WO2016021573A1 - Agent for promoting production of ceramide covalently bonded to horny cell - Google Patents

Abstract

The present invention relates to an agent for promoting the production of ceramide covalently bonded to a horny cell, which contains a sphingolipid as an active ingredient. The present invention provides: a production promoter which can promote the production of ceramide covalently bonded to a horny cell, and which can safely, simply and economically prevent, inhibit or improve the deterioration of a barrier function of skin; a composition; and a food or beverage.

Description

Production promoter for keramide covalently bound ceramide Reference to related applications

This application is based on Japanese Patent Application No. 2014-158811 (filing date: August 4, 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 a production promoter for horny cell covalently bound ceramide. More specifically, the present invention relates to an agent for promoting the production of stratum corneum cell covalently bound ceramide containing a sphingophospholipid derived from milk. 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 lipids between the stratum corneum cells surrounding them. This 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. Thereby, the barrier function which prevents the penetration | invasion of the foreign material from the outside, and the transpiration | evaporation of the water | moisture content from a body will work | function.

The epidermal keratinocytes form each layer while keratinizing in the order of basement membrane cells, spinous cells, granule cells, and stratum corneum cells. A protein constituting a cornified envelope (エ ン ベ ロ ー プ CE) is synthesized from the upper spiny layer to the granular layer. In the process of reaching the stratum corneum, the enzyme transglutaminase binds substrate proteins such as involucrin, loricrin, and cystatin to the cell membrane of epidermal keratinocytes, thereby forming insolubilized CE. Ceramide and the like are covalently bonded to the insolubilized CE to form a bonded ceramide (horny layer cell covalently bonded ceramide). These form the basis of the stratum corneum barrier function.

The stratum corneum has a structure in which a long-chain sphingosine base has a fatty acid bonded to an acid amide, and can be classified into at least 12 types depending on the type of the sphingosine base and the fatty acid (J Lipid Res 2008, Vol. 49, 1466-1476). The stratum corneum cell covalent bond ceramide is a ceramide ceramide formed by covalently bonding omega hydroxy ceramide with CE, and forming a lamellar structure with a lipid layer and a water molecule layer in the stratum corneum. Of these, ceramide is particularly bound to the stratum corneum cell membrane.

It is known that the amount of ceramide in the stratum corneum is involved as an important factor in the skin barrier function. For this reason, for example, JP-A-9-208419 discloses the use of an external composition for skin containing mevalonic acid and ceramides for the purpose of improving rough skin, improving keratin, and beautifying skin. Conventionally, it has been studied to supply ceramide to the stratum corneum by applying ceramide directly to the skin.

However, despite the fact that the amount of ceramide in the stratum corneum is normal, a phenomenon in which the barrier function of the skin is lowered under ultraviolet irradiation was observed (for example, Examples of the present application). The present inventors considered that the amount of the horny layer covalently bound ceramide is important for maintaining the skin barrier function, not the amount of the whole horny layer ceramide. From the viewpoint of promoting the formation and maturation of the cornified envelope, for example, JP 2008-303185 A discloses the use of a specific pyrrolidone derivative. As far as the present inventors know, the stratum corneum cell covalently bound ceramide is disclosed. The improvement of the barrier function of the skin by the improvement is not so much progressed. And as far as the present inventors know, among them, it has not been known so far to take orally sphingolipids.

Japanese Patent Application Laid-Open No. 2005-281257 discloses a skin-beautifying agent containing sphingomyelin as an active ingredient. By taking sphingomyelin orally, cosmetic effects such as moisturizing skin, preventing skin roughness, preventing rough skin, and preventing wrinkles are disclosed. It is disclosed that it can be obtained. Similarly, the pamphlet of International Publication No. WO2008 / 093657A discloses a skin beautifying agent containing milk-derived phospholipid containing sphingomyelin as an active ingredient.

However, these documents also describe and suggest that oral intake of milk-derived sphingolipids can promote the production of keratinocyte covalent ceramide and thereby improve the reduction of skin barrier function. It has not been.

The present inventors have now unexpectedly found that the production of stratum corneum cell covalent ceramide can be promoted by orally ingesting sphingolipids, particularly sphingomyelin. And it has also been found that the deteriorated state of the skin can be improved by this, and in particular, the decrease in the barrier function of the skin under the irradiation of ultraviolet rays can be suppressed. The present invention is based on these findings.

Therefore, an object of the present invention is to provide a production promoter that promotes the production of stratum corneum covalently bound ceramide. Another object of the present invention is to provide a composition and a food and drink for preventing, suppressing or improving the reduction of the barrier function of the skin due to irradiation of ultraviolet rays, etc. safely, simply and economically. .

