Inhibitor of decreased moisturizing function caused by glycation, skin cosmetics, and food and beverages

Abstract

To provide an inhibitor for inhibiting moisture keeping function reduction caused by saccharization, by finding a component comprising action for inhibiting reduction of moisture keeping caused by saccharization or component comprising action for inhibiting reduction of skin function caused by saccharization, out of natural product derived components having high safety property, and including the component as an active ingredient, an inhibitor for inhibiting reduction of skin function caused by saccharization, and a skin cosmetic and food / drink in which the natural product derived component is blended.SOLUTION: As an active ingredient included in an inhibitor for inhibiting reduction of moisture keeping caused by saccharization, and an inhibitor for inhibiting reduction of skin function caused by saccharization, a wild thyme extract is used. In addition, the wild thyme extract is blended in a skin cosmetic and food / drink of the invention.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an agent for suppressing a decrease in moisturizing function caused by glycation and an agent for suppressing a decrease in epidermal function caused by glycation. The present invention also relates to skin cosmetics and foods and beverages used for suppressing a decrease in moisturizing function caused by glycation. [Background technology]

[0002] Carbohydrates are extremely important as an energy source for humans and other living organisms. However, they are also known to undergo glycation reactions with proteins. Glycation is a series of reactions that begins with a non-enzymatic reaction between the carbonyl group of carbohydrates and the amino group of proteins, etc., followed by the formation of Schiff bases, Amadori compounds, and finally the formation of advanced glycation end products (AGEs). Glycation non-enzymatically modifies proteins with sugars, causing denaturation and cross-linking between proteins, ultimately impairing their function. Glycation not only causes direct damage by modifying and structurally altering extracellular matrix proteins such as collagen, but also triggers cellular responses by being recognized by receptors that use glycosylated proteins as ligands.

[0003] For example, extracellular matrix components such as collagen account for more than half of the dry weight of tissues such as skin and bone. Therefore, for example, when collagen is glycated and abnormally cross-linked, it can cause a decrease in the skin's moisturizing function and elasticity, as well as dullness due to yellowing, and in bone and cartilage tissue, it can lead to the development of osteoporosis and osteoarthritis. Furthermore, because abnormally cross-linked collagen is less susceptible to degradation by collagenases, the expression of collagenases can be induced, leading to problems such as the degradation of normal collagen. Thus, protein glycation is thought to cause functional decline in tissues such as skin, bone, and cartilage.

[0004] Therefore, if functional decline caused by glycation can be suppressed, it is expected to be useful in preventing or improving loss of skin elasticity, dullness, etc.

[0005] N-phenacylthiazolium bromide has been known as a compound with anti-glycation activity (Non-Patent Document 1). However, this compound has safety issues and is not suitable as a pharmaceutical or topical skin preparation. Furthermore, extracts from mugwort, for example, are known as naturally occurring components with anti-glycation activity (Patent Document 1). [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Japanese Patent Application Laid-Open No. 2001-122758 [Non-patent literature]

[0007] [Non-Patent Document 1] Vasan S. et al.,Nature,July 18, 1996,Vol. 82,No. 6588,p.275-278 Summary of the Invention [Problem to be solved by the invention]

[0008] The present invention aims to find, from among highly safe naturally-derived ingredients, ingredients that have the effect of suppressing a decline in moisturizing function caused by glycation or a decline in epidermal function caused by glycation, and to provide an inhibitor of a decline in moisturizing function caused by glycation or an inhibitor of a decline in epidermal function caused by glycation that contains the ingredient as an active ingredient, as well as skin cosmetics and foods and beverages that contain the naturally-derived ingredient. [Means for solving the problem]

[0009] As a result of research conducted by the present inventors to solve the above problems, they found that wild thyme extract has excellent effects in suppressing the decline in moisturizing function caused by glycation and the decline in epidermal function caused by glycation, and thus completed the present invention. Specifically, the present invention is as follows.

