Proteoglycan-related gene expression promoter, or cyclic collagen formation promoter.

Abstract

The aim is to prevent and / or improve skin conditions that negatively affect skin appearance, such as wrinkles and sagging, by promoting the formation of cyclic collagen. [Solution] Screening was conducted on the expression of proteoglycan-related genes involved in proteoglycan formation, and it was found that rosehip extract or safflower extract increases the gene expression of persican and periostin, which are proteoglycan-related genes. Thus, rosehip extract or safflower extract was identified as a promoter of cyclic collagen formation.

Description

Technical Field

[0001] The present invention relates to promoting the expression of proteoglycan-related genes involved in skin aging. Furthermore, it relates to the technical field of beauty technology for suppressing skin aging through cyclic collagen formation.

Background Art

[0002] Skin sagging has a great impact on the perceived age. Maintaining skin firmness and improving sagging are major cosmetic challenges. When skin firmness is lost, sagging or looseness appears, giving an aged impression. Skin firmness is mainly due to the thickness of the dermis layer, and firm skin contains abundant elastic fibers such as collagen and elastin that fill the dermis layer. It is known that due to the effects of aging and exposure to ultraviolet rays, the function of dermal fibroblasts decreases, the production of elastic fibers decreases, and matrix metalloproteinase is activated to decompose elastic fibers. As a result, the dermis layer becomes thinner, resulting in sagging or looseness. Although the amount of collagen tends to decrease with aging, the amount of collagen is also affected by skin care, the amount of exposure to ultraviolet rays, and diet.

[0003] On the other hand, sagging and hollowing do not necessarily depend solely on the amount of collagen in the dermis. In elderly subjects experiencing sagging, the amount of collagen is not necessarily reduced, and it is thought that the symptoms of sagging are influenced by factors other than the amount of collagen. Other known causes include weakening of facial muscles and increased subcutaneous fat. Conventional cosmetic methods have focused on increasing collagen and elastic fibers by activating dermal fibroblasts, or suppressing the breakdown of collagen and elastic fibers, in order to improve sagging. Components that have the effect of activating dermal fibroblasts, promoting collagen production, or inhibiting its breakdown have been discovered and applied to cosmetics (Patent Documents 1 and 2). In recent years, research has been conducted that focuses not on the quantity of collagen in the dermis, but on its quality. It has been reported that elastic structures are not uniformly spread in the dermis, but rather that areas with high elasticity and areas with low elasticity are scattered, and that cyclic collagen, formed in such a way that high-elasticity collagen surrounds areas with low-elasticity collagen, is related to sagging in terms of quality (Patent Document 3). The circular arrangement of collagen contracts towards the center, creating tension in the skin and preventing sagging. [Prior art documents] [Patent Documents]

[0004] [Patent Document 1] Japanese Patent Publication No. 2012-144499 [Patent Document 2] Japanese Patent Publication No. 2013-014555 [Patent Document 3] International Publication No. 2023 / 243447 [Overview of the project] [Problems that the invention aims to solve]

[0005] The aim is to prevent and / or improve skin conditions that negatively affect skin appearance, such as wrinkles and sagging, by promoting the formation of cyclic collagen. [Means for solving the problem]

[0006] The inventors of this invention conducted intensive research on the formation of cyclic collagen and realized that the formation of cyclic collagen requires the formation of proteoglycans, and that it is necessary to enhance the expression of genes involved in the formation of such proteoglycans. Therefore, by screening the expression of proteoglycan-related genes, they found that rosehip extract or safflower extract promotes the gene expression of persican and periostin, which are proteoglycan-related genes. Therefore, the present invention relates to the following. [1] An expression promoter for proteoglycan-related genes, comprising rosehip extract or safflower extract, wherein the proteoglycan-related gene is persican or periostin. [2] A cyclic collagen formation promoter, including the expression promoter described in item 1. [3] The cyclic collagen refers to collagen formed in a ring shape around the hair follicle, as described in item 2, and is a cyclic collagen formation promoter. [4] A dermal matrix formation promoter containing rosehip extract or safflower extract. [5] A cosmetic method comprising applying rosehip extract or safflower extract to the skin, comprising promoting the formation of the dermal matrix and / or controlling the formation of cyclic collagen. [Effects of the Invention]