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

<1> A keratinocyte covalently bound ceramide production promoter comprising sphingolipid as an active ingredient.

<2> In the production promoter of <1>, the sphingolipid may be a milk-derived sphingophospholipid.

<3> In the production promoter of <1> or <2>, the sphingophospholipid may be sphingomyelin.

<4> In any one of the production promoters of the above <1> to <3>, the stratum corneum covalently bound ceramide is omega hydroxy d18: 1-C28: 0, d18: 1-C30: 1, d18: 1- C30: 0, d18: 1-C32: 0, d18: 1-C32: 1, d18: 1-C34: 0, d18: 1-C34: 1, d18: 1-C36: 1, d17: 1-C32: Includes one or more molecular species selected from the group consisting of 0, d17: 1-C32: 1, d17: 1-C34: 0, d17: 1-C34: 1, and d17: 1-C36: 1 Is good.

<5> The production promoter according to any one of <1> to <4> may be for oral intake or enteral administration.

<6> A composition for preventing, suppressing or improving deterioration of skin condition, comprising the active ingredient according to any one of <1> to <5>.

<7> In the composition according to <6>, the deterioration of the skin state may be a decrease in the barrier function of the skin.

<8> In the composition according to <7>, the decrease in the barrier function of the skin may be due to a decrease in production of horny layer covalently bound ceramide in the horny layer.

<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 sphingolipid per day.

<11> A food or drink comprising the keratinocyte covalently bound ceramide production promoter according to any one of <1> to <5>.

<12> A food or drink comprising a composition for preventing, suppressing or improving deterioration of the skin condition 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 sphingolipid per package.

<14> The food or drink according to any one of <11> to <13>, which is a functional food, a health nutrition food, a supplement, a functional display food, a food for specified health use, or a food with a display for reducing disease risk Is good.

<15> A yogurt comprising the keratinocyte covalently bound ceramide production promoter according to any one of <1> to <5>.

<16> A yoghurt that promotes the production of stratum corneum cell covalently bound ceramide, comprising an additional sphingolipid.

<17> The yogurt according to <15> or <16> preferably further comprises collagen.

<18> In the yogurt according to any one of <15> to <17>, the sphingophospholipid is preferably sphingomyelin.

<15B> In the food or drink according to any one of <11> to <14>, the food or drink is preferably yogurt.

<16B> Preferably, the food or drink according to any one of <11> to <14> and <15B> further includes collagen.

<17B> In the food or drink according to any one of <11> to <14>, <15B>, and <16B>, the sphingophospholipid is preferably sphingomyelin.

<19> A composition for oral ingestion or enteral administration, comprising the keratinocyte covalently bound ceramide production promoter of any one of <1> to <5> above and yogurt.

<20> A composition for ingestion or enteral administration, comprising additional sphingolipid and yogurt.

<21> Preferably, the above <19> or <20> further comprises collagen in the composition for oral intake or enteral administration.

<22> In the composition for oral intake or enteral administration according to any one of <19> to <21>, the sphingophospholipid is preferably sphingomyelin.

<23> In the composition for oral ingestion or enteral administration according to any one of the above items <19> to <22>, it is preferably for promoting production of keratinocyte covalently bound ceramide.

<24> Sphingolipid koji for promoting the production of stratum corneum cell covalent ceramide.

<25> A method for promoting the production of stratum corneum cell covalent ceramide, comprising orally administering sphingolipid to a subject whose production of stratum corneum cell covalent bond ceramide has been reduced or enterally.

<26> A method for preventing, suppressing or ameliorating deterioration of skin condition, comprising orally administering sphingolipid to a subject with reduced production of stratum corneum cell covalent ceramide or enterally administering it.

<27> Use of sphingolipids to promote production of stratum corneum cell covalent ceramide. Preferably, the use may be the use of a sphingolipid for the production of a composition for oral ingestion or enteral administration for promoting production of horny layer covalently bound ceramide. Also here, the use can be a non-therapeutic use.

<28> Use of sphingolipids for prevention, suppression or improvement of skin condition deterioration. Preferably, the use may be the use of sphingolipid for the production of a composition for oral intake or enteral administration for prevention, suppression or amelioration of deterioration of skin condition. Also here, the use can be a non-therapeutic use.