[0010] [1] An agent for suppressing a decrease in moisturizing function caused by glycation, characterized by containing wild thyme extract as an active ingredient. [2] The agent for suppressing a decrease in moisturizing function due to glycation according to [1], which suppresses a decrease in expression due to glycation of one or more enzymes selected from the group consisting of filaggrin, serine palmitoyltransferase, and transglutaminase-1. [3] An agent for suppressing epidermal dysfunction caused by glycation, characterized by containing wild thyme extract as an active ingredient. [4] A skin cosmetic preparation containing wild thyme extract, characterized in that it is used to suppress a decrease in moisturizing function caused by glycation and / or a decrease in epidermal function caused by glycation. [5] A food or beverage containing wild thyme extract, characterized in that it is used to suppress a decrease in moisturizing function caused by glycation and / or a decrease in epidermal function caused by glycation. [Effects of the Invention]

[0011] According to the present invention, by using wild thyme extract as an active ingredient, it is possible to provide an inhibitor of moisturizing function decline due to glycation and an inhibitor of epidermal function decline due to glycation that are highly effective and safe. Furthermore, by incorporating wild thyme extract, it is possible to provide skin cosmetics and foods and beverages that are suitable for use in suppressing moisturizing function decline due to glycation and for suppressing epidermal function decline due to glycation. DETAILED DESCRIPTION OF THE INVENTION

[0012] Hereinafter, an embodiment of the present invention will be described. The inhibitor of moisturizing function decline caused by glycation and the inhibitor of epidermal function decline caused by glycation of this embodiment contain wild thyme extract as an active ingredient. In addition, the skin cosmetics and food and beverage products of this embodiment are used to suppress a decrease in moisturizing function due to glycation and / or a decrease in epidermal function due to glycation, and contain wild thyme extract.

[0013] Here, the "extract" in this embodiment includes an extract obtained using wild thyme as an extraction raw material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or any of these roughly purified or purified products.

[0014] The extraction raw material used in this embodiment is wild thyme (scientific name: Thymus serphyllum). Wild thyme (Thymus serphyllum, also known as Thymus serphyllum) is a plant belonging to the genus Thymus in the family Lamiaceae, native to Europe and Northwest Asia, where it grows widely and is easily available. Parts of the plant that can be used as raw materials for extraction include leaves, stems, flowers, aerial parts, and mixtures thereof, with aerial parts being preferred.

[0015] The wild thyme extract can be obtained by drying the raw material, pulverizing it using a crusher, or by subjecting it to extraction with an extraction solvent. Drying can be performed in the sun or using a commonly used dryer. The raw material can also be used after pretreatment, such as degreasing, with a nonpolar solvent such as hexane. Pretreatment, such as degreasing, allows for efficient extraction of the raw material with a polar solvent.

[0016] As the extraction solvent, it is preferable to use a polar solvent, such as water or a hydrophilic organic solvent, which is preferably used alone or in combination of two or more at room temperature or a temperature below the boiling point of the solvent.

[0017] Water that can be used as an extraction solvent includes pure water, tap water, well water, mineral water, hot spring water, spring water, fresh water, etc., as well as water that has undergone various treatments. Treatments that can be applied to water include, for example, purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, buffering, etc. Therefore, water that can be used as an extraction solvent in this embodiment also includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate-buffered physiological saline, etc.

[0018] Examples of hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic alcohols having 1 to 5 carbon atoms, such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones, such as acetone and methyl ethyl ketone; and polyhydric alcohols having 2 to 5 carbon atoms, such as 1,3-butylene glycol, propylene glycol, and glycerin.

[0019] When a mixture of two or more polar solvents is used as the extraction solvent, the mixing ratio can be adjusted appropriately. For example, when a mixture of water and a lower aliphatic alcohol is used as the extraction solvent, the mixing ratio of water to lower aliphatic alcohol is preferably 9:1 to 1:9 (volume ratio), and more preferably 7:3 to 2:8 (volume ratio). When a mixture of water and a lower aliphatic ketone is used, the mixing ratio of water to lower aliphatic ketone is preferably 9:1 to 2:8 (volume ratio), and when a mixture of water and a polyhydric alcohol is used, the mixing ratio of water to polyhydric alcohol is preferably 8:2 to 1:9 (volume ratio).

[0020] The extraction process is not particularly limited as long as it can dissolve the soluble components contained in the extraction raw material into the extraction solvent, and can be carried out according to conventional methods. For example, the extraction raw material is immersed in an extraction solvent in an amount (mass ratio) 5 to 15 times the amount of the extraction raw material, and the soluble components are extracted at room temperature or under reflux heating, followed by filtration to remove the extraction residue, thereby obtaining an extract. The solvent is distilled off from the obtained extract to obtain a paste-like concentrate, which is then further dried to obtain a dried product.