[0007] Rosehip extract or safflower extract improves skin wrinkles and sagging by promoting the gene expression of proteoglycans. [Brief explanation of the drawing]

[0008] [Figure 1]Figure 1A is a heatmap showing the differences in tension in the horizontal cross-section of the dermis when the skin is deformed and analyzed. Areas with high tension (orange to red in the heatmap) are indicated by dotted arrows, and areas with low tension (green to light blue in the heatmap) are indicated by solid arrows. Figure 1B shows the heatmap of the differences in tension in the horizontal cross-section of the dermis overlaid with the surrounding tissue. [Figure 2] Figure 2 shows magnified images of horizontal dermal cross-sections of facial skin specimens obtained from young and elderly subjects using X-ray-CT. [Figure 3] Figure 3 shows a graph illustrating the changes in the expression of proteoglycan-related genes (A) persican and (B) periostin when rosehip extract is added. [Figure 4] Figure 4 shows a graph illustrating the changes in the expression of proteoglycan-related genes (A) persican and (B) periostin when safflower extract is added. [Modes for carrying out the invention]

[0009] This invention relates to an expression promoter for proteoglycan-related genes, comprising rosehip extract or safflower extract. These extracts promote the expression of proteoglycan-related genes in dermal fibroblasts, enhance proteoglycan production, and promote the formation of cyclic collagen.

[0010] Cyclic collagen refers to collagen in the dermis, particularly the reticular layer of the dermis, that forms a ring-shaped structure surrounding the proteoglycan-rich dermal matrix region. When the reticular layer of the dermis is analyzed horizontally to the skin surface, the dermal matrix region is mainly located directly beneath the hair follicle in a roughly circular shape, with a diameter of 100-1,000 μm. The collagen region surrounds the proteoglycan-rich dermal matrix region, and the diameter of the cyclic structure of the collagen region is 110-1,100 μm. Cyclic collagen can be identified by detecting collagen and observing its cyclic morphology.

[0011] The dermal matrix is ​​a gel-like substance present in the dermis, composed of proteoglycans. The interstitial components, consisting of the dermal matrix and fibers such as collagen and elastin, further combine with cellular components to form the dermis. The dermal matrix alone generates almost no tension. Conventionally, there was no knowledge regarding the distribution of dermal matrix and fibers in the interstitial components, and it was thought that fibers and dermal matrix were uniformly distributed. However, our research has shown that the dermal matrix and fibers are not uniformly distributed; rather, a dermal matrix region exists directly beneath the hair follicle, and collagen is formed in a ring-like manner surrounding the dermal matrix (Figure 1). The dermal matrix can be determined by measuring the expression of genes involved in proteoglycan formation (hereinafter referred to as proteoglycan-related genes), for example, the expression of genes encoding the core protein of proteoglycans. The diameter of the cyclic collagen increases in proportion to the amount of dermal matrix.

[0012] Circularly formed collagen generates tension toward its center. Typically, beneath each hair follicle, there is one dermal matrix region and circular collagen surrounding it. The tension exerted by each circular collagen toward its center contributes to the overall firmness of the skin in the planar direction. The morphology of circular collagen is lost with age (Figure 2). Furthermore, comparing the body and face, the density of microhairs in the body is lower than in the face, and dermal collagen is present in a nearly uniformly high content in the body. In the face, while circular collagen is present surrounding the microhairs, the amount of collagen is lower. The amount of circular collagen can be measured based on the morphology and / or the collagen as a component. There is a positive correlation between the amount of circular collagen and tension, while there is an inverse correlation between the amount of circular collagen and sagging. The firmness in the planar direction differs depending on whether the collagen takes on a circular structure or is uniformly present (Figure 2). Therefore, as cyclic collagen is lost with age, the skin loses its ability to maintain firmness in the planar direction, resulting in sagging as the entire skin is pulled downwards by gravity. Furthermore, there is a correlation between the diameter of cyclic collagen and the diameter of microhairs; the larger the diameter of the microhairs, the larger the diameter of the cyclic collagen. Whether or not cyclic collagen is lost in a subject's face can be measured based on sagging, or it can be determined by measuring the amount or expression of proteoglycan glycoproteins that constitute the dermal matrix from a skin sample.

