JP6249516B2 - Transglutaminase activator - Google Patents

Description

  The present invention relates to a transglutaminase activator and a ceramide production promoter.

  The stratum corneum of the epidermis has a barrier function to prevent transpiration of moisture in the body, irritation from the outside world, and entry of foreign substances. The stratum corneum is composed of corneocytes and intercellular lipids, and the corneocytes are encapsulated in a cell membrane-like structure called a cornified envelope. The cornified envelope contributes to the construction of a stable keratinocyte structure and is an important structure for maintaining the barrier function of the skin. In the cornified envelope, keratinocytes in the basal layer of the stratum corneum are keratinized, and involucrin, loricrin, and other proteins necessary for cornification are synthesized, and then these proteins are cross-linked by activation of transglutaminase. Formed with. The activity of transglutaminase is important for the normal formation of the cornified envelope and the normal keratinization of the epidermis, and thus the maintenance and improvement of the skin moisturizing function. In this regard, it has been reported that the activation of transglutaminase leads to normal keratinization of the skin, improvement of barrier function, and improvement of rough skin (for example, see Non-Patent Documents 1 and 2 and Patent Documents 1 and 2). ).

In addition, ceramide, which is one of the sphingolipids, is a lipid that exists only in a trace amount in the entire living body. However, in the stratum corneum, the outermost layer of the skin, ceramide occupies about half of the lipid and plays an important role in the skin moisturizing and barrier mechanisms. This ceramide functions by constructing a lamellar structure between cells in the stratum corneum after being produced and secreted in epidermal cells. However, in skin diseases such as dry skin, rough skin, atopic dermatitis, senile psoriasis and psoriasis, the healthy metabolism of ceramide is hindered, the amount of ceramide in the stratum corneum is reduced, and the skin's moisture retention ability is reduced. It has been reported many that it causes a decrease, keratinization failure of the epidermis, a decrease in barrier ability, etc. (see Non-Patent Document 3). Substances that promote ceramide production can be expected to have effects such as suppression of animal cell proliferation, induction of differentiation, and induction of apoptosis. As a result, diseases caused by abnormal cell proliferation or differentiation such as inflammatory diseases and malignant tumors. It is thought that the therapeutic effect with respect to can be expected (refer nonpatent literature 4). Furthermore, ceramide has a bone resorption inhibitory action, a bone strengthening action, and an alveolar bone loss inhibitory action, and is useful for the prevention and improvement of osteoarthritis such as osteoporosis, fracture, low back pain, rheumatism (see Patent Document 3). It has also been reported that ceramide has an effect in preventing periodontal disease (see Patent Document 4), and that ceramide has an effect of imparting firmness and stiffness to the hair and improving touch (see Patent Document 5).
Thus, ceramide can be expected to have various effects, and there is a demand for a substance that can promote the production of ceramide.

On the other hand, Acanthaceae plants consist of about 250 genera and 2500 species, of which about 300 species of Justicia plants exist. Among them, kitsunenomago (Justicia procumbens) is by far used as Chinese medicine to be applied to joint pain and fever, etc., is the same Acanthaceae Genus frutescens kitsunenomago (Justicia gendarussa) the skin external analgesics and skin external anti itch agents (See, for example, Patent Documents 6 and 7). Also, a different genus of plants of the same acanthaceae Adatoda-Washika (Adhatoda vasica) are known to have a ceramide production promoting effect (e.g., see Patent Document 8). Furthermore, it is known that the physiological activity of the extract of the foxes has an inhibitory effect on melanin production and an inhibitory effect on dopa oxidase activity (see, for example, Patent Document 9).
However, it has not been known so far that the extract of foxgill activates transglutaminase, promotes the production of ceramide, is useful for maintaining or improving the skin barrier function and moisturizing function, and preventing or improving rough skin. .

JP 2004-91376 A Japanese Patent Laid-Open No. 2007-1914 JP 2001-158736 A JP 2001-158735 A Japanese Patent Laid-Open No. 10-152421 JP-A-2005-281206 JP 2007-230977 A JP 2011-79755 A International Publication No. 2013/031403 Pamphlet

Kalinin, A., et al., Journal of Cell Science, 2001, vol. 114, p. 3069-3070 Rawlings, A. V., et al., Journal of Investigative Dermatology, 2005, vol. 124, p. 1099-1110 Ishikawa, J., et al., Journal of Cosmetic Dermatology, 2013, vol. 12, p. 3-11 David E. Modrak, et al., Molecular Cancer Therapeutics, 2006, vol. 5 (2), p. 200-208

An object of the present invention is to provide a transglutaminase activator that activates transglutaminase and is useful for maintaining or improving the skin barrier function and moisturizing function, and preventing or improving rough skin.
Furthermore, an object of the present invention is to provide a ceramide production promoter that promotes the production of ceramide and is useful for maintaining or improving the skin barrier function and moisturizing function, preventing or improving rough skin, and the like.

  As a result of intensive studies in view of the above-mentioned problems, the present inventors have found that the extract of foxes activates transglutaminase, promotes the production of ceramide, increases the amount of stratum corneum, It has been found that the decrease in the amount is suppressed and the rough skin is prevented or improved. The present invention has been completed based on these findings.

