Filaggrin mRNA expression promoter

Abstract

To provide MSX-2 mRNA expression-promoting agents, excellent in MSX-2 mRNA expression promoting effect and safety; eyelash growing agents excellent in eyelash growing or external preparations for eyelash growing; filaggrin mRNA expression-promoting agents excellent in filaggrin mRNA expression promoting effect and safety; hyaluronan synthase 3 mRNA expression-promoting agents excellent in hyaluronan synthase 3 mRNA expression promoting effect and safety; and external preparations for moisturization excellent in moisturization and safety.SOLUTION: There is provided a promoting agent for MSX-2 (Muscle Segment homeobox 2) mRNA expression comprising an extract of Calendula officinalis.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a muscle segment homeobox 2 (MSX-2) mRNA expression promoter, an eyelash growth agent, and an external preparation composition for eyelash growth, as well as a filaggrin mRNA expression promoter, a hyaluronic acid synthase 3 mRNA expression promoter, a moisturizer, and an external preparation composition for moisturizing. [Background technology]

[0002] An anatomical feature of eyelashes is that the hair follicles extend to a depth of about 1.5 mm into the eyelid, but they do not have the arrector pili muscles found in hair or body hair, and are fixed within a triangle of dense connective tissue surrounded by the orbicularis oculi muscle and the lash muscles.Regarding the eyelash hair cycle, it has been reported that the growth phase is about two months, and the growth phase hair ratio is about 18% or 40%, which varies depending on age and gender.

[0003] As eye makeup becomes more popular, the use of eye makeup cosmetics such as mascara to make eyelashes appear longer, thicker, and more upward-facing is on the rise. However, damage caused by the use of mascara and other products has been pointed out, highlighting the need for eyelash care to keep eyelashes healthy.

[0004] The length of eyelashes in Japanese people is not significantly different from that of Caucasians, and it is thought that the difference in appearance is due to differences in the structure of the eyelids.It has also been reported that eyelashes in Japanese people tend to grow faster and for a shorter period of time, and that the length of the growth period has a significant impact on eyelash length.

[0005] The transcription factor MSX-2 is known to be involved in eyelash growth. Eyelash follicle cells, which are epithelial cells, are known to express various growth factors (FGF-7, ECGF, VEGF, LEF1) and transcription factor (MSX-2) that are thought to be involved in hair growth. Stevioside, which has been shown to promote eyelash growth, has also been shown to enhance MSX-2 expression in eyelash follicle cells, suggesting that MSX-2 is involved in hair growth (see, for example, Non-Patent Document 1). Therefore, promoting MSX-2 mRNA expression in epithelial cells is thought to lead to eyelash growth.

[0006] Filaggrin is a component of the skin and is thought to be involved in the barrier function of the skin, preventing the invasion of allergens, toxins, and infectious organisms. A decrease in filaggrin function due to gene mutations or other factors is known to be associated with the risk of developing atopic diseases, including atopic dermatitis (eczema, skin inflammation, skin itching, etc.), allergies, asthma, etc., and in more severe cases, can lead to skin diseases such as ichthyosis vulgaris (see, for example, Non-Patent Document 2).

[0007] Meanwhile, amino acids, the main components of natural moisturizing factors (NMFs), are produced by the degradation of filaggrin derived from keratohyalin granules in the stratum corneum. This filaggrin is expressed as profilaggrin in epidermal keratinocytes present in the granular layer just below the stratum corneum. It is then immediately phosphorylated and accumulated in keratohyalin granules, where it is dephosphorylated and hydrolyzed to filaggrin, which then migrates to the stratum corneum, where it increases the aggregation efficiency of keratin filaments and is known to be involved in the internal organization of keratinocytes (see, for example, Non-Patent Document 3). In recent years, it has been discovered that filaggrin is extremely important and essential for skin moisture retention, and that conditions such as dryness reduce the ability of filaggrin synthesis, resulting in a decrease in the amount of amino acids in the stratum corneum (see, for example, Non-Patent Document 4).

[0008] Therefore, it is believed that promoting the expression of profilaggrin in epidermal keratinocytes can prevent, treat, or ameliorate atopic diseases, including atopic dermatitis (eczema, skin inflammation, skin itching, etc.), allergies, asthma, etc. Furthermore, promoting the expression of profilaggrin, thereby increasing the amount of amino acids in the stratum corneum, is expected to essentially improve the moisture environment of the stratum corneum. Extracts of Gai Ying (Poplar Tree) (see, for example, Patent Document 1) and the like are known to have the effect of promoting profilaggrin mRNA expression.

