Composition for improving skin barrier function, composition for promoting tight junction formation, and composition for promoting synthesis of ceramide precursor in stratum corneum

Abstract

To provide compositions for improving skin barrier function, compositions for promoting tight junction formation and compositions for promoting synthesis of horny layer ceramide precursor, and an applied product thereof.SOLUTION: Provided herein is a composition for improving skin barrier function comprising an asparagus extract or fractions thereof. The composition of the invention has skin barrier function improving activity, tight junction formation enhancing activity, and horny layer ceramide precursor synthesis-promoting activity. Accordingly, the composition having activities such as dermal tissue function enhancement can prevent or improve various conditions (wrinkle, dry skin, roughened skin, itchiness, inflammation), and can be used in formulations such as pharmaceuticals, non-medical external preparation for skin such as cosmetics not applicable to pharmaceuticals, or in oral administration form such as food and supplements.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] The present invention relates to a composition for improving skin barrier function, a composition for promoting the formation of tight junctions, and a composition for promoting the synthesis of ceramide precursors in the stratum corneum, as well as applications thereof. [Background technology]

[0002] The skin is composed of three layers: the epidermis, dermis, and subcutaneous tissue, of which the epidermis is the outermost layer of the skin and is in contact with the outside world. The epidermis is composed of four layers, namely the stratum corneum, stratum granulosum, stratum spinosum, and stratum basale, from the surface side, and has a barrier function that maintains the skin's moisture content and prevents foreign substances from entering the body from the outside.

[0003] Generally, the barrier function of the skin is evaluated by transepidermal water loss (TEWL). Transepidermal water loss (TEWL) is the amount of water that evaporates from the skin, and represents the amount of water that evaporates from the body through the stratum corneum. When the barrier function of the skin is weakened, the amount of transepidermal water loss increases, and water loss from the skin increases.

[0004] Meanwhile, the barrier function of the skin consists of a barrier function based on the stratum corneum (ceramides, hyaluronic acid, etc.) and a barrier function based on the granular layer adjacent to the stratum corneum (tight junctions). If the barrier function based on the tight junctions is weakened, it also affects the barrier function based on the stratum corneum, causing problems such as dry skin, rough skin, and even skin aging. Therefore, in recent years, attention has been focused not only on the barrier function based on the stratum corneum but also on the barrier function based on the tight junctions as a barrier function of the skin.

[0005] For example, as agents that promote the formation of tight junctions, a skin barrier function improver consisting of a cactus fruit extract obtained from the fruit of a plant belonging to the genus Opuntia in the Cactaceae family (Patent Document 1), and a tight junction formation promoter containing bird's nest extract as an active ingredient (Patent Document 2) have been reported.

[0006] In addition, in terms of the barrier function based on the stratum corneum, research has been conducted on substances that promote ceramide synthesis, etc. For example, it has been reported that glucosylceramide obtained from plant materials such as rice bran and rice germ increases endogenous ceramide (stratum corneum ceramide) in the skin (Patent Document 3), and a ceramide production promoter containing one or two active ingredients selected from Echinacea purpurea and lavender (Patent Document 4). [Prior art documents] [Patent documents]

[0007] [Patent Document 1] JP 2012-121871 A [Patent Document 2] JP 2014-210748 A [Patent Document 3] JP 2012-184166 A [Patent Document 4] JP 2019-156777 A Summary of the Invention [Problem to be solved by the invention]

[0008] As disclosed in Patent Documents 1 and 2, skin barrier function improvers and tight junction formation promoters derived from plants and the like have been reported, and as disclosed in Patent Documents 3 and 4, various ceramide synthesis promoters that promote ceramide synthesis derived from plants and the like have been reported. Thus, there is an increasing demand for improving the skin barrier function, and there is a demand for the development of new components that improve the skin barrier function, promote the formation of tight junctions, and promote the synthesis of ceramide precursors in the stratum corneum.

[0009] Under these circumstances, an object of the present invention is to provide a composition for improving skin barrier function, a composition for promoting the formation of tight junctions, and a composition for promoting the synthesis of ceramide precursors in the stratum corneum, as well as applications thereof. [Means for solving the problem]

[0010] As a result of intensive research aimed at solving the above-mentioned problems, the inventors discovered that an asparagus extract or a fraction of said extract has excellent effects on improving skin barrier function, promoting the formation of tight junctions, and promoting the synthesis of ceramide precursors in the stratum corneum, and thus arrived at the present invention.

[0011] That is, the present invention relates to the following inventions. <1> A composition for improving skin barrier function, comprising an asparagus extract or a fraction of said extract. <2> The skin barrier function improvement effect is based on either the promotion of tight junction formation or the promotion of ceramide precursor synthesis in the stratum corneum. <1> The composition described in <3> The asparagus extract is a solvent extract of asparagus with one or more extraction solvents selected from water, ethanol, and butylene glycol. <1> or <2> The composition described in <4> A composition for promoting tight junction formation, comprising an asparagus extract or a fraction of said extract. <5> For promoting claudin 1 expression and / or claudin 4 expression <4> A composition for promoting tight junction formation according to claim 1. <6> A composition for promoting synthesis of ceramide precursors in the stratum corneum, comprising an asparagus extract or a fraction of said extract. <7> For promoting one or more of serine palmitoyltransferase expression, ceramide synthesase expression, and sphingomyelin synthesase expression. <6> A composition for promoting synthesis of a ceramide precursor in the stratum corneum according to claim 1. <8> For medicinal purposes, <1> from <7> 2. The composition according to claim 1 . <9> For external use on the skin (excluding medical uses), <1> from <7> 2. The composition according to claim 1 . <10> For oral use (excluding medical use); <1> from <7> 2. The composition according to claim 1 .