According to the production promoter of the present invention, it is possible to promote the production of stratum corneum cell covalent ceramide, and in particular, in the epidermis in a state in which the production of stratum corneum cell covalent bond ceramide is reduced, Production can be promoted. By promoting the production of stratum corneum covalently bound ceramide, it is possible to effectively prevent, suppress or improve the deterioration of the skin condition. In particular, the production promoter and composition of the present invention can effectively prevent, suppress or improve the deterioration of the skin barrier function under ultraviolet irradiation. 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 the graph which showed transition of the stratum corneum moisture content in Test 1 (there is a significant difference (p <0.05) between different alphanumeric characters). It is the graph which showed transition of the transdermal moisture transpiration amount in Test 1 (there is a significant difference (p <0.05) between different alphanumeric characters). It is the graph which showed transition of the amount of stratum corneum in the test 1 (there is a significant difference (p <0.05) between different alphanumeric characters). It is the graph which showed transition of the stratum corneum moisture content in Test 2 (* with significant difference (p <0.05)). It is the graph which showed transition of the amount of transdermal moisture transpiration in Test 2 (* with a significant difference (p <0.05)). It is the graph which showed transition of the stratum corneum cell covalent bond ceramide amount in Test 2 (* with a significant difference (p <0.05)). It is the graph which showed transition of the amount of stratum corneum in Test 2 (there is no significant difference). It is the graph which showed transition (for every molecular species) of the amount of stratum corneum in the test 3 (* with a significant difference (p <0.05)). It is the graph which showed transition (the total value of the value for every molecular species) of the stratum corneum amount in Test 3 (* with a significant difference (p <0.05)).

Hereinafter, embodiments of the present invention will be described.

Corneal Cell Covalent Bond Ceramide Production Promoter As described above, the stratum corneum cell covalent bond ceramide production promoter comprises sphingolipid as an active ingredient.

In the present specification, the horny layer covalently bound ceramide is a corneal ceramide in which omega hydroxy ceramide is covalently bonded to a cornified envelope (CE). Among the ceramides that form a lamellar structure by ceramide, ceramides that are bound to the horny layer cell membrane. The stratum corneum cell covalently bound ceramide is sometimes referred to as bound ceramide.

According to a preferred embodiment of the invention, the stratum corneum covalent ceramide is omega hydroxy d18: 1-C28: 0, d18: 1-C30: 1, d18: 1-C30: 0, d18: 1-C32: 0, d18: 1-C32: 1, d18: 1-C34: 0, d18: 1-C34: 1, d18: 1-C36: 1, d17: 1-C32: 0, d17: 1-C32: 1, d17: It comprises one or more molecular species selected from the group consisting of 1-C34: 0, d17: 1-C34: 1, and d17: 1-C36: 1. More preferably, the stratum corneum cell covalently bound ceramide contains three or more molecular species selected from the aforementioned group, and still more preferably contains five or more molecular species selected from the aforementioned group. It is a waste.

Sphingolipids Sphingolipids that can be used in the present invention are naturally derived, such as those derived from milk such as cow's milk, goat milk, sheep milk, and horse milk, those derived from egg yolk, rice, corn, and grains. , Konjac origin, beet origin and the like, preferably milk origin, more preferably milk origin. The sphingolipid is preferably a sphingophospholipid, more preferably a sphingomyelin. These can be prepared from natural raw materials by a conventional method, but commercially available products may be used.

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. .

According to a more preferred embodiment of the present invention, milk-derived sphingomyelin comprises tricosanylsphingosylphosphorylcholine (d18: 1-C23: 0), palmitylsphingosylphosphorylcholine (d18: 1-C16: 0), lignoceryl. 1 selected from the group consisting of sphingosylphosphorylcholine (d18: 1-C24: 0), arachidylsphingosylphosphorylcholine (d18: 1-C20: 0), and behenylsphingosylphosphorylcholine (d18: 1-C22: 0) It contains at least the above molecular species. The milk-derived sphingomyelin is more preferably one containing two or more molecular species selected from the above group, and more preferably one containing three or more molecular species selected from the above group Particularly preferred are those containing at least all five molecular species of the aforementioned group. According to a further preferred embodiment of the invention, the milk-derived sphingomyelin is a molecule of palmityl sphingosylphosphorylcholine (d18: 1-C16: 0) and / or behenylsphingosylphosphorylcholine (d18: 1-C22: 0). It contains at least seeds.

According to a preferred embodiment of the present invention, the production promoter according to the present invention is for oral intake (ie, oral intake) or enteral administration (ie, enteral administration).

The production promoter according to the present invention is preferably used for skin with a deteriorated state, skin with a reduced barrier function, or skin with a reduced production of stratum corneum cell covalent ceramide.