The present invention relates to a transglutaminase activator comprising an extract of foxes as an active ingredient.
Moreover, this invention relates to the ceramide production promoter which uses the extract of a foxglove as an active ingredient.
The present invention also relates to an epidermis keratinization improving agent comprising an extract of a foxtail as an active ingredient.
Moreover, this invention relates to the skin moisturizing function improving agent which uses the extract of a foxglove as an active ingredient.
The present invention also relates to a skin barrier function-improving agent comprising a foxtail extract as an active ingredient.
In addition, the present invention relates to a stratum corneum water content increasing agent comprising an extract of foxtail as an active ingredient.
Moreover, this invention relates to the stratum corneum water content fall inhibitor which uses the extract of a foxglove as an active ingredient.
Furthermore, this invention relates to the rough skin prevention or improvement agent which uses the extract of a foxtail egg as an active ingredient.

The transglutaminase activator of the present invention activates transglutaminase and is useful for maintaining or improving the skin barrier function and moisturizing function, and preventing or improving rough skin.
The ceramide production promoter of the present invention promotes the production of ceramide and is useful for maintaining or improving the barrier function and moisturizing function of skin, preventing or improving rough skin, and the like.

In the present specification, “improvement” refers to improvement of a disease, symptom or condition, prevention or delay of deterioration of the disease, symptom or condition, or reversal, prevention or delay of progression of the disease, symptom or condition.
Further, in the present specification, “non-therapeutic” is a concept that does not include a medical act, that is, a treatment act on the human body by therapy.
In the present specification, “prevention” means prevention or delay of the onset of a disease or symptom in an individual, or reduction of the risk of onset of a disease or symptom in an individual.
Furthermore, in this specification, “rough skin” means that the moisture retention capacity of the skin is reduced and the moisture of the skin is taken away, and the skin surface is crushed or cracked, or the skin surface roughness is increased. State. Skin in such a state is also referred to as “that skin” or “dry skin”.

The transglutaminase activator, the ceramide production promoter, the stratum corneum water content increasing agent, and the stratum corneum water content lowering inhibitor of the present invention contain an extract of a foxtail as an active ingredient. As demonstrated in the examples described later, the foxgull extract has a transglutaminase activating effect, a ceramide production promoting effect, a stratum corneum water content increasing effect, and a stratum corneum water content reducing effect.
Further, the skin keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, and rough skin preventing or improving agent of the present invention also contain a foxgull extract as an active ingredient. As described above, activation of transglutaminase, promotion of ceramide production, increase in stratum corneum water content, and suppression of decrease in stratum corneum water content are improved keratinization of skin and skin moisturizing function, maintenance of skin barrier function and Very important for preventing or improving rough skin. In addition, as demonstrated in the examples described later, the foxtail extract improves keratinization of the epidermis, improves the skin moisturizing function, improves the skin barrier function, and prevents or improves rough skin. Therefore, an extract of a fox moth having a transglutaminase activating effect, a ceramide production promoting effect, a stratum corneum water content increasing effect, and a stratum corneum water content lowering suppressing effect is used as an epidermis keratinization improving agent, skin moisturizing function improving agent, skin It can be used as an active ingredient of a barrier function improving agent and a rough skin preventing or improving agent.
As described above, ceramide is involved in the control of cell proliferation, differentiation, apoptosis, and the like. Therefore, the foxtail extract that promotes ceramide production prevents diseases caused by abnormal cell proliferation or differentiation, such as inflammatory diseases and malignant tumors, by suppressing the growth of animal cells, inducing differentiation, and inducing apoptosis. Or it is useful for the pharmaceutical for treatment, a quasi-drug, etc. Moreover, the extract of a foxtail can also be used for the prevention or improvement of osteoarthritis, bone fracture, low back pain, rheumatism, and other osteoarthritis, as well as for drugs or quasi drugs for the prevention or improvement of periodontal disease. Furthermore, the extract of foxtail is useful for applications such as quasi-drugs and cosmetics for imparting firmness and stiffness to the hair and improving the feel of the hair.

As used herein, the “fox fox” is an annual plant of the genus Justicia .
Any part of the foxtail can be used to make the foxtail extract, including whole grass, roots, tuberous roots, rhizomes, stems, branches, stems, leaves (leaf blades, petiole, etc.), bark, sap, resin, flowers (Petals, ovaries, etc.), fruits (mature fruits, immature fruits, etc.), seeds and the like can be used. A combination of these parts may also be used. Especially, it is preferable that the extract of a fox wild bean used for this invention is the extract of the whole fox wild bean.

The extract of foxes used in the present invention can be obtained by a usual extraction method used for plant extraction or the like. The extraction method can be appropriately set, and is preferably obtained by extracting the plant at room temperature or under heating or using an extraction tool such as a Soxhlet extractor.
For the preparation of an extract of a foxtail, the foxtail can be used as it is or after being dried and ground. Moreover, the steamed distillate or press thing of a foxtail can also be used, These can also use what refine | purified from essential oil etc., and can also use a commercial item. You may use a fox wild boar or its steam distillate or pressing thing individually or in combination of 2 or more types.

  The extraction solvent used for the preparation of the foxtail extract can be selected as appropriate and is usually used for extraction of plant components, such as water; alcohols such as methanol, ethanol, propanol, butanol; ethylene glycol, propylene glycol, 1 Polyhydric alcohols such as 1,2-butylene glycol, 1,3-butylene glycol, 1,4-butylene glycol, 2,3-butylene glycol; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate Chain and cyclic ethers such as tetrahydrofuran and diethyl ether; polyethers such as polyethylene glycol; halogenated hydrocarbons such as dichloromethane, dichloroethane, chloroform and carbon tetrachloride; hydrocarbons such as hexane, cyclohexane and petroleum ether Benzene, Aromatic hydrocarbons such as ruene; pyridines; supercritical carbon dioxide; fats and oils, waxes, and other oils. These may be used alone or in combination of two or more. Among these, water, ethanol, or an aqueous ethanol solution is preferable, an aqueous ethanol solution is more preferable, an aqueous ethanol solution having an alcohol content of 30% by volume or more is more preferable, and an aqueous ethanol solution having an alcohol content of 40% by volume or more is particularly preferable. Further, acid or alkali may be added during extraction to adjust the pH of the extraction solvent.

