Fibrillin production promoter and use thereof

Abstract

To provide a fibrillin production promoter containing a 3-hydroxybutyric acid component and capable of promoting production of fibrillin, a major constituent of elastic fibers.SOLUTION: A fibrillin production promoter is prepared that contains at least one 3-hydroxybutyric acid component selected from 3-hydroxybutyric acid, a salt of 3-hydroxybutyric acid, and a 3-hydroxybutyric acid alkyl ester represented by the following formula (1) (where R represents an alkyl group). The 3-hydroxybutyric acid component may include an R-form. The fibrillin may be fibrillin-1. The fibrillin production promoter may be a promoter that promotes production of fibrillin in senescent cells.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present disclosure relates to a fibrillin production promoter comprising at least one selected from 3-hydroxybutyric acid, a salt thereof, and an alkyl ester thereof, and uses thereof. [Background technology]

[0002] Human skin has a three-layer structure consisting of epidermis, dermis, and subcutaneous tissue, in that order from the surface side. The dermis, which accounts for the majority of skin tissue, mainly contains fibrous tissue composed of proteins (e.g., collagen fibers and elastin fibers), interstitial components or matrix (e.g., hyaluronic acid and glycosaminoglycan) that fill the spaces between the fibrous tissue, and fibroblasts. The fibroblasts produce the fibrous tissue and interstitial components.

[0003] Collagen fibers, the main component of the dermis, are spread throughout the skin in a mesh-like structure, and elastin fibers are entangled with the collagen fibers at regular intervals to support the mesh-like structure. The spaces (gaps) in the mesh-like structure are filled with the jelly-like, elastic interstitial components, which retain moisture in the skin and maintain the mesh-like structure.

[0004] Healthy skin that forms such an elastic mesh structure has sufficient moisture (water) and elasticity (firmness). On the other hand, in skin that has aged due to aging or exposure to ultraviolet rays, the number of fibroblasts decreases due to the destruction of the fibroblasts and a decrease in the production speed, and therefore the fibrous tissue and interstitial components cannot be produced sufficiently, and the shape of the mesh structure of the dermis cannot be maintained, resulting in wrinkles and sagging.

[0005] Furthermore, decomposition (breakage) and deterioration of elastin fibers by elastase is also known to be a cause of the breakdown of the meshwork structure of the dermis.

[0006] A glycoprotein called fibrillin is involved in the formation of elastin fibers (elastic fibers), and in humans, the main known fibrillins are fibrillin-1 and fibrillin-2. Fibrillin-1 supports the structure of elastic fibers, and fibrillin-2 is known to be synthesized simultaneously with the start of elastic fiber formation and to be involved in the initial formation of elastic fibers (Non-Patent Document 1), and fibrillin-1 and fibrillin-2 are involved in the formation of elastic fibers in a correlated manner.

[0007] Elastin fibers (elastic fibers) have a basic skeleton in which self-aggregates of tropoelastin are deposited or bonded onto microfibrils that are made up of bundles of the fibrous protein fibrillin-1.

[0008] Furthermore, factors involved in the formation of elastin fibers include factors that crosslink or self-aggregate tropoelastin to form self-aggregates [LOX (lysyl oxidase), fibulin 4, etc.], factors that promote the formation of microfibrils by bundling fibrillin-1 [LTBP-4 (Latent TGFβ binding protein 4), MFAP-4 (Microfibrillar-associated protein 4), EMILIN-1 (Elastin microfibril interfacer 1), etc.], and factors that promote the deposition of self-aggregated tropoelastin into microfibrils [Fibulin 5, etc.]. Elastin fibers are formed through the interaction of various such factors.

[0009] However, the turnover of elastin fibers is very slow, and it is believed that once degraded or deteriorated, elastin fibers have almost no ability to be regenerated, particularly in aged tissues due to aging, etc. Therefore, it is expected that this compound will promote the formation of elastin fibers, particularly in aged tissues.

[0010] Japanese Patent No. 6434180 (Patent Document 1) discloses a microfibril production promoter containing an extract of Rhodiola rosea, a perennial herb of the Crassulaceae family, as an active ingredient. In Patent Document 1, an extract using the roots of Rhodiola rosea is added to a culture system of human fibroblasts, and the expression level of genes involved in the formation of microfibrils is evaluated, demonstrating the microfibril formation promoting effect.

[0011] In addition, Japanese Patent No. 6032734 (Patent Document 2) discloses a fibrillin 1 production promoter containing as an active ingredient one or more compounds selected from the group consisting of furfural derivatives represented by the following chemical formula.

[0012] [ka]

[0013] R1 is -H, -CH 3 , -CHO, -COOH, or -CH 2 OCOCH 3 and R2 is -H or -CH 3 where R3 is -H and R4 is -CHO or -CH 2 Compound (A) is OH, R1, R2, and R3 are -H, and R4 is -CH 2 Compound (B) which is OH.

[0014] In Patent Document 2, the furfural derivatives were added to a culture system of human dermal fibroblasts, and the expression-promoting activity of Fibrillin-1 mRNA and GAPDH mRNA was measured. The production-promoting activity was evaluated as a relative amount (mRNA expression level), and it was shown that certain furfural derivatives had high production-promoting activity.

[0015] Non-Patent Document 2 discloses that retinol (ROL), which is known to promote collagen production in the dermis, also promotes the formation of elastin fibers. Specifically, Non-Patent Document 2 shows that when cultured human skin explants are locally treated with a low dose (0.04%) of ROL, the expression levels of mRNA and protein of tropoelastin and fibrillin-1, which are constituent proteins of elastin fibers, increase, and the network structure of elastin fibers increases. In other words, it is suggested that ROL not only promotes the proliferation of epidermal cells and collagen production, but also promotes the production and assembly of elastin fibers, thereby effectively acting in anti-aging. However, there are concerns about the tingling skin irritation caused by ROL, although this varies from person to person.

[0016] On the other hand, 3-hydroxybutyrate (3HB or BHB), a type of ketone body, and its salts are substances produced in the liver and found in breast milk, and have been reported to have various functions on the skin.

[0017] JP 2017-200883 A (Patent Document 3) discloses a cosmetic composition containing 3HB. This document describes the effects of the cosmetic, including a whitening effect (effect as a whitening cosmetic) evaluated by tyrosinase activity (amount of melanin produced per unit protein amount) using human pigment cells, a cell activation effect (effect as a cell activation cosmetic) evaluated by MTT reduction activity using fibroblasts and epidermal cells in a nutritionally deficient or aged state, and a moisturizing effect (effect as a moisturizing cosmetic) measured by the moisture content of the stratum corneum of the forearm.

[0018] Japanese Patent No. 6804352 (Patent Document 4) discloses an inhibitor of collagenase MMP1 and 3 production, which contains R-3-hydroxybutyric acid as an active ingredient. In this document, the inhibitory effect on collagenase MMP1 and 3 production is evaluated in normal human skin fibroblast cells and in an aging model (senescence cells) in which normal cells are treated with hydrogen peroxide.

[0019] Japanese Patent No. 6979918 (Patent Document 5) discloses a human senescent cell activator containing an alkyl ester of 3-hydroxybutyrate as an active ingredient. In this document, the human senescent cell activating effect is evaluated by measuring the collagen content of senescent cells obtained by treating normal human fibroblasts with hydrogen peroxide. [Prior art documents] [Patent documents]

[0020] [Patent Document 1] Patent No. 6434180 [Patent Document 2] Patent No. 6032734 [Patent Document 3] JP 2017-200883 A [Patent Document 4] Patent No. 6804352 [Patent Document 5] Patent No. 6979918 [Non-patent literature]

[0021] [Non-Patent Document 1] Journal of Cell Biology, Volume 129, p.1165-1176, 1995 [Non-Patent Document 2] International Journal of Cosmetic Science / Volume 33, Issue 1 / Pages 62-69 (2011) Summary of the Invention [Problem to be solved by the invention]

[0022] With growing interest in anti-aging, it is expected that the production or formation of elastin fibers involved in maintaining skin elasticity will be promoted, and compounds or compositions that act to improve the formation of elastin fibers (elastic fibers) are known, as described in Patent Documents 1 and 2 and Non-Patent Document 2. However, the ROL described in Non-Patent Document 2 is irritating to the skin, and there are cases in which it is difficult to use safely.

[0023] In addition, in recent years, it has become important that substances that come into direct contact with the skin are highly safe. As shown in Patent Documents 3 to 5, 3-hydroxybutyric acid and / or its derivatives, which are produced in the body and are highly safe, have attracted attention, and various functions on the skin have been reported.

[0024] However, it is not yet known that 3-hydroxybutyric acid and / or its derivatives are involved in the formation of elastin fibers, and in particular, that 3-hydroxybutyric acid and / or its derivatives promote or enhance the production of fibrillin.

[0025] Therefore, an object of the present disclosure is to provide a fibrillin production-promoting agent that can promote the production of fibrillin, a main component of elastin fibers, and uses thereof.

