WO2016104665A1 - Agent for inhibiting or improving decrease of loricrin, transglutaminase-3 activator, and agent for promoting formation of skin horny cell layer cornified envelope or for strengthening structure of skin horny cell layer cornified envelope - Google Patents

Abstract

The present invention relates to: an agent for inhibiting or improving the decrease of loricrin in a cornified envelope of a horny cell layer of the skin, a transglutaminase-3 activator, and an agent for promoting the formation of a cornified envelope of a horny cell layer of the skin or for strengthening the structure of a cornified envelope of a horny cell layer of the skin, each of which contains sphingomyelin as an active ingredient; a composition; and a food or beverage. According to the present invention, it becomes possible to inhibit or improve the decrease of loricrin in a cornified envelope of a horny cell layer of the skin, and it also becomes possible to activate transglutaminase-3 in a cornified envelope of a horny cell layer of the skin. Therefore, it becomes possible to promote the formation of a cornified envelope of a horny cell layer of the skin and strengthen the structure of a cornified envelope of a horny cell layer of the skin. Due to these effects, the deterioration of a barrier function of skin can be prevented, inhibited or improved in a safe, simple and economically advantageous manner.

Description

Loriculin reduction inhibitor / improving agent, transglutaminase 3 activator and skin keratokeratosis thickening film formation promoter / structure strengthening agent Reference to related applications

This application is based on Japanese Patent Application No. 2014-264578 (filing date: December 26, 2014), which is a prior Japanese patent application, and claims the benefit of its priority. The entirety is hereby incorporated by reference.

The present invention relates to an agent for inhibiting or improving the reduction of loricrin in the keratinized thickened skin layer and an activator of transglutaminase 3 in the keratinized thickened skin layer. The present invention also relates to an agent for promoting the formation of a stratum corneum thickening film and a structure reinforcing agent. They are useful for preventing, suppressing or improving the deterioration of the skin condition.

The stratum corneum of the epidermis is composed of stratum corneum cells in which epidermal keratinocytes (keratinocytes) are terminally differentiated, and stratum corneum intercellular lipids surrounding them. The intercellular lipid is composed of ceramide, cholesterol, fatty acid and the like, and a lamellar structure is formed by the lipid layer and the water molecule layer.

On the other hand, stratum corneum cells contain natural moisturizing factors such as keratin fibers and amino acids that are components related to moisturizing. It is wrapped in a very tough, insoluble backing structure by cross-linking of various proteins called keratinized thickened membranes (also called keratinous thickened membranes, or cornified envelopes (CE)). It is very stable against various stimuli. Of the proteins constituting the keratinized thickened membrane, loricrin, which is formed in particular in the granular layer, plays a role of strengthening this backing structure. In this structure, transglutaminase 3 of protein cross-linking adhesion enzyme attaches loricrin to immature keratinized thickened membrane formed by cross-linking reaction of proteins other than loricrin, and serves as a scaffold for keratinized thickened membrane. Complete by forming.

The damage (damage) of the skin due to ultraviolet rays induces an abnormal turnover of the skin, and is particularly susceptible to the outermost stratum corneum. It is known that when an incomplete keratinized thickening film is formed due to corneal cell damage or parakeratosis, the barrier function of the epidermal horny layer is reduced and the water retained in the horny layer is lost. Yes.

From the viewpoint of promoting the formation and maturation of a cornified envelope (keratinized thickened film), for example, JP 2008-303185 discloses the use of a specific pyrrolidone derivative. Here, it is considered to mature an immature conified envelope. However, as far as the present inventors know, it is possible to promote the formation of a keratinized thickening film and strengthen its structure by suppressing the decrease of loricrin due to skin damage and activating transglutaminase 3. Is not well studied. And as far as the present inventors know, it has not been known so far to take orally sphingolipids, especially sphingomyelin.

In JP 2009-084244 A, continuous oral intake of a saturated free fatty acid solution having 10 to 36 carbon atoms has an effect of improving the skin barrier function, an effect of increasing the amount of ceramide, and an effect of increasing the expression of loricrin gene. It is disclosed. However, there is no description or suggestion regarding oral intake of sphingomyelin.

Japanese Patent Application Laid-Open No. 2007-117793 discloses pharmaceuticals and foods and drinks containing sphingomyelin as an active ingredient, and various actions such as sialomucin secretion promoting action, antiallergic action and antioxidant action by sphingomyelin are disclosed. Has been. However, there is no description or suggestion about the effect of suppressing the decrease of loricrin by sphingomyelin and the activation effect of transglutaminase 3.

Hasegawa et al. (Hasegawa T et al., “Dietary glucosylceramide enhances cornified envelope formation via transglutaminase expression and involucrin production”, Lipids (2011) 46, pp.529-535) It has been reported that effects of improving the skin barrier function of mice, increasing the expression of transglutaminase 1, increasing the production of involucrin, promoting the expression of transglutaminase 1 and 3, etc. have been observed. However, it has been confirmed that glucosylceramide is structurally different from sphingomyelin and has a completely different tendency of action from the administration of sphingomyelin. That is, in the case of administration of sphingomyelin, maintenance or reduction of transglutaminase 1 and involucrin was actually confirmed (Example of the present invention), and this was a result completely opposite to the case of administration of glucosylceramide. It is.

