KR20220151196A - A composition for suppressing or improving a decrease in skin barrier function, a composition for suppressing or improving a decrease in the expression of type 4 collagen, and a screening method for a substance having an effect of suppressing or improving a decrease in skin barrier function - Google Patents

Abstract

The present invention is a composition for suppressing or improving a decrease in skin barrier function, a composition for suppressing or improving a decrease in type 4 collagen expression, and suppressing a decrease in skin barrier function, which can suppress or improve the decrease in skin barrier function. Or a method for improving skin barrier function, a method for suppressing or improving the decrease in the expression of type 4 collagen, and a method for screening a substance having an effect of suppressing or improving the decrease in skin barrier function. . The present invention is an extract of a plant of the genus Rosaceae, an extract of grape seeds and / or skin, an extract of yeast of the genus Yarrowia , an extract of a plant of the genus Camellia and camellia, an extract of a plant of the genus Rosaceae, and a plant of the genus Perilla and Mint It relates to a composition for inhibiting or improving a decrease in skin barrier function through inhibition or improvement in the expression of type 4 collagen, comprising at least one extract selected from the group consisting of plant extracts as an active ingredient.

Description

A composition for suppressing or improving a decrease in skin barrier function, a composition for suppressing or improving a decrease in the expression of type 4 collagen, and a screening method for a substance having an effect of suppressing or improving a decrease in skin barrier function

The present invention relates to a composition for inhibiting or improving a decrease in skin barrier function and a composition for inhibiting or improving a decrease in the expression of type 4 collagen. Further, the present invention relates to a method for suppressing a decrease in skin barrier function or improving a skin barrier function, and a method for suppressing a decrease in type 4 collagen expression or improving the expression. The present invention also relates to a method for screening a substance having an action of suppressing or improving a decrease in skin barrier function, and the like.

The skin is largely divided into two layers, the epidermis and the dermis, from the surface. The epidermis is composed of four layers: the deep layer, the basal layer, the spinous layer, the granular layer, and the stratum corneum, and the basal layer adheres to the dermis through the basement membrane. The basement membrane is composed of components such as type 4 collagen and type 7 collagen, and has functions such as binding the basal layer of the epidermis and the dermis, but its function declines with age.

The skin has a barrier function of preventing foreign matter from entering the living body from the outside or preventing excessive evaporation of moisture in the body from the skin to retain moisture in the skin. When the skin barrier function is lowered due to causes such as aging, it causes skin dryness and the like. Patent Document 1 describes a skin barrier function improving agent containing an extract of rooibos ( Aspalathus linearis ) and the like.

Patent Document 1: Japanese Unexamined Patent Publication No. 2009-256244

An object of the present invention is to provide a composition for suppressing or improving a decrease in skin barrier function that can suppress or improve a decrease in skin barrier function. Another object of the present invention is to provide a composition for suppressing or improving the decrease in the expression of type 4 collagen. Another object of the present invention is to provide a method for suppressing a decrease in skin barrier function or improving the function, and a method for suppressing a decrease in the expression of type 4 collagen or improving the expression. Furthermore, an object of the present invention is to provide a screening method for a substance having an effect of suppressing a decrease in skin barrier function or improving it.

As a result of research to solve the above problems, the inventors of the present invention have found that the decrease in skin barrier function is suppressed or the function is improved by suppressing the decrease in the expression of type 4 collagen in skin cells or improving the expression. . Based on these findings, the present inventors further studied and completed the present invention.

The present invention is not limited thereto, but includes the following compositions for suppressing or improving a decrease in skin barrier function, compositions for suppressing or improving a decrease in type 4 collagen expression, and the like.

[1] Rosaceae Rosaceae plant extract, grape seed and/or fruit skin extract, Yarrowia genus yeast extract, Camellia family Camellia plant extract, Rosaceae genus plant extract, and perilla family Mint plants A composition for suppressing or improving a decrease in skin barrier function through suppression or improvement in the expression of type 4 collagen, comprising at least one extract selected from the group consisting of extracts as an active ingredient.