[0026] Another object of the present disclosure is to provide a fibrillin production promoter that can promote or improve fibrillin production even in senescent cells, and uses thereof.

[0027] Still another object of the present disclosure is to provide a fibrillin production promoter that is low irritation to the skin and highly safe, and uses thereof. [Means for solving the problem]

[0028] As a result of intensive research to solve the above-mentioned problems, the inventors discovered that at least one 3-hydroxybutyric acid component selected from 3-hydroxybutyric acid, salts of 3-hydroxybutyric acid, and alkyl esters of 3-hydroxybutyric acid can promote or improve the production of fibrillin (particularly fibrillin-1 or fibrillin-1 fibers), which is closely involved in the formation of elastin fibers, and thus completed the present invention.

[0029] That is, the fibrillin production promoter according to embodiment [1] of the present disclosure comprises 3-hydroxybutyric acid, a salt of 3-hydroxybutyric acid, and a compound represented by the following formula (1):

[0030] [ka]

[0031] (wherein R represents an alkyl group). The 3-hydroxybutyric acid component includes at least one 3-hydroxybutyric acid component selected from 3-hydroxybutyric acid alkyl esters represented by the following formula:

[0032] Aspect [2] of the present disclosure is an aspect of the above aspect [1], in which the 3-hydroxybutyric acid component contains an R-form.

[0033] Aspect [3] of the present disclosure is an aspect [1] or [2] above, in which the fibrillin is fibrillin-1.

[0034] Aspect [4] of the present disclosure is any one of aspects [1] to [3] above, in which production of fibrillin in senescent cells is promoted.

[0035] Aspect [5] of the present disclosure is any one of aspects [1] to [4] above, which is applied to the skin to bring about the following effects (a) and / or (b) (or improvements):

[0036] (a) Improves the formation of elastin fibers (b) Prevent loss of skin elasticity

[0037] The present disclosure also includes, as aspect [6], a composition for promoting fibrillin production, comprising at least one 3-hydroxybutyric acid component selected from 3-hydroxybutyric acid, a salt of 3-hydroxybutyric acid, and an alkyl ester of 3-hydroxybutyric acid represented by the formula (1).

[0038] An embodiment [7] of the present disclosure is an embodiment in which the composition of the embodiment [6] is a transdermal composition (transdermal composition or topical composition).

[0039] Aspect [8] of the present disclosure is an aspect in which the composition of aspect [6] or [7] is a liquid or semi-solid composition.

[0040] An embodiment [9] of the present disclosure is an embodiment in which the composition of any one of the above embodiments [6] to [8] is a cosmetic (skin care cosmetic or skin care product).

[0041] The present disclosure also includes, as embodiment

[10] , an elastin fiber formation improver comprising at least one 3-hydroxybutyric acid component selected from 3-hydroxybutyric acid, a salt of 3-hydroxybutyric acid, and an alkyl ester of 3-hydroxybutyric acid represented by the formula (1) above.

[0042] The present disclosure also includes, as aspect

[11] , a method of promoting fibrillin production by applying (administering or transdermally) to the skin a fibrillin production promoter that is any one of aspects [1] to [5] above, and, as aspect

[12] , a method of promoting fibrillin-1 production in senescent cells by applying (administering or transdermally) to the skin a fibrillin production promoter that is any one of aspects [1] to [5] above.

[0043] In this specification and claims, the term "senescent cells (aged cells)" refers to cells that have been damaged by stress such as ultraviolet rays, drugs, and aging and are unable to divide or grow, or cells with reduced ability to divide or grow, including cells with reduced functions such as protein synthesis. Such senescent cells are not limited to the elderly, but are also found in young people and infants. Effect of the Invention

[0044] In the present disclosure, the fibrillin production promoter contains at least one selected from 3-hydroxybutyric acid (sometimes referred to as "3HB" or "BHB"), its salts, and its alkyl esters (sometimes collectively referred to as "3-hydroxybutyric acid component"), and can promote or improve the production of fibrillin (particularly fibrillin-1) in cells. In particular, the fibrillin production promoter of the present disclosure can significantly promote or improve the production of fibrillin in cells that have aged due to aging or exposure to ultraviolet light. In the present disclosure, by applying (or administering transdermally) the fibrillin production promoter to the skin, it is possible to promote the production of fibrillin not only in healthy (or normal) skin cells, but also in aged (or damaged) skin cells.

[0045] In particular, fibrillin (e.g., fibrillin-1) is a major component that forms the basis of elastin fibers, and in the present disclosure, the formation of elastin fibers can be improved by promoting the production of the fibrillin (e.g., fibrillin-1). Therefore, it is particularly useful for maintaining the mesh-like structure of the dermis against the decrease in the production ability and decomposition (or deterioration) of elastin fibers that accompanies aging, and can prevent or suppress the decrease in skin elasticity (firmness).

[0046] Furthermore, since the fibrillin production promoter disclosed herein contains a 3-hydroxybutyric acid component that is produced in the body (or converted to 3-hydroxybutyric acid in the body), it is hypoallergenic to the skin and is highly safe and can be used with confidence, even on sensitive skin or aged skin with reduced barrier function. [Brief description of the drawings]

[0047] [Figure 1] FIG. 1 is a graph showing the relationship between the concentration of 3HB-Na (the sodium salt of 3HB) and the fibrillin-1 fiber area ratio for normal human fibroblasts (NHDF) (Example 1 and Comparative Example 1) and senescence-induced fibroblasts (Example 2 and Comparative Example 2). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0048] [Fibrillin production promoter] The fibrillin production promoter of the present disclosure comprises 3-hydroxybutyric acid, a salt of 3-hydroxybutyric acid, and a compound represented by the following formula (1):

[0049] [ka]

[0050] (wherein R represents an alkyl group). The composition contains at least one 3-hydroxybutyric acid alkyl ester (3-hydroxybutyric acid component, 3HB component) selected from the 3-hydroxybutyric acid alkyl esters represented by the following formula:

[0051] The fibrillin production promoter of the present disclosure is particularly effective in improving the production of fibrillin-1, which supports the structure of elastin fibers, among the fibrillins that act on the formation of elastin fibers. Therefore, the "fibrillin production promoter" of the present disclosure is often used synonymously with the "fibrillin-1 production promoter."

[0052] (3-hydroxybutyric acid component) (3-hydroxybutyric acid) Commercially available 3-hydroxybutyric acid (3HB or BHB) may be used. Examples of commercially available products include chemically synthesized 3HB and 3HB produced by fermentation using microorganisms. Of these, fermentation-produced 3HB (fermentation-derived 3HB) is preferred because of its high purity of the R-isomer, and fermentation-produced 3HB using microorganisms from biomass raw materials (resources derived from living organisms) is particularly preferred.

[0053] The pH of 3HB (free 3HB) in a 40% by mass aqueous solution (temperature: 20° C.) may be, for example, about 1.5 to 2.5, and is preferably 1.6 to 2.3.

[0054] (salt of 3-hydroxybutyric acid) The salt of 3HB may be any salt with a physiologically or pharmacologically acceptable basic compound, and may be a salt in which the acid form of 3HB has been completely neutralized, or a salt in which the acid form of 3HB has been partially neutralized (partially neutralized salt).

[0055] The pH of a 3HB salt (including a partially neutralized salt) in a 5% by mass aqueous solution (temperature: 20°C) may be 2 or more, for example, about 2 to 8.5, preferably 3 to 8.3, further preferably 4 to 8.2, more preferably 5 to 8.1, and most preferably about 6 to 8.

[0056] Examples of the salts include alkali metal salts such as sodium salt, potassium salt, and lithium salt; alkaline earth metal salts such as magnesium salt and calcium salt; ammonium salt; amine salt; and salts with basic amino acids. These 3HB salts (3HB salts) can be used alone or in combination of two or more. Among these, alkali metal salts such as sodium salt and potassium salt; and alkaline earth metal salts such as magnesium salt and calcium salt are preferred because of their relatively low deliquescence and excellent handling properties.

[0057] (3-Hydroxybutyric acid alkyl ester) The 3-hydroxybutyric acid alkyl ester is represented by the above formula (1). The 3-hydroxybutyric acid alkyl ester is converted into 3HB and / or its salt by hydrolysis in the living body, and can be used in the same manner as 3HB and / or its salt. In addition, the 3-hydroxybutyric acid alkyl ester contains an alkyl group, and therefore has lipophilicity, which can improve the permeability into the skin.

[0058] In the formula (1), the alkyl group represented by R is, for example, a linear or branched C alkyl group such as a methyl group, an ethyl group, a propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a s-butyl group, a t-butyl group, a pentyl group, an isopentyl group, a hexyl group, an isohexyl group, an octyl group, a decyl group, a dodecyl group (lauryl group), an isododecyl group (isolauryl group), a tetradecyl group (myristyl group), an isotetradecyl group (isomyristyl group), a hexadecyl group (cetyl group or palmityl group), an isohexadecyl group (isocetyl group), an octadecyl group (stearyl group), an isooctadecyl group (isostearyl group), or an eicosyl group. 1-28 The alkyl group may be a straight or branched C 1-10 Alkyl groups, more preferably linear or branched C 1-6 Alkyl groups, more preferably linear or branched C 1-4 It is an alkyl group. The alkyl group represented by R may be appropriately selected depending on the usage form of the fibrillin production promoter of the present disclosure. These 3-hydroxybutyric acid alkyl esters can be used alone or in combination of two or more kinds.