[0009] The epidermis and dermis of the skin are composed of epidermal cells, fibroblasts, and extracellular matrices such as collagen, elastin, hyaluronic acid, etc., which are present outside these cells and support the skin structure. In young skin, fibroblasts proliferate actively, and the interaction between fibroblasts, extracellular matrix components, and other skin tissues maintains homeostasis, ensuring moisture retention, flexibility, elasticity, etc., and maintaining the skin in a firm, glossy, and moist state in appearance.

[0010] However, under the influence of certain external factors, such as ultraviolet radiation, extremely dry air, and excessive skin cleansing, or due to aging, the production of collagen, elastin, and hyaluronic acid, the major components of the extracellular matrix, decreases and undergoes degradation and alteration. As a result, the skin's moisturizing function and elasticity decline, and abnormal peeling of the keratin occurs, causing the skin to lose firmness and luster and to exhibit aging symptoms such as rough skin and wrinkles. Thus, changes associated with skin aging, such as wrinkles, dullness, changes in texture, and loss of elasticity, are associated with the reduction and denaturation of matrix components such as collagen, elastin, and hyaluronic acid. Therefore, promoting the production of hyaluronic acid and other substances is important for preventing, treating, or improving skin aging.

[0011] Among the above-mentioned extracellular matrix components, hyaluronic acid is a type of mucopolysaccharide that fills the intercellular spaces to hold cells, and also has many other functions, such as retaining moisture in the intercellular spaces, providing lubrication and flexibility to tissues, and resisting external forces such as mechanical damage. Promoting the production of hyaluronic acid is believed to prevent, treat, or improve skin aging symptoms such as rough skin, wrinkles, dullness, changes in texture, loss of elasticity, and loss of moisturizing function. Furthermore, promoting the expression of hyaluronan synthase 3 (HAS3), which is involved in promoting the production of hyaluronic acid in the epidermis, is believed to enable the prevention, treatment, or improvement of skin aging.

[0012] Hyaluronic acid is also present in connective tissues such as cartilage, synovial fluid, umbilical cord, vitreous body, and other tissues, in addition to skin tissue. Hyaluronic acid in synovial fluid coats the surface of articular cartilage, contributing to the smooth functioning of joints through its lubricating and cartilage-covering / protecting functions. Meanwhile, it is known that the concentration of hyaluronic acid in synovial fluid decreases in arthritis, such as rheumatoid arthritis. Therefore, promoting hyaluronic acid production is believed to be effective in preventing or treating arthritis, including rheumatoid arthritis, osteoarthritis, septic arthritis, gouty arthritis, traumatic arthritis, and osteoarthritis. Furthermore, granulation tissue is formed during the healing process of wounds or burns, and it is known that hyaluronic acid significantly increases in the granulation tissue. Therefore, promoting hyaluronic acid production is believed to promote wound or burn healing. Extracts from Camphor Tree (see, for example, Patent Document 2) are known to have the effect of promoting hyaluronic acid production.

[0013] However, there remains a strong demand for new materials that have a better MSX-2 expression promoting effect, are highly safe, and have excellent eyelash growth effects, as well as new materials that have at least one of a better filaggrin expression promoting effect and an excellent hyaluronic acid synthase 3 expression promoting effect, are highly safe, and have excellent moisturizing effects, and the current situation is that there is a need for their prompt development. [Prior art documents] [Patent documents]

[0014] [Patent Document 1] Japanese Patent Application Laid-Open No. 2012-219047 [Patent Document 2] Japanese Patent Application Laid-Open No. 2003-146837 [Non-patent literature]

[0015] [Non-Patent Document 1] "Journal of the Japanese Cosmetics Society", 2007, Vo3l.3, p.143-147 [Non-patent document 2] “Nat Genet.”,2006,Vol.38,No.4,p.441-446 [Non-patent document 3] "Fragrance Journal Special Issue", 2000, Vol. 17, pp. 14-19 [Non-patent document 4] “Arch. Dermatol. Res.”, 1996, Vol. 288, p. 442-446 Summary of the Invention [Problem to be solved by the invention]

