Document 3] JP 2006-083070 A [Patent Document 4] JP 2012-206984 A Summary of the Invention [Problem to be solved by the invention]

[0006] As mentioned above, various ingredients have been reported to have the effect of suppressing the production of IL-8 and collagenase, but more effective ingredients are being searched for.

[0007] Under these circumstances, an object of the present invention is to provide a novel composition having an inhibitory effect on IL-8 production or collagenase production, and applications thereof. [Means for solving the problem]

[0008] As a result of intensive research aimed at solving the above problems, the present inventors discovered that strawberry leaf extract has excellent inhibitory effects on IL-8 production and collagenase production, thereby completing the present invention.

[0009] That is, the present invention relates to the following inventions. <1> A composition for inhibiting IL-8 production and / or collagenase production, comprising strawberry leaf extract. <2> The strawberry leaf extract is a solvent extract of strawberry leaves using an extraction solvent consisting of water and / or ethanol. <1> The composition described in <3> For medicinal purposes, <1> or <2> The composition described in <4> For external use on the skin (excluding medical uses), <1> or <2> The composition described in <5> For oral use (excluding medical use); <1> or <2> The composition described in <6> A method for producing a composition for inhibiting IL-8 production and / or collagenase production, comprising an extraction step of obtaining a strawberry leaf extract by contacting strawberry leaves with an extraction solvent. Effect of the Invention

[0010] According to the present invention, there are provided novel compositions for suppressing IL-8 production and collagenase production, as well as applications thereof. [Brief description of the drawings]

[0011] [Figure 1] 1 is a graph showing the results of evaluating the effects of the strawberry leaf extracts of Examples 1 and 2 on IL-8 production in human 3-dimensionally cultured epidermis ((a): Example 1, (b): Example 2). [Diagram 2] FIG. 1 shows the results of evaluating the effects of the strawberry leaf extracts of Examples 1 and 2 on collagenase (MMP-1) production in three-dimensionally cultured human epidermis ((a): Example 1, (b): Example 2). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] The present invention will be described in detail below with reference to examples, but the present invention is not limited to the following examples, and can be modified as desired without departing from the gist of the present invention. In this specification, the expression "~" is used to include the numerical or physical values ​​before and after it. In this specification, the expression "A and / or B" includes "A only," "B only," and "both A and B."

[0013] <1. Composition for inhibiting IL-8 production and / or collagenase production> The present invention relates to a composition for suppressing IL-8 production and / or collagenase production, which contains a strawberry leaf extract (hereinafter, sometimes simply referred to as the "composition of the present invention").

[0014] The composition of the present invention exhibits excellent effects in suppressing IL-8 production and collagenase production due to the strawberry leaf extract contained therein. Therefore, based on the IL-8 production inhibitory effect and / or collagenase production inhibitory effect of the strawberry leaf extract, the composition of the present invention can be used as a "composition for suppressing IL-8 production" or a "composition for suppressing collagenase production", or even a "composition for suppressing IL-8 production and a composition for suppressing collagenase production".

[0015] Interleukin-8 (IL-8) is a proinflammatory cytokine, classified as a CXC chemokine, and exhibits chemotactic activity against leukocytes. It is a polypeptide with a molecular weight of approximately 8KDa that is produced by various cells such as fibroblasts and endothelial cells. IL-8 also has neutrophil migration activity, promotes inflammation, and promotes the degradation of elastin. IL-8 is an inflammatory cytokine that increases with age, damages epidermal stem cells, and promotes cellular senescence.

[0016] As used herein, the term "IL-8 production-inhibiting effect" refers to an effect of suppressing IL-8 production, and can be evaluated based on whether IL-8 production is suppressed or not. The IL-8 production-inhibiting effect includes not only complete suppression of IL-8 production, but also partial suppression of IL-8 production, such as suppression of excessive IL-8 production. Furthermore, the composition of the present invention has the effect of suppressing IL-8 production, and can be suitably used as a composition for suppressing IL-8 production. The inhibitory effect of the composition of the present invention on IL-8 production can be evaluated by the evaluation method (2-2.) described in the Examples below.

[0017] Collagenase is a protein or polypeptide having collagen decomposition activity, and is an enzyme that decomposes collagen produced by animal tissue cells, inflammatory cells, tumor cells, and the like. The collagenase to be treated is a matrix metalloprotease (MMP). For example, MMP-1, MMP-2, MMP-8, MMP-9, MMP-13, etc. may be used. At least one of these may be treated. Among these, MMP-1 is a particularly suitable target.