[0013] Interstitial components refer to the components produced by fibroblasts, excluding dermal cells. Interstitial components include dermal matrix such as proteoglycans and fibers such as collagen. Highly active fibroblasts increase collagen production in addition to dermal matrix production. During maturation, the produced collagen changes structure to cyclic collagen that surrounds the dermal matrix. Therefore, in an environment where dermal matrix production increases, collagen production also increases, and cyclic collagen is generated upon maturation. The amount of cyclic collagen increases in line with the increased expression of proteoglycan-related proteins, such as the core protein versican or periostin, which are part of the dermal matrix.

[0014] A proteoglycan refers to a complex of sugar and protein. A proteoglycan is formed by the binding of a glycosaminoglycan, which is a polysaccharide, and a core protein. Examples of glycosaminoglycans include hyaluronic acid and chondroitin sulfate. Known core proteins include aggrecan, versican, neurocan, brevican, decorin, biglycan, serglycin, perlecan, serglycin, syndecan, glypican, and the like. Genes encoding core proteins and genes involved in the formation of proteoglycans can be referred to as proteoglycan-related genes. Proteins expressed from proteoglycan-related genes can also be called proteoglycan constituent genes since they constitute proteoglycans.

[0015] Versican is a protein encoded by the human VCAN gene, binds to chondroitin sulfate, constitutes a proteoglycan exceeding 1000 kDa, and forms an extracellular matrix. As a proteoglycan, it is contained in the substrate part of cyclic collagen, and the formation of cyclic collagen is promoted by an increase in the amount of proteoglycan containing versican.

[0016] Periostin is a protein encoded by the human POSTN gene. Periostin is a secreted protein and a protein that constitutes the extracellular matrix. Periostin is a protein that binds to matrix proteins such as collagen I / fibronectin and tenascin C. Among these, fibronectin and tenascin C are core proteins that constitute proteoglycans, and periostin binds to these core proteins that constitute proteoglycans, thereby contributing to the structural maintenance of the extracellular matrix as a proteoglycan-related protein. In addition, periostin is known to act as a ligand for integrin, particularly αv integrin, and binds to integrin present on the cell membrane to induce signal transduction into the cell.

[0017] The plant extracts described herein can be obtained by conventional methods, for example, by immersing or refluxing the source plant with an extraction solvent at room temperature or under heating, followed by filtration and concentration. They can also be obtained by distillation, such as steam distillation. Any solvent commonly used for extraction can be used as the extraction solvent, such as aqueous solvents, such as water, physiological saline, phosphate buffer, borate buffer, or organic solvents, such as alcohols including ethanol, propylene glycol, 1,3-butylene glycol, and glycerin, aqueous alcohols, chloroform, dichloroethane, carbon tetrachloride, acetone, ethyl acetate, and hexane, each can be used alone or in combination. Preferably, a mixed solvent of water and alcohol, such as 1,3-butylene glycol, is used as the solvent. The extract obtained by extraction with the above solvent can be used as is, or concentrated by methods such as freeze-drying. If necessary, an extract obtained by adsorption, such as removing impurities using an ion exchange resin, or by adsorption using a porous polymer (e.g., Amberlite XAD-2) column, followed by elution with a desired solvent and further concentration can also be used. The plant extracts used can be commercially available extracts used as cosmetic ingredients, and these commercially available extracts can be incorporated at a predetermined concentration.