[0016] The present invention aims to solve the above-mentioned conventional problems and to achieve the following object: Namely, the present invention aims to provide an MSX-2 mRNA expression promoter that has an excellent MSX-2 mRNA expression promoting effect and is highly safe. Another object of the present invention is to provide an eyelash growth agent or an external preparation composition for eyelash growth that has an excellent eyelash growth effect and is highly safe. Another object of the present invention is to provide a filaggrin mRNA expression promoter that has an excellent filaggrin mRNA expression promoting effect and is highly safe. Another object of the present invention is to provide a hyaluronan synthase 3 mRNA expression promoter that has an excellent hyaluronan synthase 3 mRNA expression promoting effect and is highly safe. Another object of the present invention is to provide a moisturizing agent or moisturizing topical composition that has an excellent moisturizing effect and is highly safe. [Means for solving the problem]

[0017] As a result of extensive research conducted by the present inventors to solve the above problems, they discovered that calendula extract has excellent effects of promoting MSX-2 mRNA expression, filaggrin mRNA expression, and hyaluronic acid synthase 3 mRNA expression, and is useful for promoting eyelash growth and moisturizing, and thus completed the present invention.

[0018] The present invention is based on the above findings of the present inventors, and the means for solving the above problems are as follows: <1> The present invention is an MSX-2 (Muscle Segment homeobox 2) mRNA expression promoter characterized by containing a calendula extract. <2> The aforementioned <1> 1. A hair growth agent for eyelashes, characterized by containing the MSX-2 mRNA expression promoter described in 1. <3> The aforementioned <1> 1. An external composition for eyelash growth, characterized by containing the MSX-2 mRNA expression promoter described in 1. <4> A filaggrin mRNA expression promoter characterized by containing a calendula extract. <5> A hyaluronic acid synthase 3 mRNA expression promoter characterized by containing calendula extract. <6> The aforementioned <4> The filaggrin mRNA expression promoter according to <5> The moisturizing agent is characterized by containing at least one of the hyaluronic acid synthase 3 mRNA expression promoters described in 1. <7> The aforementioned <4> The filaggrin mRNA expression promoter according to <5> The present invention relates to a moisturizing external preparation composition comprising at least one of the hyaluronic acid synthase 3 mRNA expression promoters described in the above. [Effects of the Invention]

[0019] The MSX-2 mRNA expression promoter of the present invention can solve the above-mentioned problems of the prior art and achieve the above-mentioned objective, and can provide an MSX-2 mRNA expression promoter that has excellent MSX-2 mRNA expression promoting activity and is highly safe. The eyelash growth agent or topical composition for eyelash growth of the present invention can solve the above-mentioned problems of the prior art and achieve the above-mentioned object, and can provide an eyelash growth agent or topical composition for eyelash growth that has excellent eyelash growth activity and is highly safe. The filaggrin mRNA expression promoter of the present invention can solve the above-mentioned problems of the past and achieve the above-mentioned objective, and can provide a filaggrin mRNA expression promoter that has excellent filaggrin mRNA expression promoting activity and is highly safe. The hyaluronic acid synthase 3 mRNA expression promoter of the present invention can solve the above-mentioned problems of the past and achieve the above-mentioned object, and can provide a hyaluronic acid synthase 3 mRNA expression promoter that has excellent hyaluronic acid synthase 3 mRNA expression promoting activity and is highly safe. The moisturizing agent or moisturizing topical composition of the present invention can solve the above-mentioned problems of the prior art and achieve the above-mentioned object, and can provide a moisturizing agent or moisturizing topical composition that has excellent moisturizing effect and is highly safe. DETAILED DESCRIPTION OF THE INVENTION

[0020] (MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronan synthase 3 mRNA expression promoter) The MSX-2 (Muscle Segment homeobox 2) mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter of the present invention contain calendula extract, and may further contain other ingredients as needed.

[0021] Although the details of the substance contained in the calendula extract that exhibits at least one of the effects of promoting MSX-2 mRNA expression, filaggrin mRNA expression, and hyaluronic acid synthase 3 mRNA expression are unknown, it was not previously known that the calendula extract has such excellent effects and is useful as an MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter, and this is a new discovery by the present inventors.

[0022] <Calendula officinalis extract> Calendula officinalis (scientific name: Calendula officinalis) is a plant of the Asteraceae family, native to Europe, and cultivated in various parts of Japan, making it easily available in these areas.

[0023] The calendula extract may be prepared from the part of the plant used as the raw material for extraction, or may be a commercially available product.