[2] Rosaceae Rosaceae plant is rose, Yarrowia ( Yarrowia ) yeast is Yarrowia lipolytica ( Yarrowia lipolytica ), Camellia and Camellia plant is tea tree, Rosaceae turmeric plant is meadowsweet, Perilla and mint plant is peppermint Phosphorus, the composition for suppressing or improving the decrease in skin barrier function according to the above [1].

[3] Rose petals containing at least one compound selected from the group consisting of delphinidin-type anthocyanins, rosacyanin compounds, and rosadelphin compounds in Rosaceae plants, Camellia and Camellia plants in tea leaves, and plants in Rosaceae The composition for suppressing or improving a decrease in skin barrier function according to [1] or [2] above, wherein the flower of Meadowsweet, a plant of the genus Perilla and Mint, is a peppermint sheath.

[4] A composition for suppressing or improving a decrease in skin barrier function, comprising as an active ingredient an extract of a plant of the genus Pompanoaceae and/or an extract of a plant of the genus Plantain and/or Plantago.

[5] The composition for suppressing or improving a decrease in skin barrier function according to [4] above, wherein the plant of the genus Pomponaceae and the genus Pompanophyllis is Hamamelis, and the plant of the genus Plantain and Plantain is Pylori.

[6] The composition for suppressing or improving a decrease in skin barrier function according to [4] or [5] above, wherein the plant of the genus Pomonenaceae is a leaf of Hamamelis, and the plant of the genus Plantain and the genus Plantain is a seed of Pyaline.

[7] The composition for suppressing or improving the decrease in skin barrier function according to any one of [4] to [6] above, which suppresses the decrease in the skin barrier function or improves the skin barrier function by suppressing or improving the decrease in the basement membrane function. .

[8] The composition for suppressing or improving a decrease in skin barrier function according to [7], wherein the decrease in basement membrane function is suppressed or improved through suppression or improvement in expression of type 4 collagen.

[9] A composition for inhibiting or improving a decrease in skin barrier function, comprising an agent for inhibiting or improving a decrease in basement membrane function as an active ingredient.

[10] Inhibiting a decrease in skin barrier function according to [9] above, wherein the agent for inhibiting or improving a decrease in basement membrane function inhibits a decrease in basement membrane function or improves a basement membrane function by inhibiting or improving expression of type 4 collagen. Or a composition for improvement.

[11] Rosaceae Rosaceae plant extract, grape seed and / or fruit skin extract, Yarrowia ( Yarrowia ) genus yeast extract, Camellia and Camellia plant extract, Pomponaceae Pomponaceae plant extract, Rosaceae Turtle plant extract A composition for inhibiting or improving the expression of type 4 collagen, comprising, as an active ingredient, at least one extract selected from the group consisting of plant extracts of the genus Plantain and plantain, and extracts of plants belonging to the genus Perilla and Mint.

[12] Rosaceae Rosaceae plants are roses, Yarrowia ( Yarrowia ) genus yeast Yarrowia lipolytica ( Yarrowia lipolytica ), Camellia and Camellia plants are tea trees, Pomponaceae Pomponaceae plants are Hamamelis, Rosaceae Turtle grass plants The composition for suppressing or improving the decrease in the expression of type 4 collagen according to [11] above, wherein Meadowsweet, plantain and plantain plant is royal plantain, and perilla and mint plant are peppermint.

[13] Rose petals containing at least one compound selected from the group consisting of delphinidin-type anthocyanins, rosacyanin compounds, and rosadelphin compounds in Rosaceae plants, Camellia and Camellia plants in tea leaves, Pomponaceae Pomponaceae plants The expression of the type 4 collagen according to [11] or [12] above, in which the leaf of Hamamelis, the plant of the genus Rosaceae, is a flower of Meadowsweet, the plant of the genus Plantain and the genus Plantain is royal plantain seed, and the plant of the genus Perilla and peppermint is a peppermint sheath. A composition for inhibiting or improving the decrease of

[14] The composition according to any one of [1] to [13], which is an external preparation for skin.

[15] The composition according to any one of [1] to [14], which is a cosmetic.