In this way, by taking sphingomyelin orally, the decrease in loricrin is suppressed, transglutaminase 3 is activated, thereby promoting the formation of keratinized thickening film and strengthening its structure Is not known so far as the present inventors know.

The present inventors have unexpectedly found that orally taking sphingolipids, particularly sphingomyelin, suppresses the decrease of loricrin and activates transglutaminase 3. The present inventors suppress the decrease of loricrin and activate transglutaminase 3 to promote the formation of a keratinized thickened film and to strengthen the structure of an immature keratinized thickened film. Pay attention. Furthermore, it has also been found that the deteriorated state of the skin can be improved by these, and among them, it is possible to suppress a decrease in the barrier function of the skin under ultraviolet irradiation. The present invention is based on these findings.

Therefore, an object of the present invention is to provide an agent for suppressing or improving loricrin reduction in the skin keratinized thickened film and an activator of transglutaminase 3 in the skin keratinized thickened film. Another object of the present invention is to provide an agent for promoting the formation of a stratum corneum thickening film and for enhancing the structure. Another object of the present invention is to provide a composition and a food and drink for safely, easily and economically preventing, suppressing or improving the decrease in the barrier function of the skin due to ultraviolet irradiation.

That is, according to the present invention, the following inventions are provided.

<1> An agent for inhibiting or improving the reduction of loricrin in the keratinized and thickened skin keratin layer, comprising sphingomyelin as an active ingredient.

<2> An activator of transglutaminase 3 in the skin keratinized thickened membrane, containing sphingomyelin as an active ingredient.

<3> An agent for promoting the formation of a stratum corneum keratinized thickening film and for enhancing its structure, comprising sphingomyelin as an active ingredient.

<4> In the above-mentioned <1> keratinized thickening film formation promoting / structural strengthening agent, the formation promotion and structural strengthening of keratinized thickening film suppresses / improves the reduction of loricrin in the keratinized keratinized thickening film by sphingomyelin. And activation of transglutaminase 3.

<5> In the reduction suppressing / improving agent, activator or formation promoting / structural strengthening agent according to any one of <1> to <4>, the sphingomyelin may be a milk-derived sphingomyelin.

<6> The agent for suppressing or improving loliculin decrease according to <1> or <5>, the activator of transglutaminase 3 according to <2> or <5>, or any one of <3> to <5> A composition for oral ingestion or enteral administration, comprising an agent for promoting the formation of a keratinized thickening film and for enhancing the structure.

<7> The composition according to <6> may be used for preventing, suppressing, or improving deterioration of skin condition.

<8> In the composition according to <7>, the deterioration of the skin condition may be a decrease in the barrier function of the skin.

<9> In the composition according to <7> or <8>, the decrease in the barrier function of the skin may be due to irradiation with ultraviolet rays.

<10> The composition according to any one of the above items <6> to <9>, preferably containing 0.5 to 1500 mg of sphingomyelin per day.

<11> The agent for suppressing or improving loliculin decrease according to <1> or <5>, the activator of transglutaminase 3 according to <2> or <5>, or any one of <3> to <5> A food / beverage product comprising an agent for promoting the formation of a keratinized thickening film and for enhancing the structure.

<12> A food or drink comprising the composition according to any one of <6> to <9>.

<13> The food or drink according to <11> or <12> preferably includes 0.2 to 45000 mg of sphingomyelin per package.

<14> The food or drink according to any one of <11> to <13> may be a functional food, a health nutrition food, a supplement, a food for specified health use, a functional display food, or a food with a disease risk reduction display. .

<15> A method for inhibiting or improving the reduction of loricrin, comprising orally ingesting or enterally administering sphingomyelin to a subject having a reduced amount of loricrin in the keratinized thickened skin skin.

<16> Transglutaminase in a keratinized keratinized thickened skin layer, comprising orally ingesting or enterally administering sphingomyelin to a subject having reduced activity of transglutaminase 3 in a keratinized thickened skin skin 3. Activation method of 3.

<17> Sphingomyelin is orally ingested or administered enterally to a subject who wants to promote the formation and strengthening of the keratinized thick film in the horny layer of the skin. A method for promoting the formation and strengthening of the structure of the keratinized keratinized thickened skin layer, comprising suppressing or improving the decrease and activating transglutaminase 3.

According to the present invention, it is possible to suppress / improve the decrease of loricrin in the skin keratinized keratinized thickened film, and to activate transglutaminase 3 in the keratinized keratinized thickened film. By suppressing the decrease of loricrin and activating transglutaminase 3, the formation of a keratinized thickened film is promoted, and the structure of the immature keratinized thickened film is strengthened. In particular, the reduction-suppressing / improving agent, activator or formation-promoting / structural-reinforcing agent, and composition of the present invention effectively prevent, inhibit or improve the decrease in skin barrier function under ultraviolet irradiation. Can do. Furthermore, since the production promoter and composition of the present invention are taken by the oral or enteral route, and the raw materials are derived from those that have been used as conventional foods, they are safe and simple, and economical. It is also excellent in properties.

