Neutrophil elastase inhibitor and composition for prevention and / or improvement of wrinkles

Abstract

To provide a composition suitable for oral intake that can prevent and / or improve wrinkles.SOLUTION: A tomato seed extract is used as an active ingredient of a neutrophil elastase inhibitor. The neutrophil elastase inhibitor serves as an active ingredient of a composition for the prevention and / or improvement of wrinkles and is particularly effective against wrinkles around the outer corners of the eyes. In a preferred embodiment, the composition is administered orally.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a neutrophil elastase inhibitor and a composition for preventing and / or improving wrinkles. [Background technology]

[0002] Wrinkles are one of the signs of skin aging and are easily noticeable, significantly affecting the overall appearance of the face. As a result, there is a great deal of interest in wrinkles and how to prevent and improve them. In the past, moisturizing with topical skin preparations containing water-retaining polymers such as mucopolysaccharides and collagen has been used to improve wrinkles, but this alone has not been sufficient to improve wrinkles. Furthermore, recent research has revealed that in addition to aging as an important factor in skin aging symptoms, factors such as dryness, oxidation, glycation, and ultraviolet radiation also play a significant role. These factors are thought to deteriorate fibroblasts, the primary cells in the dermis, damaging the extracellular matrix and causing skin to lose elasticity, resulting in wrinkles. Elastin is one of the major components of the extracellular matrix, which forms cross-linked structures to maintain the elasticity of skin tissue. Skin irritation, such as ultraviolet exposure, is known to denature and degrade elastin by overexpressing and activating the elastase enzyme, resulting in a loss of skin firmness and elasticity and promoting wrinkle formation. Therefore, in recent years, it has been proposed to improve wrinkles by inhibiting elastase activity, with the aim of restoring skin elasticity in fibroblasts. For example, Patent Document 1 discloses the use of a compound that inhibits the enzymatic activity of neutrophil elastase as an active ingredient in a composition for preventing and / or improving wrinkles.

[0003] Recent research has revealed that the fruits and seeds of tomatoes (Solanum lycopersicum), a vegetable commonly consumed in daily meals, possess a variety of functional properties. For example, Patent Document 2 discloses that lycoperosides, saponin components inherently contained in tomato seeds, have the effect of increasing dermal thickness and improving collagen density, and proposes using lycoperosides as active ingredients in oral skin-beautifying compositions. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Patent No. 6554752 [Patent Document 2] Patent No. 7405543 Summary of the Invention [Problem to be solved by the invention]

[0005] While various studies have been conducted on technologies for preventing or improving wrinkles through transdermal administration of cosmetics and other substances, there has been insufficient research into compositions and their active ingredients that can prevent or improve wrinkles through oral ingestion. In particular, there is a great demand for prevention and improvement of wrinkles that are not caused by dryness but are primarily caused by degeneration of the extracellular matrix, such as those that occur around the corners of the eyes, and therefore there is an urgent need for research into this issue. In view of the above circumstances, an object of the present invention is to provide a composition suitable for oral ingestion that can prevent and / or improve wrinkles. [Means for solving the problem]

[0006] As a result of intensive research conducted by the present inventors to solve the above problems, it has been found that tomato seed extract, particularly the lycoperosides contained therein, is an orally ingestible component and has an effect on neutrophil elastase. The inventors have discovered that neutrophil elastase inhibitory activity can prevent and / or improve wrinkles, leading to the completion of the present invention.

