Dermal regeneration promoter

Abstract

To provide substances and compositions that are effective in improving skin elasticity and reducing wrinkles, from the perspective of maintaining or improving quality of life (QOL) in terms of both health and beauty. [Solution] The present invention relates to a composition containing components such as 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives, which can be used to promote the production of LTBP2 and / or fibrillin 1 in fibroblasts, thereby obtaining effects of improving dermal elasticity, improving dermal wrinkles, promoting the increase of dermal papillary fibers, and stabilizing dermal papillary fibers.

Description

[Technical Field]

[0001] The present invention relates to a dermal elasticity improving agent, a dermal wrinkle improving agent, a dermal papillary fiber enlargement promoting agent, a dermal papillary fiber stabilizing agent, and an LTBP2 and / or fibrillin 1 production promoting agent, and more particularly to a dermal elasticity improving agent, a dermal wrinkle improving agent, a dermal papillary fiber enlargement promoting agent, a dermal papillary fiber stabilizing agent, and an LTBP2 and / or fibrillin 1 production promoting agent characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. [Background technology]

[0002] With Japan facing a truly aging society, research and development on the mechanisms of aging are becoming increasingly active. Among these, the skin forms the outermost layer of the human body, playing a crucial role in protecting the body, and is particularly important from a cosmetic standpoint because its appearance is visually perceived. Interest in preventing skin aging has been growing in recent years from the perspective of maintaining or improving quality of life (QOL) in terms of both health and beauty.

[0003] To prevent skin aging, topical skin preparations such as cosmetics have long contained moisturizers expected to improve the skin's moisture retention capacity, as well as vitamins E and C, which are expected to have antioxidant effects. Recent research has revealed that a decrease or degeneration of matrix components in the skin is significantly involved in wrinkles, sagging, and loss of firmness associated with aging.

[0004] In particular, a decrease in collagen, a major component of the matrix, is associated with skin sagging and loss of firmness, and the degradation and denaturation of elastin caused by the expression of elastin-degrading enzymes is thought to be a cause of decreased elasticity. Therefore, collagen production enhancers, collagen-degrading enzyme inhibitors, and elastin-degrading enzyme inhibitors are included as anti-aging agents. However, while these drugs show some effect, they have not yet achieved sufficient efficacy.

[0005] Patent Document 1 discloses an anti-aging topical skin preparation that contains niacin and ubiquinone, which is effective in restoring and maintaining skin firmness and elasticity, reducing wrinkles and sagging, and reducing pigmentation, thereby restoring and maintaining a youthful skin condition.

[0006] Patent Document 2 discloses a cyclic carboxamide derivative having a specific structure as an ingredient in a whitening agent that is effective in preventing and suppressing new skin aging from the perspective of the relationship between heparanase and skin aging, and in preventing and suppressing pigmentation such as blemishes, freckles, and dullness. [Prior art documents] [Patent Documents]

[0007] [Patent Document 1] Japanese Patent Publication No. 2005-298370 [Patent Document 2] Japanese Patent Publication No. 2014-111640

[0008] [Non-Patent Document 1] VlodavskyI., et. al., Semin Cancer Biol., 2002;12(2):121-129 [Overview of the Initiative] [Problems that the invention aims to solve]

[0009] The objective of this invention is to find substances and compositions that are effective in improving skin elasticity and reducing wrinkles, from the perspective of maintaining or improving quality of life (QOL) in terms of both health and beauty. [Means for solving the problem]

[0010] As a result of diligent research, the inventors have found that heparanase activity inhibitors such as 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives promote the production of LTBP2 and / or fibrillin 1 in fibroblasts, exhibiting effects of improving dermal elasticity, improving dermal wrinkles, promoting the increase of dermal papillary fibers, and stabilizing dermal papillary fibers.

[0011] Therefore, this disclosure provides the following embodiments. (1) A dermal elasticity improving agent characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. (2) A dermal wrinkle improving agent characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. (3) A dermal papillary fiber enlargement promoting agent characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. (4) A dermal papillary fiber stabilizer characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. (5) The agent according to (3) or (4), wherein the dermal papillary fibers are dermal papillary oxytalane fibers. (6) An agent for promoting the production of LTBP2 and / or fibrillin 1, characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. (7) 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives, which are of the following general formula (I): [ka] (In general formula (I), n is an integer from 1 to 3, R 1 X is a hydrocarbon group having 1 to 6 carbon atoms, which may be substituted with a hydrogen atom or a hydroxyl group, and X is -CH2- or -N(R 2 The group is represented by )-, and R 2 (This refers to a hydrocarbon group having 1 to 6 carbon atoms, which may be substituted with a hydrogen atom or a hydroxyl group.) The agent according to any one of (1) to (6), which is a cyclic carboxamide derivative represented by (8) A cosmetic method for improving the elasticity of the dermis, characterized by increasing the production of LTBP2 and / or fibrillin-1. (9) A cosmetic method for preventing or improving wrinkles, characterized by increasing the production of LTBP2 and / or fibrillin-1. (10) A cosmetic method for preventing or improving wrinkles, characterized by local stabilization of LTBP2 and / or fibrillin-1. (11) A screening method for a dermal elasticity improving agent, using the expression of LTBP2 and / or fibrillin-1 as an index. (12) A screening method for a dermal wrinkle improving agent, using the expression of LTBP2 and / or fibrillin-1 as an index. (13) A cosmetic method for preventing or improving wrinkles, characterized by increasing the expression of PDGF-BB. (14) A screening method for a dermal elasticity improving agent, using the expression of PDGF-BB as an index. (15) A screening method for a dermal wrinkle improving agent, using the expression of PDGF-BB as an index.