It is a conceptual diagram which shows the flow of a test of an Example. It is the graph which showed the transition of the expression level of the loricrin gene in an Example, and the transition of the expression level of the transglutaminase 3 gene. Among these, FIG. 2 (A) shows the transition of the expression level of the loricrin gene, and FIG. 2 (B) shows the transition of the expression level of the transglutaminase 3 gene. In the figure, * indicates that there is a significant difference with respect to the control group (P <0.05), and # indicates that there is a significant difference with respect to day 0 (P <0.05). It is the graph which showed transition of the expression level of the transglutaminase 1 gene in an Example, and transition of the expression level of an involucrin gene. Among these, FIG. 3 (C) shows the transition of the expression level of the transglutaminase 1 gene, and FIG. 3 (D) shows the transition of the expression level of the involucrin gene. In the figure, * indicates that there is a significant difference with respect to the control group (P <0.05), and # indicates that there is a significant difference with respect to day 0 (P <0.05).

Hereinafter, embodiments of the present invention will be described.

Agent for suppressing or improving the reduction of loricrin in the keratinized thickened skin layer As described above, the agent for suppressing or improving the reduction of loricrin in the keratinized thickened skin layer of the present invention comprises sphingomyelin as an active ingredient. .
Here, the term “active ingredient” means that the agent for suppressing or improving the decrease according to the present invention exerts the action of suppressing or improving the decrease of loricrin in the keratinized thickened skin layer in the living body. It means to contain a sufficient amount (ie, effective amount) of sphingomyelin.

In addition, here, “inhibition / improvement of decrease in loricrin” means to suppress or improve the decrease of loricrin in the keratinized thickened skin layer, and to suppress or delay the decrease. It may include any meaning of stopping and maintaining the decrease, and suppressing the decrease and starting to increase. Furthermore, prevention of reduction may be included in “inhibition / improvement of reduction of loricrin”.

“Loriculin” is a major constituent protein of the lining structure of the keratinized thickening membrane of the skin. For example, as described in the examples described later, the expression of the loricrin gene is used as an index, and the horny layer angle of the skin The reduced state of loricrin in the thickened membrane can be evaluated.

Activator of transglutaminase 3 As described above, the activator of transglutaminase 3 in the skin keratinized thickened membrane of the present invention comprises sphingomyelin as an active ingredient.
Here, the term “active ingredient” means that the activator according to the present invention is used in an amount sufficient to exert the action of activating transglutaminase 3 in the skin keratinized thickened membrane in the living body (that is, , An effective amount) of sphingomyelin.

In addition, here, “activation of transglutaminase 3” means that the enzyme activity of transglutaminase 3 in the skin keratinized thickened membrane is activated and the activity is increased from the state in which the activity has decreased, the activity Is used to encompass maintaining a high state.

“Transglutaminase 3” is an enzyme having a function of attaching loricrin to a keratinized thickened membrane and strengthening the backing structure of the keratinized thickened membrane. For example, as described in Examples described later, transglutaminase 3 Using the expression state of the 3 genes as an index, the activity of transglutaminase 3 in the skin keratinized thickened membrane can be evaluated.

Formation promotion agent / structural strengthening agent for skin keratinized thickening film As described above, the formation promotion / structure strengthening agent for skin keratinized thickening film of the present invention comprises sphingomyelin as an active ingredient.
Here, the term “active ingredient” means that the formation promoting / structural strengthening agent according to the present invention can promote the formation of the keratinized and thickened skin keratinized layer and / or the effect of strengthening the structure. And a sufficient use amount (ie, an effective amount) of sphingomyelin.

In addition, here, “promoting formation / strengthening of skin keratinized thickened film” means promoting the formation of skin keratinized thickened film, maturing immature keratinized thickened film, Used to include strengthening the structure of keratinized thick films.

According to a preferred embodiment of the present invention, in the formation promoting agent / structural strengthening agent for keratinized thickening film, the formation promotion / structural strengthening of keratinized thickening film is to suppress the reduction of loricrin in the keratinized thickening skin skin by sphingomyelin. This is due to the improvement and activation of transglutaminase 3.

Sphingomyelin In the present specification, sphingomyelin is one of sphingolipids. Here, the sphingolipid is naturally derived, for example, milk, goat milk, sheep milk, horse milk and other milk-derived, egg yolk-derived, rice, corn, cereal-derived, konjac-derived, Examples include beet-derived, preferably milk-derived, and more preferably milk-derived. Therefore, the sphingomyelin that can be used in the present invention is of natural origin, preferably of milk, and more preferably of milk, similar to the sphingolipid described above. Sphingomyelin can be prepared from natural raw materials by a conventional method, but a commercially available product may be used.

According to a preferred embodiment of the present invention, the sphingomyelin is milk-derived sphingomyelin.

The milk-derived sphingomyelin that can be used in the present invention is preferably myrylstilsphingosylphosphorylcholine (d18: 1-C14: 0), tricosanylsphingosylphosphorylcholine (d18: 1-C23: 0), palmitylsphingo. Sylphosphorylcholine (d18: 1-C16: 0), tricosenoylsphingosylphosphorylcholine (d18: 1-C23: 1), stearylsphingosylphosphorylcholine (d18: 1-C18: 0), lignocerylsphingosylphosphorylcholine (d18: 1-C24: 0), arachidylsphingosylphosphorylcholine (d18: 1-C20: 0), nerbonylsphingosylphosphorylcholine (d18: 1-C24: 1), behenylsphingosylphosphorylcholine (d18: 1-C22: 0) And one or more molecular species selected from the group consisting of serotilsphingosylphosphorylcholine (d18: 1-C26: 0). More preferably, the milk-derived sphingomyelin contains three or more molecular species selected from the above group, and even more preferably contains five or more molecular species selected from the above group. .