[0007] That is, the present invention is as follows. [1] A neutrophil elastase inhibitor containing tomato seed extract. [2] The neutrophil elastase inhibitor described in [1], wherein the tomato seed extract contains lycoperosides. [3] A composition for preventing and / or improving wrinkles, comprising the neutrophil elastase inhibitor described in claim [1] or [2]. [4] The composition according to [3], wherein the wrinkles are crow's feet. [5] The composition described in [3] or [4], which is taken orally. [6] The composition described in [6], which is a functional food. [Effects of the Invention]

[0008] The present invention provides a new use of tomato seed extract for inhibiting neutrophil elastase. It also provides a neutrophil elastase inhibitor and a composition for preventing and / or improving wrinkles. The composition is suitable for oral administration and can be easily and continuously incorporated into daily life. Furthermore, the composition is particularly effective against wrinkles around the eyes, meeting consumer demand. [Brief explanation of the drawings]

[0009] [Figure 1] Graph showing the neutrophil elastase inhibitory activity of test substances (tomato seed extract or lycoperoside H), with the horizontal axis representing the amount of test substance added. [Figure 2] 1 is a graph showing the change in the wrinkle grade of the right corner of the eye as judged by image analysis in the tomato seed extract group over the test period. [Figure 3] 1 is a graph showing the change in the wrinkle grade of the left corner of the eye as judged by image analysis in the tomato seed extract group over the test period. [Figure 4]1 is a graph showing the change in the number of wrinkles (right face) in skin condition photographs taken in the tomato seed extract group over the test period. [Figure 5] 1 is a graph showing the change in VAS questionnaire results regarding crow's feet wrinkles over the test period in the tomato seed extract group. DETAILED DESCRIPTION OF THE INVENTION

[0010] The present invention is characterized in that a tomato seed extract is used as the main component (hereinafter also referred to as the active ingredient) that exerts its effects. The tomato (scientific name: Solanum lycopersicum, English: Tomato) is a plant of the Solanaceae family, genus Solanum, native to the Andes highlands of South America (Peru, Ecuador). It is a perennial plant, and its fruit is edible. It is a type of green-yellow vegetable. In Japanese, it is also known by other names such as toushi (tang persimmon), akanasu (red eggplant), banka (bastard eggplant), koganeuri (small golden gourd), and sangojunasu (coral tree eggplant). In the present invention, the active ingredient is obtained and used from the seeds of such tomato plants. Tomato seeds contain lycoperosides, which are inherent saponin components, such as lycoperoside A and lycoperoside H.

[0011] [ka]

[0012] [ka]

[0013] The tomato seed extract of the present invention can be obtained from tomato seeds by any method, such as stirring extraction, continuous extraction, immersion extraction, or countercurrent extraction, and can be extracted using any device at room temperature or under reflux. In this case, examples of extraction solvents used to obtain the extract include water, lower monohydric alcohols (methyl alcohol, ethyl alcohol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohols (glycerin, propylene glycol, 1,3-butylene glycol, etc.), lower esters (ethyl acetate, etc.), hydrocarbons (benzene, hexane, pentane, etc.), ketones (acetone, methyl ethyl ketone, etc.), ethers (diethyl ether, tetrahydrofuran, dipropyl ether, etc.), and acetonitrile, and one or more of these can be used. The tomato seed extract thus extracted generally contains lycoperosides, which are suitable as the active ingredient in the present invention.

[0014] Specific extraction methods that can extract lycoperosides at high concentrations include, but are not limited to, a method using aqueous ethanol or aqueous methanol at a concentration of 0 to 100% (v / v), extracting at room temperature or by heating for 1 to 10 hours, and then filtering.

[0015] The tomato seed extract used in the present invention may be a commercially available product, preferably one containing lycoperosides, such as "Licolift (registered trademark)" manufactured by Oryza Oil & Fat Chemical Co., Ltd.

[0016] The content of tomato seed extract in the agent of the present invention may be 0.0001 to 100% by weight in dry weight equivalent to the total weight of the agent, and may be 0.0000005 to 0.5% by weight in lycoperosides equivalent to the total weight of the agent. The content of the tomato seed extract in the composition of the present invention may be 1 to 20% by weight, calculated as dry weight, based on the total weight of the composition, and may be 0.005 to 0.1% by weight, calculated as lycoperosides, based on the total weight of the composition. By setting the content within the above range, the desired effect can be easily obtained and the degree of freedom in formulation design can be ensured. The above content can be adjusted appropriately depending on the administration route and the type of composition to be contained, as described below.