[0059] Specific examples of 3-hydroxybutyric acid alkyl esters include 3HB C, such as 3HB methyl ester and 3HB ethyl ester. 1-4 Alkyl esters and the like.

[0060] The 3-hydroxybutyric acid alkyl ester may be produced by esterifying 3-hydroxybutyric acid with an alcohol [an alcohol (R-OH) corresponding to the group R in the above formula (1)] in accordance with a commonly used method, for example, the method described in Japanese Patent No. 6979918.

[0061] (3HB ingredient characteristics) The 3-hydroxybutyric acid component (3HB component) may be an optical isomer (R or S form) or may be a racemate; however, from the standpoint of biocompatibility, safety, etc., it is preferable to contain at least the R-form of the 3HB component [R-3-hydroxybutyric acid component, (R)3HB component].

[0062] The proportion of the R-form in the 3HB component, particularly the optical purity (enantiomer or optical isomer excess), may be, for example, 50% ee or more, and is preferably 80 to 100% ee, 90 to 100% ee, 95 to 100% ee, 98 to 100% ee, 99 to 100% ee, and particularly preferably substantially 100% ee. If the optical purity is too low, there is a risk of reduced biocompatibility.

[0063] In addition, the R-form [(R)3HB component] may be used in combination with the S-form [(S)3HB component] and / or racemic form, but it is preferable that the proportion of the S-form is small, and it is particularly preferable that it is unavoidably contained. The mass proportion of the R-form in the 3HB component is preferably 10 mass% or more, even more preferably 50 mass% or more, even more preferably 90 mass% or more, and most preferably 100 mass%. When the proportion of the R-form is high, it has high biocompatibility when used as a cosmetic or pharmaceutical product, and therefore the function of the 3HB component can be efficiently expressed in the body.

[0064] The 3HB component may be any of 3HB alone in the form of an acid; a 3HB salt alone in the form of a salt; a 3HB alkyl ester alone; or a combination (mixture) of at least two or more selected from 3HB, a 3HB salt, and a 3HB alkyl ester.

[0065] The 3HB component can be selected appropriately depending on the form of use, etc. For example, when used under hydrophilic conditions, it is preferable to contain at least one selected from 3HB and a 3HB salt, and when used under lipophilic conditions, it is preferable to contain at least a 3HB alkyl ester, and in particular, it is preferable to contain only a 3HB alkyl ester.

[0066] Of the 3HB components, it is preferable to contain at least a salt of 3HB, from the viewpoint of exhibiting a weak acidity close to the pH of the skin and reducing irritation to the skin.

[0067] When the 3HB component contains at least a 3HB salt, the mass ratio of the 3HB salt to other 3HB components (other 3HB components excluding 3HB salts such as 3HB and / or 3-hydroxybutyric acid alkyl esters, particularly 3-hydroxybutyric acid alkyl esters) may be about former / latter = 100 / 0 to 5 / 95, preferably in the following stepwise manner, former / latter = 100 / 0 to 20 / 80, 100 / 0 to 50 / 50, 100 / 0 to 75 / 25, 100 / 0 to 90 / 10, and particularly preferably 100 / 0 to 95 / 5.

[0068] In particular, it is preferable that the 3HB component is a 3HB salt alone, and representative 3HB components (particularly 3HB salts) include, for example, 3HB alkali metal salts such as 3HB sodium salt and potassium salt, and 3HB alkaline earth metal salts such as 3HB magnesium salt and calcium salt.

[0069] The proportion of the 3HB component (especially the 3HB salt) in the fibrillin production promoter may be 10% by mass or more, preferably 50% by mass or more, even more preferably 80% by mass or more, even more preferably 90% by mass or more, and most preferably 100% by mass.

[0070] (Characteristics of fibrillin production promoter) The fibrillin production promoter of the present disclosure may further contain a 3HB oligomer. The average degree of polymerization of the 3HB oligomer may be 2 or more, for example, 2 to 10, preferably 2 to 5, further preferably 2 to 4, more preferably 2 to 3, and most preferably 2. The 3HB oligomer may be an oligomer that is unavoidably mixed in during the production process of 3HB, etc.

[0071] The proportion of the 3HB oligomer may be 10 parts by mass or less, preferably 5 parts by mass or less, more preferably 3 parts by mass or less, and most preferably 1 part by mass or less, per 100 parts by mass of the 3HB component (active ingredient). The fibrillin production promoter of the present disclosure may be substantially free of 3HB oligomer, and it is particularly preferred that it does not contain 3HB oligomer.

[0072] The fibrillin production promoter of the present disclosure (first fibrillin production promoter) may, if necessary, further contain another promoter that promotes fibrillin production (second fibrillin production promoter).

[0073] Examples of the second fibrillin production promoter include vitamins such as retinol (vitamin A) (e.g., retinol such as retinol palmitate and retinol acetate), vitamin C derivatives (e.g., water-soluble vitamin C derivatives such as ascorbyl phosphate and ascorbyl ethyl, oil-soluble vitamin C derivatives such as ascorbyl tetrahexyldecanoate and ascorbyl dipalmitate, and water-soluble-oil-soluble vitamin C derivatives such as ascorbyl palmitate phosphate 3Na), polyphenols such as naringin (e.g., glucosylnaringin, a glycoside with increased water solubility), ceramides (synthetic ceramides) such as human ceramide (e.g., human ceramide 1, human ceramide 2, etc.), and furfural derivatives. The second fibrillin production promoter may be contained alone or in combination of two or more types.

[0074] The total proportion of the second fibrillin production promoter in the fibrillin production promoter is less than 50% by mass, preferably 30% by mass or less, more preferably 10% by mass or less, and even more preferably 5% by mass or less. The fibrillin production promoter may not substantially contain the second fibrillin production promoter, and it is particularly preferable that it does not contain the second fibrillin production promoter.

[0075] The fibrillin production-promoting agent of the present disclosure can be used to promote the production of fibrillin in the body of a mammal, such as a human, and is preferably used to promote the production of fibrillin in fibroblasts of humans and non-human animals, such as medium-sized or large mammals (particularly humans).

[0076] The fibrillin production promoter (at least the fibrillin-1 production promoter) of the present disclosure may be applied orally (administered or given) or parenterally. Of these, parenteral application is preferred. Parenteral application methods include, for example, application by inhalation, application by injection, transdermal application, and nasal application. Of these, transdermal application is preferred because it can effectively promote the production of fibrillin (particularly fibrillin-1) in the dermis (particularly fibroblasts).

[0077] The method of percutaneous application is not particularly limited as long as the fibrillin production promoter applied to the skin surface can penetrate to the dermis. Specific methods include, for example, a method of directly applying the fibrillin production promoter to the skin by dropping or spraying, and a method of contacting the skin with a carrier (mask, cotton, sponge, film, etc. made of nonwoven fabric, etc.) impregnated with the fibrillin production promoter. The application site on human skin is not particularly limited, and can be applied to the skin of the entire body, including, for example, the head (including the face), neck, upper limbs, chest, back, lower back, buttocks, lower limbs, etc.

[0078] The application amount (or dosage) of the fibrillin production promoter of the present disclosure can be appropriately selected depending on the subject, the age and weight of the subject, the application time, the form of the fibrillin production promoter, the application route, the application method, etc. For example, when applied transdermally, the fibrillin production promoter of the present disclosure may be applied to the necessary site (such as the skin) about 1 to 10 times per day, preferably 1 to 5 times, more preferably 1 to 4 times, more preferably 1 to 3 times, and most preferably 1 to 2 times.

[0079] When the fibrillin production promoter of the present disclosure is applied to the skin, it can promote the production of fibrillin, particularly fibrillin-1, in fibroblasts contained in the dermis, and also promote the formation of elastin fibers whose main component is this fibrillin (particularly fibrillin-1). In order to promote the formation of elastin fibers and maintain the meshwork structure of the dermis, it is preferable that fibrillin, particularly fibrillin-1, is continuously produced. Therefore, the fibrillin production promoter of the present disclosure is preferably applied repeatedly intermittently (or periodically at predetermined times) for a predetermined number of days, and may be applied intermittently for, for example, 24 hours or more, preferably for 1 week or more (e.g., 1 to 20 weeks), more preferably for 4 weeks or more, more preferably for 6 weeks or more, and most preferably for 8 weeks or more.

[0080] The application concentration (final concentration) of the 3HB component of the fibrillin production promoter of the present disclosure may be an effective amount, and may be, for example, 0.0001 mM to 1 M, preferably 0.001 to 500 mM, more preferably 0.01 to 100 mM, and even more preferably 0.05 to 50 mM, calculated as 3HB. If the concentration of the 3HB component is too high, it may be an excessive amount, which may reduce economic efficiency.