As shown in the examples described later, according to the present invention, ceramide cell covalently bound ceramide production is achieved in a state of reduced skin barrier function in which the amount of ceramide in the stratum corneum does not change or the amount of ceramide increases. It can be promoted to maintain or improve skin barrier function. For this reason, the production promoter according to the present invention is more preferably a skin in which the ceramide amount in the stratum corneum does not change or the ceramide amount increases, the skin in which the state deteriorates, the skin in which the barrier function is lowered, or the stratum corneum Used for skin with reduced production of covalently bound ceramide.

According to the production promoter for horny layer covalently bound ceramide of the present invention, as will be described later, retention and increase of horny layer water content and retention and reduction of transdermal moisture transpiration amount can be expected. An agent for retaining or increasing the amount of water in the stratum corneum comprising a lipid as an active ingredient, and a agent for retaining or reducing the transdermal moisture transpiration amount comprising a sphingolipid as an active ingredient are also provided.

Use The sphingolipid, which is an active ingredient in the present invention, has an activity to promote production of keratinocyte covalently bound ceramide (Examples described later), and has an effect of preventing, suppressing or improving the deterioration of the skin condition.

Here, the promotion of production of stratum corneum cell covalent ceramide includes promotion of production of stratum corneum cell covalent bond ceramide, maintenance of production, and recovery or improvement of the lowered production state.

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 ameliorated can be achieved by promoting production of stratum corneum cell covalently bound ceramide.

Composition and Food / Beverage As described above, according to the present invention, there is provided a composition for preventing, suppressing or improving deterioration of the skin condition, comprising the active ingredient of the present invention, that is, sphingolipid. . It can also be said that this composition comprises the production promoter for horny cell covalently bound ceramide according to the present invention. Moreover, according to this invention, the food / beverage products which comprise the production promoter of the horny layer cell covalent bond ceramide by this invention are provided.

Such a composition and food and drink can be produced, for example, by a production method comprising adding the production promoter according to the present invention to the composition and the ingredients of the food and drink.

The composition for preventing, suppressing or improving the deterioration of skin condition according to 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. 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. In the case of a parenteral preparation, it can take the form of an injection or a suppository. From the viewpoint of simplicity, an oral preparation is preferable. 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 one preferred embodiment of the present invention, collagen is used as an optional component to be added to food and drink (preferably yogurt).

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 (yogurt).

Food and drink products include functional foods, health and nutrition foods, supplements, health foods, functional labeling foods, foods for specified health use, 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 functional display food, a food for specified health use, 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 according to 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. Can be provided.

According to another aspect of the present invention, there can be provided a food or drink displaying a function for preventing, suppressing or improving the deterioration of the skin condition by promoting the production of stratum corneum covalently bound ceramide.

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 Adults (body weight 60 kg) per day, sphingolipids (preferably milk-derived sphingolipids, more preferably milk-derived sphingomyelin) are 0.5-1500 mg, preferably 1-1000 mg, more preferably 5- It can be adjusted to an effective amount that can be ingested at 500 mg.

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

At this time, as the mobile phase, for example, a solution obtained by mixing 0.5% mM phosphate-citrate buffer (pH 3.0) and methanol in a ratio of 5 to 95 may be used. The measurement time is set to 20 minutes, the flow rate of the mobile phase is set to 0.6 mL / min, the column temperature is 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 it can be quantified by the area ratio.

In another aspect of the present invention, the content of the active ingredient in foods and drinks and compositions can be specified per packaging form. For example, in the case of foods and drinks, the content of sphingolipid is 5 to 1500 mg, preferably 6 In the case of a composition, the content of sphingolipid is 0.5 to 1500 mg, preferably 1 to 1000 mg, more preferably 5 to 500 mg. Can do. In addition, the amount per packaging form is not limited to a single intake, and may be an amount that includes multiple intakes or multiple daily intakes (for example, 30 days). The content of sphingolipid is 5 to 45000 mg, preferably 6 to 30000 mg, more preferably 7 to 15000 mg. In the case of the composition, the content of sphingolipid is 0.5 to 45000 mg, preferably 1 to It may contain 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 in which the content of phospholipid (particularly sphingolipid) (preferably sphingomyelin content) is intentionally increased is used.

Therefore, according to one preferred embodiment of the present invention, there is provided a yogurt having enhanced production of horny layer covalently bound ceramide containing an additional sphingolipid. The term “additional sphingolipid” as used herein is not a sphingolipid that can be originally contained in yogurt, but yogurt in which the concentration of sphingolipid is intentionally increased by adding sphingolipid separately to yogurt. A sphingolipid added separately when making.