[0024] The part of the calendula officinalis to be extracted is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include leaves, branches, stems, flowers, buds, roots, above-ground parts, and mixtures thereof. These may be used alone or in combination of two or more. Among these, flowers are preferred. The shape, structure and size of the raw material for extracting calendula officinalis are not particularly limited and can be appropriately selected depending on the purpose.

[0025] The method for preparing the Calendula officinalis extract is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include a method in which the extracted part is dried and then subjected to solvent extraction either directly or after being crushed using a crusher. The drying may be performed in the sun or using a commonly used dryer.

[0026] The calendula officinalis extract can be easily obtained by a method commonly used for plant extraction. The form of the calendula officinalis extract is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include the extract itself, a diluted extract, a concentrated extract, a dried product thereof, a roughly purified product thereof, and a purified product thereof.

[0027] The extraction method is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include a method of extracting using any extraction device at room temperature or under reflux heating, and more specifically, a method of placing the extracted part of the calendula officinalis (the raw material for extraction) into a treatment tank filled with an extraction solvent, stirring appropriately as necessary, for example, and leaving it to stand for 30 minutes to 4 hours to elute soluble components, and then filtering to remove the extraction residue, thereby obtaining an extract. The extract may be further dried by distilling off the extraction solvent. Alternatively, the extract may be used as a raw material for extraction after pre-treatment such as degreasing with a non-polar solvent such as hexane. Pre-treatment such as degreasing allows for efficient extraction with a polar solvent.

[0028] The conditions for extracting calendula officinalis (extraction time and extraction temperature), the extraction solvent, and the amount of extraction solvent used are not particularly limited and can be appropriately selected depending on the purpose.

[0029] The extraction solvent is not particularly limited and can be appropriately selected depending on the purpose. Examples thereof include water, a hydrophilic organic solvent, and a mixed solvent thereof.

[0030] The water is not particularly limited and can be appropriately selected depending on the purpose. Examples include pure water, tap water, well water, mineral water, hot spring water, spring water, fresh water, and water obtained by various treatments. Examples of treatments for water include purification, heating, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering, etc. Water that can be used as the extraction solvent also includes purified water, hot water, ion-exchanged water, saline, phosphate buffer, phosphate-buffered saline, etc. The water may be used alone or in combination of two or more types.

[0031] The hydrophilic solvent is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include lower alcohols having 1 to 5 carbon atoms, such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones, such as acetone and methyl ethyl ketone; and polyhydric alcohols having 2 to 5 carbon atoms, such as 1,3-butylene glycol, propylene glycol, and glycerin. These may be used alone or in combination of two or more.

[0032] The amount of the hydrophilic solvent used relative to the water in the mixed solvent is not particularly limited and can be appropriately selected depending on the purpose. However, when a lower alcohol is used, it is preferable to add 1 part by mass to 90 parts by mass per 10 parts by mass of water, when a lower aliphatic ketone is used, it is preferable to add 1 part by mass to 40 parts by mass per 10 parts by mass of water, and when a polyhydric alcohol is used, it is preferable to add 1 part by mass to 90 parts by mass per 10 parts by mass of water.

[0033] The temperature of the extraction solvent is not particularly limited and can be appropriately selected depending on the purpose, but it is preferably used at a temperature between room temperature and the boiling point of the solvent.

[0034] The obtained calendula extract may be subjected to treatments such as dilution, concentration, drying, purification, etc. according to conventional methods to obtain a diluted product, concentrate, dried product, roughly purified product, purified product, etc. of the calendula extract.

[0035] The method for purifying the calendula extract is not particularly limited and can be appropriately selected depending on the purpose. Examples of purification methods include liquid-liquid partition extraction, various types of chromatography, and membrane separation.

[0036] The obtained Calendula officinalis extract can be used as it is as any of the MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter, but the concentrate and the dried product are preferred in terms of ease of use. When obtaining the dried product, a carrier such as dextrin or cyclodextrin may be added to improve hygroscopicity.

[0037] The content of the calendula officinalis extract in the MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter is not particularly limited and can be appropriately adjusted depending on the physiological activity of the extract, etc. The MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter may consist solely of the calendula officinalis extract.