[16] An extract of a plant of the genus Rosaceae, an extract of grape seeds and/or skin, Yarrowia ( Use of at least one extract selected from the group consisting of a yeast extract of the genus Yarrowia , an extract of a plant of the genus Camellia and Camellia, an extract of a plant of the genus Rosaceae, and an extract of a plant of the genus Perilla and Mint.

[17] Use of an extract of a plant belonging to the genus Pomoniapsis and/or a plant belonging to the genus Plantain and/or a plantain for suppressing a decrease in skin barrier function or improving the skin barrier function.

[18] Extracts of plants of the genus Rosaceae, extracts of grape seeds and / or peels, extracts of Yarrowia genus yeast, Camellia and Camellia genus for inhibiting or improving the expression of type 4 collagen, Use of at least one extract selected from the group consisting of a plant extract, an extract of a plant of the genus Pomponaceae, an extract of a plant of the genus Rosaceae, an extract of a plant of the genus Plantain and plantain, and an extract of a plant of the genus Perilla and Mint.

[19] Rosaceae Rosaceae plant extract, grape seed and / or fruit skin extract, Yarrowia ( Yarrowia ) genus yeast extract, Camellia Camellia plant extract, Rosaceae plant extract, and Perilla and Mint plants A method for suppressing the decrease in skin barrier function or improving the skin barrier function by suppressing or improving the decrease in the expression of type 4 collagen, by administering at least one extract selected from the group consisting of extracts of.

[20] A method for suppressing a decrease in skin barrier function or improving a skin barrier function, comprising administering an extract of a plant of the genus Pompanoaceae and/or an extract of a plant of the genus Plantain and/or the genus Plantain.

[21] Rosaceae Rosaceae Plant Extract, Grape Seed and / or Fruit Skin Extract, Yarrowia ( Yarrowia ) Genus Yeast Extract, Camellia Camellia Plant Extract, Pomponaceae Pomponaceae Plant Extract, Rosaceae Turtle Plant Extract A method for suppressing or improving the expression of type 4 collagen, comprising administering at least one extract selected from the group consisting of an extract of a plant of the genus Plantain and plantain, and an extract of a plant of the genus Perilla and mint.

[22] step of culturing human epidermal keratinocytes in vitro in a medium containing a test substance or a control medium not containing the test substance, and skin cells cultured in a medium containing the test substance and control A step of comparing the expression level of type 4 collagen in control skin cells cultured in the culture medium, wherein the expression level of type 4 collagen in skin cells cultured in the medium containing the test substance is equal to the type 4 collagen expression level in the control skin cells. A method for screening a substance having an action of suppressing or improving a decrease in skin barrier function, wherein the substance to be tested is selected as a substance having an action of suppressing or improving a decrease in skin barrier function when the amount of collagen expression is higher than that of the test substance.

According to the present invention, it is possible to provide a composition for suppressing or improving a decrease in skin barrier function capable of suppressing or improving a decrease in skin barrier function. Furthermore, according to the present invention, a composition for suppressing or improving the decrease in the expression of type 4 collagen can be provided. Furthermore, according to the present invention, a method for suppressing a decrease in skin barrier function or improving the function, and a method for suppressing a decrease in the expression of type 4 collagen or improving the expression can be provided. ADVANTAGE OF THE INVENTION According to this invention, the screening method of the substance which has the action of suppressing or improving the decline of a skin barrier function can be provided.

A first aspect of the present invention is an extract of a plant of the genus Rosaceae, an extract of grape seeds and / or skin, an extract of yeast of the genus Yarrowia , an extract of a plant of the genus Camellia and Camellia, an extract of a plant of the genus Pomoniaaceae, Rosaceae A composition for inhibiting or improving the decrease in the expression of type 4 collagen, comprising, as an active ingredient, at least one extract selected from the group consisting of an extract of a plant of the genus Turtles, an extract of a plant of the genus Plantain and plantain, and an extract of a plant of the genus Perilla and Mint. . The composition for suppressing or improving the decrease in the expression of type 4 collagen of the first aspect of the present invention is also referred to as the composition of the first aspect of the present invention.