[0081] In addition, since the fibrillin production promoter of the present disclosure contains a 3HB component that is produced or converted to 3HB in the human body, it is highly biocompatible and effectively acts to promote fibrillin production even if the applied concentration (final concentration) of the 3HB component (especially a salt of 3HB) is low. In a cell-level experiment using human fibroblasts, the applied concentration (final concentration) of the 3HB component may be, for example, about 0.0001 to 5 mM in terms of 3HB, and is preferably 0.0005 to 3 mM, 0.001 to 2.5 mM, 0.005 to 2 mM, 0.007 to 1.5 mM, 0.008 to 1 mM, and 0.01 to 0.5 mM in the following stepwise manner. If the concentration of the 3HB component is too low, there is a risk that the production of fibrillin cannot be sufficiently promoted. In the above experiment, the number of cells (cells before culture) to be seeded was, for example, 1.0 x 10 3 ~1.0×10 5 cells, preferably 5.0 × 10 3 ~7.5×104 cells, and more preferably 7.5 × 10 3 ~5.0×10 4 cells.

[0082] Furthermore, when the fibrillin production promoter of the present disclosure is applied to the skin, the amount of the 3HB component to be applied is, in terms of the solid content of 3HB, 1 cm of skin. 2 For example, the amount may be about 0.001 to 10 mg, preferably 0.005 to 5 mg, further preferably 0.01 to 1 mg, more preferably 0.05 to 0.5 mg, and particularly preferably about 0.075 to 0.3 mg.

[0083] The fibrillin production promoter of the present disclosure (at least the fibrillin-1 production promoter) contains a 3HB component and can promote or improve the production of fibrillin (particularly fibrillin-1) in cells (fibroblasts). In particular, the fibrillin production promoter of the present disclosure can effectively promote or improve the production of fibrillin (particularly fibrillin-1) even in cells (fibroblasts) that have aged due to aging or exposure to ultraviolet light, and the effect of promoting fibrillin production is more pronounced in aged cells than in normal cells.

[0084] Therefore, the fibrillin production promoter of the present disclosure is preferably applied to aged skin, and is particularly preferably applied to skin that has lost elasticity due to aging for the purpose of anti-aging.

[0085] Moreover, the fibrillin production promoter of the present disclosure can improve the formation of elastin fibers containing fibrillin (e.g., fibrillin-1) as a main component by promoting the production of fibrillin (particularly fibrillin-1). Therefore, the fibrillin production promoter of the present disclosure (at least the fibrillin-1 production promoter) can also be used as an elastin fiber formation enhancer, and can improve the formation of elastin fibers and suppress or prevent a decrease in skin elasticity (firmness) even in skin in which elasticity has decreased due to a decrease in the ability to produce elastin fibers or decomposition (or deterioration) of elastin fibers caused by aging.

[0086] Moreover, since the fibrillin production promoter disclosed herein contains a 3HB component produced in the body or a 3HB component that is converted to 3HB, it is highly biocompatible, and even if applied to the skin in the form of a composition if necessary, it is low-irritation to the skin and is highly safe even for sensitive skin or aged skin with weakened barrier function.

[0087] The fibrillin production promoter and elastin fiber formation enhancer may be referred to simply as the 3HB component or "active ingredient."

[0088] Compositions for promoting fibrillin production (and enhancing elastin fiber formation) The compositions of the present disclosure are compositions for promoting fibrillin production (and enhancing the formation of elastin fibers) and contain at least a 3HB component, preferably at least an R-3HB component [(R)3HB component].

[0089] The composition of the present disclosure may be any of a liquid composition, a semi-solid composition, and a solid composition. That is, in the composition of the present disclosure, the (R)3HB component is incorporated as the fibrillin production promoter into a liquid composition, a semi-solid composition, or a solid composition.

[0090] Examples of liquid compositions include liquid preparations, drinks (beverages), suspensions, emulsions, syrups, and injections.

[0091] Examples of semi-solid compositions (semi-solid preparations) include gels, poultices, creams, slurries, pastes, and the like.

[0092] Examples of solid compositions (solid preparations) include powders, fine granules, granules, pills, tablets, flakes, cakes, gummies, nougat, films, and capsules.

[0093] Of these, because the composition of the present disclosure is particularly effective on the skin and can be easily applied or applied transdermally, topical compositions (or topical preparations), for example, liquid compositions and semi-solid compositions, are preferred, with liquid compositions being particularly preferred.

[0094] The 3HB component (active ingredient) in the composition, including preferred embodiments, is the same as the 3HB component (active ingredient) described above as the fibrillin production promoter.

[0095] The proportion of the 3HB component (active ingredient) in the composition can be selected from the range of about 0.01 to 99% by mass, preferably 0.1 to 95% by mass, and more preferably 0.5 to 90% by mass. The proportion of the 3HB component may be selected depending on the form of the composition. In the composition of the present disclosure, since the 3HB (active ingredient) has high biocompatibility, even a small amount of the 3HB component can significantly promote fibrillin production.

[0096] The method of application of the composition of the present disclosure, including preferred embodiments, is the same as the method of application described for the fibrillin production promoter. Therefore, the composition of the present disclosure is preferably a transdermal composition (transdermal composition).

[0097] The transdermal composition of the present disclosure may be a transdermal liquid composition or a transdermal semi-solid composition.

[0098] (Transdermal liquid composition) In the transdermal liquid composition (or external liquid composition), the ratio of the 3HB component is, for example, 0.01% by mass or more, preferably 0.1% by mass or more, more preferably 0.2% by mass or more, more preferably 0.3% by mass or more, particularly preferably 0.5% by mass or more, and most preferably 0.8% by mass or more in the transdermal liquid composition. If the ratio of the 3HB component is too low, there is a risk that the production of fibrillin cannot be promoted. In addition, in the transdermal liquid composition, the ratio of the 3HB component is, for example, 30% by mass or less, preferably 10% by mass or less, more preferably 5% by mass or less, even more preferably 3% by mass or less, particularly preferably 2% by mass or less, and most preferably 1.5% by mass or less in the transdermal liquid composition. The ratio of the 3HB component may be 0.1 to 5% by mass, particularly 0.3 to 3% by mass in the transdermal liquid composition. If the ratio of the 3HB component is too high, there is a risk that the handling property and the economical efficiency may be reduced.

[0099] The transdermal liquid composition of the present disclosure may further contain a liquid base, which includes a solvent, a liquid oil, and the like.

[0100] The solvent may be a lipophilic solvent, but from the viewpoint of safety, a hydrophilic solvent is preferable. Examples of hydrophilic solvents include water, lower aliphatic alcohols (e.g., C 10 solvents such as ethanol and isopropanol), 1-4 Alkyl alcohols, etc. These solvents can be used alone or in combination of two or more. Examples of liquid oils include animal and vegetable oils (e.g., jojoba oil, olive oil, palm oil, camellia oil, macadamia nut oil, avocado oil, corn oil, sesame oil, wheat germ oil, linseed oil, castor oil, etc.), mineral oils (e.g., liquid paraffin, polybutene, silicone oil, etc.), synthetic oils (e.g., synthetic ester oil, synthetic polyether oil, etc.), etc. These liquid oils can be used alone or in combination of two or more.

[0101] As the liquid base, a hydrophilic solvent such as water or ethanol may be used in combination with a liquid oil as an additive (oil component). Among these liquid bases, water, lower alcohols or mixtures thereof are preferred, and water and / or ethanol (especially water) are particularly preferred.

[0102] The proportion of the liquid base in the transdermal liquid composition is 50 to 99.9 mass %, preferably 60 to 99 mass %, further preferably 70 to 95 mass %, even more preferably 80 to 93 mass %, and most preferably 85 to 90 mass %.

[0103] The transdermal liquid composition of the present disclosure may further contain a moisturizing agent. Examples of moisturizing agents include alkylene glycols (e.g., polyalkylene glycols such as ethylene glycol, diethylene glycol, propylene glycol, dipropylene glycol, 1,3-butylene glycol, pentanediol, polyethylene glycol, polyoxyethylene-polyoxypropylene block copolymers, diethylene glycol monoethyl ether, polyoxyethylene polyoxypropylene dimethyl ether, and monoalkyl esters thereof), water-soluble vinyl polymers (e.g., polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, polyacrylic acid, polyacrylamide, and the like), polyhydric alcohols (e.g., glycerin such as concentrated glycerin, pentaerythritol, diglycerin, diglycerin propylene glycol, and the like), and the like. oxide adducts, etc.), organic acids (e.g., lactic acid, sodium lactate, sodium pyrrolidone carboxylate, etc.), amino acids (e.g., serine, glycine, threonine, alanine, etc.), sugars (e.g., sugar alcohols such as xylitol, sorbitol, maltitol, etc.; polysaccharides such as hyaluronic acid, sodium hyaluronate, sodium chondroitin sulfate, chondroitin heparin, etc.), water-soluble cellulose ethers (methylcellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxyethyl methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, etc.), proteins (e.g., vitronectin, fibronectin, keratin, elastin, royal jelly, sericin, etc.), etc.