The reduction-suppressing / improving agent, activator or formation promoting / structural strengthening agent of the present invention is preferably for oral intake (ie, oral intake agent) or enteral administration (ie, enteral administration agent).

The reduction-suppressing / improving agent, activator, or formation-accelerating / structural-strengthening agent of the present invention is preferably used for skin with deteriorated condition or skin with reduced barrier function.

As shown in the examples described below, according to the present invention, in the reduced state of the skin barrier function, such as the reduction of loricrin in the keratinized thickening film of the skin, by suppressing and improving the reduction of loricrin, The barrier function can be maintained or improved. Similarly, as shown in the examples described later, according to the present invention, transglutaminase 3 is reduced in a state where the skin barrier function is reduced such that the activity of transglutaminase 3 in the keratinized thick film of skin is reduced. By activating, skin barrier function can be maintained or improved. Furthermore, the suppression and improvement of loricrin in the stratum corneum thickened skin and the activation of transglutaminase 3 promote the formation of the keratinized thickened membrane and strengthen the structure. Can improve the reduced skin.

For this reason, the reduction-suppressing / improving agent, activator or formation-accelerating / structural strengthening agent according to the present invention is more preferably used for skin with deteriorated condition or skin with reduced barrier function.

Use Sphingomyelin, an active ingredient in the present invention, can suppress and improve the decrease of loricrin in the keratinized thickened skin skin, and also activates transglutaminase 3 in the keratinized thickened skin skin (Examples described later). Thereby, formation of a keratinized thick film can be accelerated | stimulated and the structure of the keratinized thick film in an immature state can be strengthened. Furthermore, sphingomyelin, which is an active ingredient in the present invention, has an effect of preventing, suppressing or improving deterioration of the skin condition.

Here, the deterioration of the skin condition includes, for example, a decrease in the barrier function of the skin, a drying of the skin, a skin roughness, a decrease in the amount of moisture in the stratum corneum, and atopic dermatitis. Reduction of the barrier function of the skin includes maintenance of the barrier function. Further, the reduction in the skin barrier function is preferably a reduction in the skin barrier function under the irradiation of ultraviolet rays.

Also, here, “prevention, suppression or improvement” of an exacerbated state is used in a sense encompassing adjustment, delay of progression, mitigation, prevention of onset, prevention of recurrence, etc. of such a state.

According to another aspect of the present invention, treatment, prevention, or improvement of a condition that can be treated, prevented, or improved can be achieved by suppressing or improving the decrease of loricrin or activating transglutaminase 3.

According to another preferred embodiment of the present invention, the sphingomyelin is orally ingested or enterally administered to a subject having a reduced amount of loricrin in the keratinized thickened skin keratoplasty. Methods for preventing, suppressing or ameliorating the worsening of the condition are provided.

Further, according to another preferred embodiment of the present invention, sphingomyelin is orally ingested or enterally administered to a subject whose transglutaminase 3 activity has decreased in the keratinized and thickened skin corneum. A method for preventing, suppressing or improving the deterioration of the skin condition is provided.

According to still another preferred embodiment of the present invention, sphingomyelin is orally ingested or enterally administered to a subject who wants to promote the formation of a keratinized thick film in the stratum corneum and to strengthen the structure. Deteriorating skin condition, including suppressing or improving loricrin in the keratinized thickened skin layer and activating transglutaminase 3 to promote the formation and strengthen the structure of the keratinized thickened skin layer Methods for preventing, suppressing or ameliorating are provided.

According to another aspect of the present invention, there is provided sphingomyelin for suppressing or improving the reduction of loricrin in the keratinized and thickened skin corneum.

According to another aspect of the present invention, there is provided sphingomyelin for activating transglutaminase 3 in the keratinized and keratinized thick film of skin.

According to another aspect of the present invention, a sphingomyelin is provided for promoting the formation of a keratinized thickened skin layer and / or strengthening its structure.

According to another preferred embodiment of the present invention, there is provided use of sphingomyelin for suppressing or improving the reduction of loricrin in the keratinized thickened skin skin layer. According to yet another preferred embodiment of the present invention, use of sphingomyelin in the manufacture of a composition for oral ingestion or enteral administration for suppressing or improving the reduction of loricrin in the keratinized thickened skin skin layer Is provided. Here, the use can be a non-therapeutic use.

According to another preferred embodiment of the present invention, there is provided the use of sphingomyelin for the activation of transglutaminase 3 in the keratinized keratinous thickened membrane. According to yet another preferred embodiment of the present invention, use of sphingomyelin in the manufacture of a composition for oral ingestion or enteral administration for the activation of transglutaminase 3 in a keratinized thickened skin skin layer Is provided. Here, the use can be a non-therapeutic use.

According to another preferred embodiment of the present invention, there is provided the use of sphingomyelin to promote the formation of and / or strengthen the structure of the keratinized keratinized thick film. According to yet another preferred embodiment of the present invention, a composition pharmaceutical for oral ingestion or enteral administration for promoting the formation of a keratinized thickened skin layer and / or strengthening its structure Use of sphingomyelin in the manufacture of Here, the use can be a non-therapeutic use.