The extract has an action of suppressing the decrease in the expression of type 4 collagen or improving the expression. Suppression of the decrease in the expression of type 4 collagen includes suppressing the progress of the decrease in the expression of type 4 collagen, maintaining the expression of type 4 collagen, reducing (relieving) the degree of decrease in the expression of type 4 collagen, and the like. include Improving the expression of type 4 collagen includes restoring the expression of type 4 collagen and the like. Restoration includes at least partially restoring. The expression of type 4 collagen includes the expression of a gene encoding type 4 collagen and the expression of type 4 collagen at the protein level, and either one or both of these may be used. Type 4 collagen is preferably human type 4 collagen, more preferably human type 4 collagen ?1 chain. Human type 4 collagen α1 chain is a protein encoded by the gene type 4 collagen A 1 (COL4A1). In one embodiment, the composition for suppressing or improving the decrease in the expression of type 4 collagen of the present invention suppresses the decrease in the expression of the α1 chain of human type 4 collagen or improves the expression, thereby reducing the expression of type 4 collagen in humans. It can be used to inhibit degradation or to ameliorate the expression.

The expression of type 4 collagen is reduced (decreased in the amount of expression) due to external factors such as ultraviolet rays and internal factors such as aging.

As shown in the following examples, suppressing or improving the expression of type 4 collagen in epidermal keratinocytes suppressed or improved the decrease in transepithelial electrical resistance (TEER) of the skin. This indicates that the decrease in the skin barrier function was suppressed or the skin barrier function was improved. Although the detailed mechanism by which the decrease in skin barrier function is suppressed or improved by suppressing or improving the expression of type 4 collagen is not clear, since type 4 collagen is a component of the basement membrane, the decrease in the expression of type 4 collagen By suppressing or improving the function, it is assumed that the decrease in the function of the basement membrane is suppressed or the function is improved, and as a result of maintaining the homeostasis of the skin, the decrease in the skin barrier function is suppressed or the function is improved. Therefore, it is expected that the decrease in the skin barrier function can be suppressed or the skin barrier function can be improved by suppressing the deterioration of the basement membrane function or improving the function. Since the plant or yeast extract has an excellent effect of suppressing or improving the expression of type 4 collagen, it can be used to suppress the decrease in basement membrane function or to improve the function.

From the viewpoint of suppressing or improving the expression of type 4 collagen, in the composition of the first aspect of the present invention, the extract is an extract of a plant of the genus Rosaceae, an extract of grape seeds and / or peel, and Rosaceae It is preferably at least one selected from the group consisting of plant extracts of the genus Grass.

The skin barrier function refers to a barrier function of the skin that retains moisture in the skin by preventing foreign substances from entering the living body from the outside or preventing excessive evaporation of moisture in the body from the skin. Suppression of the decrease in the skin barrier function includes suppressing the progress of the decrease in the skin barrier function, maintaining the skin barrier function, reducing (relieving) the degree of decrease in the skin barrier function, and the like. Improving the skin barrier function includes restoring the skin barrier function and the like. The decrease in the skin barrier function may be a decrease in the skin barrier function due to aging or a decrease in the skin barrier function due to exposure to ultraviolet rays.

As described above, suppressing or improving the decrease in the basement membrane function makes it possible to suppress the decrease in the skin barrier function or improve the function.

A second aspect of the present invention is a composition for suppressing or improving a decrease in skin barrier function, comprising an agent for suppressing or improving a decrease in basement membrane function as an active ingredient. The composition for suppressing or improving the decrease in skin barrier function of the second aspect of the present invention is also referred to as the composition of the second aspect of the present invention.

As a function of the basement membrane, functions such as a mechanical support connecting the epidermis and the dermis and a selective filter for material exchange between the epidermis and the dermis are known. Suppressing the decrease in expression of basement membrane components such as type 4 collagen or improving the expression is effective for suppressing or improving the decrease in basement membrane function.