Effect of the Invention

[0012] The agent according to the present disclosure can be suitably used as a cosmetic composition such as a cosmetic effective for improving dermal elasticity and / or dermal wrinkles, since it promotes the production of LTBP2 and / or fibrillin-1, thereby increasing and / or stabilizing the dermal papillary layer fibers.

Brief Description of the Drawings

[0013] [Figure 1] FIG. 1 is a graph showing the results of examining the amount of PDGFB gene expression in the epidermis of an artificial skin model (skin three-dimensional model) by quantitative PCR analysis. The results of treatment with a combination of a heparanase inhibitor (BIPBIPU) and a MMP inhibitor (CGS27023A) are shown based on the left control (Cont). [Figure 2]Figure 4 is a graph showing the changes in gene expression in fibroblasts induced by PDGF-BB. From left to right, the results for LTBP2, type V collagen (COL5A1), type III collagen (COL3A1), type I collagen (COL1A1), and fibrillin 1 (FBLN1) are shown for both the control (cont) and treatment with 10 ng / mL of PDGF-BB. [Figure 3A] Figure 3A shows images illustrating the changes in LTBP2 localization induced by heparanase inhibitors. The results of investigating changes in LTBP2 localization in organ culture models are shown. From top to bottom, the results are shown for the control group (Cont), treatment with a combination of heparanase inhibitor (BIPBIPU) and MMP inhibitor (CGS27023A), and treatment with S-173 (HEI). [Figure 3B] Figure 3B shows images illustrating the changes in LTBP2 localization induced by heparanase inhibitors. The results of investigating changes in LTBP2 localization in a three-dimensional skin model are shown. From top to bottom, the results are shown for the control group (Cont), treatment with a combination of heparanase inhibitor (BIPBIPU) and MMP inhibitor (CGS27023A), and treatment with S-173 (HEI). [Figure 4A] Figure 4A shows images illustrating the changes in the localization of LTBP2 and fibrillin 1 induced by heparanase inhibitors. The results of investigating changes in the localization of LTBP2 and fibrillin 1 in organ culture models are shown. From top to bottom, the results are shown for the control group (Cont), treatment with a combination of heparanase inhibitor (BIPBIPU) and MMP inhibitor (CGS27023A), and treatment with S-173 (HEI). [Figure 4B] Figure 4B shows images illustrating the changes in the localization of LTBP2 and fibrillin 1 induced by heparanase inhibitors. The results of investigating changes in the localization of LTBP2 and fibrillin 1 in a three-dimensional skin model are shown. From top to bottom, the results are shown for the control group (Cont), treatment with a combination of heparanase inhibitor (BIPBIPU) and MMP inhibitor (CGS27023A), and treatment with S-173 (HEI). [Figure 5]Figure 5 is a graph showing that LTBP2 and heparan sulfate bind in a concentration-dependent manner, based on the results of an investigation into the binding of heparan sulfate and LTBP2. [Figure 6] Figure 6 is a graph showing the results of measuring transepidermal water loss from both cheeks of the subject three times using a Vapometer (Delphin), stratum corneum water content five times using a Corneometer (MPA580, Courage+Khazakaelectronic GmbH), and skin elasticity three times using a Cutometer. [Figure 7] Figure 7 is a graph showing the results of measuring the thickness of the dermal papillary layer using an acoustic microscope. [Figure 8] Figure 8 is a graph showing the change in wrinkle area percentage (%) with the use of a cream formulation containing 1.5% HEI (Active) and a cream formulation without HEI (Placebo). The results are shown at 0 months (0M), 1 month (1M), and 2 months (2M) after the start of application. [Modes for carrying out the invention]

[0014] Dermal elasticity improving agent One embodiment of the present disclosure relates to a dermal elasticity improving agent characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone (HEI) or a derivative thereof as an active ingredient. In this specification, 1-(2-hydroxyethyl)-2-imidazolidinone (HEI) may also be referred to as S-173 or S173.

[0015] 1-(2-hydroxyethyl)-2-imidazolidinone (HEI) is a compound having the following structure: [ka]

[0016] Derivatives of 1-(2-hydroxyethyl)-2-imidazolidinone may be compounds represented by the following general formula (I): [ka] (In general formula (I), n is an integer from 1 to 3, R 1 X is a hydrocarbon group having 1 to 6 carbon atoms, which may be substituted with a hydrogen atom or a hydroxyl group, and X is -CH2- or -N(R 2 The group is represented by )-, and R 2 (This refers to a hydrocarbon group having 1 to 6 carbon atoms, which may be substituted with a hydrogen atom or a hydroxyl group.)

[0017] 1-(2-hydroxyethyl)-2-imidazolidinone and its derivatives are known to act as heparanase activity inhibitors (see Japanese Patent Publication No. 2014-111640 (Patent Document 2)). Heparanase is an enzyme present in various cells that specifically degrades the heparan sulfate chains of various heparan sulfate proteoglycans. In the skin, it is produced by epidermal keratinocytes that make up the epidermis, as well as fibroblasts and vascular endothelial cells in the dermis. It is also known that production is elevated in various cancer cells, and a correlation with the malignancy of cancer has been suggested. High heparanase production in cancer cells is associated with high metastasis and high angiogenesis induction (see Vlodavsky I., et. al., Semin Cancer Biol., 2002;12(2):121-129 (Non-Patent Document 1)).

[0018] Heparan sulfate proteoglycans are heparan sulfate-binding growth factors (bFGF, HGF, It plays a role in accumulating VEGF, HB-EGF, etc., extracellularly. Perlecan, a type of heparan sulfate proteoglycan, is also present in the epidermal basement membrane at the boundary between the epidermis and dermis. In the skin, it controls the movement of growth factors between the epidermis and dermis by binding heparan sulfate-binding growth factors to the epidermal basement membrane. Furthermore, perlecan present in the epidermal basement membrane also regulates the action of growth factors on epidermal basal cells bound to the basement membrane, and it has been shown to be essential for healthy epidermal proliferation and differentiation.