In the milk beverage and fermented milk of the present invention, when lipid is contained in a normal amount (for example, 3% by weight of lipid), phospholipid is contained at 1 to 300 mg / g, preferably 1.5 to 250 mg / g. More preferably 2 to 200 mg / g, still more preferably 2.5 to 150 mg / g. Further, in the milk beverage and fermented milk of the present invention, when lipid is contained in a normal amount, sphingomyelin is contained at 0.2 to 60 mg / g, preferably 0.25 to 50 mg / g, more preferably 0.3. It can be contained at -40 mg / g, more preferably 0.35-30 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), phospholipid is contained at 0.5 to 300 mg / g, preferably 0 .6 to 250 mg / g, more preferably 0.7 to 200 mg / g, still more preferably 0.8 to 150 mg / g. In the milk beverage and fermented milk of the present invention, when lipid is contained in a small amount, sphingomyelin is contained at 0.05 to 60 mg / g, preferably 0.1 to 50 mg / g, more preferably 0.15 to It can be contained at 40 mg / g, 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), phospholipid is contained at 0.2 to 300 mg / g. Preferably, it can be contained at 0.3 to 250 mg / g, more preferably 0.4 to 200 mg / g, and still more preferably 0.5 to 150 mg / g. Further, in the milk beverage and fermented milk of the present invention, when almost no lipid is contained, sphingomyelin is contained at 0.02 to 60 mg / g, preferably 0.04 to 50 mg / g, more preferably 0.06 to It can be contained at 40 mg / g, more preferably 0.08-30 mg / g.

According to another aspect of the present invention, the production of stratum corneum cell covalent ceramide comprising sphingolipid being orally ingested or enterally administered to a subject with reduced production of stratum corneum cell covalent ceramide. A promotion method is provided.

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

The method preferably comprises keratinocyte covalent ceramide for a subject having reduced production of horny cell covalent ceramide, wherein the sphingolipid is continuously orally ingested for 2 days or more or enterally administered. It can be a production promotion method.

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

As the sphingolipid used, a phospholipid concentrate (PC700, Fontera) containing 16.0% by weight of sphingomyelin derived from milk was used.

Table 1 below shows the composition of the sphingomyelin molecular species of the phospholipid concentrate actually used.
The sphingomyelin molecular species composition was measured as follows.

Measurement of sphingomyelin molecular species composition The phospholipid concentrate was dissolved in Tris buffer (pH 7.4) containing 2% Triton X-100. 1 U sphingomyelinase (Sigma) was added and reacted at 37 ° C. for 2 hours to hydrolyze sphingomyelin to ceramide. Hydrolyzed ceramide molecular species were extracted with a chloroform / methanol mixture (2: 1, v / v).
The extracted ceramide molecular species were used as a column using ACQUITY UPLC BEH C18 (2 mm × 100 mm, particle size 1.7 μm, Waters) using a liquid chromatography mass spectrometer LC / MS / MS (Quatro premier XE (Waters)). ) And measured.
At this time, ammonium acetate (5 mM) / acetonitrile was used as mobile phase A, and ammonium acetate (5 mM) / methanol (95%) was used as mobile phase B. Then, mobile phase A was started from 100%, and after 30 minutes, mobile phase B was gradiented to 100%, then mobile phase B was made 100% and held for 2 minutes, and after another 3 minutes, The mobile phase A was switched to 100%. That is, the measurement time for one sample of LC / MS / MS was set to 35 minutes. The mobile phase flow rate was set to 0.4 mL / min, the column temperature was set to 40 ° C., and detection was performed using electrospray ionization in the positive mode. As LC / MS / MS analysis variables, capillary voltage is 3000V, source temperature is 120 ° C, solvent removal temperature is 400 ° C, solvent removal gas flow rate is 850L / h, cone gas flow rate is 50L / h, cone The voltage was set to 30 eV / h.
The sphingomyelin molecular species amount was calculated from the molar concentration of the obtained ceramide molecular species.

 

Test 1: Effect of sphingolipids on dry skin (dry skin) Hairless mice were fed a diet that induces dry skin (dry skin), and the effect of oral intake of phospholipid concentrate was examined.

(1-1) Induction of dry skin Feed that induces dry skin after acclimating hairless mice (Hos: HR-1, female, 4 weeks old) for 1 week (special feed HR-AD, Nippon Agricultural Industrial Co., Ltd.) Made freely) in 8 weeks.

(1-2) Composition of Evaluation Group Using a dry skin model animal test system, the effect of oral intake of phospholipid concentrate (PC700) on skin condition was evaluated. At this time, there were 10 mice in each group.
The relationship between the group configuration of animal experiments (configuration of evaluation group) and the content of sphingomyelin is shown in Table 2 below.

 

(1-3) Evaluation Test Method For each experimental animal group, the stratum corneum water content, transdermal water transpiration (TEWL), and stratum corneum cell covalently bound ceramide amount were measured, and the skin barrier function was evaluated from these. .