  Normal conditions can be applied as the extraction conditions. For example, the foxtail is 0 ° C. or higher (preferably 4 ° C. or higher) and 100 ° C. or lower (preferably 80 ° C. or lower, more preferably 40 ° C. or lower). , More preferably 1 day or more) 50 days or less (preferably 30 days or less) soaking or heating under reflux. In order to increase the extraction efficiency, stirring may be performed together or homogenization treatment may be performed in a solvent. The amount of the extraction solvent to be used is not less than 1 time (preferably not less than 5 times) and not more than 100 times (preferably not more than 50 times, more preferably not more than 40 times) with respect to the weight of foxtail (in terms of dry matter) It is.

In the present invention, the foxgull extract may be used as it is, or a fraction having high activity may be fractionated and used by an appropriate separation means such as gel filtration, chromatography, precision distillation or the like. Further, the obtained foxtail extract can be diluted, concentrated or lyophilized, and then prepared and used in the form of powder or paste. In addition, the extract obtained by the above method can be used by being dissolved in a solvent different from the extraction solvent.
In the present invention, the extract includes various solvent extracts obtained by the extraction method as described above, diluted solutions thereof, concentrated solutions thereof, purified fractions thereof, dried powders thereof or transferred solutions thereof.

  Transglutaminase activator, ceramide production promoter, epidermis keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, horny layer moisture content increasing agent, horny layer moisture content depressing agent, rough skin inhibitory or The form of the improving agent and the rough skin preventing or improving agent can be appropriately selected, and for example, it can be a pharmaceutical composition, a cosmetic composition or a food composition, or can be contained in these.

  When preparing a pharmaceutical composition, it is usually prepared as a preparation containing the active ingredient and preferably a pharmaceutically acceptable carrier. A pharmaceutically acceptable carrier generally refers to an inert, non-toxic, solid or liquid, bulking agent, diluent or encapsulating material that does not react with the active ingredient, eg, water. , Ethanol, polyols (for example, propylene glycol, butylene glycol, glycerin, and polyethylene glycol), suitable mixtures thereof, solvents or dispersion media such as vegetable oils, and the like.

  The pharmaceutical composition is orally, parenterally, e.g., in the oral cavity, in the skin, subcutaneously, in the mucosa, intravenously, in the artery, in the muscle, in the abdominal cavity, in the vagina, in the lungs. Administered intracerebrally, ocularly and intranasally. Examples of the preparation for oral administration include tablets, granules, fine granules, powders, capsules, chewables, pellets, syrups, solutions, suspensions and inhalants. As parenteral preparations, suppositories, retention enemas, drops, eye drops, nasal drops, pessaries, injections, mouth washes, ointments, creams, gels, controlled release patches and patches Skin external preparations, and the like. The pharmaceutical compositions may be administered parenterally in the form of sustained release subcutaneous implants or in the form of targeted delivery systems (eg, monoclonal antibodies, vector delivery, ion implantation, polymer matrices, liposomes and microspheres). .

  The pharmaceutical composition may further contain additives conventionally used in the pharmaceutical field. Such additives include, for example, excipients, binders, disintegrants, lubricants, antioxidants, colorants, flavoring agents, and the like, and can be used as necessary. In order to achieve sustained release so that it can act for a long time, it can also be coated with a known retarder or the like. Examples of excipients include sodium carboxymethylcellulose, agar, light anhydrous silicic acid, gelatin, crystalline cellulose, sorbitol, talc, dextrin, starch, lactose, sucrose, glucose, magnesium metasilicate magnesium phosphate, calcium hydrogen phosphate, etc. Can be used. Examples of the binder include gum arabic, sodium alginate, ethyl cellulose, sodium caseinate, sodium carboxymethyl cellulose, agar, purified water, gelatin, starch, tragacanth, and lactose. Examples of the disintegrant include carboxymethyl cellulose, carboxymethyl cellulose sodium, carboxymethyl cellulose calcium, crystalline cellulose, starch, hydroxypropyl starch and the like. Examples of the lubricant include stearic acid, calcium stearate, magnesium stearate, talc, hydrogenated oil, sucrose fatty acid ester, waxes and the like. Examples of the antioxidant include tocopherol, gallic acid ester, dibutylhydroxytoluene (BHT), butylhydroxyanisole (BHA), ascorbic acid and the like. Other additives and drugs as required, such as antacids (sodium bicarbonate, magnesium carbonate, precipitated calcium carbonate, synthetic hydrotalcite, etc.), gastric mucosa protective agents (synthetic aluminum silicate, sucralfate, copper chlorophyllin sodium, etc.) ) May be added.

  When preparing a cosmetic composition, the form thereof can be selected as appropriate, such as solution, emulsion, powder, water-oil two-layer system, water-oil-powder three-layer system, gel, tablet and other solids, aerosol, It can be in any form such as mist, capsule, and sheet. In addition, the product form of the cosmetic composition is also arbitrary. For example, skin care cosmetics such as face wash, makeup remover, lotion, cosmetic liquid, pack, milky lotion, cream and sunscreen, foundation, makeup base, lipstick, eye Makeup cosmetics such as shadow, eyeliner, mascara, eyebrow, blusher and nail enamel, hair cosmetics such as hair shampoo, hair rinse, hair styling, hair dye and hair restorer, soap, body soap, deodorant cosmetic and bath preparation And other body cleansing agents, oral cosmetics such as dentifrices and mouthwashes, and aromatic cosmetics such as perfumes. Moreover, this cosmetic may belong to either cosmetics or quasi-drugs in accordance with Japanese Pharmaceutical Affairs Law.