[0012] <1a> A method for producing a composition for improving skin barrier function, comprising an extraction step of obtaining an asparagus extract by contacting asparagus with an extraction solvent. <2a> A method for producing a composition for promoting tight junction formation, comprising an extraction step of obtaining an asparagus extract by contacting asparagus with an extraction solvent. <3a> A method for producing a composition for promoting synthesis of a ceramide precursor in the stratum corneum, comprising an extraction step of obtaining an asparagus extract by contacting asparagus with an extraction solvent. Effect of the Invention

[0013] According to the present invention, there are provided a composition for improving skin barrier function, a composition for promoting the formation of tight junctions, and a composition for promoting the synthesis of ceramide precursors in the stratum corneum, as well as applications thereof. [Brief description of the drawings]

[0014] [Figure 1] FIG. 1 is an explanatory diagram showing the synthetic pathway of stratum corneum ceramide in the epidermis. [Diagram 2] FIG. 1 shows the results of evaluating the effect of the asparagus extract of Example 1 on transepidermal water loss (TEWL) of three-dimensionally cultured human epidermis ((a): cultured for 2 days, (b): cultured for 8 days). [Diagram 3] FIG. 1 shows the results of evaluating the effect of the asparagus extract of Example 1 on the formation of tight junctions in three-dimensionally cultured human epidermis ((a): Claudin 1 gene expression, (b): Claudin 4 gene expression). [Figure 4] FIG. 1 shows the results of evaluating the effect of the asparagus extract of Example 1 on the synthesis of ceramide precursors in the stratum corneum of 3D cultured human epidermis ((a): CerS3 gene expression, (b): SMS1 gene expression, (c): SMS2 gene expression, (d): SPT1 gene expression, (e): SPT2 gene expression). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] The present invention will be described in detail below with reference to examples, but the present invention is not limited to the following examples, and can be modified as desired without departing from the gist of the present invention. In this specification, the expression "~" is used to include the numerical or physical values ​​before and after it. In this specification, the expression "A and / or B" includes "A only," "B only," and "both A and B."

[0016] <1. Composition for improving skin barrier function of the present invention> The present invention relates to a composition for improving skin barrier function containing an asparagus extract or a fraction of said extract (hereinafter, may be referred to as "the composition for improving skin barrier function of the present invention").

[0017] An asparagus extract or a fraction of said extract has the effect of improving the skin barrier function, and can be used as an active ingredient of a composition for improving skin barrier function. In the present invention, "improving skin barrier function" not only includes maintaining normal skin barrier function, improving normal skin barrier function, and restoring impaired skin barrier function, but also includes suppressing impairment of skin barrier function and preventing impairment of skin barrier function. The composition for improving skin barrier function of the present invention is useful for at least one of preventing, treating, and improving a disease or symptom (wrinkles, rough skin, itchiness, inflammation, sensitive skin, etc.) caused by a decrease in skin barrier function. That is, "prevention" includes suppressing and delaying the onset of the disease or symptom. Furthermore, "treatment" includes improving and relieving the pathology of the disease or symptom, and suppressing the progression of the disease or symptom.

[0018] In addition, the asparagus extract or a fraction thereof has an effect of improving the skin barrier function and therefore has a skin moisturizing effect, so the composition of the present invention can be used as a skin moisturizing composition.

[0019] In the present invention, the "skin barrier function" can be determined using the transepidermal water loss (TEWL) as an index. TEWL increases when the skin barrier function is reduced, and decreases when the skin barrier function is increased. The method for evaluating TEWL will be described later in <2-2. Evaluation by measuring transepidermal water loss (TEWL)> in the Examples.

[0020] The skin barrier function improving effect is based on either one of the effects of promoting the formation of tight junctions and the synthesis of ceramide precursors in the stratum corneum, which are attributable to the asparagus extract or a fraction of the extract contained therein. These effects may be either one or both.

[0021] In addition, the asparagus extract or a fraction thereof has an effect of promoting the formation of tight junctions present in the granular layer, and therefore the asparagus extract or a fraction thereof can be used as an active ingredient of a composition for promoting the formation of tight junctions.

[0022] That is, the composition for promoting tight junction formation of the present invention contains an asparagus extract or a fraction of said extract as an active ingredient. Asparagus extract or a fraction of said extract promotes the formation of tight junctions, which are intercellular bonds localized in the granular layer just below the stratum corneum, thereby inhibiting the evaporation of water from the body, thereby improving the skin barrier function.

[0023] The effect of promoting the formation of tight junctions can be determined by evaluating the expression of tight junction-related genes, and specifically, can be evaluated by the evaluation method described in the Examples below (2-3-1. Evaluation of the effect of promoting the formation of tight junctions).

[0024] The composition for promoting tight junction formation of the present invention may have an effect of promoting the formation of at least one of tight junction-associated genes. Examples of the tight junction-associated genes that are the targets of the promotion of tight junction formation include claudin, occludin, and ZO (Zonula Occludens). Among these, Claudin 1 and Claudin 4 are particularly preferred targets. That is, the composition for promoting tight junction formation of the present invention preferably promotes the expression of Claudin 1 and / or Claudin 4, and particularly preferably promotes the expression of both of these.