[0021] [Inhibitor of moisturizing function decline caused by glycation, inhibitor of epidermal function decline caused by glycation] The extract from wild thyme obtained as described above (hereinafter sometimes simply referred to as "wild thyme extract") has excellent inhibitory effects on the decline in moisturizing function caused by glycation and the decline in epidermal function caused by glycation, and can therefore be used as an active ingredient in an inhibitor of a decline in moisturizing function caused by glycation and an inhibitor of a decline in epidermal function caused by glycation. The inhibitor of a decline in moisturizing function caused by glycation and the inhibitor of a decline in epidermal function caused by glycation according to this embodiment can be used in a wide range of applications, such as pharmaceuticals, quasi-drugs, and cosmetics. The inhibitor of moisturizing function decline caused by glycation and the inhibitor of epidermal function decline caused by glycation according to this embodiment may be simply referred to as an "inhibitor of moisturizing function decline" and an "inhibitor of epidermal function decline," respectively. The same applies to their actions and uses.

[0022] Here, the effect of wild thyme extract in inhibiting the decline in moisturizing function is exerted, for example, based on its effect of inhibiting the decline in expression of moisturizing-related factors caused by glycation. Examples of moisturizing-related factors whose expression is decreased by glycation and whose decline in expression is inhibited by wild thyme extract include filaggrin, serine palmitoyltransferase (SPT), transglutaminase-1 (TGM1), etc. However, the effect of wild thyme extract in inhibiting the decline in moisturizing function is not limited to the effect exerted based on its effect of inhibiting the decline in expression of the moisturizing-related factors.

[0023] Furthermore, the effect of wild thyme extract in inhibiting the decline in epidermal function is exerted, for example, by its ability to inhibit the decline in expression of epidermal function-related factors due to glycation. Examples of epidermal function-related factors whose expression is decreased by glycation and whose decline in expression is inhibited by wild thyme extract include filaggrin, serine palmitoyltransferase (SPT), transglutaminase-1 (TGM1), etc. However, the effect of wild thyme extract in inhibiting the decline in epidermal function is not limited to the effect exerted based on its ability to inhibit the decline in expression of the epidermal function-related factors.

[0024] Wild thyme extract may be used as an active ingredient for an inhibitor of decreased filaggrin expression caused by glycation, an inhibitor of decreased SPT expression caused by glycation, or an inhibitor of decreased TGM1 expression caused by glycation, taking advantage of its inhibitory effect on decreased expression of moisturizing-related factors or epidermal function-related factors caused by glycation, particularly its inhibitory effect on decreased filaggrin expression caused by glycation, SPT expression caused by glycation, or TGM1 expression caused by glycation.

[0025] The moisturizing function decline inhibitor and epidermal function decline inhibitor of the present embodiment may consist of wild thyme extract alone, or may be a formulation of wild thyme extract.

[0026] The moisturizing function decline inhibitor or epidermal function decline inhibitor of this embodiment can be formulated into any dosage form, such as powder, granules, tablets, or liquid, using a pharmaceutically acceptable carrier such as dextrin or cyclodextrin, or any other auxiliary agent, according to a conventional method. In this case, examples of auxiliary agents that can be used include excipients, binders, disintegrants, lubricants, stabilizers, and flavoring and odor masking agents. The moisturizing function decline inhibitor and epidermal function decline inhibitor can be incorporated into other compositions (e.g., skin cosmetics) and used, and can also be used as ointments, topical liquids, patches, etc.

[0027] When the moisturizing function decline inhibitor or epidermal function decline inhibitor of this embodiment is formulated, the content of wild thyme extract is not particularly limited and can be set appropriately depending on the purpose.

[0028] In addition, the moisturizing function decline inhibitor or epidermal function decline inhibitor of this embodiment can be used as an active ingredient by blending other natural extracts, etc. that have the effect of inhibiting moisturizing function decline or epidermal function decline together with the wild thyme extract, if necessary.

[0029] The method of administering the moisturizing function decline inhibitor or epidermal function decline inhibitor of this embodiment to a patient includes transdermal administration, oral administration, etc., and a method suitable for prevention, treatment, etc. may be selected appropriately depending on the type of disease. Furthermore, the dosage of the moisturizing function decline inhibitor or epidermal function decline inhibitor of this embodiment may be increased or decreased appropriately depending on the type and severity of the disease, individual differences between patients, administration method, administration period, etc.