[0104] These moisturizing agents can be used alone or in combination of two or more. Among these moisturizing agents, alkylene glycols such as dipropylene glycol, 1,3-butylene glycol, pentanediol (pentylene glycol), and polyethylene glycol, polyhydric alcohols such as glycerin, and sugars such as sodium hyaluronate are preferred.

[0105] The proportion of the moisturizer is, for example, 1 to 100 parts by mass, preferably 3 to 80 parts by mass, further preferably 5 to 50 parts by mass, more preferably 8 to 30 parts by mass, and most preferably 10 to 20 parts by mass, relative to 100 parts by mass of the liquid base.

[0106] The transdermal liquid composition of the present disclosure may further contain an emollient. Examples of the emollient include higher fatty acids or oils (e.g., oleic acid, isostearic acid, oxystearic acid, oleyl alcohol, stearyl alcohol, isopropyl stearate, octyl oxystearate, glycerin oxystearate, decyl oleate, octyldodecyl 12-stearoyl stearate, polyoxyethylene oleyl ether, polyoxyethylene stearyl ether, polyoxyethylene hydrogenated castor oil, triethylhexanoin, dimethicone, etc.), waxes (e.g., lanolin, lanolin alcohol, lanolin oil, polyoxyethylene lanolin, polyoxyethylene propylene lanolin, honeysuckle, etc.), and the like. wax, polyoxyethylene beeswax, etc.), sugars (e.g., polyoxyethylene methyl glycoside, methyl sesquistearate glycoside, sorbitan monooleate, sorbitan monolaurate, polyoxyethylene sorbitan oleate, polyoxyethylene sucrose oleate, polyoxyethylene methyl dioleate glycoside, polyoxyethylene methyl glycoside sesquistearate, etc.), ceramides, squalane, squalene, honey, emulsions of oily components (e.g., emulsions obtained by emulsifying oily components such as triglyceride oil, squalane, ester oil, etc. with a nonionic emulsifier such as monoglyceride, etc.), etc.

[0107] These emollients can be used alone or in combination of two or more. Among these emollients, preferred are higher alcohols such as cetearyl alcohol, oils and fats such as triethylhexanoin and dimethicone, polyoxyethylene group-containing sugars such as polyoxyethylene methyl glycoside, oils and fats such as glycerin oxystearate and polyoxyethylene hydrogenated castor oil, and waxes such as beeswax and lanolin.

[0108] The proportion of the emollient may be, for example, 50 parts by mass or less, and is preferably 0.01 to 30 parts by mass, more preferably 0.1 to 20 parts by mass, even more preferably 0.5 to 20 parts by mass, and most preferably 1 to 10 parts by mass, relative to 100 parts by mass of the liquid base.

[0109] The transdermal liquid composition of the present disclosure may further contain a surfactant. The surfactant may be used as a solubilizer or emulsifier. The surfactant may be any of anionic surfactants, nonionic surfactants, cationic surfactants, and amphoteric surfactants.

[0110] In this specification and claims, "POE" means polyoxyethylene (polyethylene oxide) and "POP" means polyoxypropylene (polypropylene oxide).

[0111] Examples of anionic surfactants include glycerin fatty acid esters such as glyceryl monostearate (glyceryl stearate); higher fatty acid soaps such as sodium laurate; higher alkyl sulfates such as sodium lauryl sulfate; alkyl ether sulfates such as sodium POE-lauryl sulfate and triethanolamine POE-lauryl sulfate; N-acyl sarcosine salts such as sodium N-lauroyl sarcosine; sodium N-stearoyl-N-methyl taurine, sodium N-myristoyl-N-methyl taurine, and sodium N-stearoyl-N-methyl taurine. Examples of such salts include higher fatty acid amide sulfonates such as sodium lauryl-N-methyl taurate; phosphate salts such as sodium POE-oleyl ether phosphate; sulfosuccinates such as sodium di-2-ethylhexyl sulfosuccinate; alkylbenzene sulfonates such as sodium dodecylbenzene sulfonate and triethanolamine dodecylbenzene sulfonate; N-acyl glutamates such as sodium N-lauroyl-L-glutamate and disodium N-stearoyl-L-glutamate; and sulfated oils (e.g., turmeric oil, etc.).

[0112] Examples of nonionic surfactants include POE-sorbitan fatty acid esters such as POE-sorbitan monooleate, POE-sorbitan monostearate, POE-sorbitan monooleate, and POE-sorbitan tetraoleate; POE-sorbitan fatty acid esters such as POE-sorbitan monolaurate, POE-sorbitan monooleate, POE-sorbitan pentaoleate, and POE-sorbitan monostearate; sucrose fatty acid esters such as sucrose laurate; POE-glycerin monostearate, POE -POE-glycerin fatty acid esters such as glycerin monoisostearate and POE-glycerin triisostearate; POE-fatty acid esters (e.g., POE-distearate, POE-monodioleate, ethylene glycol distearate, etc.); POE-alkyl ethers (e.g., POE-lauryl ether, POE-oleyl ether, POE-stearyl ether, POE-behenyl ether, POE-2-octyldodecyl ether, POE-cholestanol ether, etc.); Pluronic types (e.g., Pluronic POE·POP-alkyl ethers (e.g., POE·POP-cetyl ether, POE·POP-2-decyltetradecyl ether, POE·POP-monobutyl ether, POE·POP-hydrogenated lanolin, POE·POP-glycerin ether, etc.); tetraPOE·tetraPOP-ethylenediamine condensates (e.g., Tetronic, etc.); POE-castor oil hydrogenated castor oil derivatives (e.g., POE-castor oil, POE-hydrogenated castor oil, POE-hydrogenated castor oil monoisostearate, POE-hydrogenated castor oil triisostearate, etc.); stearates, POE-hydrogenated castor oil monopyroglutamic acid monoisostearate diester, POE-hydrogenated castor oil maleic acid, etc.); POE-beeswax and lanolin derivatives (e.g., POE-sorbitol beeswax, etc.); alkanolamides (e.g., coconut oil fatty acid diethanolamide, lauric acid monoethanolamide, fatty acid isopropanolamide, etc.); POE-propylene glycol fatty acid esters; POE-alkylamines; POE-fatty acid amides; alkylethoxydimethylamine oxides; trioleyl phosphate, etc.

[0113] Examples of the cationic surfactant include stearyltrimethylammonium chloride; cetylpyridinium chloride; benzalkonium chloride; and benzethonium chloride.

[0114] Examples of amphoteric surfactants include phospholipid-based amphoteric surfactants such as hydrogenated lecithin; imidazoline-based amphoteric surfactants; and betaine-based surfactants.

[0115] Silicone surfactants can also be used as the surfactant, such as polyether-modified silicone (an adduct of at least ethylene oxide with polydimethylsiloxane).

[0116] These surfactants can be used alone or in combination of two or more. Among these surfactants, anionic surfactants such as glyceryl stearate, sodium lauryl sulfate, triethanolamine lauryl ether sulfate, and polyoxyethylene disodium lauryl sulfosuccinate, nonionic surfactants such as POE-hydrogenated castor oil, and amphoteric surfactants such as hydrogenated lecithin are preferred.

[0117] The proportion of the surfactant is, for example, 0.01 to 5 parts by mass, preferably 0.03 to 3 parts by mass, further preferably 0.05 to 1 part by mass, even more preferably 0.1 to 0.5 parts by mass, and most preferably 0.2 to 0.3 parts by mass, relative to 100 parts by mass of the liquid base.

[0118] The transdermal liquid composition of the present disclosure may further contain a preservative. The preservative includes an antiseptic, a bactericide or an antibacterial agent, an antioxidant, an ultraviolet absorbing agent or an ultraviolet scattering agent, and the like.

[0119] Examples of preservatives include phenoxyethanol, chlorphenesin, benzoic acid, salicylic acid, sorbic acid, and alkyl paraoxybenzoates (parabens).

[0120] Bactericides or antibacterial agents include, for example, sodium benzoate.

[0121] Examples of antioxidants include inorganic acids such as phosphoric acid and sodium pyrosulfite; organic acids such as malonic acid, succinic acid, ascorbic acid, maleic acid, fumaric acid, and edetic acid; phosphorus compounds such as cephalin and phytic acid; and polyphenols such as anthocyanin.

[0122] Examples of ultraviolet absorbing agents or ultraviolet scattering agents include tocopherol acetate, ethylhexyl methoxycinnamate, ethylhexyl 4-(N,N-dimethylamino)benzoate, t-butyl methoxydibenzoylmethane, oxybenzone, zinc oxide, and titanium oxide.

[0123] These preservatives may be used alone or in combination of two or more. Among these preservatives, preservatives such as phenoxyethanol are preferred.