[0038] <Other ingredients> The other components are not particularly limited and can be appropriately selected depending on the use form of the MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter, and examples thereof include excipients, moisture-proofing agents, preservatives, strengthening agents, thickeners, emulsifiers, antioxidants, sweeteners, acidulants, seasonings, coloring agents, fragrances, whitening agents, moisturizers, oily components, UV absorbers, surfactants, thickeners, alcohols, powder components, coloring agents, aqueous components, water, skin nutrients, etc. These may be used alone or in combination of two or more.

[0039] The content of the other components in the MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter is not particularly limited and can be selected appropriately depending on the purpose.

[0040] <Application> The uses of the MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter are not particularly limited and can be selected appropriately depending on the purpose, and examples include pharmaceuticals, quasi-drugs, cosmetics, etc. The MSX-2 mRNA expression promoter has an excellent MSX-2 mRNA expression promoting effect and is highly safe, and therefore can be suitably used, for example, as an active ingredient in eyelash growth agents and topical compositions for eyelash growth. The filaggrin mRNA expression promoter and hyaluronic acid synthase 3 mRNA expression promoter have excellent filaggrin mRNA expression promoting activity or excellent hyaluronic acid synthase 3 mRNA expression promoting activity, and are highly safe, so they can be suitably used, for example, as active ingredients in moisturizers and moisturizing topical compositions.

[0041] The MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter of the present invention are preferably applied to humans, but can also be applied to animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as their respective effects are achieved.

[0042] The method of use of the MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter is not particularly limited, and can be appropriately selected according to purpose, and can be, for example, orally, parenterally, externally, etc. Among these, externally is preferred.

[0043] The dosage forms of the MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter are not particularly limited and can be selected appropriately depending on the purpose. Examples include oral administration preparations such as tablets, powders, capsules, granules, extracts, and syrups; parenteral administration preparations such as injections, drips, and suppositories; and external preparations such as lotion, emulsion, cream, ointment, serum, lotion, pack, jelly, lip balm, lipstick, foundation, bath additives, soap, body soap, astringent, hair tonic, hair lotion, hair cream, hair liquid, pomade, shampoo, rinse, and conditioner. There are no particular limitations on the method for producing each of the dosage forms of the MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter, and any known method can be appropriately selected.

[0044] There are no particular limitations on the amount used, duration of use, and other usage methods of the MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter, and these can be selected appropriately depending on the purpose.

[0045] In addition, the MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter of the present invention can also be used as reagents for research into the mechanism of action of the MSX-2 mRNA expression promoting action, filaggrin mRNA expression promoting action, or hyaluronic acid synthase 3 mRNA expression promoting action.

[0046] (Eyelash growth agent and topical composition for eyelash growth) The eyelash growth agent and the external preparation composition for eyelash growth of the present invention contain the MSX-2 mRNA expression promoter of the present invention, and may further contain other ingredients as required.

[0047] The eyelash growth agent and the eyelash growth topical composition have an effect of promoting MSX-2 mRNA expression. In the present invention, "eyelash hair growth" is a concept that includes, in addition to promoting the hair growth, growth or normality of eyelashes, an increase in the length and thickness of eyelashes, and a firmness and resilience of eyelashes.

[0048] <MSX-2 mRNA expression promoter> The MSX-2 mRNA expression promoter is the MSX-2 mRNA expression promoter of the present invention described above.

[0049] The content of the MSX-2 mRNA expression promoter in the eyelash hair growth agent and the external preparation composition for eyelash hair growth is not particularly limited and can be appropriately adjusted according to the form of the eyelash hair growth agent and the external preparation composition for eyelash hair growth and the physiological activity of the Primula sieboldii extract, etc. However, in terms of the Primula sieboldii extract, 0.0001% by mass to 20% by mass is preferable, and 0.0001% by mass to 10% by mass is more preferable. The eyelash hair growth agent and the external preparation composition for eyelash hair growth may consist only of the MSX-2 mRNA expression promoter.

[0050] <Other components> The other components in the eyelash hair growth agent and the external preparation composition for eyelash hair growth are not particularly limited and can be appropriately selected according to the usage form of the eyelash hair growth agent and the external preparation composition for eyelash hair growth. For example, those described in the items of the above MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronan synthase 3 mRNA expression promoter, as well as the main agents, auxiliaries or other components usually used in the production of cosmetics, etc. can be mentioned. Examples of the main agents, auxiliaries or other components usually used in the production of cosmetics include astringents, bactericidal / antibacterial agents, whitening agents, ultraviolet absorbers, moisturizing agents, cell activators, anti-inflammatory / anti-allergy agents, antioxidants / reactive oxygen scavengers, oils, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, fragrances, etc. These may be used alone or in combination of two or more.