[0018] As used herein, the term "collagenase production inhibitory effect" refers to an effect of inhibiting collagenase production, and can be evaluated based on whether or not gene expression of the target collagenase is inhibited. The collagenase production inhibitory effect includes not only complete inhibition of collagenase production, but also partial inhibition of collagenase production, such as inhibition of collagenase overproduction. Furthermore, the composition of the present invention has the effect of inhibiting collagenase production and can be suitably used as a composition for inhibiting collagenase production. The collagenase production inhibitory effect of the composition of the present invention can be evaluated by the evaluation method (2-3) described in the Examples below.

[0019] The composition of the present invention has an inhibitory effect on IL-8 production and collagenase production. Based on this, it has an inhibitory effect on inflammation and the like, and can prevent or improve various symptoms, and can be used in pharmaceutical and non-pharmaceutical cosmetic and other external skin applications, and in oral forms such as foods and supplements.

[0020] The composition of the present invention will now be described in further detail.

[0021] The "strawberry" used as the raw material for the composition of the present invention is a plant of the genus Fragaria in the family Rosaceae, cultivated for its edible fruit. In the present invention, the "strawberry leaf" includes all parts of the strawberry except the root and the receptacle, i.e., the leaf, stem, bud, runner, and crown.

[0022] The variety, brand, and place of origin of the raw strawberry are not particularly limited, and any strawberry can be used. Specific examples include, but are not limited to, white strawberry, Ichigosan, Sagahonoka, Toyonoka, Tochiotome, Amaou, Akihime, Asukarubee, Aiberry, Tochihime, Red Pearl, Sachinoka, Benihoppe, and Momoichigo. These may be used alone or in combination of two or more.

[0023] Strawberry leaves are usually discarded after fruit harvest, so in the present invention, strawberry leaves after fruit harvest may be collected and used. Strawberry leaves can be used as they are for the solvent extract without prior treatment, but in order to increase the extraction efficiency, they are usually used in the form of fine particles by slicing, crushing, squeezing, grating, etc.

[0024] In the present invention, the term "extract" refers to a general concept of a form in which the content of active ingredients has been increased by compressing or solvent extraction of the target raw plant, or a product thereof that has been pretreated (dried or chopped) as necessary.

[0025] The strawberry leaf extract includes any of the extract obtained from strawberry leaves as an extraction raw material, a diluted or concentrated solution of the extract, or a crude or purified product thereof. In addition, a dried product obtained by drying the extract is also considered to be an extract.

[0026] When the strawberry leaf extract is obtained by solvent extraction, the extraction solvent may be any solvent that can extract the components contained in strawberry leaves that exhibit the effect of inhibiting IL-8 production or collagenase production. Specific examples include water, ethanol, isopropyl alcohol, acetone, glycerin, ethylene glycol, butylene glycol, propylene glycol, methanol, dichloromethane, dichloroethane, chloroform, hexane, ethyl acetate, etc. The water used as the extraction solvent may be any water as long as it does not impair the effects of the present invention, and examples of the water include tap water, distilled water, mineral water, etc. These extraction solvents can be used alone or as a mixture of two or more of them. When a mixture of two or more solvents is used as the extraction solvent, the mixing ratio can be appropriately adjusted.

[0027] The extraction solvent may contain additives within the range that does not impair the effects of the present invention, such as a pH adjuster and a viscosity adjuster.

[0028] From the standpoint of extraction efficiency, safety to the environment and living organisms, and production costs, a suitable example of a strawberry leaf extract is a solvent extract of strawberry leaves using an extraction solvent consisting of water and / or ethanol. By using water or a mixed solvent of water and ethanol as the extraction solvent, a strawberry leaf extract having excellent IL-8 production inhibitory activity and collagenase production inhibitory activity can be obtained.

[0029] When a mixed solvent of water and ethanol is used as the extraction solvent, the mixing ratio of water and ethanol may be appropriately adjusted within a range in which the components that exhibit the IL-8 production inhibitory effect and collagenase production inhibitory effect contained in strawberry leaves can be extracted. For example, when the mixed solvent of water and ethanol is 100% by volume, the ratio of ethanol is 0.1 to 80% by volume.

[0030] The amount of extraction solvent relative to the raw material strawberry leaves is not particularly limited, but is usually about 1 to 100 times the amount of strawberry leaves by weight.