[0124] The proportion of the preservative is, for example, 0.01 to 10 parts by mass, preferably 0.05 to 5 parts by mass, further preferably 0.1 to 3 parts by mass, more preferably 0.2 to 1 part by mass, and most preferably 0.3 to 0.5 parts by mass, relative to 100 parts by mass of the liquid base.

[0125] The transdermal liquid composition of the present disclosure may further contain a fragrance. The fragrance may be a natural fragrance or a synthetic fragrance.

[0126] Examples of natural flavorings include fruit essences or oils (vanillin, lemon oil, etc.) such as strawberry, blueberry, apple, plum, orange, lemon, lime, vanilla, and pepper; fruit peel essences or oils such as orange, white grape, grapefruit, and lemon; bark essences or oils such as cinnamon; bark powders such as cinnamon powder; root vegetable essences or oils such as ginger; root vegetable powders such as ginger powder; seed powders such as vanilla beans and cocoa powder; branch and leaf essences or oils such as peppermint, spearmint, rosemary, and shiso; branch and leaf powders such as peppermint powder; and flower essences or oils such as jasmine, lavender, rose, rosemary, and hyacinth.

[0127] Examples of synthetic flavors include benzyl acetate, linalyl acetate, citral, citronellal, citronellol, cis-jasmine, cis-3-hexenol, and menthol.

[0128] These fragrances may be used alone or in combination of two or more.

[0129] The proportion of the flavoring is, for example, 0.001 to 5 parts by mass, preferably 0.003 to 1 part by mass, further preferably 0.005 to 0.5 parts by mass, even more preferably 0.01 to 0.1 parts by mass, and most preferably 0.015 to 0.05 parts by mass, relative to 100 parts by mass of the liquid base.

[0130] The transdermal liquid composition of the present disclosure may further contain a pH adjuster. Examples of pH adjusters include bases such as sodium bicarbonate; acids such as citric acid, sodium citrate, and sodium monohydrogen phosphate; and borax. These pH adjusters can be used alone or in combination of two or more. Among these, organic acids such as citric acid and sodium citrate are preferred. The proportion of the pH adjuster can be appropriately selected depending on the target pH.

[0131] The transdermal liquid composition of the present disclosure may further comprise a physiologically or pharmacologically active ingredient.

[0132] Examples of physiologically active ingredients (or pharmacologically active ingredients) include cell activators (e.g., riboflavin, pyridoxine, nicotinic acid, pantothenic acid, α-tocopherol or derivatives thereof; plant extracts such as saxifrage extract, etc.), skin roughness prevention agents (e.g., vitamins such as vitamin A, vitamin B, vitamin C, vitamin D, vitamin E, vitamin K, etc.), skin whitening agents (e.g., ascorbic acid or its derivatives, cysteine, placenta extract, arbutin, kojic acid, rucinol, ellagic acid, chamomile extract, etc.), and blemish and freckle inhibitors (e.g., , tyrosinase activity inhibitors, melanin reducing agents, etc.), acne inhibitors (for example, keratin softeners such as sulfur, anti-inflammatory agents, adrenal cortical hormones, sebum secretion inhibitors, etc.), skin softeners (for example, salicylic acid or its derivatives, urea, etc.), anti-inflammatory agents (for example, allantoin, guaiazulene, glycyrrhizic acid or its salts, glycyrrhetinic acid or its salts, ε-aminocaproic acid, tranexamic acid, ibuprofen, indomethacin, zinc oxide or their derivatives; plant extracts such as arnica extract, etc.), skin itching inhibitors, blood circulation promoters, etc. These bioactive ingredients can be used alone or in combination of two or more.

[0133] The proportion of the physiologically active ingredient is, for example, 0.001 to 10 parts by mass, preferably 0.01 to 5 parts by mass, further preferably 0.03 to 3 parts by mass, more preferably 0.05 to 2 parts by mass, and most preferably 0.1 to 1 part by mass, per 100 parts by mass of the liquid base.

[0134] The liquid composition of the present disclosure may further contain other ingredients.

[0135] Other ingredients include, for example, astringents or antiperspirants (e.g., oxyacids such as lactic acid, tartaric acid, or salts thereof; aluminum compounds such as aluminum chloride; zinc compounds such as zinc sulfate and zinc sulfonate; proanthocyanidins; extracts of tannin-containing plants such as witch hazel and white birch; chinese laurel extract, rhubarb extract, and horsetail extract), coenzymes (e.g., coenzyme Q10, etc.), amino acids (e.g., tryptophan, etc.), cooling agents (e.g., menthol or its derivatives, camphor, thymol, etc.), inorganic salts (e.g., sodium sulfate, potassium chloride, etc.), fibers (e.g., synthetic fibers such as nylon fibers, natural natural fibers, etc.), binders (e.g., carboxymethylcellulose, sodium carboxymethylcellulose, carrageenan, etc.), chelating agents or sequestering agents (e.g., gluconic acid, citric acid, succinic acid, ascorbic acid, sodium edetate, sodium 1-hydroxyethane-1,1-diphosphonate, phosphoric acid, sodium hexametaphosphate, etc.), reducing agents (e.g., thioglycolic acid or a salt thereof), enzymes (e.g., lipase, protease, etc.), thickening agents (e.g., gelatin, gluten, fish protein, etc.), basic agents, oxidizing agents, cleansing agents, cooling agents, colorants, opacifying agents, solidifying agents, plasticizers, etc.

[0136] These other components may be used alone or in combination of two or more.

[0137] The proportion of other components is, for example, 0.001 to 30 parts by mass, preferably 0.01 to 20 parts by mass, further preferably 0.03 to 10 parts by mass, more preferably 0.05 to 5 parts by mass, and most preferably 0.1 to 3 parts by mass, relative to 100 parts by mass of the liquid base.

[0138] The pH of the transdermal liquid composition of the present disclosure is, for example, 3 to 9, preferably 4 to 8, further preferably 5 to 7, more preferably 5.5 to 6.5, and most preferably 5.8 to 6.2.

[0139] When the transdermal liquid composition of the present disclosure contains a liquid base, it can be produced by mixing a 3HB component (active ingredient), the liquid base, and, as necessary, ingredients other than the 3HB component and the liquid base. A production method in which the 3HB component is dissolved or dispersed in the liquid base is preferred.

[0140] (Semi-solid composition for transdermal application) In the semi-solid composition for transdermal use (or semi-solid composition for external use), the ratio of the 3HB component is 0.01% by mass or more in the semi-solid composition for transdermal use, preferably 0.1% by mass or more, more preferably 0.2% by mass or more, more preferably 0.3% by mass or more, particularly preferably 0.5% by mass or more, and most preferably 0.8% by mass or more. If the ratio of the 3HB component is too low, there is a risk that the production of fibrillin cannot be promoted. In addition, in the semi-solid composition for transdermal use, the ratio of the 3HB component is, for example, 30% by mass or less in the semi-solid composition for transdermal use, preferably 10% by mass or less, more preferably 5% by mass or less, even more preferably 3% by mass or less, particularly preferably 2% by mass or less, and most preferably 1.5% by mass or less. The ratio of the 3HB component may be 0.1 to 5% by mass, particularly 0.3 to 3% by mass, in the semi-solid composition for transdermal use. If the ratio of the 3HB component is too high, there is a risk that the handling property is reduced.

[0141] The semi-solid composition for transdermal application of the present disclosure may further contain a liquid base. Examples of the liquid base include the liquid bases exemplified in the section on the liquid composition for transdermal application. The liquid bases may be used alone or in combination of two or more. The preferred embodiments are also the same as those of the liquid composition for transdermal application.

[0142] The proportion of the liquid base in the transdermal semi-solid composition is 10 to 95% by mass, preferably 30 to 90% by mass, further preferably 50 to 85% by mass, even more preferably 60 to 80% by mass, and most preferably 65 to 75% by mass.

[0143] The semi-solid composition for transdermal application of the present disclosure may further contain an emollient. Examples of the emollient include the emollients exemplified in the section on the liquid composition for transdermal application. The emollients may be used alone or in combination of two or more. The preferred embodiments are the same as those of the liquid composition. In particular, in the semi-solid composition for transdermal application, a plurality of emollients may be combined, for example, a combination of oils, a higher fatty acid or a derivative thereof, and a wax.

[0144] The proportion of the emollient may be, for example, 1 part by mass or more, for example, 1 to 1000 parts by mass, preferably 5 to 100 parts by mass, further preferably 10 to 80 parts by mass, more preferably 15 to 50 parts by mass, and most preferably 20 to 30 parts by mass, relative to 100 parts by mass of the liquid base. If the proportion of the emollient is too low, there is a risk that the semi-solid form cannot be maintained, and if it is too high, there is a risk that the handling properties may be reduced.

[0145] The semi-solid composition for transdermal application of the present disclosure may further contain a gelling agent. Examples of gelling agents include water-soluble or water-swellable polysaccharides such as dextran, pullulan, agar, pectin, alginic acid, sodium alginate, carrageenan, galactomannan, glucomannan, curdlan, gum arabic, tragacanth gum, gellan gum, karaya gum, guar gum, xanthan gum, locust bean gum, tara gum, tamarind seed gum, and psyllium seed gum; proteins such as collagen, casein, albumin, gelatin, and royal jelly; and inorganic water-soluble polymers such as bentonite, organically modified bentonite, magnesium aluminum silicate, and anhydrous silicic acid. These gelling agents can be used alone or in combination of two or more. Of these gelling agents, water-swellable polysaccharides such as xanthan gum are preferred.