[0051] The content of the other ingredients in the eyelash growth agent and the composition for an external preparation for eyelash growth is not particularly limited and can be appropriately selected depending on the purpose.

[0052] <Application> The uses of the eyelash growth agent and the topical composition for eyelash growth are not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include pharmaceuticals, quasi-drugs, cosmetics, etc. The eyelash growth agent and topical composition for eyelash growth of the present invention can be used daily, and the active ingredient, calendula extract, can extremely effectively exert various physiologically active actions, including eyelash growth.

[0053] The eyelash growth agent and the topical composition for eyelash growth of the present invention are preferably applied to humans, but can also be applied to animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as the respective functional effects are exerted.

[0054] The method of use of the eyelash growth agent is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include oral administration, parenteral administration, external administration, etc. Among these, external administration is preferred. The eyelash growth topical composition is used externally.

[0055] The formulation and manufacturing method of the eyelash growth agent and the topical composition for eyelash growth are not particularly limited and can be appropriately selected depending on the purpose. Examples include those similar to those described above in the sections on MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter.

[0056] The amount of the eyelash growth agent and the topical composition for eyelash growth, the duration of use, etc. are not particularly limited and can be appropriately selected depending on the purpose.

[0057] As described above, the eyelash growth agent and topical composition for eyelash growth of the present invention have excellent eyelash growth effects. Therefore, the present invention also relates to a method for growing eyelashes, which comprises administering the eyelash growth agent or topical composition for eyelash growth to an individual.

[0058] The eyelash growth agent and the external preparation composition for eyelash growth of the present invention can also be used as a reagent for research into the mechanism of action of eyelash growth.

[0059] (Moisturizing agent and moisturizing topical composition) The moisturizing agent and moisturizing topical composition of the present invention contain at least one of the filaggrin mRNA expression promoter and the hyaluronic acid synthase 3 mRNA expression promoter of the present invention, and further contain other ingredients as necessary.

[0060] The moisturizing agent and the moisturizing topical composition have at least one of an action of promoting filaggrin mRNA expression and an action of promoting hyaluronic acid synthase 3 mRNA expression.

[0061] <At least one of a filaggrin mRNA expression promoter and a hyaluronic acid synthase 3 mRNA expression promoter> The filaggrin mRNA expression promoter and hyaluronan synthase 3 mRNA expression promoter are the filaggrin mRNA expression promoter and hyaluronan synthase 3 mRNA expression promoter of the present invention described above.

[0062] The total content of at least one of the filaggrin mRNA expression promoter and the hyaluronic acid synthase 3 mRNA expression promoter in the moisturizer and moisturizing topical composition is not particularly limited and can be adjusted appropriately depending on the form of the moisturizer and moisturizing topical composition and the physiological activity of the calendula extract, but is preferably 0.0001% to 20% by mass, and more preferably 0.0001% to 10% by mass, of the calendula extract. The moisturizer and moisturizing topical composition may consist solely of at least one of the filaggrin mRNA expression promoter and the hyaluronic acid synthase 3 mRNA expression promoter. Furthermore, the moisturizer and moisturizing topical composition may contain either one of the filaggrin mRNA expression promoter and the hyaluronic acid synthase 3 mRNA expression promoter, or both.

[0063] <Other ingredients> The moisturizers and other ingredients in the moisturizing topical composition are not particularly limited and can be selected appropriately depending on the intended use of the moisturizers and moisturizing topical composition. Examples include those listed above for the MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter, as well as main ingredients, auxiliary agents, and other ingredients typically used in the production of cosmetics. Examples of main ingredients, auxiliary agents, and other ingredients typically used in the production of cosmetics include astringents, bactericides / antibacterial agents, whitening agents, UV absorbers, moisturizers, cell activators, anti-inflammatory / antiallergic agents, antioxidants / active oxygen scavengers, oils and fats, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, and fragrances. These may be used alone or in combination of two or more.

[0064] The content of the other components in the moisturizing agent and the moisturizing topical preparation composition is not particularly limited and can be appropriately selected depending on the purpose.