[0030] The moisturizing function decline inhibitor of this embodiment can suppress the decline in expression of moisturizing-related factors caused by glycation through the action of wild thyme extract as an active ingredient. Such moisturizing-related factors include, in particular, filaggrin, SPT, and TGM1. By the action of the wild thyme extract, the moisturizing function decline inhibitor of this embodiment can suppress the decline in skin's moisturizing function, preventing, treating, or ameliorating symptoms caused by a decline in moisturizing function due to glycation, such as dry skin, rough skin, fine wrinkle formation, and reduced stratum corneum flexibility, and ultimately preventing, treating, or ameliorating skin aging symptoms caused by glycation, such as wrinkle formation and reduced elasticity. However, in addition to these uses, the moisturizing function decline inhibitor of this embodiment can also be used for all uses in which it is meaningful to suppress the decline in expression of moisturizing-related factors caused by glycation.

[0031] The epidermal function decline inhibitor of this embodiment can suppress the decline in expression of epidermal function-related factors caused by glycation through the action of wild thyme extract as an active ingredient. Such epidermal function-related factors include, in particular, filaggrin, SPT, and TGM1. By the action of the wild thyme extract, the epidermal function decline inhibitor of this embodiment can suppress the decline in skin epidermal function and prevent, treat, or improve symptoms caused by glycation-induced decline in epidermal function, such as dry skin, rough skin, fine wrinkle formation, and reduced stratum corneum flexibility, and ultimately prevent, treat, or improve skin aging symptoms caused by glycation, such as wrinkle formation and reduced elasticity. However, in addition to these uses, the epidermal function decline inhibitor of this embodiment can also be used for all uses in which it is meaningful to inhibit the decline in expression of epidermal function-related factors caused by glycation.

[0032] The moisturizing function decline inhibitor or epidermal function decline inhibitor of this embodiment, or the aforementioned agent for inhibiting filaggrin expression decline, inhibits the decline in filaggrin expression caused by glycation in epidermal keratinocytes through the filaggrin expression decline inhibitory effect of wild thyme extract, thereby increasing the amount of amino acids in the stratum corneum and essentially improving the moisture environment of the stratum corneum. This reduces the risk of developing atopic diseases, including atopic dermatitis (eczema, skin inflammation, skin itching, etc.), allergies, and asthma, and can prevent or treat these diseases.

[0033] The moisturizing function decline inhibitor or epidermal function decline inhibitor of this embodiment, or the aforementioned TGM1 expression decline inhibitor or SPT expression decline inhibitor, can suppress the decline in skin barrier function caused by glycation through the TGM1 expression decline inhibitory effect or SPT expression decline inhibitory effect possessed by wild thyme extract, and can prevent, treat, or ameliorate dry skin diseases (e.g., atopic dermatitis, psoriasis, ichthyosis, etc.).

[0034] Furthermore, the moisturizing function decline inhibitor and epidermal function decline inhibitor of the present embodiment have excellent effects in inhibiting the moisturizing function decline or the epidermal function decline, respectively, and are therefore suitable for incorporation into, for example, external skin preparations. In this case, the wild thyme extract may be incorporated as is, or a moisturizing function decline inhibitor or epidermal function decline inhibitor formulated from the wild thyme extract may be incorporated.

[0035] Here, the topical skin preparation is not limited to any particular category, and includes a wide range of products such as skin cosmetics, which will be described later, as well as quasi-drugs and pharmaceuticals that are used transdermally.

[0036] Furthermore, since the moisturizing function decline inhibitor or epidermal function decline inhibitor of this embodiment has excellent moisturizing function decline inhibitory effect or epidermal function decline inhibitory effect, it can also be suitably used as a reagent for research into the mechanisms of these actions.

[0037] [Skin cosmetics] Wild thyme extract has an excellent effect of suppressing the decline in moisturizing function or epidermal function, and is therefore suitable for incorporation into skin cosmetics. In this case, the wild thyme extract may be incorporated as is, or a moisturizing function decline inhibitor or epidermal function decline inhibitor formulated from the wild thyme extract may be incorporated.

[0038] By blending wild thyme extract or the above-mentioned inhibitor of moisturizing function decline, or inhibitor of epidermal function decline, a skin cosmetic suitable for inhibiting the decline in moisturizing function or the decline in epidermal function can be produced.

[0039] The types of skin cosmetics that can be blended with wild thyme extract, or the above-mentioned moisturizing function decline inhibitor, or epidermal function decline inhibitor are not particularly limited, and examples include ointments, creams, emulsions, skin lotions, lotions, gels, cosmetic oils, packs, foundations, etc.