The cosmetic composition includes other components commonly used in cosmetics, quasi drugs and pharmaceuticals, such as powder components, liquid fats and oils, solid fats and oils, waxes, hydrocarbons, higher fatty acids, higher alcohols, esters, silicones, anions. Surfactant, cationic surfactant, amphoteric surfactant, nonionic surfactant, humectant, water-soluble polymer, thickener, film agent, UV absorber, sequestering agent, lower alcohol, polyhydric alcohol , Sugar, amino acids, organic amines, polymer emulsions, pH adjusters, skin nutrients, vitamins, antioxidants, antioxidant auxiliaries, fragrances, water, etc. may be blended as necessary and manufactured by conventional methods. it can.
Examples of other components that can be incorporated into the cosmetic composition include preservatives (ethyl paraben, butyl paraben, etc.), anti-inflammatory agents (eg, glycyrrhizic acid derivatives, glycyrrhetinic acid derivatives, salicylic acid derivatives, hinokitiol, zinc oxide, allantoin, etc. ), Whitening agents (for example, ascorbic acid and its derivatives, placenta extract, saxifrage extract, arbutin, etc.), various extracts (for example, buckwheat, auren, shikon, peonies, assembly, birch, sage, loquat, carrot, aloe , Mallow, iris, grape, yokuinin, loofah, lily, saffron, senkyu, ginger, hypericum, onionis, garlic, capsicum, chimpi, red snapper, seaweed, etc.), activator (eg royal jelly, photosensitizer, cholesterol derivative, etc.) , Blood circulation promoter For example, nonyl acid valenylamide, nicotinic acid benzyl ester, nicotinic acid β-butoxyethyl ester, capsaicin, gingerone, cantalis tincture, ictamol, tannic acid, α-borneol, nicotinic acid tocopherol, inositol hexanicotinate, cyclandrate, cinnarizine , Trazoline, acetylcholine, verapamil, cephalanthin, γ-oryzanol, etc.), antiseborrheic agents (eg, sulfur, thianthol, etc.), anti-inflammatory agents (eg, tranexamic acid, thiotaurine, hypotaurine, etc.) and fungicides (eg, triclosan, Cetylpyridinium chloride, thymols, benzalkonium chloride, etc.).

The pharmaceutical composition and cosmetic composition can be applied in the form of an oral composition, an external composition, an internal use composition, etc., and is preferably used in the form of a skin external composition.
When used in the form of a composition for external use on the skin, in addition to the foxtail extract, components used in normal composition for external use on the skin, such as surfactants, oily substances, polymer compounds, preservatives, various medicinal ingredients, powders UV absorbers, dyes, fragrances, emulsion stabilizers, pH adjusters, and the like can be appropriately blended. As a medicinal component, in the case of an epidermis keratinization improving agent or a skin moisturizing function improving agent, for example, vitamin D3, a sphingosine derivative, oleanolic acid, clofibric acid, oleethanolamide may be mentioned.

  When preparing a food composition, the form can be selected as appropriate, and beverages are also included. In addition to general foods, treatment or prevention of diseases or conditions that can be treated, prevented or ameliorated by activating transglutaminase or promoting production of ceramide, such as improving keratinization of the epidermis or improving or maintaining skin moisturizing function or barrier function Or the food / beverage products which displayed that on the basis of the improvement etc., ie, health food, functional food, food for sick people, food for specified health, etc. are also included. Health foods, functional foods, foods for patients and foods for specified health use are specifically formulated in various forms such as fine granules, tablets, granules, powders, capsules, syrups, liquids, and liquid foods. Can be used for these formulations. A food composition in the form of a preparation can be produced in the same manner as a pharmaceutical preparation. The active ingredient and a carrier acceptable as a food, for example, an appropriate excipient (eg, starch, processed starch, lactose, glucose, water, etc.) ) And the like, and then can be produced using conventional means. In addition, food compositions include soups, juices, milk beverages, tea beverages, coffee beverages, cocoa beverages, jelly-like beverages and other liquid food compositions, pudding, yogurt and other semi-solid food compositions, breads, udon Such as noodles such as cookies, chocolate, candy, gum, rice crackers and the like, spreads such as sprinkles, butter, jam and the like. The food also includes feed.

  Food compositions include various food additives such as antioxidants, fragrances, various esters, organic acids, organic acid salts, inorganic acids, inorganic acid salts, inorganic salts, pigments, emulsifiers, preservatives, seasonings Additives such as additives, sweeteners, acidulants, fruit juice extracts, vegetable extracts, nectar extracts, pH adjusters, and quality stabilizers may be used alone or in combination.

Transglutaminase activator, ceramide production promoter, epidermis keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, stratum corneum water content increasing agent, stratum corneum water content lowering inhibitor, or rough skin prevention Alternatively, the administration target of the improving agent is preferably a warm-blooded vertebrate, more preferably a mammal. As used herein, mammals include, for example, humans and non-human mammals such as monkeys, mice, rats, rabbits, dogs, cats, cows, horses, and pigs. Transglutaminase activator, ceramide production promoter, epidermis keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, stratum corneum water content increasing agent, stratum corneum water content lowering inhibitor, or rough skin prevention Alternatively, the improving agent is suitable for administration to humans, primates such as monkeys, particularly humans.
The extract used in the present invention, as well as the transglutaminase activator, ceramide production promoter, epidermis keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, horny layer water content increasing agent, horny layer of the present invention Water content lowering inhibitor, or rough skin prevention or improvement agent, prevention or treatment of keratinization failure of the epidermis, skin moisturization, improvement of skin barrier function, increase in stratum corneum water content, suppression of decrease in stratum corneum water content, Or it can apply to the subject who desires prevention or improvement of rough skin. The extract or agent is preferably applied under necessary conditions (preferably under low humidity and dry conditions). The extract or agent is preferably applied to the skin.