[0025] In addition, the asparagus extract or a fraction thereof has an effect of promoting the synthesis of stratum corneum ceramide precursors present in the stratum spinosum and stratum granulosum, which are components of the skin, and therefore the asparagus extract or a fraction thereof can also be used as an active ingredient of a composition for promoting the synthesis of stratum corneum ceramide precursors.

[0026] That is, the composition for promoting synthesis of stratum corneum ceramide precursors of the present invention contains asparagus extract or a fraction of said extract as an active ingredient. The asparagus extract or a fraction of said extract promotes synthesis of stratum corneum ceramide precursors present in the granular layer and the spinous layer of the four layers (stratum corneum, stratum granulosum, stratum spinosum, and stratum basale) that constitute the epidermis, thereby improving the moisture retention ability of the skin and thereby improving the skin barrier function.

[0027] In the present invention, the "stratum corneum ceramide precursor" includes those synthesized in a stage prior to the synthesis of stratum corneum ceramide in the synthetic pathway of stratum corneum ceramide described below. That is, the stratum corneum ceramide precursor includes one or more selected from dihydrosphingosine, ceramide, sphingomyelin, and glycosylceramide, which are synthesized in the process of synthesizing stratum corneum ceramide. In the following description, the "ceramide" synthesized in the stage prior to the synthesis of stratum corneum ceramide is referred to as "ceramide (outside the stratum corneum)" to distinguish it from "stratum corneum ceramide".

[0028] The synthetic pathway of stratum corneum ceramide in the epidermis will be explained below with reference to FIG. Various enzymes are involved in the synthesis of ceramide in the stratum corneum of the epidermis. First, from the basal layer to the spinous layer, the first step in the synthesis of ceramide in the stratum corneum is the condensation of serine and palmitoyl-CoA by the action of serine palmitoyltransferase (SPT), resulting in the synthesis of dihydrosphingosine. Then, ceramide (outside the stratum corneum) is formed from dihydrosphingosine by N-acylation of fatty acids by ceramide synthase (CerS).

[0029] There are two pathways by which ceramide (outside the stratum corneum) is synthesized into stratum corneum ceramide. In one pathway, ceramide (outside the stratum corneum) is converted to glucosylceramide by the action of glucosylceramide synthesase (GCS) and accumulated in the granular layer. After that, stratum corneum ceramide is synthesized in the stratum corneum by β-glucocerebrosidase (GBA) (pathway on the right side of Figure 1). In the other pathway, ceramide (outside the stratum corneum) is converted to sphingomyelin by the action of sphingomyelin synthase (SMS), which accumulates in the granular layer, and is then synthesized into sphingomyelin in the stratum corneum by acid sphingomyelinase (SMase) (left pathway in Figure 1).

[0030] The composition for promoting synthesis of a ceramide precursor in the stratum corneum of the present invention promotes synthesis of a ceramide precursor in the stratum corneum. Among ceramide precursors in the stratum corneum, the composition for promoting synthesis of a ceramide precursor in the stratum corneum of the present invention is preferably one that promotes synthesis of dihydrosphingosine, ceramide (outside the stratum corneum), or sphingomyelin.

[0031] The composition for promoting synthesis of a stratum corneum ceramide precursor of the present invention may have the effect of promoting the synthesis of at least one of the stratum corneum ceramide precursor-related genes. Examples of stratum corneum ceramide precursor-related genes that are targets for promoting the synthesis of stratum corneum ceramide precursors include serine palmitoyltransferase (SPT), ceramide synthesase (CerS), sphingomyelin synthesase (SMS), and glucosylceramide synthesase (GCS). Among these, SPT (SPT1, SPT2), CerS3, and SMS (SMS1, SMS2) are particularly preferred targets. That is, the composition for promoting synthesis of a stratum corneum ceramide precursor of the present invention preferably promotes the expression of one or more selected from SPT (SPT1, SPT2), CerS3, and SMS (SMS1, SMS2), and it is particularly preferred that the expression of all of these be promoted.

[0032] The effect of promoting the synthesis of stratum corneum ceramide precursors can be determined by evaluating the expression of the target stratum corneum ceramide precursor-related gene, and specifically, it can be evaluated by the evaluation method described in the Examples below, <2-3-2. Evaluation of the effect of promoting the synthesis of stratum corneum ceramide precursors>.

[0033] The composition for improving skin barrier function of the present invention, the composition for promoting the formation of tight junctions of the present invention, and the composition for promoting the synthesis of ceramide precursors in the stratum corneum of the present invention will be described in more detail below. Note that, hereinafter, the "composition for improving skin barrier function of the present invention," the "composition for promoting the formation of tight junctions of the present invention," and the "composition for promoting the synthesis of ceramide precursors in the stratum corneum of the present invention" are collectively referred to as the "composition of the present invention."

[0034] (asparagus) "Asparagus" used as a raw material for the composition of the present invention is a concept including the stem, cut portion and pseudo-leaves of Asparagus officinalis, which is generally used for food. The "stalk" is the young stem of asparagus and is the edible part. The "cut part" is the base part of asparagus that does not meet the standard, usually about 2 cm from the bottom. These are usually discarded. The "pseudo-leaves" are stems that branch out after the stem has grown further, and are the part that produces nutrients through photosynthesis in place of leaves.

[0035] Asparagus is used either unprocessed or processed into a suitable form depending on the part to be used. That is, the stems, cut parts and pseudo-leaves of asparagus can be used as they are, sliced, crushed, squeezed or grated into fine particles.