[0040] When wild thyme extract, or the above-mentioned moisturizing function decline inhibitor, or epidermal function decline inhibitor is incorporated into a skin cosmetic, the amount incorporated can be adjusted as appropriate depending on the type of skin cosmetic. A suitable incorporation rate is approximately 0.0001 to 10% by mass, and a particularly suitable incorporation rate is approximately 0.001 to 1% by mass, calculated as a standard extract.

[0041] The skin cosmetic of this embodiment can be used in combination with main ingredients, auxiliaries, or other ingredients typically used in the manufacture of skin cosmetics, such as astringents, bactericides / antibacterial agents, whitening agents, UV absorbers, moisturizers, cell activators, anti-inflammatory / antiallergic agents, antioxidants / active oxygen scavengers, oils and fats, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, fragrances, etc., as long as the effects of the wild thyme extract in inhibiting a decline in moisturizing function or epidermal function are not hindered. Such combinations can result in more versatile products, and the synergistic effects of the combined active ingredients can bring about effects superior to those normally expected.

[0042] The skin cosmetic of this embodiment can, through the effect of wild thyme extract in inhibiting a decline in moisturizing function and / or the effect of inhibiting a decline in epidermal function, prevent, treat, or improve a decline in moisturizing function caused by glycation; prevent, treat, or improve a decline in epidermal function caused by glycation; prevent, treat, or improve rough skin, dry skin, and dry skin diseases (e.g., atopic dermatitis, psoriasis, ichthyosis, etc.); and the like.

[0043] [Food and beverages] Wild thyme extract has an excellent effect of suppressing the decline in moisturizing function or epidermal function, and is therefore suitable for incorporation into foods and beverages. In this case, the wild thyme extract may be incorporated as is, or a moisturizing function decline inhibitor or epidermal function decline inhibitor formulated from the wild thyme extract may be incorporated. By incorporating wild thyme extract or the above-mentioned inhibitor of moisturizing function decline or inhibitor of epidermal function decline, a food or drink suitable for use in inhibiting the decline in moisturizing function or the decline in epidermal function can be produced.

[0044] Here, food and beverage products refer to products that are ingested orally or by gastrointestinal administration in normal social life and are unlikely to pose a risk to human health. They are not limited to administrative classifications such as food, medicine, or quasi-drug. Therefore, in this embodiment, "food and beverage products" broadly encompasses orally ingested general foods, health foods (functional food and beverage products), health functional foods (foods for specified health uses, foods with nutrient functions), quasi-drugs, medicines, etc. Food and beverage products according to this embodiment are preferably food and beverage products that can display the favorable effects of wild thyme extract on the food and beverage product or its packaging, and are particularly preferably health functional foods (foods for specified health uses, foods with functional claims, foods with nutrient functions), quasi-drugs, and medicines.

[0045] When wild thyme extract, or a moisturizing function decline inhibitor or epidermal function decline inhibitor formulated from wild thyme extract, is incorporated into a food or beverage, the amount of active ingredient can be varied as appropriate taking into account the intended use, symptoms, gender, etc., but taking into account the general intake of the food or beverage to which it is to be added, it is preferable to adjust the daily intake of the extract to about 1 to 1000 mg per adult. When the food or beverage to which it is to be added is in the form of granules, tablets, or capsules, the amount of wild thyme extract, or a moisturizing function decline inhibitor or epidermal function decline inhibitor added is usually 0.1 to 100% by mass, and preferably 5 to 100% by mass, of the food or beverage to which it is to be added.

[0046] The food and drink of this embodiment may be any food and drink containing wild thyme extract that does not interfere with its activity, or may be a nutritional supplement containing wild thyme extract as its main ingredient.

[0047] When producing the food and beverage of this embodiment, any auxiliary agent can be added, such as sugars such as dextrin and starch; proteins such as gelatin, soy protein and corn protein; amino acids such as alanine, glutamine and isoleucine; polysaccharides such as cellulose and gum arabic; and fats and oils such as soybean oil and medium-chain fatty acid triglycerides, to form the food and beverage into any shape.

[0048] The foods and beverages that can incorporate wild thyme extract are not particularly limited, and specific examples include beverages such as soft drinks, carbonated drinks, energy drinks, fruit drinks, and lactic acid drinks (including concentrated concentrates and powders for adjusting these beverages); frozen desserts such as ice cream, ice sherbet, and shaved ice; noodles such as soba, udon, vermicelli, gyoza wrappers, shumai wrappers, Chinese noodles, and instant noodles; sweets such as candy, chewing gum, candy, chewing gum, chocolate, tablet candy, snacks, biscuits, jelly, jam, cream, and baked goods; processed seafood and livestock foods such as kamaboko, ham, and sausage; dairy products such as processed milk and fermented milk; oils and fats and oil-based foods such as salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, and dressing; condiments such as sauces and dressings; soups, stews, salads, side dishes, and pickles; and various other forms of health and nutritional supplements; tablets, capsules, and energy drinks. When wild thyme extract is blended into these foods and beverages, commonly used auxiliary raw materials and additives can be used in combination.