[0019] Furthermore, 1-(2-hydroxyethyl)-2-imidazolidinone (HEI) is known to act as an inhibitor of matrix metalloproteinases (MMPs). HEI inhibits MMP-9, for example, but does not affect the activity of MMP-1 and MMP-2.

[0020] As detailed in the Examples section, the inventors have now discovered that 1-(2-hydroxyethyl)-2-imidazolidinone has the effect of improving human skin elasticity. Skin elasticity can be measured using, for example, a skin measuring instrument such as a Cutometer®.

[0021] The dermal elasticity improving agent according to this disclosure can also be described as a composition for improving dermal elasticity, containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. Such a composition may be used particularly for non-therapeutic cosmetic purposes. From another perspective, one embodiment according to this disclosure relates to the use of 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof in improving dermal elasticity. In such use, 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof may be used in the form of a composition in combination with other ingredients. From yet another perspective, one embodiment according to this disclosure relates to the use of 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof in the manufacture of a dermal elasticity improving agent. From yet another perspective, one embodiment according to this disclosure relates to a method for improving dermal elasticity, which includes administering 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof to a subject (e.g., a human).

[0022] LTBP2 and / or fibrillin 1 production promoters One embodiment of the present disclosure relates to an LTBP2 and / or fibrillin 1 production promoter characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. As detailed in the Examples section, the inventors have now found that 1-(2-hydroxyethyl)-2-imidazolidinone has the effect of promoting the production of LTBP2 and / or fibrillin 1 in human fibroblasts. The increase or decrease in the production amount of LTBP2 and / or fibrillin 1 can be investigated, for example, by quantitative PCR analysis.

[0023] The LTBP2 and / or fibrillin 1 production promoters relating to this disclosure may also be described as compositions for use in promoting the production of LTBP2 and / or fibrillin 1, containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. Such compositions may be used particularly in non-therapeutic cosmetic applications. In another view, one embodiment relating to this disclosure relates to the use of 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof in promoting the production of LTBP2 and / or fibrillin 1. In such use, 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof may be used in the form of a composition in combination with other ingredients. In yet another view, one embodiment relating to this disclosure relates to the use of 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof in the manufacture of LTBP2 and / or fibrillin 1 production promoters. From yet another perspective, one embodiment of the present disclosure relates to a method for promoting the production of LTBP2 and / or fibrillin 1, comprising administering 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof to a subject (e.g., a human). From yet another perspective, one embodiment of the present disclosure relates to a cosmetic method for improving dermal elasticity, characterized by increasing the production of LTBP2 and / or fibrillin 1. From yet another perspective, one embodiment of the present disclosure relates to a cosmetic method for preventing or improving wrinkles, characterized by increasing the production of LTBP2 and / or fibrillin 1. From yet another perspective, one embodiment of the present disclosure relates to a cosmetic method for preventing or improving wrinkles (particularly dermal wrinkles), characterized by stabilizing the localization of LTBP2 and / or fibrillin 1 directly beneath the basement membrane. Furthermore, as shown in the embodiments of this application, PDGF-BB promotes the production of LTBP2 and fibrillin 1. Therefore, one embodiment of this disclosure relates to a cosmetic method for preventing or improving wrinkles (especially dermal wrinkles) characterized by increasing the expression of PDGF-BB. It should be noted that the cosmetic method of this disclosure is a non-therapeutic method and does not include so-called medical procedures.

[0024] Dermal papillary layer fiber enlargement promoter One embodiment of the present disclosure relates to a dermal papillary fiber growth promoter characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. As detailed in the Examples section, the inventors have now found that 1-(2-hydroxyethyl)-2-imidazolidinone has the effect of promoting the growth of dermal papillary fibers. The growth of dermal papillary fibers can be evaluated, for example, by measuring the thickness of the dermal papillary layer with an acoustic microscope (for example, AMS-50SI manufactured by Honda Electronics Co., Ltd.).

[0025] Some embodiments relate to a dermal papillary fiber enlargement promoter characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient, wherein the dermal papillary fibers are dermal papillary oxytalane fibers.

[0026] The dermal papillary fiber enlargement promoter according to this disclosure can also be described as a composition for promoting dermal papillary fiber enlargement, containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. Such compositions may be used particularly for non-therapeutic cosmetic purposes. From another perspective, one embodiment according to this disclosure relates to the use of 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof in promoting dermal papillary fiber enlargement. In such use, 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof may be used in the form of a composition in combination with other components. From yet another perspective, one embodiment according to this disclosure relates to the use of 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof in the manufacture of a dermal papillary fiber enlargement promoter. From yet another perspective, one embodiment of the present disclosure relates to a method for promoting dermal papillary fiber growth, comprising administering 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof to a subject (e.g., a human).

[0027] Dermal papillary layer fiber stabilizer One embodiment of the present disclosure relates to a dermal papillary fiber stabilizer characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. As detailed in the Examples section, the inventors have now found that 1-(2-hydroxyethyl)-2-imidazolidinone has the effect of promoting the increase of dermal papillary fibers. The stabilization of dermal papillary fibers can be evaluated, for example, by measuring the thickness of the dermal papillary layer with an acoustic microscope (for example, AMS-50SI manufactured by Honda Electronics Co., Ltd.).

[0028] Some embodiments relate to a dermal papillary fiber enlargement promoter characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient, wherein the dermal papillary fibers are dermal papillary oxytalane fibers.