  Transglutaminase activator, ceramide production promoter, epidermis keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, stratum corneum water content increasing agent, stratum corneum water content lowering inhibitor, or rough skin prevention Or the dosage of the said active ingredient in an improving agent can be suitably determined by an individual's condition, body weight, sex, age, activity of a material, administration or intake route, administration or intake schedule, formulation form, or other factors. For example, based on the mass of the active ingredient, it is preferably 0.001 mg or more, 1 g or less, or preferably 0.001 to 1 mg per kg of body weight per day. The active ingredient can be taken or administered once a day to several times a day, or at an arbitrary period and interval.

  Transglutaminase activator, ceramide production promoter, epidermis keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, stratum corneum water content increasing agent, stratum corneum water content lowering inhibitor, or rough skin prevention Or content of the said active ingredient in an improving agent can be suitably determined so that the said dosage may be achieved. For example, the transglutaminase activator of the present invention, ceramide production promoter, epidermis keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, increase in stratum corneum water content, suppression of decrease in stratum corneum water content, Alternatively, in the rough skin preventing or improving agent, the content of the active ingredient is preferably 0.00001% by mass or more, more preferably 0.0001% by mass or more, further preferably 0.0005% by mass or more, preferably 20% by mass or less, and 10% by mass or less. Is more preferably 5% by mass or less, preferably 0.00001 to 20% by mass, more preferably 0.0001 to 10% by mass, and further preferably 0.0005 to 5% by mass.

  In relation to the above-described embodiment, the present invention further includes the following transglutaminase activator, ceramide production promoter, epidermis keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, horny layer moisture content increasing agent, horny layer Disclosed are water content lowering inhibitors, rough skin prevention or improvement agents, production methods, methods and uses.

<1> A transglutaminase activator comprising an extract of foxes as an active ingredient.
<2> A ceramide production promoter comprising an extract of foxes as an active ingredient.
<3> An epidermis keratinization improving agent comprising an extract of foxglove as an active ingredient.
<4> A skin moisturizing function-improving agent comprising an extract of foxtail as an active ingredient.
<5> A skin barrier function-improving agent comprising an extract of foxes as an active ingredient.
<6> A stratum corneum water content increasing agent comprising an extract of foxtail as an active ingredient.
<7> A stratum corneum moisture content inhibitor comprising, as an active ingredient, an extract of a foxglove.
<8> An agent for preventing or improving rough skin, comprising an extract of a foxglove as an active ingredient.

<9> The transglutaminase activator, the ceramide production promoter, and the improvement of skin keratinization according to any one of the above <1> to <8>, wherein the extract of foxtail is a whole plant extract of foxtail Agent, skin moisturizing function improving agent, skin barrier function improving agent, stratum corneum moisture content increasing agent, stratum corneum moisture content lowering inhibitor, or rough skin preventing or improving agent.
<10> The foxgull extract was obtained by extracting a foxglove with an ethanol aqueous solution (preferably an ethanol aqueous solution having an alcohol content of 30% by volume or more (more preferably 40% by volume or more)) as an extraction solvent. <1> to <9>, wherein the transglutaminase activator, ceramide production promoter, epidermis keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, horny layer water content increase Agent, stratum corneum moisture reduction inhibitor, or rough skin prevention or improvement agent.
<11> The content of the active ingredient is 0.00001% by mass or more (preferably 0.0001% by mass or more, more preferably 0.0005% by mass or more), 20% by mass or less (preferably 10% by mass or less, more preferably 5% by mass or less). The transglutaminase activator, ceramide production promoter, epidermal keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, horn of any one of the above <1> to <10> Layer moisture content increasing agent, stratum corneum moisture content decreasing inhibitor, or rough skin preventing or improving agent.

<12> Use of an extract of foxes as a transglutaminase activator or a ceramide production promoter.
<13> Use of an extract of a foxes for the production of a transglutaminase activator or a ceramide production promoter.
<14> A method of using an extract of a foxtail as a transglutaminase activator or a ceramide production promoter.
<15> A method for activating transglutaminase or a method for promoting ceramide production, using an extract of a foxglove.
<16> The method according to <15>, wherein the extract is applied to a subject who wants to prevent or treat keratinization failure of the epidermis, moisturize skin, improve skin barrier function, or prevent or improve rough skin. .
<17> The method according to <15> or <16>, wherein the extract is applied under conditions that require application of the extract (preferably under dry conditions with low humidity).
<18> The method according to any one of <15> to <17>, wherein the extract is applied to skin.
<19> The use or method according to any one of the above <12> to <18>, wherein the extract of the foxtail is a whole plant extract of the foxtail.
<20> The foxgull extract was obtained by extracting foxglove with an aqueous ethanol solution (preferably an ethanol aqueous solution having an alcohol content of 30% by volume or more (more preferably 40% by volume or more)) as an extraction solvent. The use or method according to any one of <12> to <19>.
<21> The content of the foxgull extract in the transglutaminase activator or ceramide production promoter is 0.00001% by mass or more (preferably 0.0001% by mass or more, more preferably 0.0005% by mass or more) 20% by mass or less (preferably Is 10% by mass or less, more preferably 5% by mass or less). The use or method according to any one of <12> to <20>.