[0036] Asparagus can be used for the solvent extraction as is without prior treatment, but is usually used in fine particles such as by slicing, crushing, squeezing or grating to increase the efficiency of the extraction.

[0037] In the present invention, the term "extract" refers to a general concept of a form in which the content of active ingredients has been increased by compressing or solvent extraction of the target raw plant, or a product thereof that has been pretreated (dried or chopped) as necessary.

[0038] The asparagus extract includes any of an extract obtained from asparagus as an extraction raw material, a diluted or concentrated solution of the extract, or a crude or purified product thereof. In addition, a dried product obtained by drying the extract is also considered to be an extract.

[0039] When an asparagus extract is obtained by solvent extraction, the extraction solvent should be one that can extract the components contained in asparagus that have the effect of improving skin barrier function (in particular, the effect of promoting the formation of tight junctions and the synthesis of stratum corneum ceramide precursors). Specific examples include water, ethanol, isopropyl alcohol, acetone, glycerin, ethylene glycol, butylene glycol, propylene glycol, methanol, dichloromethane, dichloroethane, chloroform, hexane, ethyl acetate, etc. As the extraction solvent, cold water, room temperature water, warm water, hot water, etc. can be used as long as the effects of the present invention are not impaired. These extraction solvents can be used alone or as a mixture of two or more of them. When a mixture of two or more solvents is used as the extraction solvent, the mixing ratio can be appropriately adjusted.

[0040] The extraction solvent may contain additives within the range that does not impair the effects of the present invention, such as a pH adjuster and a viscosity adjuster.

[0041] From the standpoint of extraction efficiency, safety to the environment and living organisms, and production costs, a suitable example of an asparagus extract is a solvent extract of asparagus with one or more extraction solvents selected from water, ethanol, and butylene glycol. By using one or more extraction solvents selected from water, ethanol, and butylene glycol as the extraction solvent, it is possible to obtain an asparagus extract having excellent skin barrier function improving effects (particularly, the effect of promoting the formation of tight junctions and the synthesis of stratum corneum ceramide precursors).

[0042] When a mixed solvent of water and ethanol is used as the extraction solvent, the mixing ratio of water and ethanol may be appropriately adjusted within a range in which the components contained in asparagus that exhibit the skin barrier function improving effect (particularly the effect of promoting the formation of tight junctions and the effect of promoting the synthesis of stratum corneum ceramide precursors) can be extracted. For example, when the mixed solvent of water and ethanol is 100% by volume, the ratio of ethanol is 0.1 to 95% by volume.

[0043] There are no particular limitations on the amount of extraction solvent relative to the amount of asparagus used as a raw material, but it is usually about 1 to 100 times the amount of asparagus by weight.

[0044] The method for extracting a solvent extract from asparagus may be any method that can extract the components contained in asparagus that have the effect of improving skin barrier function (in particular, the effect of promoting the formation of tight junctions and the synthesis of stratum corneum ceramide precursors). A suitable example of a method for producing an asparagus extract includes contacting asparagus with an extraction solvent and stirring for a certain period of time until the active ingredients are sufficiently extracted from the asparagus in the extraction solvent. The extraction time is appropriately selected taking into consideration the type of extraction solvent and the ratio of asparagus to the extraction solvent, but when the extraction solvent is a mixed solvent of water and ethanol, the extraction time can be, for example, 1 to 24 hours.

[0045] The extraction process is carried out at room temperature to avoid deterioration of quality, but may be carried out under heated conditions to increase the efficiency of extraction. If necessary, the solvent may be removed by vacuum concentration or freeze-drying to increase the proportion of active ingredients in the asparagus extract.

[0046] In addition, filtration or centrifugation may be performed to remove residues contained in the extract. The obtained extract may be used as is, or may be subjected to treatments such as dilution, concentration, drying, purification, etc. according to conventional methods. If necessary, the solvent may be removed by concentration / drying such as vacuum concentration, freeze drying, spray drying, etc.

[0047] The active ingredient of the composition of the present invention may be a fraction of an extract obtained from the raw asparagus. In the present invention, the "fraction of the extract" can be obtained by a fractionation method such as filtration, centrifugation, or purification / separation treatment that is commonly used for asparagus extracts. Furthermore, the fraction of the obtained extract only needs to contain at least one component contained in asparagus that exhibits the effect of improving skin barrier function (particularly the effect of promoting the formation of tight junctions and the effect of promoting the synthesis of stratum corneum ceramide precursors), and may also contain other components.

[0048] The fractionation method may be any method that can separate the components contained in asparagus that exhibit the effect of improving skin barrier function (particularly the effect of promoting the formation of tight junctions and the effect of promoting the synthesis of ceramide precursors in the stratum corneum). As a fractionation method, various types of chromatography can be used, for example, gel filtration column chromatography, silica gel column chromatography, thin layer chromatography, high performance liquid chromatography, etc. These fractionation methods can be used alone or in combination of two or more of them.

[0049] <2. Application of the composition of the present invention> The composition of the present invention may contain the asparagus extract of the present invention or a fraction of said extract as is, but is usually used in combination with other ingredients that do not impair the effects of the present invention. The other ingredients may be any ingredients that can be used safely by humans, and are appropriately determined taking into consideration the form of use (medicinal use, topical use on the skin, oral use) described below. The blending ratio of the other components can be appropriately selected and determined depending on the purpose as long as it does not impair the effect of the present invention.