[0029] The dermal papillary fiber stabilizer according to this disclosure can also be described as a composition for use in stabilizing dermal papillary fiber, containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. Such compositions may be used particularly for non-therapeutic cosmetic applications. From another perspective, one embodiment according to this disclosure relates to the use of 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof in stabilizing dermal papillary fiber. In such use, 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof may be used in the form of a composition in combination with other components. From yet another perspective, one embodiment according to this disclosure relates to the use of 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof in the manufacture of a dermal papillary fiber stabilizer. From yet another perspective, one embodiment of the present disclosure relates to a method for stabilizing dermal papillary fibers, comprising administering 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof to a subject (e.g., a human).

[0030] Dermal wrinkle improvement agent One embodiment of the present disclosure relates to a dermal wrinkle improving agent characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. As detailed in the Examples section, the inventors have now found that 1-(2-hydroxyethyl)-2-imidazolidinone has the effect of improving human dermal wrinkles. Fine wrinkles caused by dryness are commonly called epidermal wrinkles, while deep wrinkles around the eyes are defined as deep wrinkles accompanied by changes in the dermis and fat layer. The technology disclosed in this application is groundbreaking in that, in addition to improving fine epidermal wrinkles, it also improves deep dermal wrinkles by promoting the regeneration of the papillary layer (elastin fibers of the papillary layer). Until now, the theory among those skilled in the art has been that dermal wrinkles are improved by inhibiting the breakdown of general matrix components such as collagen, elastin, and hyaluronic acid in the dermis, or simply by promoting the production of collagen, elastin, and hyaluronic acid. In the technology disclosed herein, treatment with 1-(2-hydroxyethyl)-2-imidazolidinone stimulates the production of PDGF-BB from the epidermis, which acts on PDGFRβ-positive fibroblasts in the papillary layer to enhance collagen and elastin production. Meanwhile, 1-(2-hydroxyethyl)-2-imidazolidinone suppresses the degradation of the basement membrane, promoting the regeneration of a healthy basement membrane microstructure. A key point is that the matrix, whose production is enhanced by PDGF-BB, interacts with the basement membrane formed by the suppression of degradation, enabling the construction of a normal matrix in the papillary layer. The improvement of dermal wrinkles can be evaluated, for example, using an image analysis device.

[0031] The dermal wrinkle-improving agent according to this disclosure can also be described as a composition for use in improving dermal wrinkles, containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient. Such compositions may be used particularly for non-therapeutic cosmetic purposes. From another perspective, one embodiment according to this disclosure relates to the use of 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof in the improvement of dermal wrinkles. In such use, 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof may be used in the form of a composition in combination with other ingredients. From yet another perspective, one embodiment according to this disclosure relates to the use of 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof in the manufacture of a dermal wrinkle-improving agent. From yet another perspective, one embodiment according to this disclosure relates to a method for improving dermal wrinkles, which includes administering 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof to a subject (e.g., a human).

[0032] Screening method One embodiment of the present disclosure relates to a screening method for dermal elasticity improving agents using the expression of LTBP2 and / or fibrillin 1 as an indicator. Another embodiment of the present disclosure relates to a screening method for dermal wrinkle improving agents using the expression of LTBP2 and / or fibrillin 1 as an indicator. Such a screening method includes, for example, the steps of contacting cells with a test compound, measuring the expression of LTBP2 and / or fibrillin 1, and determining whether there is an increase in the expression of LTBP2 and / or fibrillin 1 due to the presence or absence of the compound. The increase in the expression of LTBP2 and / or fibrillin 1 can be evaluated by measuring the amount of mRNA or protein using known methods. Compounds to be screened may be, for example, those contained in a compound library or a library of extracts from microorganisms, plants, animals, etc.

[0033] Furthermore, one embodiment of the present disclosure relates to a dermal elasticity improving agent and a screening method that uses PDGF-BB expression as an indicator. Such a screening method includes, for example, the steps of contacting cells with a test compound, measuring the expression of PDGF-BB, and determining whether there is an increase in PDGF-BB expression due to the presence or absence of the compound. An increase in PDGF-BB expression can be evaluated by measuring the amount of mRNA or protein using known methods. Compounds to be screened may, for example, be those contained in a compound library or a library of extracts from microorganisms, plants, animals, etc.

[0034] Furthermore, one embodiment of the present disclosure relates to a screening method for dermal wrinkle-improving agents using PDGF-BB expression as an indicator. Such a screening method includes, for example, the steps of contacting cells with a test compound, measuring the expression of PDGF-BB, and determining whether there is an increase in PDGF-BB expression due to the presence or absence of the compound. An increase in PDGF-BB expression can be evaluated by measuring the amount of mRNA or protein using known methods. Compounds to be screened may be, for example, those contained in a compound library or a library of extracts from microorganisms, plants, animals, etc.

[0035] compounding 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives can be synthesized by known methods or readily available commercially.

[0036] Furthermore, 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives can be prepared as inorganic or organic salts by known methods. The salts used in the embodiments of this disclosure are not particularly limited, but examples of inorganic salts include hydrochloride, sulfate, phosphate, hydrobromide, sodium, potassium, magnesium, calcium, and ammonium salts. Examples of organic salts include acetate, lactate, maleate, fumarate, tartrate, citrate, methanesulfonate, p-toluenesulfonate, triethanolamine, diethanolamine, and amino acid salts.

[0037] In the agents according to this disclosure, 1-(2-hydroxyethyl)-2-imidazolidinone and its derivatives having heparanase activity inhibitory effect and / or MMP inhibitory effect, particularly those having both heparanase activity inhibitory effect and MMP inhibitory effect (particularly MMP-9 inhibitory effect), can be preferably used. The agents according to this disclosure may contain only one of 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives alone, or may contain two or more of the above compounds or salts thereof in any combination and ratio.