[0049] The moisturizing function decline inhibitor, epidermal function decline inhibitor, skin cosmetics, and food and beverage products of this embodiment are suitable for use on humans, but can also be applied to animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as their respective effects are achieved. [Example]

[0050] The present invention will be described in detail below with reference to test examples, but the present invention is not limited to the following examples. In these test examples, a freeze-dried product (Sample 1) of Wild Thyme Extract BG (Maruzen Pharmaceutical Co., Ltd.) was used as the test sample.

[0051] [Test Example 1] Inhibitory effect of glycated collagen on the decrease in moisturizing-related gene expression in keratinocytes The wild thyme extract (Sample 1) was tested for its inhibitory effect on the decrease in moisturizing-related gene expression in keratinocytes on glycated collagen as follows.

[0052] 500 μL of collagen acidic solution I-AC (native collagen, bovine dermis, 3 mg / mL, manufactured by Koken Co., Ltd.) was added to a 35 mm dish and air-dried and allowed to stand for 1 hour. After that, 500 μL of glyceraldehyde (GA) solution (4.5 mg / mL) prepared with PBS was added and the dish was left to stand in an incubator at 37°C for 4 days to glycate type I collagen.

[0053] After washing three times with PBS, 1.5 × 10 keratinocytes (HaCaT) were added to the dish. 5 After culturing for 1 day, a test sample (Sample 1, final concentration: 400 μg / mL) dissolved in DMSO was added and cultured for 5 days. After culturing, the medium was removed, and total RNA was extracted from the cells using an RNeasy Mini Kit (QIAGEN).

[0054] This total RNA was used as a template for One Step SYBR (r) PrimeScript TM RT-PCR Kit II (Takara Bio Inc.) and real-time PCR device (ABI PRISM (r)The mRNA expression levels of filaggrin (FLG), serine palmitoyltransferase 2 (SPTLC2), and the internal standard β-actin (ACTB) were measured using a 7900HT Fast Real-Time PCR System (Applied Biosystems). The primers used were purchased from QIAGEN and are shown below. The expression levels of FLG and SPTLC2 mRNA were normalized by the expression level of ACTB mRNA. FLG:RT 2 qPCR Primer Assay for Human FLG SPTLC2:RT 2 qPCR Primer Assay for Human SPTLC2 ACTB:RT 2 qPCR Primer Assay for Human ACTB

[0055] From the corrected values ​​obtained, the rate of suppression (%) of the decrease in expression of each mRNA was calculated according to the following formula. Suppression rate of decrease in mRNA expression (%)={(AB)-(AC)} / (AB)×100 The terms in the formula represent the following: A: Corrected value without GA treatment (GA-) B: Correction value for no sample added and GA treatment (control) C: Correction value after addition of test sample and GA treatment The results are shown in Table 1.

[0056] [Table 1]

[0057] As shown in Table 1, it was confirmed that the wild thyme extract (sample 1) had an excellent inhibitory effect on the decrease in moisturizing-related gene expression in keratinocytes exposed to glycated collagen.

[0058] [Test Example 2] Inhibitory effect of glycated basement membrane extract on the decrease in moisturizing-related gene expression in keratinocytes The wild thyme extract (sample 1) was tested for its inhibitory effect on the decrease in moisturizing-related gene expression in keratinocytes on a glycated basement membrane extract in the following manner.

[0059] Basement membrane extract (Cultrex) diluted 10 times with PBS (r) Reduced Growth Factor BME,PathClear (r) 500 μL of the basement membrane extract (R&D Systems) was added to a 35 mm dish, and the dish was allowed to air dry and stand for 1 hour. After that, 500 μL of glyceraldehyde (GA) solution (0.09 mg / mL) prepared using PBS was added, and the dish was left to stand in an incubator at 37°C for 4 days, thereby glycating the basement membrane extract.

[0060] After washing three times with PBS, 1.5 × 10 keratinocytes (HaCaT) were added to the dish. 5 After culturing for 1 day, a test sample (Sample 1, final concentration: 400 μg / mL) dissolved in DMSO was added and cultured for 2 days. After culturing, the medium was removed, and total RNA was extracted from the cells using an RNeasy Mini Kit (QIAGEN).