[0065] <Application> The uses of the moisturizing agent and the moisturizing topical composition are not particularly limited and can be appropriately selected depending on the purpose. Examples include pharmaceuticals, quasi-drugs, and cosmetics. The moisturizing agent and moisturizing topical composition of the present invention can be used on a daily basis, and the active ingredient, calendula extract, can extremely effectively exert various physiologically active effects, including moisturizing action.

[0066] The moisturizing agent and moisturizing topical composition of the present invention are preferably applied to humans, but can also be applied to animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as their respective functional effects are achieved.

[0067] The method of use of the moisturizing agent is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include oral administration, parenteral administration, external administration, etc. Among these, external administration is preferred. The moisturizing external preparation composition is used externally.

[0068] The formulation and manufacturing method of the moisturizer and moisturizing topical composition are not particularly limited and can be selected appropriately depending on the purpose. For example, they can be similar to those described above in the sections on MSX-2 mRNA expression promoter, filaggrin mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter.

[0069] The amount of the moisturizing agent and the moisturizing topical composition used, the period of use, etc. are not particularly limited and can be appropriately selected depending on the purpose.

[0070] As described above, the moisturizing agent and the moisturizing topical composition of the present invention have excellent moisturizing effects. Therefore, the present invention also relates to a moisturizing method, which comprises administering the moisturizing agent and the moisturizing topical composition to an individual.

[0071] The moisturizing agent and moisturizing topical composition of the present invention can also be used as a reagent for research into the mechanism of moisturizing action. [Example]

[0072] Test examples and formulation examples of the present invention will be explained below, but the present invention is not limited to these test examples and formulation examples.

[0073] In the following test examples and formulation examples, the calendula extract used was "Calendula Extract BG-J" (extracted part: flowers, extraction solvent: mixed solvent of water and 1,3-butylene glycol) manufactured by Maruzen Pharmaceutical Co., Ltd.

[0074] (Test Example 1: MSX-2 mRNA expression promoting effect test) The calendula extract was used as a test sample and its promoting effect on MSX-2 (muscle segment homeobox 2) mRNA expression was tested by the following test method.

[0075] Normal human neonatal epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) and then harvested by trypsinization. 30 × 10 cells were cultured in a 35 mm dish using KGM. 4 The cells were seeded at 2 mL each and cultured overnight at 37°C under 5% CO2. After culture, the medium was replaced with growth factor-free medium (KBM). After 24 hours, the culture medium was discarded, and 2 mL of the test sample dissolved in KBM to the required concentration (see Table 1 below for sample concentrations) was added to each dish, and the cells were cultured at 37°C under 5% CO2 for 24 hours. After culture, the culture medium was discarded, and total RNA was extracted using ISOGEN II (NIPPON GENE; Cat. no. 311-07361). The amount of RNA in each sample was measured using a spectrophotometer, and total RNA was adjusted to 200 ng / μL. Using this total RNA as a template, the expression levels of MSX-2 and GAPDH mRNA (an internal standard) were measured. Detection was performed using a real-time PCR device, Thermal Cycler Dice® Real Time System III (TaKaRa), with TaKaRa SYBR® PrimeScript TM This was performed by real-time two-step RT-PCR using the RT-PCR Kit (Perfect Real Time) (code No. RR063A). The expression level of MSX-2 mRNA was calculated by correcting it with the expression level of GAPDH mRNA. The test was also carried out in the same manner without adding the test sample. The method for calculating the MSX-2 mRNA expression promotion rate is as follows. MSX-2 mRNA expression promotion rate (%) = A / B x 100 A: Correction value when test sample is added B: Corrected value when no test sample is added

[0076] [Table 1]

[0077] The results in Table 1 confirm that the Calendula officinalis extract has an excellent effect of promoting MSX-2 mRNA expression.

[0078] (Test Example 2: FLG mRNA expression promoting effect test) The calendula extract was used as a test sample and tested for its FLG (filaggrin) mRNA expression promoting effect by the following test method.