[0038] The content of 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives or salts in the agent according to this disclosure is not particularly limited as long as it is in an amount sufficient to effectively exert the desired effect, and can be appropriately selected depending on the application of the agent. However, generally, it is preferable that the ratio of 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives or salts to the total agent is usually 0.0001% by mass or more, more preferably 0.0001% by mass or more, and usually 5% by mass or less, more preferably 1.5% by mass or less. When using two or more types of 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives or salts, it is sufficient that their total amount satisfies the above range.

[0039] Furthermore, the agent relating to this disclosure may contain any other components in addition to 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives or salts, provided that the effects of 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives or salts are not substantially impaired. Examples of other components include other compounds having heparanase activity inhibitory effects or other activities (such as MMP inhibitory activity, particularly MMP-9 inhibitory activity) (e.g., CGS27023A, BIPBIPU), and pharmaceutically acceptable carriers and / or adjuvants. Examples of other components include, but are not limited to, crude drugs that act as heparanase inhibitors, such as valerian extract, lily extract, long-life grass extract, soapberry extract, and citrus peel extract (see Japanese Patent Application No. 2014-165586), and crude drugs that act as MMP inhibitors, such as mangosteen extract, turmeric extract, and (tormentilla extract) (see Japanese Patent Application No. 2018-545781). Such other components may be used individually or in any combination and ratio of two or more. Accordingly, some aspects of the present invention relate to agents such as dermal elasticity improving agents, dermal wrinkle improving agents, dermal papillary fiber increasing agents, dermal papillary fiber stabilizing agents, and LTBP2 and / or fibrillin 1 production promoting agents, which contain as active ingredients a compound having both heparanase inhibitory activity and MMP (particularly MMP-9) inhibitory activity, and / or a combination of a heparanase inhibitor and an MMP (particularly MMP-9) inhibitor.

[0040] The agent relating to this disclosure can be manufactured in accordance with conventional methods, and as a component of the topical skin preparation, it can be prepared using one or more of 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives or salts alone. However, ingredients commonly used in topical skin preparations such as cosmetics and pharmaceuticals, including quasi-drugs, such as oils, surfactants, powders, colorants, water, alcohols, thickeners, chelating agents, silicones, antioxidants, UV absorbers, moisturizers, fragrances, various pharmaceutically active ingredients, preservatives, pH adjusters, and neutralizing agents may be appropriately blended as needed.

[0041] The administration route and dosage form of the agent relating to this disclosure are not limited and may be appropriately selected according to their intended use. Examples of administration routes include topical administration (such as external skin application), oral administration, and parenteral administration (such as intravenous administration and intraperitoneal administration). However, when used as an agent relating to this disclosure, such as a dermal regeneration promoter, it is preferable to use it as a topical skin preparation. Examples of dosage forms for topical administration (topical skin preparations) include solutions, solubilization, emulsification, powder dispersion, water-oil two-layer systems, and water-oil-powder three-layer systems, which can be made into patches, ointments, creams, lotions, lotions, gels, aerosols, etc. Examples of dosage forms for oral administration include solid preparations such as tablets, coated tablets, sugar-coated tablets, granules, powders, and capsules (e.g., hard or soft gelatin capsules), as well as liquid preparations (solutions, suspensions) such as oral solutions and syrups. Examples of parenteral administration include injectable solutions.

[0042] Furthermore, the agent relating to this disclosure may contain, in addition to 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives or salts relating to this disclosure, one or more other components such as carriers and / or auxiliary agents, provided that the effects of 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives or salts thereof are not substantially impaired. The other components are not particularly limited and may be appropriately selected depending on the use, dosage form, administration method, etc., of the pharmaceutical composition, but examples include pharmaceutically acceptable carriers and / or auxiliary agents. Examples of auxiliary agents include diluents, binders, disintegrants, thickeners, dispersants, reabsorption enhancers, flavoring agents, buffers, surfactants, solubilizers, preservatives, emulsifiers, isotonic agents, stabilizers, pH adjusters, etc.

[0043] As a specific example, when the agent relating to this disclosure is used as a topical skin preparation, ingredients commonly used in topical preparations, such as whitening agents, moisturizers, antioxidants, oily components, UV absorbers, surfactants, thickeners, alcohols, powder components, colorants, aqueous components, water, various skin nutrients, etc., can be appropriately blended as needed. Furthermore, metal ion sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid; preservatives such as methylparaben, ethylparaben, and butylparaben; caffeine, tannins, verapamil, tranexamic acid and its derivatives; licorice extract, glabridin, hot water extract of quince fruit, various herbal medicines; drugs such as tocopherol acetate, glycyrrhizic acid and its derivatives or salts; whitening agents such as vitamin C, magnesium ascorbate phosphate, ascorbate glucoside, arbutin, and kojic acid; sugars such as glucose, fructose, mannose, sucrose, and trehalose; and vitamin A derivatives such as retinoic acid, retinol, retinyl acetate, and retinyl palmitate can also be added as appropriate.

[0044] The above ingredients are examples only and are not limited to these. Furthermore, these ingredients can be combined and formulated as appropriate according to the desired form.

[0045] The dosage form of the topical skin preparation relating to this disclosure is not particularly limited, and any dosage form can be used, for example, a solution system, a solubilized system, an emulsified system, a powder dispersion system, a two-layer water-oil system, a three-layer water-oil-powder system, an ointment, a gel, an aerosol, etc. Furthermore, the form of use is not particularly limited, and any form can be used, for example, a lotion, emulsion, cream, essence, jelly, gel, ointment, pack, mask, foundation, etc.