(a) Water content of stratum corneum The water content of the stratum corneum was measured 5 times / animal using a Corneometer (Corneometer, manufactured by Courage and Khazaka Electronic GmbH). Among these, the average of three measurement values excluding the maximum value and the minimum value was adopted to obtain the stratum corneum moisture content.

(b) Transdermal moisture transpiration (TEWL)
The transdermal moisture transpiration was measured using a tevameter (Tewemeter MPA580, Courage and Khazaka Electronic GmbH) with a probe kept at 29 ° C. for 20 seconds. About the obtained measurement result, the average of 3 times was employ | adopted and it was set as the amount of transdermal moisture transpiration.

(c) Amount of stratum corneum covalently bound ceramide The amount of stratum corneum covalently bound ceramide was measured using a liquid chromatography mass spectrometer LC / MS / MS (Quatro Premier XE (Waters)) as a column as ACQUITY UPLC BEH C18. (2 mm × 100 mm, particle size 1.7 μm, Waters) was used for measurement.
At this time, ammonium acetate (5 mM) / acetonitrile was used as mobile phase A, and ammonium acetate (5 mM) / methanol (95%) was used as mobile phase B. Then, mobile phase A was started from 100%, and after 30 minutes, mobile phase B was gradiented to 100%, then mobile phase B was made 100% and held for 2 minutes, and after another 3 minutes, The mobile phase A was switched to 100%. That is, the measurement time for one sample of LC / MS / MS was set to 35 minutes. The mobile phase flow rate was set to 0.4 mL / min, the column temperature was set to 40 ° C., and detection was performed using electrospray ionization in the positive mode. As LC / MS / MS analysis variables, capillary voltage is 3000V, source temperature is 120 ° C, solvent removal temperature is 400 ° C, solvent removal gas flow rate is 850L / h, cone gas flow rate is 50L / h, cone The voltage was set to 30 eV / h.

(d) Amount of horny layer ceramide The amount of horny layer ceramide was measured using a liquid chromatography mass spectrometer LC / MS / MS (Quatro Premier XE (Waters)) as a column and ACQUITY UPLC BEH C18 (2 mm × 100 mm, particles Measurement was performed using a diameter of 1.7 μm (Waters).
At this time, ammonium acetate (5 mM) / acetonitrile was used as mobile phase A, and ammonium acetate (5 mM) / methanol (95%) was used as mobile phase B. Then, mobile phase A was started from 100%, and after 30 minutes, mobile phase B was gradiented to 100%, then mobile phase B was made 100% and held for 2 minutes, and after another 3 minutes, The mobile phase A was switched to 100%. That is, the measurement time for one sample of LC / MS / MS was set to 35 minutes. The mobile phase flow rate was set to 0.4 mL / min, the column temperature was set to 40 ° C., and detection was performed using electrospray ionization in the positive mode. As LC / MS / MS analysis variables, capillary voltage is 3000V, source temperature is 120 ° C, solvent removal temperature is 400 ° C, solvent removal gas flow rate is 850L / h, cone gas flow rate is 50L / h, cone The voltage was set to 30 eV / h.

The stratum corneum ceramide levels are palmitic sphingosine (d18: 1-C16: 0), stearyl sphingosine (d18: 1-C18: 0), arachidyl sphingosine (d18: 1-C20: 0), behenyl sphingosine (d18: 1 -C22: 0), Tricosanyl sphingosine (d18: 1-C23: 0), Tricocenoyl sphingosine (d18: 1-C23: 1), Lignoceryl sphingosine (d18: 1-C24: 0), Nerbonyl sphingosine The total of (d18: 1-C24: 1) and serotilsphingosine (d18: 1-C26: 0) was determined. The standard substance of each ceramide was Avanti Polar Lipids, and was quantified by the area ratio.

(1-4) Evaluation results The results were as follows.

(a) stratum corneum water content The results were as shown in FIG.
In FIG. 1, the stratum corneum moisture content of the HD-AD group was lower than that of the Normal group. On the other hand, the stratum corneum water content of the L-MPL group and the H-MPL group was higher than that of the HD-AD group. From this, it can be said that the deterioration of the barrier function of the skin was suppressed in the L-MPL group and the H-MPL group.

(b) Transdermal moisture transpiration (TEWL)
The result was as shown in FIG.
In FIG. 2, the TEWL of the HD-AD group was higher than the TEWL of the Normal group. On the other hand, the TEWL of the L-MPL group and the H-MPL group was lower than that of the HD-AD group. From this, it can be said that the deterioration of the barrier function of the skin was suppressed in the L-MPL group and the H-MPL group.