[0031] The method for extracting a solvent extract from strawberry leaves may be any method that can extract the components contained in strawberry leaves that exhibit IL-8 production inhibitory activity and collagenase production inhibitory activity. An example of a suitable method for producing a strawberry leaf extract includes contacting strawberry leaves with an extraction solvent and stirring for a certain period of time until the active ingredients are sufficiently extracted from the strawberry leaves into the extraction solvent. The extraction time is appropriately selected taking into consideration the type of extraction solvent and the ratio of strawberry leaves to the extraction solvent, and when the extraction solvent is a mixed solvent of water and ethanol, the extraction time can be, for example, 1 to 24 hours.

[0032] The extraction process is carried out at room temperature to avoid deterioration of quality, but it can be carried out under heated conditions to increase the extraction efficiency. If necessary, the solvent may be removed by vacuum concentration or freeze-drying to increase the proportion of active ingredients in the strawberry leaf extract.

[0033] In addition, filtration or centrifugation may be performed to remove residues contained in the extract. The obtained extract may be used as is, or may be subjected to treatments such as dilution, concentration, drying, purification, etc. according to conventional methods. If necessary, the solvent may be removed by concentration / drying such as vacuum concentration, freeze drying, spray drying, etc.

[0034] <2. Application of the composition of the present invention> In the composition of the present invention, the strawberry leaf extract of the present invention may be used as is, but is usually used in combination with other ingredients that do not impair the effects of the present invention. The other ingredients may be any ingredients that can be used safely by humans, and are appropriately determined taking into consideration the form of use (medicinal use, topical use on the skin, oral use) described below. The blending ratio of the other components can be appropriately selected and determined depending on the purpose as long as it does not impair the effect of the present invention.

[0035] Since the composition of the present invention has an inhibitory effect on IL-8 production and / or collagenase production, it can be used as a "composition for inhibiting IL-8 production" or a "composition for inhibiting collagenase production." In addition, the composition of the present invention can be used in any form depending on the purpose. That is, it can be used as a pharmaceutical composition, a composition for external application to the skin (such as a cosmetic composition) that does not fall under the category of a pharmaceutical composition, an oral composition (such as a supplement or functional food), etc.

[0036] When the composition of the present invention is used to suppress IL-8 production, the amount of strawberry leaf extract contained is appropriately determined within a range such that the required amount can be ingested, taking into consideration the type and severity of symptoms involving IL-8 production, the form of the composition of the present invention, and the method of use.

[0037] Furthermore, when the composition of the present invention is used to inhibit collagenase production, the amount of strawberry leaf extract contained is appropriately determined within a range such that the required amount can be ingested, taking into consideration the type and severity of symptoms involving collagenase production, the form of the composition of the present invention, and the method of use.

[0038] Pharmaceutical Composition One suitable form of the composition of the present invention is a pharmaceutical composition (hereinafter sometimes referred to as the "pharmaceutical composition of the present invention") containing an effective amount of strawberry leaf extract formulated together with a pharma- ceutical acceptable base material.

[0039] As used herein, the term "pharmaceutical composition" refers to a drug that is useful for at least one of preventing, treating, and improving symptoms of a target disease. In addition, as used herein, the term "pharmaceutical composition" includes not only pharmaceutical products but also quasi-drugs.

[0040] The pharmaceutical composition of the present invention may contain, in addition to the strawberry leaf extract, other drugs or any pharmacologically acceptable ingredients within a range that does not impair the efficacy of the composition.

[0041] The pharmaceutical composition of the present invention is useful for at least one of preventing, treating, and improving symptoms of diseases involving IL-8 production or collagenase production. That is, "prevention" includes suppressing and delaying the onset of the disease or symptoms. Furthermore, "treatment" includes improving and relieving the pathology of the disease or symptoms, and suppressing the progression of the disease or symptoms.

[0042] There is no limitation on the diseases or symptoms involving IL-8 production or collagenase production that are the subject of the pharmaceutical composition of the present invention, and specific examples include rheumatism, chronic inflammation, osteoarthritis, decreased elasticity of organs, myocardial infarction, periodontitis, etc.

[0043] The pharmaceutical composition of the present invention may be in any form as long as it contains strawberry leaf extract. The pharmaceutical composition of the present invention may be in the form of tablets, pills, granules, powders, liquids, suspensions, syrups, or capsules. The pharmaceutical composition of the present invention may be in the form of solids or liquids, or may be a combination of solids and liquids, and is not particularly limited. The administration method may be oral or parenteral. Suitable embodiments of the pharmaceutical composition of the present invention include oral agents and skin topical agents.