Composition and Food / Beverage As described above, according to the present invention, the composition for oral intake or enteral administration, comprising the reduction inhibitor / ameliorator, activator or formation promoter / structure enhancer of the present invention. Is provided. Moreover, according to this invention, the food / beverage products which comprise the reduction | decrease suppression and improvement agent of this invention, an activator, or a formation promotion and structure strengthening agent are provided.

Such compositions and foods and drinks include, for example, adding the reduction suppressing / improving agent, activator or formation promoting / structural strengthening agent of the present invention according to the present invention to the composition and material components of the food and drinks. It can manufacture with the manufacturing method which consists of these.

The composition for oral intake or enteral administration of the present invention is preferably used for prevention, suppression or improvement of skin condition deterioration. That is, according to the present invention, a composition for preventing, suppressing or improving deterioration of the skin condition is provided.

The composition for preventing, suppressing or improving deterioration of the skin condition of the present invention is a pharmaceutical composition as one preferred embodiment.

In the present invention, the pharmaceutical composition is prepared as an oral preparation or a parenteral preparation according to a conventional method using additives that are acceptable for formulation. From the viewpoint of simplicity, an oral preparation is preferable. For oral preparations, take the form of solid preparations such as tablets, powders, fine granules, granules, capsules, pills, sustained-release preparations, and liquid preparations such as solutions, suspensions, and emulsions. Can do. Additives that are acceptable for formulation include, for example, excipients, stabilizers, preservatives, wetting agents, emulsifiers, lubricants, sweeteners, colorants, fragrances, buffers, antioxidants, pH Examples thereof include regulators.

Arbitrary ingredients can be added to the food and drink of the present invention as required. As such optional components that can be added, there are no particular restrictions, but usually ingredients to be blended in foods and drinks, sweeteners, acidulants, vegetable and fruit juices and their extracts, vitamins, minerals, Nutrients such as amino acids, useful microorganisms such as lactic acid bacteria, bifidobacteria and propionic acid bacteria and their cultures, functional sugars such as oligosaccharides, royal jelly, collagen, ceramide, glucosamine, astaxanthin and polyphenols A raw material, a fragrance | flavor, a pH adjuster, an excipient | filler, a sour agent, a coloring agent, an emulsifier, a preservative, etc. can be mix | blended.

In the present invention, the food and drink are other than the pharmaceutical composition and are not particularly limited as long as they are ingestible forms such as solutions, suspensions, emulsions, powders, and solid molded products. Specifically, for example, dairy products such as milk drinks, yogurts, lactic acid bacteria drinks, fermented milk, ice creams, creams, cheeses; soft drinks, fruit juice drinks, vegetable drinks, soy milk drinks, coffee drinks, tea drinks Jelly drinks, cocoa, smoothie powdered drinks and sports powdered drinks, nutrition-enriched powdered drinks, cosmetic powdered foods, powdered soup, steamed bread, concentrated drinks, alcoholic drinks, etc .; bread, pasta , Flour products such as noodles, cake mix, fried flour, bread crumbs; confectionery such as chocolate, gum, candy, cookies, gummi, snacks, Japanese confectionery, jelly, pudding, etc .; processed seasonings, flavor seasonings, Seasoning such as cooking mix; curry, busta sauce, potof, stew, Japanese food retort food; processed oil, butter, margarine, spray Oils and fats such as dough and mayonnaise; Instant foods such as freeze-dried foods; Agricultural processed products such as canned agricultural products, jams and marmalades, pickles, boiled beans, cereals, miscellaneous foods; processed marine products; processed livestock products; Frozen foods such as gratin, sugar beet, and fries; liquid foods, animal feeds, tablets, and cosmetics used in the oral cavity.

According to one preferred embodiment of the present invention, the food and drink are milk drinks (which may include processed milk), fermented milk, soft drinks, jelly drinks, tablets, cosmetic powdered foods, powdered drinks, liquid foods, liquids. According to a more preferred embodiment, the food or drink is a milk beverage, fermented milk, soft drink, jelly drink, tablet, powdered food for beauty, and according to a more preferred embodiment, the food or drink is Milk beverage (which may include processed milk), fermented milk.

In addition, functional foods, health nutrition foods, supplements, health foods, foods for specified health use, functional indication foods, nutritional functional foods, foods for the sick, infant formulas, milk powders for pregnant and lactating women Or foods and beverages with a disease risk reduction label are included. According to one preferred embodiment of the present invention, the food or drink is a functional food, a health nutrition food, a supplement, a food for specified health use, a functional display food, or a food with a disease risk reduction display.

Here, the indication of disease risk reduction is the indication of food and drink that may reduce the disease risk, and is based on the standards established by the FAO / WHO Joint Food Standards Committee (Codex Committee). , Or with reference to the standard, it can be a defined or recognized display.

Therefore, the food and drink of the present invention are, for example, foods suitable for consumers who expect improvement or alleviation of skin condition deterioration, foods suitable for improvement of skin condition deterioration, that is, so-called foods for specific health use or functions It can be provided as sex indication food.

According to another aspect of the present invention, a food or drink on which a function for preventing, suppressing, or improving deterioration of the skin state is displayed can be provided.

Since the content of the active ingredient in foods and drinks and compositions can be changed depending on the type and form of the composition, the purpose of prevention and improvement, etc., it is difficult to define uniformly, but in the composition of the present invention The sphingomyelin can be adjusted to an effective amount that can be ingested by an adult (weight: 60 kg) per day at 0.5 to 1500 mg, preferably 1 to 1000 mg, more preferably 5 to 500 mg.