[0061] This total RNA was used as a template for One Step SYBR (r) PrimeScript TM RT-PCR Kit II (Takara Bio Inc.) and real-time PCR device (ABI PRISM (r) The mRNA expression levels of filaggrin (FLG), serine palmitoyltransferase 2 (SPTLC2), transglutaminase-1 (TGM1), and the internal standard β-actin (ACTB) were measured using a 7900HT Fast Real-Time PCR System (Applied Biosystems). The primers used were purchased from QIAGEN and are shown below. The expression levels of FLG, SPTLC2, and TGM1 mRNA were normalized by the expression level of ACTB mRNA. FLG:RT 2 qPCR Primer Assay for Human FLG SPTLC2:RT 2 qPCR Primer Assay for Human SPTLC2 TGM1:RT 2 qPCR Primer Assay for Human TGM1 ACTB:RT 2 qPCR Primer Assay for Human ACTB

[0062] From the corrected values ​​obtained, the rate of suppression (%) of the decrease in expression of each mRNA was calculated according to the following formula. Suppression rate of decrease in mRNA expression (%)={(AB)-(AC)} / (AB)×100 The terms in the formula represent the following: A: Corrected value without GA treatment (GA-) B: Correction value for no sample added and GA treatment (control) C: Correction value after addition of test sample and GA treatment The results are shown in Table 2.

[0063] [Table 2]

[0064] As shown in Table 2, it was confirmed that the wild thyme extract (sample 1) had an excellent inhibitory effect on the decrease in moisturizing-related gene expression in keratinocytes on the glycated basement membrane extract.

[0065] [Test Example 3] Inhibitory effect of glycated basement membrane extract on the decrease in moisturizing-related protein expression in keratinocytes The wild thyme extract (sample 1) was tested for its inhibitory effect on the decrease in moisturizing-related protein expression in keratinocytes on a glycated basement membrane extract in the following manner.

[0066] Basement membrane extract (Cultrex) diluted 10 times with PBS (r) Reduced Growth Factor BME,PathClear (r)500 μL of the basement membrane extract (R&D Systems) was added to a 35 mm dish, and the dish was allowed to air dry and stand for 1 hour. After that, 500 μL of glyceraldehyde (GA) solution (0.09 mg / mL) prepared using PBS was added, and the dish was left to stand in an incubator at 37°C for 4 days, thereby glycating the basement membrane extract.

[0067] After washing three times with PBS, 1.5 × 10 keratinocytes (HaCaT) were added to the dish. 5 After culturing for 1 day, a test sample (Sample 1, final concentration: 400 μg / mL) dissolved in DMSO was added and the cells were cultured for 3 days. After culturing, the medium was removed, and proteins were extracted from the cells using Laemmli Buffer supplemented with a protease inhibitor (Protease Inhibitor Cocktail Tablets, manufactured by Roche).

[0068] Ten micrograms of the resulting protein was electrophoresed using anti-FLG antibody (Cloud-Clone), anti-SPTLC2 antibody (GeneTex), and anti-TGM1 antibody (Proteintech Group) as primary antibodies, and color development was performed using a Horseradish Peroxidase-labeled secondary antibody (ECL anti-rabbit IgG, GE Healthcare). After detecting each band, the band intensity was quantified using densitometry, and the expression levels of FLG, SPTLC2, and TGM1 proteins were calculated.

[0069] From the expression level of each protein thus obtained, the inhibition rate (%) of the decrease in expression of each protein was calculated according to the following formula. Protein expression reduction inhibition rate (%) = {(AB) - (AC)} / (AB) × 100 The terms in the formula represent the following: A: Protein expression level in GA-untreated (GA-) B: Protein expression level in the control group without sample addition and with GA treatment C: Protein expression level after addition of test sample and GA treatment The results are shown in Table 3.

[0070] [Table 3]

[0071] As shown in Table 3, it was confirmed that the wild thyme extract (sample 1) had an excellent inhibitory effect on the decrease in the expression of moisturizing-related proteins in keratinocytes on the glycated basement membrane extract.