[0044] The pharmaceutical composition of the present invention can also be applied to animals other than humans, and therefore can also be made into a pharmaceutical composition for animals.

[0045] The amount of intake of the pharmaceutical composition of the present invention is not particularly limited as long as it is an amount that can exert the effects of the present invention, and is appropriately determined depending on the sex, weight, health condition, etc. of the subject to be ingested. The intake amount may be taken once or may be taken in several divided doses per day. In addition, from the viewpoint of prevention, treatment, and symptom improvement of the target disease, it is preferable to take the pharmaceutical composition of the present invention continuously.

[0046] <Forms other than pharmaceutical compositions> As mentioned above, the strawberry leaf extract may be incorporated into products other than pharmaceuticals and quasi-drugs. Among such forms, preferred examples include compositions for external application to the skin and compositions for oral administration (excluding pharmaceutical compositions).

[0047] (Composition for external use on the skin) One suitable form of the composition of the present invention is a composition for external application to the skin containing strawberry leaf extract (hereinafter, sometimes referred to as "the composition for external application to the skin of the present invention").

[0048] A cosmetic composition, which is a typical example of the composition for external application to the skin of the present invention, will be described below; however, the composition for external application to the skin of the present invention is not limited to a cosmetic composition.

[0049] The composition for external application to the skin of the present invention has excellent IL-8 production inhibitory activity and collagenase production inhibitory activity, and as described above, can prevent or improve symptoms associated with IL-8 production and collagenase production, and can provide cosmetic effects such as reducing skin firmness and elasticity, moisturizing the skin, wrinkle formation, and redness of the skin.

[0050] The skin external composition of the present invention can be made into a desired formulation by appropriately blending a conventional cosmetic base material. The form is not particularly limited, and examples thereof include lotion, milky lotion, gel, cream, pack, hair tonic, hair cream foundation, aqueous ointment, spray, etc. In addition, in the present invention, the cosmetic composition is a concept that also includes bath compositions such as bath additives, body soap, and shampoo.

[0051] In addition, any optional ingredient used in ordinary cosmetics, skin medicines, and bath products can be added to the composition for external use on the skin of the present invention, so long as it does not impair the effects of the present invention. Specific examples of such optional ingredients include thickening / gelling agents, antioxidants, ultraviolet absorbing agents, coloring agents, sequestering agents, preservatives, pH adjusting agents, fragrances, beeswax, etc. The blending ratio of these optional ingredients can be appropriately selected and determined depending on the purpose.

[0052] The ratio of strawberry leaf extract to be blended in the composition for external application to the skin is arbitrary, but the blending ratio is selected within a range that can contribute to at least one of the effects of inhibiting IL-8 production and collagenase production.

[0053] The composition for external application on the skin of the present invention can also be applied to animals other than humans, and therefore can also be made into a composition for external application on the skin for animals.

[0054] (Oral Composition) One suitable form of the composition of the present invention is an oral composition containing strawberry leaf extract (hereinafter, sometimes referred to as "oral composition of the present invention"). The oral composition of the present invention has excellent IL-8 production inhibitory effects and collagenase production inhibitory effects, and is therefore suitable for use in functional foods, supplements, and the like.

[0055] Hereinafter, functional foods, supplements, and food additives, which are typical examples of the oral composition of the present invention, will be described; however, the oral composition of the present invention is not limited to functional foods, supplements, and food additives.

[0056] The amount of intake of the oral composition of the present invention is not particularly limited as long as it is an amount that can exert the effects of the present invention, and is appropriately determined depending on the form of the oral composition of the present invention, the sex, weight, health condition, etc. of the subject to be ingested. The intake amount may be taken once or may be taken in several divided doses per day. In addition, in order to obtain the action or efficacy of the oral composition of the present invention, it is preferable to continuously take the oral composition of the present invention.

[0057] The composition of the present invention can be mixed with various foods and beverages to form functional foods that can be easily taken orally on a daily basis, and can also be easily taken over a long period of time. The term "functional food" as used herein means, in addition to general foods, foods and / or beverages that are taken for the purpose of maintaining health, and is a concept that includes health functional foods (specified health foods, nutrient functional foods, or functional food), health foods, nutritional supplements, nutritional insurance foods, etc. Among these, health functional foods, such as specified health foods, nutrient functional foods, and functional food, are preferred forms of functional foods. When commercializing functional foods, various additives used in foods, specifically colorants, preservatives, thickening stabilizers, antioxidants, bleaching agents, antibacterial and antifungal agents, acidulants, seasonings, emulsifiers, strengthening agents, manufacturing agents, flavors, etc. may be added.