<22> Skin keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, stratum corneum water content increasing agent, stratum corneum water content decreasing inhibitor, or skin rot extract as a rough skin preventing or improving agent use.
<23> Epidermis keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, stratum corneum moisture content increasing agent, stratum corneum moisture content reducing agent, or skin rough preventing or improving agent Use of extract.
<24> The foxglove extract is used as an epidermis keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, stratum corneum water content increasing agent, stratum corneum water content decreasing inhibitor, or rough skin preventing or improving agent. how to.
<25> Epidermis keratinization improving method, skin moisturizing function improving method, skin barrier function improving method, stratum corneum moisture content increasing method, stratum corneum moisture content suppressing method, or rough skin prevention or improving method, applying the extract of <25> Fox .
<26> Prevention or treatment of keratinization failure of the epidermis, moisturizing skin, improvement of skin barrier function, increase of stratum corneum moisture, suppression of decrease in stratum corneum, or prevention or improvement of rough skin The method according to <24> or <25> above, which is applied to a desired subject.
<27> The method according to any one of <24> to <26>, wherein the extract is applied under conditions that require application of the extract (preferably under dry conditions with low humidity).
<28> The method according to any one of <24> to <27>, wherein the extract is applied to skin.
<29> For prevention or treatment of skin epikeratinization failure, skin moisturizing function improving method, skin barrier function improving method, stratum corneum water content increasing method, stratum corneum water content decrease suppressing method, or rough skin preventing or improving method An extract of the fox wild egg
<30> Preparation of preventive or therapeutic agent for cutaneous epidermal keratinization, skin moisturizing function improving agent, skin barrier function improving agent, stratum corneum moisture increase agent, stratum corneum moisture decrease inhibitor, or rough skin preventing or improving agent For the use of the extract of fox wild eggs.
<31> Use of an extract of a foxglove used for a non-therapeutic treatment method of skin epikeratinization failure, skin moisturizing function, skin barrier function, stratum corneum water content, or rough skin.
<32> The use according to <31> above, wherein the extract of foxglove is applied in the form of a pharmaceutical composition or a cosmetic composition.
<33> Use according to <32> above, wherein the extract of foxtail is applied in the form of a composition for external use.
<34> The use according to <31> above, wherein the extract of foxtail is applied in the form of food or beverage.
<35> The use or method according to any one of the above <22> to <34>, wherein the extract of the foxtail is a whole plant extract of the foxtail.
<36> The foxgull extract was obtained by extracting foxglove with an aqueous ethanol solution (preferably an ethanol aqueous solution having an alcohol content of 30% by volume or more (more preferably 40% by volume or more)) as an extraction solvent. The use or method according to any one of <22> to <35>.
<37> An extract of the foxglove in the above-mentioned skin keratinization improving agent, skin moisturizing function improving agent, skin barrier function improving agent, stratum corneum moisture content increasing agent, stratum corneum moisture content reducing agent, or rough skin preventing or improving agent. The content is 0.00001% by mass or more (preferably 0.0001% by mass or more, more preferably 0.0005% by mass or more) and 20% by mass or less (preferably 10% by mass or less, more preferably 5% by mass or less). The use or method according to any one of> to <36>.

  EXAMPLES Hereinafter, although this invention is demonstrated further in detail based on an Example, this invention is not limited to this.

Preparation Example 1
To 80 g of the whole foxglove (manufactured by Shinwa Bussan), 800 mL of a 50% by volume aqueous ethanol solution was added and extracted at room temperature for 7 days. Thereafter, a crude extract was obtained by filtration, and then concentrated to dryness to obtain 6.6 g of an extracted solid. This extracted solid was dissolved in a 50% by volume ethanol aqueous solution so that the evaporation residue was 1.0% (w / v) to prepare a 50% by volume ethanol extract of foxtail.

Preparation Example 2
To 50 g of the whole foxtail plant (manufactured by Shinwa Bussan Co., Ltd.), 500 mL of a 95% by volume ethanol aqueous solution was added, followed by extraction at room temperature for 7 days. Then, after filtering and obtaining a crude extract, it concentrated and dried and obtained 897 mg of extraction solid content. This extracted solid was dissolved in a 95% by volume ethanol aqueous solution so as to have an evaporation residue of 1.0% (w / v) to prepare a 95% by volume ethanol extract of a foxtail.

Test Example Measurement of transglutaminase activity Human epidermal keratinocyte cell line HEKn (manufactured by KURABO) was seeded at 4 × 10 ⁴ cells / well in a 12-well plate and cultured. As the medium, commercially available EpiLife-KG2 manufactured by Kurabo Industries was used. After culturing for one day at 37 ° C. and 5% CO ₂ , the medium was replaced with a medium containing no growth factors (BPE, EGF), and 1.0% (w / v) in terms of solid content prepared in Production Examples 1 and 2. The concentration of the foxroot extract thus obtained was added so that the final concentration would be the value shown in Table 1. Further, instead of the foxtail extract, 50% ethanol aqueous solution or 95% ethanol aqueous solution as the extraction solvent was used as a control at a final concentration of 0.1% (v / v), and CaCl ₂ was used as a positive control at a final concentration of 1 Each was added at 5 mM. CaCl ₂ is known to promote keratinization and was used as a positive control. All of these were further cultured at 37 ° C. for 3 days.
After completion of the culture, the culture solution was removed, washed twice with PBS (−), and 150 μL of extraction buffer (10 mM Tris-HCl buffer, pH 7.4 containing 0.5 mM EDTA, 1% TritonX-100, and protease inhibitors). The cells were collected using a cell scraper to obtain a cell disruption solution by ultrasonic treatment. The supernatant obtained by centrifugation (15,000 rpm, 10 minutes) was used for evaluation as a lysate. Enzyme activity was measured using Transglutaminase Colorimetric Microassay Kit (trade name) according to the manufacturer's instructions. The protein concentration was quantified using BCA Protein Assay Kit (trade name, manufactured by Thermo Scientific) according to the manufacturer's instructions.