[0046] The agent relating to this disclosure can be used in cosmetic procedures to prevent the formation of dermal wrinkles and / or to reduce or eliminate existing dermal wrinkles when applied to the skin. The usage and dosage of the topical skin agent relating to this disclosure in such cosmetic procedures are not particularly limited and will be appropriately determined depending on the dosage form and the condition of the dermal wrinkles being treated. Typically, however, it is applied several times a day, for example, 1 to 5 times, in an appropriate amount, for example, 1 square cm. 2 You can apply 0.1ml to 1ml per serving by rubbing it directly into the skin, or you can soak an appropriate amount into gauze and then apply it to the skin.

[0047] Although the present invention has been described with specific examples above, these examples are merely illustrative, and the present invention can be implemented with any modifications without departing from the scope of the claims. The various features and embodiments of the present invention mentioned in the above sections may be applied to other sections of the description with the necessary modifications as appropriate. Accordingly, features specified in one embodiment may be combined with functions specified in other embodiments as appropriate. All references cited herein, including patents, patent applications, papers, textbooks, and sequence accession numbers, and the references cited therein, are incorporated herein by reference in their entirety. If one or more incorporated documents and similar materials differ from or contradict the present application in terms including, but not limited to, defined terms, usage of terms, and techniques described, the description of the present application shall prevail.

[0048] 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. [Examples]

[0049] 1. Materials and Methods Skin model culture A three-dimensional skin model (EFT-400) purchased from MatTeK was used with the MMP-9 inhibitor CGS27023A (Reference 1) (final concentration 10 -5 M) BIPBIPU (Reference 2), a heparanase inhibitor (final concentration 10-5 M), a self-developed component with the effect of inhibiting both MMP-9 and heparanase, was cultured in a dedicated medium (EFT400-ASY) supplemented with 1-(2-hydroxyethyl)-2-imidazolidinone (Reference 3) (hereinafter referred to as HEI, final concentration 0.01%). As a control, the cells were cultured in a medium to which an equal amount of DMSO solvent without these inhibitors was added. The medium was changed every two days, and tissue pieces were collected on the 4th and 7th days.

[0050] Culture of fresh human skin Fresh skin samples from the abdomen of subjects (aged 20s to 30s) who had given informed consent were purchased from KAC Co., Ltd. Using the same method as described in Reference 4, the obtained samples were irradiated with 50 mJ / cm 2 of UVB, and cultured in William's E medium (Thermo Fisher Science, Waltham, MA) containing CGS27023A (final concentration 10 -5 M), BIPBIPU (final concentration 10 -5 M), and HEI (final concentration 0.01%). As a control, the cells were cultured using a culture medium to which DMSO solvent was added in an equal amount without adding CGS27023A, BIPBIPU, or HEI. The medium was changed daily, and skin pieces were collected on the 5th day.

[0051] Paraffin block, sectioning The collected skin models and fresh human skin were dehydrated and fixed with cold acetone according to the AMeX method, then replaced with acetone, methyl benzoate, and xylene in that order, and embedded in paraffin. Sections were prepared with a thickness of 3 μm for tissue staining.

[0052] Various immunostaining Paraffin sections prepared with a thickness of 3 μm were deparaffinized with xylene and then hydrated with EtOH. Antibodies against type V collagen (Acris, AM10159PU-N, V13F6, mouse monoclonal antibody) and antibodies against cytokeratin 14 (K-14) (Fitzgerald, (20R-CP002, guniea pig polyclonal antibody), antibody against fibrillin-1 (Abcam, 11C1.3, mouse monoclonal antibody), antibody against α6 integrin (Santa Cruz, GOH-3, sc-19622, rat monoclonal antibody), antibody against LTBP2 (Protein-tech Group, 17708-1-AP, rabbit) Immunostaining was performed using a polyclonal antibody.

[0053] Gene expression of PDGFB (PDGF-BB) Only the epidermis of the collected skin model was peeled off with tweezers, and the epidermis was placed in a 1 mL Trizol solution containing zirconia balls. The epidermis was then disrupted by vibrating a tissue disruptor for 3 minutes. RNA was extracted using chloroform and isopropanol, purified using the RNeasy mini kit (QIAGEN), and the RNA concentration was measured using NanoDrop. cDNA was then synthesized using SuperScript VILO (Invitrogen). Quantitative PCR analysis was then performed using the synthesized cDNA with platinum SYBER green (Invitrogen). The primers used for the genes are shown in Table 1.

[0054] [Table 1]

[0055] LTBP2 gene expression Fibroblasts derived from the abdominal skin of informed consent-given subjects (ages 20-30 and 50-60) were purchased from KAC Corporation. They were cultured in DMEM medium containing 10% serum. 250,000 cells / well were seeded into 6-well plates, and mRNA was extracted two days later using the RNeasy mini kit (QUIAGEN). cDNA was then synthesized using SuperScript VILO (invitrogen). Quantitative PCR analysis was performed using the synthesized cDNA with platinum SYBER green (invitrogen). The primers used for the genes are shown in Table 2.

[0056] [Table 2]

[0057] LTBP2-heparan sulfate binding assay Heparan sulfate purchased from Seikagaku Kogyo was subjected to a ring-opening reaction with sodium hypoiodate and added to a carbobind plate (Corning) to solidify the heparan sulfate (the method for solidifying biotinylated heparan sulfate onto the plate was the same as the method described in Reference 5). Various concentrations of recombinant were added to this heparan sulfate plate. Human LTBP2 (abcam) was added and the mixture was reacted at room temperature for 2 hours. After washing three times with PBS, HRP-labeled anti-LTBP2 antibody was added and the mixture was reacted at room temperature for 2 hours, followed by three washes with PBS. After a color reaction with TMB, the reaction was stopped with 1N HCl, and the amount of bound LTBP2 was calculated by measuring the OD475nm.