[0018] Safflower extract is an extract of plants belonging to the genus Carthamus in the Asteraceae family, particularly the plant body of Carthamus tinctorius. Also, in the display name used for the full ingredient label of cosmetics defined by the Japan Cosmetic Industry Association and the international display name according to the INCI name (International Nomenclature of Cosmetic Ingredients), the display name / INCI name is: Safflower Flower Extract / CARTHAMUS TINCTORIUS (SAFFLOWER) FLOWER EXTRACT. The plant body may be leaves, stems, flowers, roots, or fruits, but flowers are particularly used. Safflower is native to North Africa and has been cultivated in various regions since ancient times. Safflower extract can be obtained by directly pressing and extracting with a solvent the plant body of safflower, particularly the flowers, either as they are or after drying or fermentation. Solvent extraction can be prepared by extracting using water, alcohol, or a mixed solution thereof. As the alcohol, ethanol, glycerol, propylene glycol, or butylene glycol is used. More preferably, it can be extracted with a mixed solution of water and alcohol, for example, a mixed solution of any ratio of 1,3-butylene glycol, such as a mixed solution of 10:90 to 90:10, preferably 30:70 to 70:30, and more preferably 50:50. Safflower extract is formulated at a final concentration of 0.001% to 10%, and 0.01 to 1%, particularly 0.05% to 0.5% is more preferable.

[0019] Wild rose extract is an extract obtained from the plant body of a plant belonging to the genus Rosa. In particular, it is an extract from the plant body of Rosa canina. Furthermore, in the display name used in the full ingredient list of cosmetics as defined by the Japan Cosmetic Industry Association and the international display name according to the INCI (International Nomenclature for Cosmetic Ingredients), it is represented as Display Name / INCI Name: Rosa Canina Fruit Extract / ROSA CANINA FRUIT EXTRACT. The plant body may include leaves, stems, flowers, roots, or fruits, but the fruit is particularly used. Rosa canina is distributed in Europe, North Africa, and West Asia. Wild rose extract is obtained by pressing the fruit of Rosa canina, either as is, or after drying or fermentation, and then extracting it with a solvent. Solvent extraction can be prepared by using water, alcohol, or a mixture thereof. As alcohol, ethanol, glycerol, propylene glycol, or butylene glycol can be used. More preferably, the extract can be obtained with a mixture of water and alcohol, such as 1,3-butylene glycol, in any proportion, for example, a mixture of 10:90 to 90:10, preferably 30:70 to 70:30, and even more preferably 50:50. The rosehip extract is formulated at a final concentration of 0.001% to 10%, with 0.01% to 1%, and particularly 0.05% to 0.5% being more preferable.

[0020] The proteoglycan-related gene expression promoter or cyclic collagen formation promoter of the present invention can be optionally selected in concentration and dosage form from the viewpoint of producing the desired effect, namely improvement of wrinkles and sagging. The proteoglycan-related gene expression promoter or cyclic collagen formation promoter can be incorporated into foods, cosmetics, pharmaceuticals, or quasi-drugs. When incorporated into foods, it may be incorporated into nutritional supplements such as supplements and energy drinks, or into foods with functional claims. When incorporated into cosmetics, it can be incorporated into facial or body cosmetics such as lotions, emulsions, serums, creams, packs, essences, and gels, as well as makeup cosmetics such as foundations, makeup bases, and concealers, and even bath additives. When incorporated into pharmaceuticals, it may be administered orally or parenterally, for example, transdermally. When administered transdermally, it can be formulated into a topical skin preparation. By using cosmetics, foods, pharmaceuticals, and quasi-drugs containing proteoglycan-related gene expression promoters or cyclic collagen formation promoters, the production of proteoglycans in the dermis can be promoted, thereby preventing and / or improving wrinkles and sagging of the skin through the formation of cyclic collagen. The proteoglycan-related gene expression promoter or cyclic collagen formation promoter can be administered over a long period of time as a food, cosmetic, or pharmaceutical of the present invention. From the viewpoint of improving wrinkles and sagging, it may be administered for several days or more, one week or more, two weeks or more, one month or more, three months or more, or six months or more. There is no particular upper limit, but it may be several years or less, for example, one year or less.