  The evaluation results are shown in Table 1. The transglutaminase activity of each extract sample is the relative value when the transglutaminase activity of the control obtained by adding 50% by volume ethanol aqueous solution is 1 for the extract obtained in Preparation Example 1, and is obtained in Preparation Example 2. The extract was shown as a relative value when the transglutaminase activity of a control to which a 95% by volume aqueous ethanol solution was added was 1.

As can be seen from Table 1, in the system to which the foxgull extract was added, the transglutaminase activity was significantly increased to the same level or higher than that of the positive control.
Furthermore, transglutaminase activity is related to maintenance of skin barrier function, maintenance or improvement of moisturizing function, and prevention or improvement of rough skin, and activation of transglutaminase prevents keratinization failure of the epidermis and moisturizing function of skin. Improvement and prevention or improvement of rough skin are possible. Therefore, an extract of a fox moth having a transglutaminase activating effect can be used as an active ingredient of an epidermis keratinization improving agent, a skin moisturizing function improving agent, a skin barrier function improving agent, and a rough skin preventing or improving agent.

Test Example 2 Verification of ceramide production promotion effect Normal human epidermal keratinocytes (trade name: NHEK (F), manufactured by KURABO) in a culture solution (trade name: EpiLife-KG2, manufactured by KURABO) using a culture plate ) At 37 ° C. and 5% CO ₂ .
Thereafter, the culture solution was changed to EpiLife-KG2 from which growth factors such as epidermal growth factor were removed, and the foxtail extract prepared in Preparation Example 2 was adjusted so that the concentration was 1 w / v% in terms of solid content. , 0.01%, 0.05%, or 0.1%. Also, instead of the foxtail extract, a 50% ethanol by volume as a control at a final concentration of 0.1% v / v and a Eucalyptus globulus extract prepared as shown below as a positive control. The final concentrations shown in Table 2 were added respectively. Eucalyptus extract is known to have an effect of promoting ceramide production and was used as a positive control.
After culturing for 3 days, each cell was collected together with 1 well.

The organic phase from which the lipid was extracted from the collected cells by the Bligh and Dyer method was transferred to a glass tube, solidified with nitrogen, and then redissolved with chloroform and methanol to obtain a lipid sample.
In addition, 0.1N NaOH, 1% SDS aqueous solution is added to the cells after lipid extraction, and the protein is solubilized by heating at 60 ° C. for 2 hours, cooled to room temperature, and then neutralized by adding 2N HCl. The amount of protein was quantified by the BCA method.

The prepared lipid sample was horizontally developed by thin film chromatography (TLC) twice with chloroform: methanol: acetic acid = 190: 9: 1. The copper sulfate solution was sprayed and sprayed with a hot plate to detect ceramide, and the amount was determined. The amount of ceramide was expressed as a relative value, assuming that the amount of ceramide when ethanol of 50% by volume was added was 1.
The results are shown in Table 2.

(Reference example)
40 g of Eucalyptus globules Labillardiere (manufactured by Shinwa Bussan) was cut into small pieces , 400 mL of 50% by volume ethanol was added, and extraction was performed for 7 days under room temperature and standing conditions. Then, it filtered and 291 mL of eucalyptus extracts were obtained. When the evaporation residue was calculated for the obtained extract, the evaporation residue was 3.16% (w / v). This was diluted with a 50% by volume aqueous ethanol solution to prepare a 1.0% (w / v) extract.

As is apparent from Table 2, the ceramide yield was increased in the system added with the foxgull extract compared to the control system. Therefore, it turns out that the ceramide production promoter of this invention which uses a foxglove extract as an active ingredient can promote ceramide production.
Furthermore, ceramide is related to maintaining or improving the skin barrier function, moisturizing function, and preventing or improving rough skin, and promotes the production of ceramide, thereby preventing keratinization failure of the epidermis and improving skin moisturizing function. , And prevention or improvement of rough skin are possible. Therefore, an extract of foxtail having an effect of promoting ceramide production can be used as an active ingredient of an epidermis keratinization improving agent, a skin moisturizing function improving agent, a skin barrier function improving agent, and a rough skin preventing or improving agent.

Test example 3
About 5 g of the sunflower (Lot. SB-3436) obtained from Affinity Products Co., Ltd. was soaked in lukewarm water, and the plant was identified by a specialist under a microscope. As a result, flower spikes were contained, the sepals were thin, had a white transparent border, and the presence of long hair was confirmed. From these characteristics, Lot. The plant obtained as SB-3436 was identified to be a foxtail but not a sunflower.