[0146] The proportion of the gelling agent is, for example, 0.01 to 100 parts by mass, preferably 0.03 to 10 parts by mass, further preferably 0.05 to 1 part by mass, more preferably 0.08 to 0.5 parts by mass, and most preferably 0.1 to 0.3 parts by mass, relative to 100 parts by mass of the liquid base.

[0147] The semi-solid composition for transdermal application of the present disclosure may further contain a moisturizing agent. Examples of the moisturizing agent include the moisturizing agents exemplified in the section on the liquid composition for transdermal application. The moisturizing agents may be used alone or in combination of two or more. The preferred embodiments are the same as those of the liquid composition for transdermal application.

[0148] The proportion of the moisturizer is, for example, 1 to 100 parts by mass, preferably 3 to 80 parts by mass, further preferably 5 to 50 parts by mass, more preferably 8 to 30 parts by mass, and most preferably 10 to 20 parts by mass, relative to 100 parts by mass of the liquid base.

[0149] The semi-solid composition for transdermal application of the present disclosure may further contain a surfactant. Examples of the surfactant include those exemplified in the section on the liquid composition for transdermal application. The surfactant may be used alone or in combination of two or more. The preferred embodiments are the same as those of the liquid composition for transdermal application.

[0150] The proportion of the surfactant is, for example, 0.01 to 30 parts by mass, preferably 0.1 to 20 parts by mass, further preferably 0.5 to 15 parts by mass, more preferably 1 to 10 parts by mass, and most preferably 3 to 5 parts by mass, relative to 100 parts by mass of the liquid base.

[0151] The semi-solid composition for transdermal application of the present disclosure may further contain a preservative. Examples of the preservative include the preservatives exemplified in the section on the liquid composition for transdermal application. The preservatives may be used alone or in combination of two or more. The preferred embodiments are the same as those of the liquid composition for transdermal application.

[0152] The proportion of the preservative is, for example, 0.01 to 10 parts by mass, preferably 0.05 to 5 parts by mass, further preferably 0.1 to 3 parts by mass, more preferably 0.2 to 1 part by mass, and most preferably 0.3 to 0.5 parts by mass, relative to 100 parts by mass of the liquid base.

[0153] The semi-solid composition for transdermal application of the present disclosure may further contain a fragrance. Examples of the fragrance include the fragrances exemplified in the section on the liquid composition for transdermal application. The fragrances may be used alone or in combination of two or more. The preferred embodiments are also the same as those of the liquid composition for transdermal application.

[0154] The proportion of the flavoring is, for example, 0.001 to 5 parts by mass, preferably 0.003 to 1 part by mass, further preferably 0.005 to 0.5 parts by mass, even more preferably 0.01 to 0.1 parts by mass, and most preferably 0.015 to 0.05 parts by mass, relative to 100 parts by mass of the liquid base.

[0155] The semi-solid composition for transdermal application of the present disclosure may further contain a pH adjuster. Examples of the pH adjuster include the pH adjusters exemplified in the section on the liquid composition for transdermal application. The pH adjuster may be used alone or in combination of two or more. The preferred embodiments are the same as those of the liquid composition for transdermal application. The proportion of the pH adjuster may be appropriately selected according to the desired pH.

[0156] The semi-solid composition for transdermal application of the present disclosure may further contain a physiologically active ingredient. Examples of the physiologically active ingredient include the physiologically active ingredients exemplified in the section on the liquid composition for transdermal application. The physiologically active ingredient may be used alone or in combination of two or more kinds. The preferred embodiments are also the same as those of the liquid composition for transdermal application.

[0157] The proportion of the physiologically active ingredient is, for example, 0.001 to 10 parts by mass, preferably 0.01 to 5 parts by mass, further preferably 0.03 to 3 parts by mass, more preferably 0.05 to 2 parts by mass, and most preferably 0.1 to 1 part by mass, per 100 parts by mass of the liquid base.

[0158] The semi-solid composition for transdermal application of the present disclosure may further contain other components. Examples of the other components include the other components exemplified in the section on the liquid composition for transdermal application. The components can be used alone or in combination of two or more. The preferred embodiments are also the same as those of the liquid composition for transdermal application.

[0159] The proportion of other components is, for example, 0.001 to 30 parts by mass, preferably 0.01 to 20 parts by mass, further preferably 0.03 to 10 parts by mass, more preferably 0.05 to 5 parts by mass, and most preferably 0.1 to 3 parts by mass, relative to 100 parts by mass of the liquid base.

[0160] The pH of the semi-solid transdermal composition of the present disclosure is, for example, 3 to 9, preferably 4 to 8, further preferably 5 to 7, more preferably 5.5 to 6.5, and most preferably 5.8 to 6.2.

[0161] When the semi-solid transdermal composition of the present disclosure contains an emollient, it can be produced by mixing the 3HB component (active ingredient), the emollient, and, if necessary, ingredients other than the 3HB component and the emollient.

[0162] (Use of transdermal composition) The transdermal composition (topical composition or topical agent) of the present disclosure can be used as various cosmetic materials, medicines, or quasi-drugs. The term "cosmetics" is used to include cosmetics.

[0163] Examples of the cosmetics include moisturizing cosmetics, whitening cosmetics, UV care cosmetics, cleansing agents, scrubbing agents, and the like.

[0164] Cosmetics include skin care products, makeup products, UV care products, body products, hair care products, etc.

[0165] Examples of basic cosmetics include skin lotions, beauty essences, milky lotions, creams, facial cleansers, and cleansing cosmetics (oil-, gel-, cream-, lotion-, and milk-based cosmetics).

[0166] Examples of skin care cosmetics (skin care products or cosmetics for the skin) include lotions, milky lotions, gel creams, skin care creams, and massage lotions.

[0167] Examples of makeup cosmetics include makeup bases, BB creams, foundations (powder foundations, liquid foundations, cream foundations, emulsion foundations, etc.), lipsticks, lip glosses, eye shadows, eyeliners, eyebrow pencils, and mascaras.

[0168] Examples of UV care cosmetics include emulsions, lotions, gels, creams, and sprays of sunscreen cosmetics.

[0169] Examples of body cosmetics include body washes, body lotions, body milks, body creams, body powders, and body massage products.

[0170] Examples of hair care cosmetics include shampoos, conditioners, treatments, after-bath treatments, tonics, hair styling products, hair growth products, and hair creams.

[0171] Examples of pharmaceuticals or quasi-drugs include medicated cosmetics, liquids, aerosols, creams, gels (jelly), ointments, poultices, and liniments.

[0172] Among these uses, in view of the ease with which the effects of the transdermal composition of the present disclosure are exhibited, preferred are cosmetics (including beauty products), pharmaceuticals, and quasi-drugs that are used in a manner that allows the composition to penetrate into the skin without being washed off immediately after use, more preferred are cosmetics and quasi-drugs, and especially preferred are cosmetics.

[0173] Furthermore, the transdermal composition of the present disclosure is particularly preferably a skin care cosmetic (skin care product or skin cosmetic) for improving skin elasticity (firmness). The skin care cosmetic may be a transdermal liquid composition such as a skin lotion, beauty essence, lotion, milk, emulsion, or mist, or a transdermal semi-solid composition such as a cream, gel, paste (balm), or foam (bubble). The transdermal liquid composition and transdermal semi-solid composition may be a cosmetic composition or a cosmetic composition (cosmetics), or may be an anti-aging care product for the purpose of anti-aging. EXAMPLES

[0174] The present disclosure will be described in more detail below based on examples, but the present disclosure is not limited to these examples. The raw materials and the method for producing senescence-induced fibroblasts used in the examples are as follows.

[0175] [Raw materials] 3HB-Na: Crystalline (R) 3HB sodium, "OHALOS (registered trademark)" manufactured by Osaka Gas Chemicals Co., Ltd.

[0176] [Preparation of senescence-induced fibroblasts and culture conditions] Normal human fibroblasts (Kurabo Industries, Ltd., hereafter referred to as "NHDF") were cultured in a 10 cm petri dish at 6.0 × 10 cells using a growth medium [Dulbecco's Modified Eagle Medium (Sigma-Aldrich, Ltd.), hereafter referred to as "DMEM"] containing 5% FBS. 5 The cells were seeded at a density of 100 cells / dish and cultured at 37°C for 24 hours.

[0177] Next, NHDF was dissolved in hydrogen peroxide (H 2 O 2 To treat NHDF, the growth medium in which the NHDFs were grown was diluted with 25 μM hydrogen peroxide (H 2 O 2 The medium was replaced with phenol red-free serum-free DMEM (Sigma-Aldrich), and the cells were cultured at 37°C for 30 minutes. 2 O 2The medium containing was replaced with fresh DMEM and cultured at 37°C for 24 hours.