[0058] Gene expression analysis of fibroblasts treated with PDGF-BB Fibroblasts from informed consent subjects (14 months old) were purchased from KAC Corporation. They were cultured in DMEM medium containing 10% serum. 250,000 cells / well were seeded into a 6-well plate, and after 1 day, a 10 ng / mL recombinant solution was added. Human PDGF-BB (R&D Systems) was added, and one day later, mRNA was extracted using the RNeasy mini kit (QUIAGEN). cDNA was then synthesized using SuperScript VILO (Invitrogen). Quantitative PCR analysis was then performed using the synthesized cDNA with platinum SYBER green (Invitrogen). The primers used for the genes are shown in Table 3.

[0059] [Table 3]

[0060] Human trials to verify the effects on skin elasticity Thirty healthy Japanese women aged 30-50 used a serum containing 1.5% HEI on one half of their face and a serum without HEI on the other half for four weeks. At 0, 2, and 4 weeks after the start of application, they visited the testing facility for acclimatization in a constant temperature and humidity chamber. Afterward, the transepidermal water loss on both cheeks was measured three times using a Vapometer (Delphin), and the stratum corneum moisture content was measured using a Corneometer (MPA580, Courage+Khazaka). Skin elasticity was measured five times using an electronic GmbH instrument, three times using a Cutometer, and the thickness of the dermal papillary layer was measured using an acoustic microscope AMS-50SI (Honda Electronics Co., Ltd.).

[0061] Human trials to verify the effectiveness of wrinkle improvement Thirty-four healthy Japanese women aged 30-50 used either a cream formulation containing 1.5% HEI or a cream formulation without HEI, applying 0.1 mL each to the outer corner of each eye twice daily for two months. At 0, 1, and 2 months after the start of application, participants came to the testing facility for acclimatization in a temperature and humidity controlled room. After that, a replica agent (SILFLO (Flexico, England)) was applied to the outer corner of the eye in a 4 cm circular area, and replicas were taken. The acquired replicas were analyzed using the ASA-03RXD reflective replica analysis system and reflective three-dimensional skin analysis software. A 10 mm square was used as the analysis area, with a point 5 mm away from the outer corner of the eye as the starting point, and the wrinkle area percentage (%) was calculated. After calculating the wrinkle area percentage (%) for both the left and right eyes at 0, 1, and 2 months, the participants were divided into active and placebo groups based on an allocation table, and the measurements before use (0 months), after 1 month, and after 2 months were compared for the active and placebo groups.

[0062] 2. Results and Discussion Promotion of PDGF-BB production by heparanase inhibitors and MMP inhibitors Figure 1 shows the results of quantitative PCR analysis of PDGFB gene expression levels in the epidermis of an artificial skin model (three-dimensional skin model). The results in Figure 1 indicate that treatment with a heparanase inhibitor (BIPBIPU) and an MMP inhibitor (CGS27023A) increases PDGFB production.

[0063] Changes in fibroblast gene expression due to PDGF-BB Since treatment with heparanase inhibitors and MMP inhibitors increased the amount of PDGFB genes in the epidermis, it is presumed that the amount of PDGF-BB protein in the epidermis also increased. Because PDGF-BB acts on fibroblasts in the dermis by passing through the basement membrane, the effects of PDGF-BB on fibroblasts were investigated next. Previous studies have shown that PDGF-BB enhances the production of type V collagen when applied to fibroblasts, but its effects on the expression of fibrillin 1 and LTBP2, which are involved in elastin fiber formation, have not been clarified. Therefore, Figure 2 shows the results of investigating the effects of PDGF-BB on the gene expression of fibrillin and LTBP2 when applied to fibroblasts. The results in Figure 2 show that PDGF-BB also promotes the production of LTBP2 and fibrillin 1.

[0064] LTBP2 localization changes due to heparanase inhibitors Based on the above results, it is possible that simultaneous application of a heparanase inhibitor (BIPBIPU) and an MMP inhibitor (CGS27023A), or application of HEI, an inhibitor that simultaneously inhibits the enzymatic activity of both heparanase and MMP, increases the amount of PDGF-BB in the epidermis and enhances the production of fibrillin and LTBP2 by fibroblasts in the dermal papillary layer. Furthermore, it is hypothesized that application of heparanase inhibitors (BIPBIPU, HEI) suppresses the degradation of heparan sulfate chains in the basement membrane component perlecan, allowing LTBP2, which has a heparan sulfate binding domain, to adhere to the basement membrane. Accordingly, the changes in LTBP2 localization in organ culture human skin models and three-dimensional skin models are shown in Figures 3A and 3B. The results in Figures 3A and 3B show that treatment with a heparanase inhibitor (BIPBIPU), an MMP inhibitor (CGS27023A), and a compound (HEI) that inhibits both MMP and heparanase enzymes increases LTBP2 production, allowing it to bind to heparan sulfate and stably localize to the basement membrane.

[0065] Heparanase inhibitor-induced changes in LTBP2 / Fibrillin-1 localization Figures 4A and 4B show the results of investigating changes in fibrillin 1 and LTBP2 localization in organ culture human skin models and three-dimensional skin models. The results in Figures 4A and 4B indicate that fibrillin 1 can be stably localized directly beneath the basement membrane via LTBP2 by treatment with a heparanase inhibitor (BIPBIPU), an MMP inhibitor (CGS27023A), or a compound that inhibits both heparanase and MMP enzymes (HEI). Furthermore, these results indicate that treatment with heparanase inhibitors (BIPBIPU, HEI) affects oxytalan fibers in the dermal papillary layer.

[0066] The binding of heparan sulfate and LTBP2 Figure 5 shows the results of an investigation into the binding of heparan sulfate and LTBP2 using a plate with heparan sulfate as the solid phase. The results in Figure 5 indicate that LTBP2 and heparan sulfate bind in a concentration-dependent manner.