The sphingomyelin content can be measured by a conventional method. For example, it can be measured by using liquid chromatography and using AQUASIL SP100 (4.6 × 250 mm, Senshu Kagaku) as a column.

At this time, as the mobile phase, for example, a solution in which 0.5 mM phosphate-citrate buffer (pH 3.0) and methanol are mixed at a ratio of 5 to 95 may be used. The measurement time can be set to 20 minutes, the flow rate of the mobile phase can be set to 0.6 mL / min, the column temperature can be set to 40 ° C., and the absorbance can be detected at 205 nm. As the standard substance, for example, sphingomyelin (derived from milk, manufactured by Nagara Science Co., Ltd.) can be used, and can be quantified by the area ratio.

In another aspect of the present invention, the content of active ingredients in foods and drinks and compositions can be specified per packaging form. For example, in the case of foods and drinks, the content of sphingomyelin is 5 to 1500 mg, preferably 6 In the case of the composition, the sphingomyelin content is 0.5 to 1500 mg, preferably 1 to 1000 mg, more preferably 5 to 500 mg. Can do. In addition, the amount (content) per packaging form is not limited to a single intake, and may include intakes for multiple doses or multiple days (for example, for 30 days). The sphingomyelin content is 5 to 45000 mg, preferably 6 to 30000 mg, more preferably 7 to 15000 mg. In the case of the composition, the sphingomyelin content is 0.5 to 45000 mg, preferably 1 It can be contained in an amount of ˜30000 mg, more preferably 5 to 15000 mg.

The conventional general milk drink and fermented milk contain 20 mg of phospholipid per gram of lipid, whereas the milk drink and fermented milk of the present invention contain 25 to 3000 mg of phospholipid per gram of lipid. , Preferably 30 to 2500 mg, more preferably 40 to 2000 mg, and still more preferably 50 to 1500 mg. In addition, the conventional general milk drink and fermented milk contain 6 mg of sphingomyelin per gram of lipid, while the milk drink and fermented milk of the present invention contain 7 to 1500 mg of sphingomyelin per gram of lipid. Contained, preferably 8 to 1250 mg, more preferably 9 to 1000 mg, and still more preferably 10 to 750 mg.

That is, in one preferred embodiment of the present invention, a milk drink or fermented milk with an intentionally increased sphingomyelin content can be used.

In the milk beverage and fermented milk of the present invention, lipid is contained in a normal amount (for example, lipid is contained at 3% by weight), sphingomyelin is contained at 0.2 to 60 mg / g, preferably 0.25 to 50 mg / g. g, more preferably 0.3 to 40 mg / g, and still more preferably 0.35 to 30 mg / g.

On the other hand, in the milk beverage and fermented milk of the present invention, when lipid is contained in a small amount (for example, low fat containing 1% by weight of lipid), sphingomyelin is contained at 0.05 to 60 mg / g, preferably 0. 0.1 to 50 mg / g, more preferably 0.15 to 40 mg / g, and still more preferably 0.2 to 30 mg / g.

In the milk beverage, fermented milk, soft drink, jelly drink, tablet, and cosmetic powder food of the present invention, when containing almost no lipid (non-fat), sphingomyelin is contained at 0.02 to 60 mg / g. Preferably, it can be contained at 0.04 to 50 mg / g, more preferably 0.06 to 40 mg / g, and still more preferably 0.08 to 30 mg / g.

Therefore, according to one more preferable aspect of the present invention, a milk drink or fermented milk containing additional sphingomyelin is provided, and a composition or food or drink containing such a milk drink or fermented milk is further provided. Is done. Examples of such a composition or food and drink include yogurt.

For example, in the case of yogurt, a yogurt containing additional sphingomyelin can be provided, which is not only a sphingomyelin that can be naturally contained in yogurt, but additionally adding sphingomyelin to yogurt, Yogurt that intentionally increases the concentration of sphingomyelin. In this case, the term “additional sphingomyelin” is not a sphingomyelin that can be naturally contained in yogurt, but a sphingo added separately when adding yogurt intentionally with increased sphingomyelin concentration. Myelin.

Therefore, according to another preferred embodiment of the present invention, the agent for suppressing or improving loricrin reduction according to the present invention, the activator of transglutaminase 3 according to the present invention, or the agent for promoting the formation of keratinized thickening film or the structure reinforcing agent according to the present invention. A yogurt comprising is provided.

According to another preferred embodiment of the present invention, there is provided a yogurt having an effect of suppressing or improving the reduction of loricrin, comprising additional sphingomyelin.
According to another preferred embodiment of the present invention, a yogurt having an activation effect on transglutaminase 3 is provided, which comprises additional sphingomyelin.
According to another preferred embodiment of the present invention, there is provided a yogurt containing an additional sphingomyelin and having an action of promoting the formation of a keratinized thick film and strengthening the structure.

According to another aspect of the present invention, sphingomyelin is orally administered or administered enterally to a subject having a reduced amount of loricrin in the keratinized thickened skin keratinized layer. An improvement method is provided.
Here, the amount of loricrin means the amount of loricrin present in the skin keratinized thickened membrane, and can be evaluated by, for example, the expression level of the loricrin gene.