[0017] As shown in the Examples below, tomato seed extract and lycoperosides have the activity of inhibiting neutrophil elastase activity. Here, "inhibition of enzyme activity" means that enzyme activity is reduced by preferably 90% or less, more preferably 80% or less, even more preferably 70% or less, and particularly preferably 60% or less when the active ingredient is applied compared to when the active ingredient is not applied. Tomato seed extract and lycoperosides have neutrophil elastase inhibitory activity and can therefore be effective ingredients for preventing and / or improving skin wrinkles. It is known that overexpression or activation of elastase in the skin leads to the denaturation and breakdown of elastin, promoting wrinkle formation. Inhibiting elastase activity can prevent the denaturation and breakdown of elastin. Here, "prevention" includes preventing or delaying the onset of wrinkles in a subject to which the composition is applied. Furthermore, "improvement" includes improving wrinkles, preventing or delaying the worsening of wrinkles, and reversing, preventing, or delaying the progression of wrinkles. It is known that pressure on the skin attracts neutrophils and increases neutrophil elastase activity, which accelerates the denaturation and breakdown of elastin and promotes wrinkle formation (Reference: https: / / www.pola-rm.co.jp / pdf / release_20191004_2.pdf). Therefore, wrinkles caused by neutrophil elastase activity are likely to occur in areas prone to facial pressure, such as the corners of the eyes. Therefore, the agents and compositions of the present invention, which can prevent and / or improve wrinkles through neutrophil elastase inhibitory activity, are particularly suitable for use in areas such as the corners of the eyes. In other words, the wrinkle-preventing and / or improving compositions of the present invention are suitable for use on wrinkles caused by neutrophil elastase activity and can be distinguished from their use on wrinkles caused by different factors, such as dryness.

[0018] The route of administration of the agent and composition of the present invention is not particularly limited and may be oral, transdermal, nasal, intravenous injection, etc., but oral administration or transdermal administration is more preferred. Here, "administration" may be substituted with "ingestion." The dosage is not particularly limited, but considering the desired effect and safety, it is generally recommended that adults take 0.5 to 5,000 mg / day in dry weight equivalent of tomato seed extract or 0.0025 to 25 mg / day in lycoperosides equivalent, in one or several divided doses. In addition to single doses, continuous or intermittent administration over several weeks to several months is also preferred.

[0019] When the agent or composition of the present invention is orally administered, it is preferably formulated as a food or drink. When the agent or composition of the present invention is in the form of a food or drink, ingredients that are usually used in the production of food or drink can be optionally blended therein. Examples of such optional ingredients include proteins, carbohydrates, fats, nutrients, seasonings, and flavorings. Examples of carbohydrates include monosaccharides, such as glucose and fructose; disaccharides, such as maltose, sucrose, and oligosaccharides; and polysaccharides, such as dextrin and cyclodextrin, and sugar alcohols, such as xylitol, sorbitol, and erythritol. Examples of flavorings that can be used include natural flavorings (thaumatin, stevia extract, and the like) and synthetic flavorings (saccharin, aspartame, and the like). Other additives commonly used in pharmaceutical compositions and commonly added to foods may also be used.

[0020] The form of the food or drink may be liquid, paste, solid, powder, granules, etc. Tablets, liquid food, feed, etc. are also included in the form of the food or drink.