[0067] Human trials to verify the effects on skin elasticity Thirty healthy Japanese women aged 30-50 used a serum containing 1.5% HEI on one half of their face and a serum without HEI on the other half for four weeks. At 0, 2, and 4 weeks after the start of application, they visited the testing facility for acclimatization in a constant temperature and humidity chamber. Afterward, the transepidermal water loss on both cheeks was measured three times using a Vapometer (Delphin), and the stratum corneum moisture content was measured using a Corneometer (MPA580, Courage+Khazaka). Skin elasticity was measured five times using an electronic GmbH device, and skin elasticity was measured three times using a cutometer. The results are shown in Figure 6. The results in Figure 6 show that treatment with a serum containing 1.5% HEI, a compound that inhibits the activity of both heparanase and MMP enzymes, improves dermal elasticity. In addition, the thickness of the dermal papillary layer was measured using an acoustic microscope AMS-50SI (Honda Electronics Co., Ltd.). The results are shown in Figure 7. The results in Figure 7 show that treatment with HEI promotes the increase of dermal papillary layer fibers and stabilizes the dermal papillary layer fibers.

[0068] Human trials to verify the effectiveness of wrinkle improvement Thirty-four healthy Japanese women aged 30-50 applied either a cream formulation containing 1.5% HEI (active) or a cream formulation without HEI (placebo) to the corners of their eyes. Replicas were taken, and the wrinkle area percentage (%) was calculated. The results are shown in Figure 8. Figure 8 shows that the wrinkle area percentage (%) was significantly reduced with the active formulation containing HEI and the treatment. On the other hand, no change was observed in the group treated with the placebo formulation. Since a decrease was observed on the active side before and after use, a Mann-Whitney U test was performed, and a significant decrease in the wrinkle area percentage (%) on the active side was confirmed before and after use.

[0069] References 1) Pan W, Miao HQ, Xu YJ, Navarro EC, Tonra JR, Corcoran E, et al. 1-[4-(1H-Benzoimidazol-2-yl)-phenyl]-3-[4-(1H-benzoimidazol-2-yl)-phenyl]- urea derivatives as small molecule heparanase inhibitors. Bioorganic & medicinalchemistry letters. 2006;16(2):409-12. 2) MacPherson LJ, Bayburt EK, Capparelli MP, Carroll BJ, Goldstein R, Justice MR, et al. Discovery of CGS 27023A, a non-peptidic, potent, and orally active stromelysin inhibitor that blocks cartilage degradation in rabbits. Journal of medicinal chemistry. 1997;40(16):2525-32. 3) Iriyama S, Yamanishi H, Kunizawa N, Hirao T, Amano S. 1-(2-Hydroxyethyl)-2-imidazolidinone, a heparanase and matrix metalloproteinase inhibitor, improves epidermal basement membrane structure and epidermal barrier function. Experimental dermatology. 2019;28(3):247-53. 4) Iriyama S, Yasuda M, Nishikawa S, Takai E, Hosoi J, Amano S. Decrease of laminin-511 in the basement membrane due to photoaging reduces epidermal stem / progenitor cells. Scientific reports. 2020;10(1):12592. 5) Farhad Behzad, Paul EC Brenchley. A multiwell format assay for heparinase. Anal Biochem. 2003 Sep 15;320(2):207-13. [Industrial applicability]

[0070] The agent relating to this disclosure promotes the production of PDGF-BB, LTBP2, and / or fibrillin 1, thereby increasing and / or stabilizing dermal papillary fibers, and can therefore be suitably used as a cosmetic composition such as a cosmetic product that is effective in improving dermal elasticity and / or dermal wrinkles.

Claims

1. A dermal elasticity improving agent characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient.

2. A dermal wrinkle-improving agent characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient.

3. A dermal papillary fiber enlargement promoting agent characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient.

4. A dermal papillary fiber stabilizer characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient.

5. The agent according to claim 3 or 4, wherein the dermal papillary fibers are dermal papillary oxytalane fibers.

6. An LTBP2 and / or fibrillin 1 production promoter characterized by containing 1-(2-hydroxyethyl)-2-imidazolidinone or a derivative thereof as an active ingredient.

7. 1-(2-hydroxyethyl)-2-imidazolidinone or its derivatives are defined by the following general formula (I): 【Chemistry 1】 (In the general formula (I), n is an integer from 1 to 3, R 1 X is a C1-C6 hydrocarbon group which may be substituted with a hydrogen atom or a hydroxyl group, and X is -CH 2 - or -N(R 2 ) is a group represented by -, R 2 (This refers to a hydrocarbon group having 1 to 6 carbon atoms, which may be substituted with a hydrogen atom or a hydroxyl group.) The agent according to any one of claims 1 to 6, which is a cyclic carboxamide derivative represented by .

8. A cosmetic method for improving dermal elasticity, characterized by increasing the production of LTBP2 and / or fibrillin 1.

9. A cosmetic method for preventing or improving wrinkles, characterized by increasing the production of LTBP2 and / or fibrillin 1.

10. A cosmetic method for preventing or improving wrinkles, characterized by the localization stabilization of LTBP2 and / or fibrillin 1.

11. A screening method for dermal elasticity improving agents using LTBP2 and / or fibrillin 1 expression as indicators.

12. A screening method for dermal wrinkle-improving agents using LTBP2 and / or fibrillin 1 expression as indicators.

13. A cosmetic method for preventing or improving wrinkles, characterized by increasing the expression of PDGF-BB.

14. A screening method for dermal elasticity improving agents using PDGF-BB expression as an indicator.

15. A screening method for dermal wrinkle-improving agents using PDGF-BB expression as an indicator.

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