(c) Amount of stratum corneum covalently bound ceramide The results were as shown in Table 3 below.

 

Table 3 shows the content ratio of stratum corneum covalently bound ceramide. The content ratio of the stratum corneum cell covalent bond ceramide is defined as the amount of each stratum corneum cell covalent bond ceramide in the Normal group at the end of the test (after 8 weeks of intake of the test meal), and the amount of each other stratum corneum cell covalent bond ceramide. Is shown as a relative value.
Here, the content ratio of each stratum corneum cell covalent bond ceramide in the HD-AD group was lower than the content ratio of the stratum corneum cell covalent bond ceramide in the Normal group. On the other hand, the content ratio of each stratum corneum cell covalent ceramide in the H-MPL group was higher than the content ratio of each stratum corneum cell covalent ceramide in the HD-AD group. From this, it can be said that the deterioration of the barrier function of the skin was suppressed in the L-MPL group and the H-MPL group.

(d) Amount of stratum corneum The results were as shown in FIG.
As can be seen from the results, the amount of each horny layer ceramide in the HD-AD group did not change or increased as compared with the amount of the horny layer ceramide in the Normal group. In contrast, as described above, the content ratio of each stratum corneum cell covalent ceramide in the HD-AD group was lower than the content ratio of the stratum corneum cell covalent bond ceramide in the Normal group.
That is, even if the skin condition deteriorates, the amount of stratum corneum ceramide does not decrease, it increases rather than the normal value, whereas the amount of stratum corneum covalently bound ceramide tends to be independent, It was found that the amount decreased as the skin condition worsened.

(E) Others When the skin surface of each group of mice was visually observed, the HD-AD group clearly showed rough skin caused by dry skin, but the H-MPL group had the same level as the Normal group. The skin condition was good.

Further, according to the histological findings obtained by collecting the skin surface tissue of each group, staining with HD, and observing with a microscope, the H-MPL group had a good skin state at the same level as the Normal group.

Test 2: Effect of sphingolipids on deterioration of skin barrier function under UV irradiation Effect of oral intake of phospholipid concentrate on this condition by reducing skin barrier function by irradiating hairless mice with UV light I investigated.

(2-1) Induction of reduced skin barrier function by UV irradiation After acclimatization of hairless mice (Hos: HR-1, female, 4 weeks old) for 1 week, UV irradiation was performed at 20 mJ / cm2. (UV-B (GL20SE, Sankyo Electric Co., Ltd.)) was irradiated.

(2-2) Configuration of Evaluation Group Using a dry skin model animal test system, the effect of sphingomyelin (purity: 98%, manufactured by Nagara Science Co., Ltd.) on the skin condition was evaluated. . At this time, there were 10 mice in each group. The relationship between the group configuration of animal experiments (configuration of evaluation group) and the content of sphingomyelin is shown below.

Control group: Group in which normal feed is consumed SM group: Sphingomyelin (purity: 98%) is orally administered at 146 mg / kg / day

(2-3) Evaluation Test Method For each experimental animal group, the amount of stratum corneum water, the amount of transdermal water transpiration (TEWL), the amount of stratum corneum covalently bound ceramide, And the amount of ceramide ceramide was measured, and the barrier function of skin was evaluated from these.

(2-4) Evaluation results The results were as follows.

(a) The stratum corneum moisture content The results were as shown in FIG.
On the second and third days of the test, the stratum corneum moisture in the SM group was higher than the stratum corneum moisture in the Control group.

(b) Transdermal moisture transpiration (TEWL)
The result was as shown in FIG.
Here, on the second and third days of the test, the SM group TEWL was lower than the Control group TEWL.

(c) Amount of stratum corneum covalently bound ceramide The results were as shown in FIG.
Here, the transition of the stratum corneum cell covalently bound ceramide amount (d18: 1-C34: 1) is shown. The amount of stratum corneum covalently bound ceramide was expressed as a relative value, with the amount of stratum corneum covalently bound ceramide in the Control group at the start of the test being 100, and the amount of other stratum corneum covalently bound ceramides being expressed. Here, the amount of stratum corneum covalently bound ceramide in the SM group was higher than the amount of stratum corneum covalently bound ceramide in the Control group.

(d) Amount of stratum corneum The results were as shown in FIG.
Under the irradiation of ultraviolet rays, the amount of stratum corneum ceramide does not decrease and increases to a normal value, but rather increases, whereas the amount of stratum corneum covalently bound ceramide tends to be independent of the amount of skin condition. It was found that the amount decreased with deterioration.