[0058] There is no particular limit to the foods and beverages that can be used as functional foods. For example, food products include dried powders, sausages, hams, processed seafood products, jellies, candies, chewing gums, and other foods. Beverages include various types of tea, green juice drinks, soft drinks, alcoholic beverages, and nutritional drinks.

[0059] The oral composition of the present invention can also be used in the form of a supplement. The form of the supplement is not particularly limited, and may be any form such as a tablet, powder, granule, capsule, sugar-coated tablet, film, lozenge, chewable, solution, emulsion, suspension, etc. The supplement of the present invention may contain any ingredient that is usually used as a supplement, in addition to the composition of the present invention. Such ingredients include, for example, amino acids, peptides, vitamins such as vitamin E, vitamin C, vitamin A, vitamin B, and folic acid, minerals, sugars, inorganic salts, citric acid or its salts, tea extract, oils and fats, tonic ingredients such as propolis, royal jelly, and taurine, herbal extracts such as ginger extract and ginseng extract, and herbs such as collagen.

[0060] The strawberry leaf extract may be incorporated into functional foods or supplements at any ratio, but the ratio is selected within a range that can contribute to at least one of the IL-8 production inhibitory effect and collagenase production inhibitory effect.

[0061] When the oral composition of the present invention is used as a health food, the action or efficacy brought about by the strawberry leaf extract of the present invention may be indicated on the product, for example, on the main body, packaging, container, package, or other storage items of the product of the present invention, or on its advertisements.

[0062] In addition, examples of claims regarding effects or efficacy that may be added to a product when it is commercialized include "maintains skin elasticity," "improves skin firmness," "prevents loss of skin firmness," "prevents wrinkles," "retains skin moisture," "maintains blood vessel flexibility," "improves redness associated with inflammation," "maintains joint health," "maintains dental health," "maintains gum health," etc.

[0063] In addition, the composition of the present invention can be used as a food additive either by itself or by adding other ingredients to it. There is no particular limitation on the other ingredients as long as they can be used as food and beverage additives. There is no particular limitation on the beverages and foods to which the food additive is added.

[0064] The composition of the present invention may be mixed into a food additive at any ratio, but the ratio is selected within a range that can contribute to at least one of the effects of inhibiting IL-8 production and the effect of inhibiting collagenase production.

[0065] The oral composition of the present invention can also be applied to animals other than humans, and can therefore be used as an oral composition for animals, such as pet food. EXAMPLES

[0066] The present invention will be described in more detail below with reference to examples, but the present invention is not limited thereto. In addition, the "%" shown in the mixed solvent in the examples indicates "volume %" unless otherwise specified. In addition, when the extraction solvent is a mixed solvent, the balance of water is omitted. For example, when "50% ethanol" is written, it means a mixed solvent of 50 volume % ethanol and 50 volume % water.

[0067] 1. Preparation of Strawberry Leaf Extract The strawberry leaf extracts of Examples 1 and 2 were produced by the following method.

[0068] Example 1 The raw material used was strawberry leaves (all parts except roots and receptacles (leaves, stems, buds, runners, crowns, etc.)) of "White Jewel (White Strawberry)" (registered trademark) (provided by Teshima Farm, Karatsu City). First, strawberry leaves were washed with water, dried, and powdered. 50% (v / v) aqueous ethanol was added to the obtained strawberry leaf powder, and the mixture was stirred at room temperature for 4 hours. The supernatant was collected by centrifugation and filtered under suction to obtain an extract from which insoluble matter was removed. This extract was dried using an evaporator and a freeze dryer, and the obtained solid matter was used as a strawberry leaf extract. The strawberry leaf extract was dissolved in 50% ethanol before use to obtain the strawberry leaf extract of Example 1.

[0069] Example 2 The strawberry leaf extract of Example 2 was obtained in the same manner as in Example 1, except that "Ichigosan" (registered trademark) strawberry leaves (provided by the Saga Prefectural Agricultural Research Center) were used as the raw material.