[0021] It may also be contained in other general foods and beverages, for example, wheat flour products such as bread, macaroni, spaghetti, noodles, cake mix, fried chicken flour, and breadcrumbs; instant foods such as instant noodles, cup noodles, retort and cooked foods, canned foods, microwave foods, instant soups and stews, instant miso soup and clear soups, canned soups, and freeze-dried foods; processed agricultural products such as canned agricultural products, canned fruit, jams and marmalades, pickles, boiled beans, dried agricultural goods, and cereals (processed grain products); processed marine products such as canned seafood, fish ham and sausage, fish paste products, marine delicacies, and tsukudani (simmered fish paste); processed livestock products such as canned livestock products and pastes, and livestock ham and sausage; milk and dairy products such as processed milk, milk drinks, yogurt, lactic acid bacteria drinks, cheese, ice cream, modified milk powder, cream, and other dairy products; butter, marmalade, basic seasonings such as soy sauce, miso, sauces, processed tomato seasonings, mirin, vinegars, etc.; complex seasonings and foods such as cooking mixes, curry bases, sauces, dressings, noodle soup, spices, and other complex seasonings; frozen foods such as frozen ingredients, semi-cooked frozen foods, and cooked frozen foods; sweets such as caramel, candy, chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese sweets, rice snacks, bean snacks, and dessert sweets; beverages such as carbonated drinks, natural fruit juices, fruit juice drinks, soft drinks with fruit juice, fruit pulp drinks, fruit drinks with fruit particles, vegetable drinks, soy milk, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks, sports drinks, nutritional drinks, and alcoholic drinks; and foods other than these, the agent or composition of the present invention may be added.

[0022] The food and drink of the present invention may be in the form of ordinary foods, beverages, functional foods such as functional foods and foods for specified health uses, supplements, etc., with functional foods being particularly preferred. When the food or drink of the present invention is used to inhibit neutrophil elastase or to prevent and / or improve wrinkles and promote their production, the product may be labeled with information about its usefulness and functionality when it is commercialized. Such "indication" acts include all acts for informing consumers of the above-mentioned uses, and any expression that can evoke or infer uses such as "prevents wrinkles," "improves wrinkles," or "for those concerned with wrinkles around the eyes" falls under the category of "indication" acts of the present invention, regardless of the purpose of the indication, the content of the indication, the object or medium on which it is displayed, etc. Furthermore, it is preferable that the "labeling" be done in an expression that allows consumers to directly recognize the above-mentioned uses. Specific examples include acts of transferring, delivering, displaying for transfer or delivery, or importing food and beverage products or product packaging, containers, etc., on which the above-mentioned uses are written; displaying or distributing product advertisements, price lists, catalogs, pamphlets, POP, and other sales promotion materials at the point of sale, or transaction documents, or providing information containing the above-mentioned uses via electromagnetic means (such as the Internet). Furthermore, if the food and beverage of the present invention is approved under various government systems for health claim foods and is implemented under that approval, it is preferable to label it in a manner consistent with that approval.

[0023] Preferred embodiments of transdermal administration of the agent or composition of the present invention are topical skin preparations such as cosmetics, quasi-drugs, and pharmaceuticals. The formulation of topical skin compositions is not particularly limited, and examples include lotion formulations, emulsion formulations such as lotions and creams, oil formulations, gel formulations, and pack formulations.

[0024] When the agent or composition of the present invention is in the form of an external preparation for skin, the content of tomato seed extract is 0.0001 to 30% by weight in terms of dry weight based on the total amount of the composition, or lycoperosides When the amount is 0.0000005 to 0.15% by weight, the desired effect can be easily obtained and the degree of freedom in formulation design can be ensured.