It was suggested that oral intake of sphingomyelin suppresses deterioration of the skin barrier function based on ultraviolet rays. Furthermore, it was suggested that as the sphingomyelin is continuously ingested over 2 days, the deterioration of the skin barrier function is more likely to be suppressed.

Test 3: Effects of sphingolipids on humans with dry skin The effects of food intake containing phospholipid concentrates on women with subjective dry symptoms were examined.

(3-1) Preparation of test food Nonfat dry milk 8.8 parts, sugar 5.3 parts, milk protein 1.7 parts, collagen peptide 1.7 parts, modified starch 0.2 parts, thickening polysaccharide 0.1 Part, 0.04 part phospholipid concentrate, 0.05 part fragrance, 0.01 part acidulant, 0.01 part arginine, 0.0001 part sweetener, and 82.0 parts water, at 95 ° C. Sterilized under conditions of 1 minute. A mixed starter of Lactobacillus bulgaricus and Streptococcus thermophilus was added to this raw milk as a yogurt starter at 0.15% and fermented at 37 to 45 ° C. to produce yogurt as a test food.
75 g of this test food contains 1000 mg of collagen (Nitta Gelatin) and phospholipid concentrate (PC700, Fontera) (containing 10 mg of sphingomyelin).

(3-2) Evaluation Test Method The test food containing sphingomyelin was ingested for 75 weeks (10 mg as sphingomyelin) for 4 weeks to 32 women (panelists) who had subjective dry symptoms.

(3-3) Evaluation index Before and after ingestion, a total of 3 stratum corneum was collected from the same site using a PPS tape (PPS film adhesive tape, Teraoka Seisakusho) from the inner side of the forearm, and the amount of stratum corneum covalently bound ceramide (d18: 1-C28: 0, d18: 1-C30: 0, d18: 1-C30: 1, d18: 1-C32: 0, d18: 1-C32: 1, d18: 1-C34: 0, d18: 1- C34: 1, d18: 1-C36: 0, d18: 1-C36: 1, and their sum).

(3-4) Evaluation results The results were as follows.

The results of the amount of ceramide covalently bound to horny layer cells are as shown in FIGS.

The total amount of stratum corneum covalently bound ceramide was higher after ingestion than before ingestion. In addition, the amount of each horny cell covalently bound ceramide was also higher after ingestion than before ingestion. From this, it was shown that stratum corneum cell covalent binding ceramide increases by continuously ingesting food containing sphingomyelin.

Claims (18)

1. A stratum corneum cell covalent ceramide production promoter containing sphingolipid as an active ingredient.
2. The production promoter according to claim 1, wherein the sphingolipid is a sphingophospholipid derived from milk.
3. The production promoter according to claim 1 or 2, wherein the sphingophospholipid is sphingomyelin.
4. The stratum corneum covalent ceramide is omega hydroxy d18: 1-C28: 0, d18: 1-C30: 1, d18: 1-C30: 0, d18: 1-C32: 0, d18: 1-C32: 1, d18: 1-C34: 0, d18: 1-C34: 1, d18: 1-C36: 1, d17: 1-C32: 0, d17: 1-C32: 1, d17: 1-C34: 0, d17: The production promoter according to any one of claims 1 to 3, which comprises one or more molecular species selected from the group consisting of 1-C34: 1 and d17: 1-C36: 1.
5. The production promoter according to any one of claims 1 to 4, which is for oral intake or enteral administration.
6. A composition for preventing, suppressing or improving deterioration of skin condition, comprising the active ingredient according to any one of claims 1 to 5.
7. The composition according to claim 6, wherein the deterioration of the skin condition is a decrease in the barrier function of the skin.
8. The composition according to claim 7, wherein the decrease in the barrier function of the skin is due to a decrease in production of stratum corneum cell covalent ceramide in the stratum corneum.
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 sphingolipid that can be ingested from 0.5 to 1500 mg per day.
11. A food or drink comprising the keratinocyte covalently bound ceramide production promoter according to any one of claims 1 to 5.
12. A food or drink comprising the composition for preventing, suppressing or improving deterioration of the skin condition according to any one of claims 6 to 9.
13. The food or drink according to claim 11 or 12, comprising an amount of sphingolipid that can be ingested by 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 functional indication food, a food for specified health use, or a food with a disease risk reduction indication.
15. A yogurt comprising the keratinocyte covalently bound ceramide production promoter according to any one of claims 1 to 5.
16. Yogurt with enhanced production of stratum corneum covalent ceramide containing additional sphingolipids.
17. The yogurt according to claim 15 or 16, further comprising collagen.
18. The yogurt according to any one of claims 15 to 17, wherein the sphingophospholipid is sphingomyelin.
    

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