[0050] The composition of the present invention exhibits an excellent effect on improving skin barrier function due to the asparagus extract or a fraction of said extract contained therein, and therefore can be used as a "composition for improving skin barrier function."

[0051] The composition of the present invention exhibits excellent effects in promoting the formation of tight junctions and the synthesis of ceramide precursors in the stratum corneum due to the asparagus extract or a fraction of said extract contained therein. Therefore, in addition to being able to be used as a composition for improving skin barrier function as described above, the composition can also be used individually as a "composition for promoting the formation of tight junctions" and a "composition for promoting the synthesis of ceramide precursors in the stratum corneum."

[0052] The composition of the present invention can be used in any form depending on the purpose. That is, it can be used as a pharmaceutical composition, a composition for external application to the skin (such as a cosmetic composition) that does not fall under the category of a pharmaceutical composition, an oral composition (such as a supplement or functional food), etc.

[0053] When the composition of the present invention is used for improving skin barrier function, the amount of asparagus extract or a fraction of said extract contained therein is appropriately determined within a range such that the required amount can be ingested, taking into consideration the type and severity of symptoms involving skin barrier function, the form of the composition of the present invention, and the method of use.

[0054] Furthermore, when the composition of the present invention is used for promoting the formation of tight junctions, the amount of asparagus extract or a fraction of said extract contained therein is appropriately determined within a range such that the required amount can be ingested, taking into consideration the type and severity of symptoms in which tight junction formation is involved, the form of the composition of the present invention, and the method of use.

[0055] Furthermore, when the composition of the present invention is used to promote the synthesis of stratum corneum ceramide precursors, the amount of asparagus extract or a fraction of said extract contained is appropriately determined within a range such that the required amount can be ingested, taking into consideration the type and severity of symptoms in which stratum corneum ceramide precursor synthesis is involved, the form of the composition of the present invention, and the method of use.

[0056] Pharmaceutical Composition One suitable form of the composition of the present invention is a pharmaceutical composition comprising an effective amount of an asparagus extract or a fraction of said extract formulated together with a pharma- ceutical acceptable carrier (hereinafter, sometimes referred to as the "pharmaceutical composition of the present invention").

[0057] As used herein, the term "pharmaceutical composition" refers to a drug that is useful for at least one of preventing, treating, and improving symptoms of a target disease. In addition, as used herein, the term "pharmaceutical composition" includes not only pharmaceutical products but also quasi-drugs.

[0058] The pharmaceutical composition of the present invention may contain, in addition to the asparagus extract or a fraction of said extract, any other drug or pharmacologically acceptable ingredient within a range that does not impair the efficacy of the composition.

[0059] The pharmaceutical composition of the present invention is useful for at least one of preventing, treating, and improving symptoms of diseases involving skin barrier function. That is, "prevention" includes suppressing and delaying the onset of the disease or symptoms. Furthermore, "treatment" includes improving and relieving the pathology of the disease or symptoms, and suppressing the progression of the disease or symptoms.

[0060] There are no limitations on the diseases or symptoms involving skin barrier function that are the target of the pharmaceutical composition of the present invention, and specific examples include atopic dermatitis, inflammatory dermatitis, acne inflammation, allergic diseases, eczema, psoriasis, etc.

[0061] The pharmaceutical composition of the present invention may be in any form as long as it contains an asparagus extract or a fraction of the extract. The pharmaceutical composition of the present invention may be in the form of a tablet, pill, granule, powder, liquid, suspension, syrup, or capsule. The pharmaceutical composition of the present invention may be in the form of a solid or liquid, or a solid and a liquid, and is not particularly limited. The administration method may be oral or parenteral. Suitable embodiments of the pharmaceutical composition of the present invention include oral preparations and topical preparations for skin.

[0062] The pharmaceutical composition of the present invention can also be applied to animals other than humans, and therefore can also be made into a pharmaceutical composition for animals.

[0063] The amount of intake of the pharmaceutical composition of the present invention is not particularly limited as long as it is an amount that can exert the effects of the present invention, and is appropriately determined depending on the sex, weight, health condition, etc. of the subject to be ingested. The intake amount may be taken once or may be taken in several divided doses per day. In addition, from the viewpoint of prevention, treatment, and symptom improvement of the target disease, it is preferable to take the pharmaceutical composition of the present invention continuously.

[0064] <Forms other than pharmaceutical compositions> As mentioned above, the asparagus extract or a fraction thereof may be incorporated into products other than pharmaceuticals and quasi-drugs. Among such forms, preferred examples include compositions for topical application to the skin and compositions for oral administration (excluding pharmaceutical compositions).

[0065] (Composition for external use on the skin) One suitable form of the composition of the present invention is a composition for external application to the skin containing an asparagus extract or a fraction of said extract (hereinafter, sometimes referred to as the "composition for external application to the skin of the present invention").

[0066] A cosmetic composition, which is a typical example of the composition for external application to the skin of the present invention, will be described below; however, the composition for external application to the skin of the present invention is not limited to a cosmetic composition.

[0067] The composition for external use on the skin of the present invention has an excellent effect of improving the skin barrier function (particularly, the effect of promoting the formation of tight junctions and the synthesis of ceramide precursors in the stratum corneum), and as described above, can prevent or improve symptoms associated with the skin barrier function. For example, it can provide cosmetic effects against loss of firmness and elasticity of the skin, moisturizing of the skin, formation of wrinkles on the skin, etc.