[0025] When the agent or composition of the present invention is in the form of a topical skin preparation, it may optionally contain, in addition to tomato seed extract, ingredients that are typically incorporated into topical skin compositions, as long as the effects of the present invention are not impaired. Such ingredients include, for example, oils and waxes such as macadamia nut oil, avocado oil, corn oil, olive oil, rapeseed oil, sesame oil, castor oil, safflower oil, cottonseed oil, jojoba oil, coconut oil, palm oil, liquid lanolin, hydrogenated coconut oil, hydrogenated oil, Japan wax, hydrogenated castor oil, beeswax, candelilla wax, carnauba wax, Ibota wax, lanolin, reduced lanolin, hard lanolin, and jojoba wax; liquid paraffin, squalane, pristane, ozokerite, paraffin, Hydrocarbons such as ceresin, petrolatum, and microcrystalline wax; higher fatty acids such as oleic acid, isostearic acid, lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, and undecylenic acid; higher alcohols such as cetyl alcohol, stearyl alcohol, isostearyl alcohol, behenyl alcohol, octyldodecanol, myristyl alcohol, and cetostearyl alcohol; cetyl isooctanoate, isopropyl myristate, hexyldecyl isostearate, diisopropyl adipate, di-2-ethylhexyl sebacate, cetyl lactate, diisostearyl malate, ethylene glycol di-2-ethylhexanoate, neopentyl glycol dicaprate, di-2-heptylundecanoate, glycerin tri-2-ethylhexanoate, glycerin tri-2-ethylhexanoate, trimethylolpropane tri-2-ethylhexanoate, trimethylolpropane triisostearate, and pentane erythritol tetra-2-ethylhexanoate. synthetic ester oils such as ritt; chain polysiloxanes such as dimethylpolysiloxane, methylphenylpolysiloxane, and diphenylpolysiloxane; cyclic polysiloxanes such as octamethylcyclotetrasiloxane, decamethylcyclopentasiloxane, and dodecamethylcyclohexanesiloxane; oils such as silicone oils and modified polysiloxanes such as amino-modified polysiloxanes, polyether-modified polysiloxanes, alkyl-modified polysiloxanes, and fluorine-modified polysiloxanes;

[0026] Anionic surfactants such as fatty acid soaps (sodium laurate, sodium palmitate, etc.), potassium lauryl sulfate, and alkyl sulfate triethanolamine ether; cationic surfactants such as stearyltrimethylammonium chloride, benzalkonium chloride, and laurylamine oxide; imidazoline-based amphoteric surfactants (2-cocoyl-2-imidazolinium hydroxide-1-carboxyethyloxy disodium salt, etc.), and betaine-based surfactants (alkyl betaine, amido betaine, sulfobetaine, etc.). ), amphoteric surfactants such as acylmethyl taurine; sorbitan fatty acid esters (sorbitan monostearate, sorbitan sesquioleate, etc.), glycerin fatty acids (glycerin monostearate, etc.), propylene glycol fatty acid esters (propylene glycol monostearate, etc.), hydrogenated castor oil derivatives, glycerin alkyl ether, POE sorbitan fatty acid esters (POE sorbitan monooleate, polyoxyethylene sorbitan monostearate, etc.), POE sorbit fatty acid esters ( POE-sorbitol monolaurate, etc.), POE glycerin fatty acid esters (POE-glycerin monoisostearate, etc.), POE fatty acid esters (polyethylene glycol monooleate, POE distearate, etc.), POE alkyl ethers (POE 2-octyldodecyl ether, etc.), POE alkyl phenyl ethers (POE nonylphenyl ether, etc.), Pluronic types, POE·POP alkyl ethers (POE·POP 2-decyltetradecyl ether, etc.), Tetronics, POE hydroxypropyl ethers Nonionic surfactants such as castor oil and hydrogenated castor oil derivatives (POE castor oil, POE hydrogenated castor oil, etc.), sucrose fatty acid esters, and alkyl glucosides; polyhydric alcohols such as polyethylene glycol, glycerin, 1,3-butylene glycol, erythritol, sorbitol, xylitol, maltitol, propylene glycol, dipropylene glycol, diglycerin, isoprene glycol, 1,2-pentanediol, 2,4-hexanediol, 1,2-hexanediol, and 1,2-octanediol;