[0021] The proteoglycan-related gene expression promoter or cyclic collagen formation promoter according to the present invention is preferably applied to the skin and can be incorporated into topical skin preparations. From the viewpoint of incorporation into topical skin preparations, safflower extract and rosehip extract can each be incorporated in concentrations of 0.001% to 10%. Safflower extract and rosehip extract may be incorporated individually or in combination. From the viewpoint of fully exhibiting the effect, they can preferably be incorporated in a topical skin preparation at a concentration of 0.01% or more, and more preferably at a concentration of 0.05% or more. From the viewpoint of economic efficiency, they can preferably be incorporated at a concentration of 1% or less, and more preferably at a concentration of 0.5% or less.

[0022] Topical skin preparations are not particularly limited as long as they can be applied to the skin, and any dosage form can be used, such as solution, emulsion, solid, semi-solid, powder, powder dispersion, water-oil two-layer separation, water-oil-powder three-layer separation, ointment, gel, aerosol, mousse, stick, etc. When formulated as a topical skin preparation, bases and excipients commonly used in topical skin preparations, such as preservatives, emulsifiers, and pH adjusters, may be used.

[0023] Another aspect of the present invention may relate to a beauty method comprising applying a cosmetic containing rosehip extract or safflower extract to the skin, particularly the skin of the face. Such a beauty method may be used on the skin of a person who is troubled by sagging skin or wrinkles, or who wishes to prevent sagging skin or wrinkles. Mechanical stimulation, such as vibration, or electrical stimulation, such as iontophoresis, may be applied with the intention that the components of the extract reach the skin, particularly the dermis. The beauty method according to the present invention may be offered as a service in beauty salons, beauty clinics, etc., or may be offered together with the sale of cosmetics in cosmetic retail stores. The beauty method is a non-therapeutic or non-medical method that can be distinguished from treatment or preventive methods performed by doctors or medical professionals, and is provided by a practitioner who is not a doctor or other medical professional.

[0024] All references made herein are incorporated herein by reference in their entirety.

[0025] The embodiments of the present invention described below are for illustrative purposes only and do not limit the technical scope of the invention. The technical scope of the invention is limited solely by the claims. Modifications to the invention, such as additions, deletions, and substitutions of constituent elements of the invention, can be made without departing from the spirit of the invention. [Examples]

[0026] Example 1: Human fibroblasts were cultured in Dulbecco's modified Eagle medium (DMEM) (containing 10% fetal bovine serum (FBS)) at a rate of 1 × 10⁶ 4 Seeds were seeded at a density of 1 cell / well (24-well plate) and cultured at 37°C under a 5% CO2 atmosphere. The day after seeding, EcoFarm Rose B extract (Ichimaru Falcos) and Safflower B extract (Ichimaru Falcos) were added to a final concentration of 0.1%, and culture was carried out. In the control group (cont), only the same amount of solvent (50% butylene glycol aqueous solution) as the extract was added instead. After 2 days of culture, the cells were harvested, RNA was extracted using the RNeasy mini kit (QIAGEN, Valencia, CA), and cDNA was prepared. Gene expression levels of the prepared cDNA were measured by real-time PCR using LightCycler 480 (Roche Applied Science, Indianaporis, IN) with the following primers. [Table 1] The results are shown in Figures 3 and 4. When rosehip extract and safflower extract were added, the expression of versican and periostin, respectively, was enhanced.

Claims

1. An expression promoter for proteoglycan-related genes, comprising rosehip extract or safflower extract, wherein the proteoglycan-related gene is persican or periostin.

2. A cyclic collagen formation promoter comprising the expression promoter described in claim 1.

3. The cyclic collagen formation promoter according to claim 2, wherein the cyclic collagen refers to collagen formed in a ring shape around the hair follicle.

4. A dermal matrix formation promoter containing rosehip extract or safflower extract.

5. A cosmetic method comprising applying rosehip extract or safflower extract to the skin, comprising promoting the formation of dermal matrix and / or controlling the formation of cyclic collagen.

Images