[0068] The skin external composition of the present invention can be made into a desired formulation by appropriately blending a conventional cosmetic base material. The form is not particularly limited, and examples thereof include lotion, milky lotion, gel, cream, pack, hair tonic, hair cream foundation, aqueous ointment, spray, etc. In addition, in the present invention, the cosmetic composition is a concept that also includes bath compositions such as bath additives, body soap, and shampoo.

[0069] In addition, any optional ingredient used in ordinary cosmetics, skin medicines, and bath products can be added to the composition for external use on the skin of the present invention, so long as it does not impair the effects of the present invention. Specific examples of such optional ingredients include thickening / gelling agents, antioxidants, ultraviolet absorbing agents, coloring agents, sequestering agents, preservatives, pH adjusting agents, fragrances, beeswax, etc. The blending ratio of these optional ingredients can be appropriately selected and determined depending on the purpose.

[0070] The asparagus extract or a fraction of said extract may be blended in a composition for topical application to the skin at any ratio, but the ratio is selected within a range that can contribute to improving the skin barrier function (in particular, promoting the formation of tight junctions and promoting the synthesis of stratum corneum ceramide precursors).

[0071] The composition for external application on the skin of the present invention can also be applied to animals other than humans, and therefore can also be made into a composition for external application on the skin for animals.

[0072] (Oral Composition) One suitable form of the composition of the present invention is an oral composition containing an asparagus extract or a fraction of said extract (hereinafter, sometimes referred to as the "oral composition of the present invention"). The oral composition of the present invention has an excellent effect of improving skin barrier function (particularly, the effect of promoting the formation of tight junctions and the effect of promoting the synthesis of ceramide precursors in the stratum corneum), and is therefore suitable for use in functional foods, supplements, and the like.

[0073] Hereinafter, functional foods, supplements, and food additives, which are typical examples of the oral composition of the present invention, will be described; however, the oral composition of the present invention is not limited to functional foods, supplements, and food additives.

[0074] The amount of intake of the oral composition of the present invention is not particularly limited as long as it is an amount that can exert the effects of the present invention, and is appropriately determined depending on the form of the oral composition of the present invention, the sex, weight, health condition, etc. of the subject to be ingested. The intake amount may be taken once or may be taken in several divided doses per day. In addition, in order to obtain the action or efficacy of the oral composition of the present invention, it is preferable to continuously take the oral composition of the present invention.

[0075] The composition of the present invention can be mixed with various foods and beverages to form functional foods that can be easily taken orally on a daily basis, and can also be easily taken over a long period of time. The term "functional food" as used herein means, in addition to general foods, foods and / or beverages that are taken for the purpose of maintaining health, and is a concept that includes health functional foods (specified health foods, nutrient functional foods, or functional food), health foods, nutritional supplements, nutritional insurance foods, etc. Among these, health functional foods, such as specified health foods, nutrient functional foods, and functional food, are preferred forms of functional foods. When commercializing functional foods, various additives used in foods, specifically colorants, preservatives, thickening stabilizers, antioxidants, bleaching agents, antibacterial and antifungal agents, acidulants, seasonings, emulsifiers, strengthening agents, manufacturing agents, flavors, etc. may be added.

[0076] There is no particular limit to the foods and beverages that can be used as functional foods. For example, foods include sausages, hams, seafood products, jellies, candies, chewing gums, and other foods. Beverages include various types of tea, green juice drinks, soft drinks, alcoholic beverages, and nutritional drinks.

[0077] The oral composition of the present invention can also be used in the form of a supplement. The form of the supplement is not particularly limited, and may be any form such as a tablet, powder, granule, capsule, sugar-coated tablet, film, lozenge, chewable, solution, emulsion, suspension, etc. The supplement of the present invention may contain any ingredient that is generally used as a supplement, in addition to the composition of the present invention.Such ingredients include, for example, amino acids, peptides, vitamins such as vitamin E, vitamin C, vitamin A, vitamin B, and folic acid, minerals, sugars, inorganic salts, citric acid or its salts, tea extract, oils and fats, tonic ingredients such as propolis, royal jelly, and taurine, herbal extracts such as ginger extract and ginseng extract, and herbs: collagen, etc.

[0078] The asparagus extract or a fraction of said extract may be incorporated into functional foods or supplements at any ratio, but the ratio is selected within a range that contributes to improving skin barrier function (in particular, promoting the formation of tight junctions and the synthesis of stratum corneum ceramide precursors).

[0079] When the oral composition of the present invention is used as a health food, the product may display the action or efficacy brought about by the asparagus extract or a fraction of said extract of the present invention, for example, on the product itself, packaging, container, packaging, or other storage items, or on advertisements, etc.

[0080] In addition, examples of claims regarding the action or efficacy that are added to products when they are commercialized include "for skin that can withstand UV rays," "for moisturized skin," "retains moisture in the skin," "provides a skin barrier," "protects the skin," "prevents loss of skin elasticity," "prevents wrinkles," "retains moisture in the skin," etc.

[0081] In addition, the composition of the present invention can be used as a food additive either by itself or by adding other ingredients to it. There is no particular limitation on the other ingredients as long as they can be used as food and beverage additives. There is no particular limitation on the beverages and foods to which the food additive is added.

[0082] The composition of the present invention may be blended into a food additive at any ratio, but the blending ratio is selected within a range that can contribute to improving skin barrier function (particularly, promoting the formation of tight junctions and promoting the synthesis of stratum corneum ceramide precursors).