[0027] Moisturizing ingredients such as sodium pyrrolidone carboxylate, lactic acid, sodium lactate, etc.; powders such as mica, talc, kaolin, synthetic mica, calcium carbonate, magnesium carbonate, anhydrous silicic acid (silica), aluminum oxide, barium sulfate, etc., which may be surface-treated; inorganic pigments such as red iron oxide, yellow iron oxide, black iron oxide, cobalt oxide, ultramarine, Prussian blue, titanium oxide, and zinc oxide, which may be surface-treated; pearlizing agents such as titanium mica, fish phosphorus foil, and bismuth oxychloride, which may be surface-treated; Red No. 202, Red No. 228, Red No. 226, Yellow No. 4, and Blue No. 404, which may be laked. Organic dyes such as Yellow No. 5, Red No. 505, Red No. 230, Red No. 223, Orange No. 201, Red No. 213, Yellow No. 204, Yellow No. 203, Blue No. 1, Green No. 201, Purple No. 201, and Red No. 204; organic powders such as polyethylene powder, polymethyl methacrylate, nylon powder, and organopolysiloxane elastomers; para-aminobenzoic acid-based UV absorbers; anthranilic acid-based UV absorbers; salicylic acid-based UV absorbers; cinnamic acid-based UV absorbers; benzophenone-based UV absorbers; sugar-based UV absorbers; 2-(2'-hydroxy-5'-t-octylphenyl)benzotriazole, 4-methoxy ... UV absorbers such as 4'-t-butyldibenzoylmethane;

[0028] Lower alcohols such as ethanol and isopropanol; vitamin A or its derivatives, vitamin B6 hydrochloride, vitamin B6 tripalmitate, vitamin B6 dioctanoate, vitamin B2 or its derivatives, vitamin B 12 , Vitamin B 15or their derivatives; vitamin E such as α-tocopherol, β-tocopherol, γ-tocopherol, vitamin E acetate, vitamin D, vitamin H, pantothenic acid, pantethine, pyrroloquinoline quinone, etc.; antibacterial agents (preservatives) such as methylparaben, ethylparaben, butylparaben, phenoxyethanol, etc.; anti-inflammatory agents such as glycyrrhizic acid derivatives, glycyrrhetinic acid derivatives, salicylic acid derivatives, hinokitiol, zinc oxide, allantoin, etc.; various extracts (e.g., Phellodendron bark, Oenothera biennis, etc.) Examples of effective ingredients include: Lithospermum root, peony, Swertia japonica, birch, sage, loquat, carrot, aloe, mallow, iris, grape, Job's tears, loofah, lily, saffron, Cnidium rhizome, Angelica acutiloba, St. John's wort, Ononis, garlic, chili pepper, tangerine peel, Angelica acutiloba, seaweed, etc.); activators such as royal jelly, photosensitizers, cholesterol derivatives, etc.; blood circulation promoters such as nonylic acid valenylamide, capsaicin, zingerone, tannic acid, etc.; antiseborrheic agents such as sulfur and thianthol; water-soluble polymers such as collagen and hyaluronic acid, etc.

[0029] When the agent or composition of the present invention is in the form of a topical skin preparation, it may optionally contain components capable of exerting cosmetic effects that are incorporated into typical topical skin compositions, as long as the effects of the present invention are not impaired. Examples of such components capable of exerting cosmetic effects include other wrinkle-reducing components, whitening components, and anti-inflammatory components. Other wrinkle-reducing ingredients include, for example, vitamin A or its derivatives such as retinol, retinal, retinoic acid, tretinoin, isotretinoin, tocopherol retinoate, retinol palmitate, and retinol acetate, as well as ursolic acid benzyl ester, ursolic acid phosphate, betulinic acid benzyl ester, benzilic acid phosphate, trifluoroisopropyloxopropylaminocarbonylpyrrolidinecarbonylmethylpropylaminocarbonylbenzoylaminoacetic acid sodium, niacinamide, tocopherol, and farnesyl acetate.