[0178] This H 2 O 2 The treatment was carried out four times (four days) in succession, and the final H 2 O 2 The cells were harvested 72 hours after the treatment and used as senescence-induced fibroblasts for the study.

[0179] Example 1 and Comparative Example 1 In Example 1, steady-state human fibroblasts (NHDF) were cultured in DMEM supplemented with 3HB-Na to the final concentration shown in Table 1, and fibrillin-1 fibrillogenesis was evaluated by the following method. In Comparative Example 1, NHDF were cultured in DMEM without 3HB-Na (3HB-Na 0 mM), and fibrillin-1 fibrillogenesis was evaluated.

[0180] [Evaluation of fibrogenic potential] (Preparation of cell samples) Normal human fibroblasts (NHDF) were cultured in a 96-well plate at 2.0 × 10 4 The cells were seeded at a density of 1000 cells / well and cultured at 37°C for 24 hours.

[0181] Next, the DMEM after culture was replaced with DMEM to which 3HB-Na had been added to reach the final concentration shown in Table 1, and the cells were further cultured for 7 days at 37°C. After culture, the cells were treated with 4% (w / v) formaldehyde to fix the cells, and a blocking treatment was performed using a 1% (w / v) bovine serum albumin solution at 37°C for 1 hour.

[0182] The cells were treated with a primary antibody, anti-fibrillin-1 antibody (manufactured by Thermo Fisher Scientific), and cultured overnight at 4° C. Then, the cells were treated with a secondary antibody ("Alexa Fluor (registered trademark)" manufactured by Cell Signaling Technology) that specifically binds to the primary antibody, and cultured at room temperature for 1 hour to prepare a cell sample.

[0183] (Image analysis of cell samples) The prepared cell samples were treated with a fluorescent staining agent ["Hoechest (registered trademark) 33342" (manufactured by Thermo Fisher Scientific)] to perform nuclear staining, and images were analyzed using a fluorescence microscope ("BZ-X800" manufactured by Keyence Corporation).

[0184] [Evaluation results of fibrogenic potential] For normal human fibroblast (NHDF) cell samples, the fiber area ratio calculated from images observed under a fluorescent microscope is shown as the evaluation result of fibrogenic ability in Table 1. The fiber area ratio (%) was determined as the fiber area ratio (%) per unit area from images observed under a fluorescent microscope using Corneo Cytometry 2 (manufactured by CIEL Co., Ltd.).

[0185] [Table 1]

[0186] In Table 1, "SD" means standard deviation, and the fiber area ratio of Example 1 is a relative value when the fiber area ratio of Comparative Example 1 is set to 100%.

[0187] As is clear from the results in Table 1, when normal human fibroblasts were cultured in medium containing 3HB-Na, the fiber area ratio increased or was the same as when they were cultured in medium not containing 3HB-Na, and at a 3HB-Na concentration of 0.02 mM, the fiber area ratio increased by approximately 1.07-fold, and at 0.2 mM, the fiber area ratio increased by approximately 1.02-fold, indicating that 3HB-Na promoted the production of fibrillin-1 in normal human fibroblasts.

[0188] Example 2 and Comparative Example 2 Example 2 was carried out in the same manner as Example 1, except that senescence-induced fibroblasts were used instead of NHDF. That is, in Example 2, senescence-induced fibroblasts were cultured in DMEM to which 3HB-Na was added to achieve the final concentration shown in Table 2, in the same manner as in Example 1, and fibrillin-1 fibrillogenesis ability was evaluated.

[0189] In Comparative Example 2, the senescence-induced fibroblasts prepared by the above-mentioned method were cultured in DMEM containing no 3HB-Na (0 mM 3HB-Na) and the fibrillin-1 fibrillogenesis ability was evaluated.

[0190] [Evaluation results of fibrogenic potential] For cell samples of senescence-induced fibroblasts prepared in the same manner as in Example 1, the fiber area ratios calculated from images observed under a fluorescent microscope are shown in Table 2 as evaluation results of the fibrogenesis ability. The fiber area ratios were determined in the same manner as in Example 1.

[0191] [Table 2]

[0192] The fiber area ratios in Example 2 and Comparative Example 2 are relative values ​​when the fiber area ratio of NHDF cultured in DMEM without 3HB-Na (Comparative Example 1) was taken as 100%.

[0193] In addition, in Table 2, "SD" means standard deviation, "*" means that the p-value for NHDF not containing 3HB-Na is less than 0.05 (p<0.05), "#" means that the p-value for senescence-induced fibroblasts not containing 3HB-Na is less than 0.05 (p<0.05), and "##" means that the p-value for senescence-induced fibroblasts not containing 3HB-Na is less than 0.01 (p<0.01).

[0194] In Comparative Example 2, the fiber area ratio of senescence-induced fibroblasts not containing 3HB-Na was H 2 O 2 When the fiber area ratio of untreated NHDF was taken as 100%, the fiber area ratio was 69.0±6.0% (p value less than 0.05 compared to NHDF not containing 3HB-Na), confirming that the senescence-induced fibroblasts prepared were suitable as a senescence-induced fibroblast model.

[0195] Furthermore, as is clear from the results in Table 2, when senescence-induced fibroblasts were cultured in a medium containing 3HB-Na, the fiber area ratio increased statistically significantly by approximately 1.39-fold at a 3HB-Na concentration of 0.02 mM and approximately 1.32-fold at 0.2 mM compared to when the cells were cultured in a medium not containing 3HB-Na (Comparative Example 2), demonstrating that 3HB-Na greatly promoted the production of fibrillin-1 in senescence-induced fibroblasts.

[0196] FIG. 1 summarizes the results of Examples 1 and 2 and Comparative Examples 1 and 2, and is a graph showing the relationship between 3HB-Na concentration (3HB addition concentration) and fibrillin-1 fiber area ratio for NHDF and senescence-induced fibroblasts.

[0197] As is clear from Figure 1, when cultured in a medium containing 3HB-Na, the fiber area ratio increased and fibrillin-1 production was promoted in both NHDF and senescence-induced fibroblasts, but the increase in fiber area ratio was clearly greater in senescence-induced fibroblasts than in NHDF, and the increase in fiber area ratio in senescence-induced fibroblasts relative to the increase in fiber area ratio in NHDF was (96.0-69.0) / (106.6-100) = approximately 4.1 / 1 at a 3HB-Na concentration of 0.02 mM, and (91.2-69.0) / (101.6-100) = approximately 13.9 / 1 at 0.2 mM.

[0198] Thus, it was shown that 3HB-Na has the effect of promoting the production of fibrillin-1 in NHDF and senescence-induced fibroblasts, and the effect of improving the formation of elastin fibers whose main component is fibrillin-1. [Industrial Applicability]

[0199] The fibrillin production promoter of the present disclosure can promote the production of fibrillin in fibroblasts, and can therefore be used in various applications such as beauty products or cosmetics, foods, medicines, quasi-drugs, etc. In particular, the fibrillin production promoter of the present disclosure can promote the production of fibrillin, thereby improving the formation of elastin fibers that maintain the mesh-like structure of the dermis, and can prevent or suppress a decrease in skin elasticity (firmness), and can therefore be suitably used in applications such as cosmetics and quasi-drugs, particularly in cosmetics, for example, skin care products such as anti-aging care products.

Claims

1. 3-Hydroxybutyric acid, a salt of 3-hydroxybutyric acid, and a compound represented by the following formula (1) 【Chemistry 1】 (wherein R represents an alkyl group). A fibrillin production promoter comprising at least one 3-hydroxybutyric acid component selected from 3-hydroxybutyric acid alkyl esters represented by the following formula:

2. The fibrillin production promoter according to claim 1, wherein the 3-hydroxybutyric acid component contains an R-isomer.

3. The fibrillin production promoter according to claim 1 or 2, wherein the fibrillin is fibrillin-1.

4. The fibrillin production promoter according to claim 1 or 2, which promotes the production of fibrillin in senescent cells.

5. 3. The fibrillin production promoter according to claim 1 or 2, which is applied to the skin and has the following effects (a) and / or (b): (a) Improves the formation of elastin fibers (b) Prevents loss of skin elasticity

6. A composition for promoting fibrillin production, comprising at least one 3-hydroxybutyric acid component selected from 3-hydroxybutyric acid, a salt of 3-hydroxybutyric acid, and an alkyl ester of 3-hydroxybutyric acid represented by formula (1) according to claim 1.

7. The composition according to claim 6, which is a transdermal composition.

8. 8. The composition according to claim 6 or 7, which is a liquid or semi-solid composition.

9. The composition according to claim 6 or 7, which is a cosmetic.

10. 1. An elastin fiber formation enhancer comprising at least one 3-hydroxybutyric acid component selected from 3-hydroxybutyric acid, a salt of 3-hydroxybutyric acid, and an alkyl ester of 3-hydroxybutyric acid represented by formula (1) according to claim 1.

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