[0083] The oral composition of the present invention can also be applied to animals other than humans, and can therefore be used as an oral composition for animals, such as pet food. EXAMPLES

[0084] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples. In addition, the "%" shown in the mixed solvent in the examples indicates "volume %" unless otherwise specified. In addition, when the extraction solvent is a mixed solvent, the balance of water is omitted. For example, when it is written "50% ethanol", it means a mixed solvent of 50 volume % ethanol and 50 volume % water.

[0085] 1. Preparation of Asparagus Extract The asparagus extract of Example 1 was produced in the following manner.

[0086] <1-1. Preparation of dried asparagus> Asparagus (Asparagus officinalis) from Saga Prefecture was used as the raw asparagus. The raw asparagus was cut 25 cm from the tip (top) and the lower part (stalk and cut part) was used as the raw asparagus. The raw asparagus was heat-treated with steam at 100°C for 7 minutes using a steam convection oven and then freeze-dried. The resulting dried product was pulverized to obtain dried pulverized asparagus.

[0087] <1-2. Preparation of asparagus extract> To the dried and ground asparagus obtained in the above <1-1. Preparation of dried asparagus>, 20 times the weight of 80% (v / v) ethanol solution was added, and the mixture was stirred and extracted at room temperature for 4 hours. After that, the mixture was centrifuged (12,000×g, 30 minutes, 4° C.), and the resulting supernatant was used as the asparagus extract, and the asparagus extract of Example 1 was obtained.

[0088] 2. Evaluation method <2-1. Test using human 3D cultured epidermis> Transfer the culture cup of the LabCyte EPI-MODEL24 (J-TEC) to an assay plate containing 500 μL of assay medium (J-TEC) and incubate in CO. 2 After culturing for 18 hours in an incubator, the culture cup was transferred to an assay plate to which the assay medium or the assay medium containing the asparagus extract of Example 1 was dispensed, and the culture cup was incubated in a CO 2 After 4 hours of incubation, the cells were exposed to UVB (280-320 nm, lamp: Toshiba SH1002MA, filter: Asahi spectra SH0325) at 150 mJ / cm 2 The culture cup was transferred to an assay plate into which 1.0 mL of the assay medium or the assay medium containing the asparagus extract of Example 1 had been dispensed, and the culture cup was then exposed to CO. 2 The epidermis was cultured in an incubator for 2 or 8 days. As a control, the assay medium was added instead of the asparagus extract and the epidermis was not exposed to UVB. The epidermis cultured for 2 or 8 days was used for transepidermal water loss (TEWL) and gene analysis by RT-qPCR. Although not shown, an MTT assay (cell viability assay) was performed using three-dimensionally cultured human epidermis, and it was confirmed that there was no effect on cell viability in all groups.

[0089] <2-2. Evaluation in Transepidermal Water Loss (TEWL) Measurement> Transepidermal water loss (TEWL) was measured using human 3D cultured epidermis LabCyte EPI-MODEL24. For the measurement, cultured epidermis after 2 or 8 days of culture was used.

[0090] After 2 or 8 days of incubation, the LabCyte EPI-MODEL24 was placed on a HIENAI MAT (Cosmo Bio) maintained at 32°C with the lid open for 5 minutes. After that, a Tewitro TW24 (Courage+Khazaka electronic GmbH) was set and covered with a box (W340mm x H320mm x D275mm), and the transepidermal water loss was measured for 900 seconds. The transepidermal water loss for 20 seconds from 881 to 900 seconds, when the measured value stabilized, was used for analysis. These operations were performed in a sterile environment using a table coach (Koken).

[0091] The transepidermal water loss of the control was set at 100, and the relative value of the transepidermal water loss of the test group to which the asparagus extract of Example 1 was added was calculated. The results are shown in Figures 2(a) and 2(b). In Figure 2, * indicates p<0.05 (both based on the Tukey-Kramer test).

[0092] As shown in Figures 2(a) and 2(b), TEWL increased in the UVB-irradiated samples compared to the non-irradiated samples (control), whereas the increase in TEWL tended to be suppressed in the test group to which the asparagus extract (200 μg / mL) of Example 1 was added. In particular, as shown in Figure 2(b), it was confirmed that the increase in TEWL caused by UVB irradiation was significantly suppressed by adding the asparagus extract of Example 1 in the human 3D cultured epidermis after 8 days of culture. Therefore, it was demonstrated that the asparagus extract is useful for suppressing an increase in transepidermal water loss (TEWL).

[0093] <2-3. Genetic analysis using RT-qPCR method> Genetic analysis was performed using RT-qPCR method using human 3D cultured epidermis LabCyte EPI-MODEL24. After 2 days of culture, the cultured epidermis was disrupted using a BioMasher II. After disruption, total RNA was extracted using TRIzol Reagent (Thermo).

[0094] cDNA synthesis was performed using PrimeScript TM PrimeScript was used according to the manufacturer's protocol using RT Master Mix (Takara). TM RT Master Mix, 500 ng of total RNA, and sterilized ultrapure water were mixed to prepare a 10 μL reaction solution, and reverse transcription was performed. Real-time PCR was performed using TB Green Premix EX Taq TM The primer sets used for real-time PCR are shown in Table 1.