[0030] Examples of whitening ingredients include 4-n-butylresorcinol, ascorbic acid glucoside, 3-O-ethyl ascorbic acid, tranexamic acid, arbutin, 1-triphenylmethylpiperidine, 1-triphenylmethylpyrrolidine, 2-(triphenylmethyloxy)ethanol, 2-(triphenylmethylamino)ethanol, 2-(triphenylmethyloxy)ethylamine, triphenylmethylamine, triphenylmethanol, triphenylmethane, and aminodiphenylmethane, N-(o-toluoyl)cysteic acid, N-(m-toluoyl)cysteic acid, N-(p-toluoyl)cysteic acid, N-(p-methoxybenzoyl)cysteic acid, N-benzoyl-serine, and N-(p-methyl)-2-methyl-2-methyl-1 ... Examples of such benzoylserine include N-(p-ethylbenzoyl)serine, N-(p-methoxybenzoyl)serine, N-(p-fluorobenzoyl)serine, N-(p-trifluoromethylbenzoyl)serine, N-(2-naphthoyl)serine, N-(4-phenylbenzoyl)serine, N-(p-methylbenzoyl)serine methyl ester, N-(p-methylbenzoyl)serine ethyl ester, N-(2-naphthoyl)serine methyl ester, N-benzoyl-O-methylserine, N-(p-methylbenzoyl)-O-methylserine, N-(p-methylbenzoyl)-O-acetylserine, N-(2-naphthoyl)-O-methylserine, dexpanthenol W, and niacinamide.

[0031] Anti-inflammatory ingredients include kurarinone, glabridin, glycyrrhizinic acid and its salts, alkyl glycyrrhetinate and its salts, glycyrrhetinic acid and its salts, pantothenyl alcohol, niacinamide, tranexamic acid, thiotaurine, hypotaurine, and the like. [Example]

[0032] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples as long as it does not depart from the gist of the invention.

[0033] <Test Example 1> Measurement of neutrophil elastase activity inhibition rate Human neutrophil elastase (NEUTROPHIL ELASTASE ENZYME; obtained from Enzo Life Sciences) at a final concentration of 2.2 μU / mL and the test substance were added to each well of a 96-well plate and incubated at 37°C for 30 minutes. Wells without the test substance were also incubated in the same manner. Then, a peptide (Suc(OMe)-Ala-Ala-Pro-Val-MCA; Peptide Institute), a substrate for human neutrophil elastase, was added to a final concentration of 25 μg / mL. Immediately after the addition of the substrate, the amount of the degradation product, 7-amino-4-methylcoumarin, was measured by fluorescence intensity measurement at an excitation wavelength of 360 nm and an emission wavelength of 460 nm using an Infinite M Plex (TECAN). The residual activity was calculated using the following formula (1) and used as an evaluation of the neutrophil elastase activity inhibition rate. Residual activity rate (%) = (1 - (measured value without test substance added - measured value with test substance) / (measured value without inhibitor added - blank)) × 100 Equation (1) The test substances used were tomato seed extract and lycoperoside H, added in amounts of 0.24 to 1000 μg / mL, with the amount of lycoperoside H added being a concentration equivalent to the content in the tomato seed extract.

[0034] The neutrophil elastase inhibitory activity of each test substance is shown in Figure 1. Both tomato seed extract and lycoperoside H exhibited concentration-dependent neutrophil elastase inhibitory activity.

[0035] <Test Example 2> Human double-blind parallel group comparative study A double-blind, parallel-group comparative study was conducted in humans to confirm the effects of oral ingestion of tomato seed extract on skin quality. Voluntary participants were divided into two groups (Table 1) and, under a double-blind condition, were asked to orally ingest either a test food (Licolift (registered trademark); manufactured by Oryza Oil & Fat Chemical Co., Ltd.) containing 50 mg of tomato seed extract per tablet, or a placebo food, two tablets per day, once daily for 12 weeks. The tomato seed extract intake was 100 mg / day in the tomato seed extract group and 0 mg / day in the placebo group. The test food or placebo food was taken with water or lukewarm water after breakfast (or in the morning if breakfast was not eaten). If participants forgot to take the morning intake, they could take it at any time on the same day. If they realized they had missed the intake the next day, they could take the previous day's intake all at once. We decided not to do this and to ask participants to maintain the recommended daily intake.