[0079] Normal human neonatal epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) and then harvested by trypsinization. 30 × 10 cells were cultured in a 35 mm dish using KGM. 4 The cells were seeded at 2 mL each and cultured overnight at 37°C under 5% CO2. After culture, the medium was replaced with growth factor-free medium (KBM). After 24 hours, the culture medium was discarded, and 2 mL of the test sample dissolved in KBM to the required concentration (see Table 2 below for sample concentrations) was added to each dish, and the cells were cultured at 37°C in 5% CO2 for 24 hours. After culture, the culture medium was discarded, and total RNA was extracted using ISOGEN II (NIPPON GENE; Cat. no. 311-07361). The amount of RNA in each sample was measured using a spectrophotometer, and total RNA was adjusted to 200 ng / μL. Using this total RNA as a template, the expression levels of FLG and GAPDH mRNA (an internal standard) were measured. Detection was performed using a real-time PCR device, Thermal Cycler Dice® Real Time System III (TaKaRa), with TaKaRa SYBR® PrimeScript TM This was performed by real-time two-step RT-PCR using the RT-PCR Kit (Perfect Real Time) (code No. RR063A). The expression level of FLG mRNA was calculated by correcting it with the expression level of GAPDH mRNA. The test was also carried out in the same manner without adding the test sample. The FLG mRNA expression promotion rate was calculated as follows. FLG mRNA expression promotion rate (%) = A / B × 100 A: Correction value when test sample is added B: Corrected value when no test sample is added

[0080] [Table 2]

[0081] The results in Table 2 confirm that the Calendula officinalis extract has an excellent effect of promoting FLG mRNA expression.

[0082] (Test Example 3: HAS3 mRNA expression promoting effect test) The calendula extract was used as a test sample and its promoting effect on HAS3 (hyaluronic acid synthase 3) mRNA expression was tested by the following test method.

[0083] Normal human neonatal epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) and then harvested by trypsinization. 30 × 10 cells were cultured in a 35 mm dish using KGM. 4 The cells were seeded at 2 mL each and cultured overnight at 37°C under 5% CO2. After culture, the medium was replaced with growth factor-free medium (KBM). After 24 hours, the culture medium was discarded, and 2 mL of the test sample dissolved in KBM to the required concentration (see Table 3 below for sample concentrations) was added to each dish, and the cells were cultured at 37°C in 5% CO2 for 24 hours. After culture, the culture medium was discarded, and total RNA was extracted using ISOGEN II (NIPPON GENE; Cat. no. 311-07361). The amount of RNA in each sample was measured using a spectrophotometer, and total RNA was adjusted to 200 ng / μL. Using this total RNA as a template, the expression levels of HAS3 mRNA and GAPDH mRNA (an internal standard) were measured. Detection was performed using a real-time PCR device, Thermal Cycler Dice® Real Time System III (TaKaRa), with TaKaRa SYBR® PrimeScript TM This was performed by real-time two-step RT-PCR using the RT-PCR Kit (Perfect Real Time) (code No. RR063A). The expression level of HAS3 mRNA was calculated by correcting it with the expression level of GAPDH mRNA. The test was also carried out in the same manner without adding the test sample. The method for calculating the HAS3 mRNA expression promotion rate is as follows. HAS3 mRNA expression promotion rate (%) = A / B × 100 A: Correction value when test sample is added B: Corrected value when no test sample is added

[0084] [Table 3]

[0085] The results in Table 3 confirm that the Calendula officinalis extract has an excellent effect of promoting HAS3 mRNA expression.

[0086] (Combination example 1) An eyelash beauty serum having the following composition was prepared by a conventional method. Calendula officinalis extract 0.05% by mass 1,3-butylene glycol 15.0% by mass Alkyl-modified carboxyvinyl polymer 0.5% by mass Sodium hydroxide 0.1% by mass Hydroxyethyl cellulose 0.5% by mass Phenoxyethanol 0.5% by mass · EDTA-4Na 0.05% by mass Remaining purified water

[0087] (Combination example 2) A cream having the following composition was prepared by a conventional method. Calendula officinalis extract 0.05% by mass Sophora root extract 0.1% by mass Scutellaria root extract 0.1% by mass Liquid paraffin 5.0% by mass White beeswax 4.0% by mass Squalane 10.0% by mass Cetyl alcohol 3.0% by mass Lanolin 2.0% by mass Stearic acid 1.0% by mass Polyoxyethylene sorbitan oleate (20E.O.) 1.5% by mass Glyceryl monostearate 3.0% by mass Oil-soluble licorice extract 0.1% by mass 1,3-butylene glycol 6.0% by mass Methyl parahydroxybenzoate 1.5% by mass · Fragrance 0.1% by mass Remaining purified water

Claims

[Claim 1] A filaggrin mRNA expression promoter comprising an extract of the flower parts of Calendula officinalis using water, a hydrophilic organic solvent, or a mixed solvent thereof.

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