According to another aspect of the present invention, the sphingomyelin is orally ingested or enterally administered to a subject having reduced activity of transglutaminase 3 in the keratinized thickened skin keratin layer. A method for activating transglutaminase 3 in a keratinized thickened membrane is provided.
Here, the activity of transglutaminase 3 can be evaluated by, for example, the expression level of the transglutaminase 3 gene.

According to another aspect of the present invention, the sphingomyelin is orally ingested or enterally administered to a subject where it is desired to promote the formation and strengthening of the keratinized thick film in the horny layer of the skin. There is provided a method for promoting the formation and strengthening of the keratinized keratinized thickened skin layer, comprising suppressing and improving the decrease of loricrin in the keratinized thickened membrane and activating transglutaminase 3.

Note that, preferably, the above method excludes medical use. The subject is preferably a human or a non-human mammal.

And the method preferably reduces loricrin for a subject with reduced amount of loricrin in the cutaneous keratinized thickened membrane, preferably continuously ingesting or enterally administering sphingomyelin for more than 2 days Inhibition / improvement method, method of activating transglutaminase 3 in skin keratinized thickened film for subjects with decreased activity of transglutaminase 3 in skin keratinized thickened film, or skin keratinized thickened film It can be a formation promotion / structure strengthening method.

Hereinafter, the present invention will be described in detail with reference to the following examples, but the present invention is not limited thereto.

Test: Effect of sphingomyelin on deterioration of skin barrier function under ultraviolet irradiation Hairless mice were irradiated with ultraviolet light (GL20SE, Sankyo Electric Co., Ltd.) to deteriorate the skin barrier function, and continuous oral administration of sphingomyelin. The effect of ingestion was evaluated.

<Experiment method>
(1) Deterioration of skin barrier function by ultraviolet irradiation Hairless mice (Hos: HR-1, female, 8 weeks old) were grouped, and one week later, under the condition of 20 mJ / cm ² , ultraviolet rays (UV -B) was irradiated.
In the test, individuals were required for each measurement and skin collection, so a total of 48 mice were used, and each group was divided into 8 mice.

(2) Group composition and test procedure Using the test system for UV-irradiated model animals prepared in (1) above, gene expression in the skin by oral intake (oral administration) of sphingomyelin (purity 98%, manufactured by Nagara Science) The impact on was evaluated.

Each group subjected to the test was subjected to the following conditions.
Oral administration was performed for 9 days from the day of grouping.
(i) Control day 0 group (ii) Control day 1 group (iii) Control day 2 group (iv) Sphingomyelin intake day 0 group (v) Sphingomyelin intake day 1 group (vi) Sphingomyelin intake Second day group

In the control 0, 1, and 2 day groups, normal feed was ingested, and a solvent (5% aqueous solution of ethanol) was orally administered at 10 mL / kg-BW / day. In addition, in the 0th, 1st and 2nd day groups of sphingomyelin intake, sphingomyelin (purity: 98%, Nagara Science): 146 mg / 10 mL-solvent (ethanol 5% aqueous solution) was orally administered at 10 mL / kg-BW / day did.

Specifically, in the test, the expression of loricrin gene and transglutaminase 3 gene was determined for a predetermined group before irradiation (day 0), on irradiation day 1, and on irradiation day 2, in accordance with the flow of FIG. The amount was quantified. At this time, the expression levels of the transglutaminase 1 gene and the involucrin gene were also quantified.

(Quantification method for expression levels of loricrin, transglutaminase 3, transglutaminase 1 and involucrin genes)
Total RNA was extracted from the skin sample collected from each individual using RNeasy Fibrous Tissue Mini kit (QIAGEN), and 1.5 μg equivalent of total RNA extracted from the skin was used for the RiverAid First Strand cDNA Synthesis Kit (Thermo ICIC TherM IC). ) Was used for complementary DNA (cDNA) synthesis and reverse transcription reaction.

The synthesized cDNA was subjected to RT-PCR, the primer was TaqmanGene Expression Assay (Applied Biosystems), and 7500 real-time PCR® System (Applied Biosystems) was used as the target gene for each sample, Loricrin (Lorrisrin). ID: Mm01962650_s1), transglutaminase 3 (Transglutaminase 3) (Assay ID: Mm00436999_m1), transglutaminase 1 (Transglutaminase 1) (Assay ID: Mm00498375_m1), mol 52 The NA expression level was measured.

Simultaneously, the mRNA expression level of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Assay ID: Mm9999999_g1) was measured as an internal standard.

Quantification was performed by the ΔΔCt method, and the ratio of the target gene expression level of each group to the target gene expression level of the control day 0 group was calculated as the relative expression level.

Specifically, the relative expression level was calculated using the following formula.

ΔCt = (Ct value of target gene of each individual sample) − (Ct value of GAPDH gene of each individual sample)

ΔΔCt = (ΔCt value of target gene of each individual sample) − (ΔCt average value of control day 0 group)

Gene expression level = 2−ΔΔCt

Relative expression level = (2-ΔΔCt of target gene of each individual sample) / (2-ΔΔCt average value of control day 0 group)

(3) Test results Changes in gene expression related to the promotion of keratinized thickening film formation and structural enhancement were evaluated based on the expression levels of loricrin gene and transglutaminase 3 gene.

Here, when the expression level of loricrin gene is high in the 0, 1, 2 day group of sphingomyelin intake compared to the control 0, 1, 2 day group, the decrease in loricrin expression is suppressed. means. Moreover, if the expression level of the transglutaminase 3 gene is higher in the 0, 1, 2 day group of the sphingomyelin intake compared to the control 0, 1, 2 day group, the increase in the expression of the transglutaminase 3 gene is increased. It means being promoted.