[0095] [Table 1]

[0096] PCR reactions were performed using BIO-RAD and c1000 Touch TM Thermal Cycler, CFX 96 TM The analysis was performed using the Real-Time System and the BIO-RAD CFX Manager version 3.1, using the Comparative CT method with GAPDH as an internal standard.

[0097] <2-3-1. Evaluation of the effect of promoting tight junction formation> The effect of the asparagus extract of Example 1 on tight junction formation was investigated using human 3D cultured epidermis LabCyte EPI-MODEL24. RT-qPCR was performed as described above in <2-3. Genetic analysis by RT-qPCR> to analyze the expression of a gene (Claudin) involved in tight junction formation. The formation of tight junctions was evaluated using Claudin 1, Claudin 4, and GAPDH shown in Table 1.

[0098] The expression level of the Claudin 1 gene or Claudin 4 gene in the control was set to 100, and the relative expression level of the Claudin 1 gene or Claudin 4 gene in the test group to which the asparagus extract of Example 1 was added was calculated. The results are shown in Figure 3(a) and Figure 3(b). In Figure 3, a significant difference test was performed on the samples to which the assay medium was added instead of the asparagus extract and which were irradiated with UVB, and * indicates p<0.05 and ** indicates p<0.01 (both by Tukey-Kramer test).

[0099] As shown in FIG. 3(a), the expression level of Claudin 1 was decreased in UVB-irradiated human 3D cultured epidermis, whereas the decrease in the expression level was suppressed in the case of the epidermis cultured in a medium supplemented with the asparagus extract of Example 1. In addition, as shown in FIG. 3(b), the decrease in the expression level of Claudin 4 was also suppressed in the case of the epidermis cultured in a medium supplemented with the asparagus extract of Example 1. Therefore, it was shown that the asparagus extract is useful for maintaining and improving tight junctions by suppressing the decrease in the expression level of Claudin or enhancing the expression level.

[0100] <2-3-2. Evaluation of the effect of promoting synthesis of ceramide precursors in the stratum corneum> The effect of the asparagus extract of Example 1 on the synthesis of ceramide precursors in the stratum corneum was investigated using human 3D cultured epidermis LabCyte EPI-MODEL24. RT-qPCR was performed as described above in <2-3. Genetic analysis by RT-qPCR> to analyze the expression of genes involved in the synthesis of stratum corneum ceramide precursors (CerS, SMS, SPT). The synthesis of ceramide precursors in the stratum corneum was evaluated using CerS3, SMS1, SMS2, SPT1, SPT2, and GAPDH shown in Table 1.

[0101] The expression levels of the CerS3 gene, SMS1 gene, SMS2 gene, SPT1 gene or SPT2 gene in the control were set at 100, and the relative values ​​of the expression levels of the CerS3 gene, SMS1 gene, SMS2 gene, SPT1 gene or SPT2 gene in the test group to which the asparagus extract of Example 1 was added were calculated. The results are shown in Figures 4(a) to (e). In Figure 4, a significant difference test was performed on the samples to which the assay medium was added instead of the asparagus extract and which were irradiated with UVB, with * indicating p<0.05 and ** indicating p<0.01 (both by Tukey-Kramer test).

[0102] As shown in Figures 4(a) to (e), when comparing human 3D cultured epidermis irradiated with UVB, the expression levels of genes involved in the synthesis of stratum corneum ceramide precursors (CerS3, SMS1, SMS2, SPT1, SPT2) increased in a concentration-dependent manner in the cultured epidermis cultured in a medium containing the asparagus extract of Example 1. In particular, as shown in Figures 4(a), 4(c), and 4(e), the expression levels of the CerS3 gene, SMS2 gene, and SPT2 gene increased significantly. These results indicate that the asparagus extract is useful for maintaining and improving stratum corneum ceramide precursors in the body by suppressing the decrease in the expression levels of genes involved in the synthesis of stratum corneum ceramide precursors, such as the CerS3 gene, SMS2 gene, and SPT2 gene, or by enhancing the expression levels. [Industrial Applicability]

[0103] The composition of the present invention has excellent effects of improving skin barrier function, promoting the formation of tight junctions, and promoting the synthesis of stratum corneum ceramide precursors, and is therefore effective for improving skin firmness, elasticity, wrinkles, moisturizing, etc. In addition, the composition of the present invention can be suitably used in various pharmaceutical compositions, compositions for external use on the skin, compositions for oral administration, etc.

Claims

1. Contains asparagus extract as an active ingredient. The asparagus extract is a solvent extract of asparagus with one or more extraction solvents selected from water, ethanol and butylene glycol; A composition for improving skin barrier function using transepidermal water loss as an indicator (excluding cases where it is used in combination with sphingolipids).

2. A composition comprising asparagus extract as an active ingredient, A composition for promoting claudin 1 expression and / or claudin 4 expression, wherein the asparagus extract is a solvent extract of asparagus using one or more extraction solvents selected from water, ethanol and butylene glycol.

3. A composition comprising asparagus extract as an active ingredient, The asparagus extract is a solvent extract of asparagus with one or more extraction solvents selected from water, ethanol and butylene glycol; A composition for promoting one or more of serine palmitoyltransferase expression, ceramide synthesase expression, and sphingomyelin synthesase expression.

4. The composition according to any one of claims 1 to 3, which is for pharmaceutical use.

5. 4. The composition according to claim 1, which is for external application to the skin (excluding cases where it is for pharmaceutical use).

6. The composition according to any one of claims 1 to 3, which is for oral use (excluding cases where it is for pharmaceutical use).

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