  Lot. 600 g of the foxtail obtained as SB-3436 was chopped, 6000 mL of a 99.5% by volume aqueous ethanol solution was added, and extraction was performed for 7 days under room temperature and standing conditions. Thereafter, filtration was performed to obtain 4504 mL of a foxglove extract. After 2,000 mL of hexane was added to 4000 mL of the resulting extract and stirred, 6000 mL of water was further added. After liquid-liquid partitioning, 8367 mL of the lower layer was extracted. 1000 mL of 1,3-butylene glycol was added to 2500 mL of the obtained extract and concentrated with an evaporator to remove ethanol and water. After adding 1500 mL of water to this, it was starched at 5 ° C. for 4 days. After removing insolubles by filtration, 40 volume% 1,3-butylene glycol aqueous solution was added to 2403 L of the obtained filtrate to prepare 3266 mL of an extract. When the evaporation residue was calculated for the obtained extract, the evaporation residue was 0.02% (w / v).

  To each of the left and right cheeks and lower leg outer parts of 10 healthy men and women, the lotion lotion or placebo lotion having the formulation shown in Table 3 was applied twice a day for 4 consecutive weeks. After applying lotion for 4 weeks, application of these lotions was stopped.

  Moisture content and skin surface roughness of cheek and crus outer side before start of applying lotion (0W), after continuous application of lotion for 4 weeks (4W), and 1 week after stop of application of lotion (5W) Was measured by the following method to evaluate skin properties.

(Measurement of stratum corneum moisture content)
On the day of measurement, the measurement site was washed and acclimated for 20 minutes in an environment set to 20 ° C. and humidity 40%.
Using Corneometer CM825MP (trade name, Courage + Khazaka electronic GmbH), capacitance was measured 5 times per site, and the water content of the stratum corneum was evaluated using the average value of 5 times as the capacitance value of the measurement site of each subject. . The results are shown in Table 4 (cheek part) and Table 5 (crus outer part). In addition, when the capacitance value before the start of applying lotion (0W) is 0, the amount of change in the capacitance value after 4 weeks of continuous application of lotion (4W) and after 1 week of application of lotion (5W) The average value of 10 subjects was taken as the Δ capacitance value at that site.

(Measurement of skin surface roughness)
On the day of measurement, the measurement site was washed and acclimated for 20 minutes in an environment set to 20 ° C. and humidity 40%.
Take two images per part using Visioscan VC98 (trade name, Courage + Khazaka electronic GmbH), calculate SELS parameter (SEr) per fixed area with the attached analysis software, and calculate the average value of the two images. The skin surface roughness was evaluated as the SEr value of the measurement site of the subject. The results are shown in Table 6 (cheek part) and Table 7 (crus outer part). In addition, when the SELS parameter before the start of applying lotion (0W) is set to 0, the amount of change in the SELS parameter after 4 weeks of continuous application of lotion (4W) and 1 week after application of lotion (5W) The average value of 10 subjects was defined as ΔSEr of the site.

  As is clear from Table 4, the amount of change from the initial value (0 W) of the stratum corneum water content represented by the Δ capacitance value at the time of 5 W in the cheeks is greater than that of the placebo lotion application part. The lotion application part was larger. That is, the results in Table 4 indicate that the foxroot extract has an effect of increasing the stratum corneum moisture content.

  As is apparent from Table 5, in the outer part of the lower leg, the amount of change from the initial value of the stratum corneum water content at the time of 4W and 5W, the foxtail extract extract-containing lotion application part rather than the placebo lotion application part. It can be seen that the amount of water decrease is smaller. That is, the results in Table 5 indicate that the foxroot extract has an action of suppressing a decrease in stratum corneum moisture content.

  As is apparent from Table 6, the skin surface roughness of the placebo lotion application part is large in the cheek part from the value of the amount of change from the initial value of the skin surface roughness indicated by ΔSEr at the time of 4W and 5W. On the other hand, it can be seen that the roughness of the skin surface is reduced in the application area of the lotion applied with the foxroot extract. That is, the results in Table 6 indicate that the foxglove extract has an action of preventing or improving rough skin in the cheek.

  As is clear from Table 7, the skin surface roughness of the placebo lotion application part is determined from the value of the amount of change from the initial value of the skin surface roughness indicated by ΔSEr at the time of 4 W and 5 W at the outer side of the lower leg. On the other hand, it can be seen that the occurrence of roughness of the skin surface is suppressed in the application area of the lotion application mixture containing the foxroot extract. That is, the results in Table 7 indicate that the foxglove extract has an effect of preventing or improving rough skin on the outer side of the lower leg.

  As described above, the foxgull extract has an action of increasing the stratum corneum moisture content, an action of suppressing a decrease in the stratum corneum moisture content, and an action of preventing or improving rough skin. Therefore, the extract of foxtail is used as an active ingredient in an epidermis keratinization improving agent, a skin moisturizing function improving agent, a skin barrier function improving agent, a stratum corneum moisture content increasing agent, a stratum corneum moisture content decreasing inhibitor, and a rough skin preventing or improving agent. It can be.

Claims (8)

A transglutaminase activator comprising as an active ingredient an extract of foxtail ( Justicia procumbens ). A ceramide production promoter comprising as an active ingredient an extract of foxtail ( Justicia procumbens ). An epidermis keratinization improver comprising as an active ingredient an extract of foxtail ( Justicia procumbens ). A skin moisturizing function improver comprising as an active ingredient an extract of foxtail ( Justicia procumbens ). A skin barrier function improving agent comprising an extract of foxtail ( Justicia procumbens ) as an active ingredient. A stratum corneum moisture-increasing agent comprising an extract of foxtail ( Justicia procumbens ) as an active ingredient. A stratum corneum water content lowering inhibitor comprising, as an active ingredient, an extract of foxtail ( Justicia procumbens ). An agent for preventing or improving rough skin, comprising an extract of foxtail ( Justicia procumbens ) as an active ingredient.