[0070] 2. Evaluation method 2-1. Test using human 3D cultured epidermis (evaluation of cytotoxicity) Human 3D cultured epidermis LabCyte EPI-MODEL24 (J-TEC) was pre-cultured in assay medium (J-TEC). After 18 hours, the culture cup was transferred to assay medium (control), assay medium containing the strawberry leaf extract of Example 1 or Example 2, and cultured for 4 hours in a CO2 incubator. After that, UVA (30 J / cm 2 ; Lamp TOSHIBA SH1002MA, Filter ASAHI SPECTRA SH0385 LU0325), and the culture cup was transferred to a new assay medium or an assay medium containing the strawberry leaf extract of Example 1 or Example 2, and cultured in a CO2 incubator. After 48 hours of culture, the IL-8 concentration in the culture medium was measured by enzyme antibody method using IL-8 Human Uncoated ELISA Kit (Invitrogen). Collagenase (MMP-1) in the culture medium was detected by Western blotting using mouse anti-human MMP-1 antibody (Santa cruz) and HRP-labeled goat anti-mouse IgG antibody (Bethyl). Although not shown, MTT assay (cell survival assay) was performed using human 3D cultured epidermis, and it was confirmed that there was no effect on cell survival rate in all groups.

[0071] 2-2. Evaluation of IL-8 production inhibitory effect (enzyme antibody method) The effects of the strawberry leaf extracts of Examples 1 and 2 on IL-8 production were investigated using human 3-dimensional cultured epidermis LabCyte EPI-MODEL24. After 48 hours of culture, the IL-8 concentration in the culture medium in each well of the culture plate was measured using an IL-8 Human Uncoated ELISA Kit (Invitrogen) according to the manufacturer's protocol. The absorbance of each well was measured at wavelengths of 450 nm and 570 nm, and the IL-8 concentration was calculated from the obtained measurement results.

[0072] 2-3. Evaluation of collagenase production inhibitory effect (Western blotting method) Next, the effects of the strawberry leaf extracts of Examples 1 and 2 on collagenase (MMP-1) production were examined by Western blotting. The culture medium from each well of the culture plate was collected, and collagenase (MMP-1) was detected by Western blotting using mouse anti-human MMP-1 antibody and HRP-labeled goat anti-mouse IgG antibody.

[0073] 3. Evaluation results 3-1.Effect of strawberry leaf extract on IL-8 production The results are shown in Figures 1(a) and 1(b). As shown in Figure 1(a), it was found that the addition of the strawberry leaf extract of Example 1 reduced IL-8 production induced by UVA irradiation in a concentration-dependent manner. Also, as shown in Figure 1(b), it was found that the addition of the strawberry leaf extract of Example 2 reduced IL-8 production induced by UVA irradiation in a concentration-dependent manner. From this, it was revealed that the strawberry leaf extract suppresses IL-8 production and exhibits an anti-inflammatory effect on human 3D cultured epidermis.

[0074] 3-2.Effect of strawberry leaf extract on collagenase production The results are shown in Figures 2(a) and 2(b). As shown in Figure 2(a), it was confirmed that collagenase production was significantly reduced by adding the strawberry leaf extract of Example 1. Also, as shown in Figure 2(b), it was confirmed that collagenase production was significantly reduced when the strawberry leaf extract of Example 2 was added. This demonstrated that the strawberry leaf extract inhibits collagenase production and inhibits the onset of collagen degradation. [Industrial Applicability]

[0075] The composition of the present invention has excellent IL-8 production inhibitory effects and collagenase production inhibitory effects, and is therefore effective for improving skin firmness and elasticity, wrinkles, moisturizing, etc. In addition, the composition of the present invention can be suitably used in various pharmaceutical compositions, compositions for external application to the skin, compositions for oral administration, etc.

Claims

1. A composition for inhibiting collagenase production, comprising a strawberry leaf extract.

2. The composition according to claim 1, wherein the strawberry leaf extract is a solvent extract of strawberry leaves using an extraction solvent consisting of water and / or ethanol.

3. The composition according to claim 1 or 2, which is for pharmaceutical use.

4. 3. The composition according to claim 1 or 2, which is for external application to the skin (excluding cases where it is for pharmaceutical use).

5. The composition according to claim 1 or 2, which is for oral use (excluding cases where it is for pharmaceutical use).

6. A method for producing a composition for inhibiting collagenase production, comprising an extraction step of obtaining a strawberry leaf extract by contacting strawberry leaves with an extraction solvent.

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