[0072] [Formulation example 1] An emulsion was prepared in a conventional manner according to the following composition. Wild thyme extract 0.01g Jojoba oil 4.00g 1,3-butylene glycol 3.00g Arbutin 3.00g Polyoxyethylene cetyl ether (20E.O.) 2.50g 2.00g olive oil Squalane 2.00g Cetyl alcohol 2.00g Glyceryl monostearate 2.00g Polyoxyethylene sorbitan oleate (20E.O.) 2.00g Methyl parahydroxybenzoate 0.15g Stearyl glycyrrhetinate 0.10g Phellodendron bark extract 0.10g Dipotassium glycyrrhizinate 0.10g Ginkgo biloba extract 0.10g Conchiolin 0.10g Phellodendron bark extract 0.10g Chamomile extract 0.10g Fragrance 0.05g Purified water Rest (total amount is 100g)

[0073] [Formulation example 2] A cream having the following composition was prepared by a conventional method. Wild thyme extract 0.05g Sophora root extract 0.1g Scutellaria root extract 0.1g Liquid paraffin 5.0g White beeswax 4.0g Squalane 10.0g Cetanol 3.0g Lanolin 2.0g Stearic acid 1.0g Polyoxyethylene sorbitan oleate (20E.O.) 1.5g Glyceryl monostearate 3.0g Oil-soluble licorice extract 0.1g 1,3-butylene glycol 6.0g Methyl parahydroxybenzoate 1.5g Fragrance 0.1g Purified water Rest (total amount is 100g)

[0074] [Formulation example 3] A cosmetic essence having the following composition was prepared by a conventional method. Wild thyme extract 0.01g Chamomile extract 0.1g Carrot extract 0.1g Xanthan gum 0.3g Hydroxyethyl cellulose 0.1g Carboxyvinyl polymer 0.1g 1,3-butylene glycol 4.0g Dipotassium glycyrrhizinate 0.1g Glycerin 2.0g Potassium hydroxide 0.25g Fragrance 0.01g Preservative (methyl parahydroxybenzoate) 0.15g Ethanol 2.0g Purified water Rest (total amount is 100g)

[0075] [Formulation example 4] Tablets having the following composition were prepared by a conventional method. Wild thyme extract 5.0mg Dolomite (contains 20% calcium and 10% magnesium) 83.4mg Casein phosphopeptide 16.7mg Vitamin C 33.4mg Maltitol 136.8mg Collagen 12.7mg Sucrose fatty acid ester 12.0mg

[0076] [Formulation example 5] An oral liquid preparation having the following composition was prepared by a conventional method. <Composition in 1 ampoule (100 mL)> Wild thyme extract 0.3% by mass Sorbitol 12.0% by mass Sodium benzoate 0.1% by mass Fragrance 1.0% by mass Calcium sulfate 0.5% by mass Purified water remainder (100% by mass) [Industrial Applicability]

[0077] The inhibitor of a decrease in moisturizing function caused by glycation, the inhibitor of a decrease in epidermal function caused by glycation, the skin cosmetic, and the food and beverage product according to the present invention can make a significant contribution to the prevention, treatment, or amelioration of a decrease in moisturizing function caused by glycation; the prevention, treatment, or amelioration of a decrease in epidermal function caused by glycation; the prevention, treatment, or amelioration of rough skin, dry skin, and other dry skin diseases (e.g., atopic dermatitis, psoriasis, ichthyosis, etc.); and the like.

Claims

1. Wild thyme extract is the active ingredient. Used to suppress the decrease in expression of serine palmitoyltransferase and / or transglutaminase-1 caused by glycation An agent for suppressing a decrease in moisturizing function caused by glycation.

2. Wild thyme extract is the active ingredient. Used to suppress the decrease in expression of serine palmitoyltransferase and / or transglutaminase-1 caused by glycation 1. An agent for suppressing epidermal dysfunction caused by glycation.

3. Contains wild thyme extract, Used to suppress the decrease in expression of serine palmitoyltransferase and / or transglutaminase-1 caused by glycation A skin cosmetic for inhibiting a decrease in moisturizing function caused by glycation.

4. Contains wild thyme extract, Used to suppress the decrease in expression of serine palmitoyltransferase and / or transglutaminase-1 caused by glycation A skin cosmetic for inhibiting a decline in epidermal function caused by glycation.

5. Contains wild thyme extract, Used to suppress the decrease in expression of serine palmitoyltransferase and / or transglutaminase-1 caused by glycation A food or drink for inhibiting a decrease in moisturizing function caused by glycation.

6. Contains wild thyme extract, Used to suppress the decrease in expression of serine palmitoyltransferase and / or transglutaminase-1 caused by glycation A food or drink for inhibiting a decline in epidermal function caused by glycation.

Images