On the other hand, if the expression level of the transglutaminase 1 gene is lower in the 0, 1 and 2 day groups of sphingomyelin intake than the control 0, 1 and 2 day groups, This means that the increase in the expression of the transglutaminase 1 gene was suppressed. In addition, in the sphingomyelin intake 0, 1 and 2 day groups, when the involucrin gene expression level was low, the increase in involucrin gene expression was suppressed compared to the control 0, 1 and 2 day groups. Means that.

The results were as shown in FIGS.

As can be seen from FIG. 2 (A), the expression level of the loricrin gene was higher in the first day group of sphingomyelin intake than in the first day group of control (1 day after irradiation with ultraviolet rays).
As can be seen from FIG. 2 (B), compared to the control day 1 group (1 day after UV irradiation) and the control day 2 group (2 days after UV irradiation), the first day group ingested the sphingomyelin group and the sphingomyelin group In the ingestion day 2 group, the expression level of the transglutaminase 3 gene was high.

As can be seen from FIG. 3 (A), compared to the control 1, 2 and 3 day group (control group) (after UV irradiation 1, 2 and 3 days), in the sphingomyelin intake 1, 2 and 3 day group, In each case, the expression level of the transglutaminase 1 gene was low.
As can be seen from FIG. 3 (B), compared to the control 1, 2 and 3 day group (SM group) (after UV irradiation 1, 2 and 3 days), in the sphingomyelin intake 1, 2 and 3 day group, Each of the involucrin gene expression levels were low.

From these results, by taking sphingomyelin orally, the decrease in the expression of loricrin gene by ultraviolet irradiation is suppressed, the increase in the expression of transglutaminase 3 gene is promoted, and the formation of keratinized thickening film is promoted. It was suggested that the structure was strengthened.

Moreover, since the increase in the expression of transglutaminase 1 gene and involucrin gene was not promoted by ingestion of sphingomyelin, oral ingestion of sphingomyelin promotes the formation of keratinized thickening film and strengthens the structure. It was suggested that it contributes specifically to the promotion of expression of related specific genes (here, loricrin gene and transglutaminase 3 gene).

Claims (17)

1. An agent for suppressing or improving the reduction of loricrin in the keratinized thickened skin layer, comprising sphingomyelin as an active ingredient.
2. Activator of transglutaminase 3 in the keratinized and thickened skin keratin layer, containing sphingomyelin as an active ingredient.
3. An agent that promotes the formation of a stratum corneum keratinized thickening film and contains a sphingomyelin as an active ingredient.
4. The keratinized thickening according to claim 3, wherein the formation promotion and structural strengthening of the keratinized thickening film are due to the suppression and improvement of loricrin in the keratinized thickened skin layer by sphingomyelin and the activation of transglutaminase 3. Film formation promoter and structural strengthening agent.
5. The reduction inhibitor / improver, activator or formation promoter / structure enhancer according to any one of claims 1 to 4, wherein the sphingomyelin is milk-derived sphingomyelin.
6. The agent for suppressing or improving loricrin reduction according to claim 1 or 5, the activator of transglutaminase 3 according to claim 2 or 5, or the keratinized thickening film according to any one of claims 3 to 5. A composition for oral ingestion or enteral administration, comprising an agent for promoting the formation of and for enhancing the structure.
7. The composition according to claim 6, which is used for prevention, suppression or improvement of deterioration of skin condition.
8. The composition according to claim 7, wherein the deterioration of the skin condition is a decrease in the barrier function of the skin.
9. The composition according to claim 7 or 8, wherein the skin barrier function is reduced by irradiation with ultraviolet rays.
10. The composition according to any one of claims 6 to 9, comprising an amount of sphingomyelin that can be ingested from 0.5 to 1500 mg per day.
11. The agent for suppressing or improving loricrin reduction according to claim 1 or 5, the activator of transglutaminase 3 according to claim 2 or 5, or the keratinized thickening film according to any one of claims 3 to 5. Food / beverage products comprising a formation promoting / structural reinforcing agent.
12. A food or drink comprising the composition according to any one of claims 6 to 9.
13. The food or drink according to claim 11 or 12, comprising an amount of sphingomyelin in an amount of 0.2 to 45000 mg per package.
14. The food or drink according to any one of claims 11 to 13, which is a functional food, a health nutrition food, a supplement, a food for specified health use, a functional indication food or a food with a disease risk reduction indication.
15. A method for suppressing or improving the reduction of loricrin, comprising orally ingesting or enterally administering sphingomyelin to a subject with a reduced amount of loricrin in the keratinized thickened skin skin layer.
16. Activity of transglutaminase 3 in keratinized keratinized thickened skin, including ingestion or enteral administration of sphingomyelin to subjects with reduced activity of transglutaminase 3 in keratinized keratinized thickened skin Method.
17. Sphingomyelin is orally ingested or administered enterally to a subject who wants to promote the formation of stratum corneum in the skin stratum corneum and to strengthen the structure, thereby suppressing the decrease in loricrin in the stratum corneum thickening skin. A method for promoting the formation and strengthening of the skin keratinized keratinized thickened film, comprising improving and activating transglutaminase 3.
    
    

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