[0036] [Table 1]

[0037] Before the intake period, and after 4, 8, or 12 weeks of intake, digital photographs were taken and wrinkle grades were determined based on the photographs, and skin condition photographs and analysis, as well as VAS questionnaires, and other evaluations were conducted. After the test period, the blinding was lifted and statistical analysis was performed for each group.

[0038] The wrinkle grade was determined by the following procedure. The corners of the left and right eyes of the subject were photographed using a digital camera. Specifically, the subject was seated in a chair, their chin and forehead were fixed on a support stand, and with their eyes lightly closed, the corners of the left and right eyes were photographed from a 45-degree angle in front. Based on the photographs taken, the wrinkle grade was determined according to the following criteria published in the Journal of the Japanese Society of Cosmetic Chemistry, Vol. 30, No. 4, pp. 316-322. did. (Wrinkle grade standard) 0: No wrinkles 1: Slightly indistinct shallow wrinkles are observed 2: Slightly clear, shallow wrinkles are visible 3: Clear, shallow wrinkles are visible 4: A few slightly deeper wrinkles are visible among the clearly visible shallow wrinkles. 5: Somewhat deep wrinkles are observed 6: Clear, deep wrinkles are visible 7: Significantly deep wrinkles are observed

[0039] The skin condition was photographed and analyzed using the following procedure. The subject's face was photographed using a skin measurement device, VISIA-Evolution (registered trademark) (Canfield Scientific Inc.). The subject was wearing a gray cape and photographed with their eyes closed, accompanied by a color chart for image correction (CASMATCH). Three photographing modes were used: standard light, UV, and cross-polarization. One photograph was taken from the front, one from the left, and one from the right, in each mode. Based on the photographs obtained, a skin condition score, such as the number of wrinkles, was calculated using the device's analysis program.

[0040] The VAS questionnaire was conducted by asking the subjects to mark the position on a 10 cm line indicating the degree to which they felt wrinkles at the corners of their eyes, with the left end being "not felt at all" and the right end being "felt strongly."

[0041] Table 2 and Figures 2 and 3 show the results of the wrinkle grade assessment, Table 3 and Figure 4 show the results of the skin condition photographs, and Table 4 and Figure 5 show the results of the VAS questionnaire. During the test food intake period, significant improvement in wrinkles was observed after intake compared to before intake in all evaluations (Figures 2-5). Additionally, although no significant difference was observed between the tomato seed extract group and the placebo group, there was a tendency for wrinkles to improve in the tomato seed extract group.

[0042] [Table 2]

[0043] [Table 3]

[0044] [Table 4] [Industrial Applicability]

[0045] The present invention provides a new use of tomato seed extract, namely, inhibition of neutrophil elastase. Furthermore, the tomato seed extract can be contained as an active ingredient in a composition for preventing and / or improving wrinkles. Such a composition is suitable for oral administration and provides a wrinkle prevention and / or improvement composition that can be easily and continuously incorporated into daily life, making it extremely useful industrially.

Claims

1. Neutrophil elastase inhibitor containing tomato seed extract.

2. 2. The neutrophils of claim 1, wherein the tomato seed extract contains lycoperosides. Elastase inhibitors.

3. A composition for preventing and / or improving wrinkles, comprising the neutrophil elastase inhibitor according to claim 1 or 2.

4. The composition according to claim 3, wherein the wrinkles are crow's feet.

5. The composition of claim 3 , which is taken orally.

6. The composition according to claim 6, which is a food with functional claims.

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