The agent for suppressing or improving a decrease in basement membrane function is not particularly limited as long as it has an action for suppressing or improving a decrease in basement membrane function, for example, an action for suppressing or improving expression of components of a basement membrane. The agent for suppressing or improving the basement membrane function is preferably one that suppresses or improves the basement membrane function by suppressing or improving the expression of type 4 collagen. As an agent for suppressing or improving a decrease in basement membrane function, for example, an extract of the above plant or yeast having an effect of suppressing or improving the expression of type 4 collagen can be used. In one embodiment, as an agent for inhibiting or improving a decrease in basement membrane function, an extract of a plant of the genus Rosaceae, an extract of grape seeds and/or skin, an extract of yeast of the genus Yarrowia , an extract of a plant of the genus Camellia and Camellia, Pomponaceae At least one extract selected from the group consisting of a plant extract of the genus Rosaceae, an extract of a plant of the genus Rosaceae, an extract of a plant of the genus Plantain and plantain, and an extract of a plant of the genus Perilla and Mint may be used.

Among the above extracts, at least one selected from the group consisting of extracts of plants of the genus Rosaceae, extracts of grape seeds and/or rind, and extracts of plants of the genus Rosaceae is preferred as an agent for inhibiting or improving a decrease in basement membrane function.

A third aspect of the present invention is an extract of a plant of the genus Rosaceae, an extract of grape seeds and/or skin, an extract of yeast of the genus Yarrowia , an extract of a plant of the genus Camellia and a plant of the genus Rosaceae, and , A composition for suppressing or improving a decrease in skin barrier function through suppression or improvement in the expression of type 4 collagen, comprising at least one extract selected from the group consisting of extracts of plants belonging to the genus Perilla and Mint as an active ingredient. The composition for suppressing or improving the decrease in skin barrier function according to the third aspect of the present invention is also referred to as the composition according to the third aspect of the present invention.

In the composition of the third aspect of the present invention, the extract is an extract of grape seeds and / or skin, an extract of yeast of the genus Yarrowia , an extract of a plant of the genus Camellia and camellia, and an extract of a plant of the genus Rosaceae It is preferably at least one selected from the group consisting of.

A fourth aspect of the present invention is a composition for suppressing or improving a decrease in skin barrier function, comprising as an active ingredient an extract of a plant of the genus Pomponaceae and/or an extract of a plant of the genus Plantain and/or of the genus Plantain. The composition for suppressing or improving the decrease in skin barrier function of the fourth aspect of the present invention is also referred to as the composition of the fourth aspect of the present invention. The composition of the fourth aspect of the present invention preferably contains, as an active ingredient, an extract of a plant of the genus Pomponaceae.

The extract used in the composition of the third aspect and the composition of the fourth aspect of the present invention has an action of suppressing or improving the expression of type 4 collagen. This makes it possible to suppress or improve the degradation of the basement membrane function. In one aspect, the composition for inhibiting or improving the decrease in skin barrier function of the third and fourth aspects of the present invention suppresses the decrease in skin barrier function or improves the skin barrier function through suppression or improvement of basement membrane function can be used to do In addition, the composition for suppressing or improving the decrease in skin barrier function of the fourth aspect of the present invention suppresses or improves the decrease in basement membrane function through suppression or improvement in the expression of type 4 collagen, thereby suppressing the decrease in skin barrier function or to improve skin barrier function. In one aspect, the composition for suppressing or improving the decrease in skin barrier function of the fourth aspect of the present invention can be used to suppress or improve the decrease in skin barrier function by suppressing or improving the decrease in the expression of type 4 collagen. .

Hereinafter, the composition for suppressing or improving the decrease in the expression of type 4 collagen of the first aspect of the present invention and the composition for suppressing or improving the decrease in skin barrier function of the second, third and fourth aspects of the present invention are collectively present. Also referred to as the composition of the invention. In the composition of the present invention, two or more of the above extracts may be used as active ingredients in combination.

In the present invention, a plant of the genus Rosaceae (Rosaceae Rosa) is a rose of the genus Rosaceae represented by Rosa hybrida (scientific name). As a plant of the genus Rosaceae, a rose containing at least one compound selected from the group consisting of delphinidin-type anthocyanins, rosacyanin compounds, and rosadelphin compounds is preferred, and delphinidin-type anthocyanins, rosacyanin compounds, and rosadelphin compounds in petals Roses containing at least one compound selected from the group consisting of are more preferred. Among them, roses containing delphinidin-type anthocyanin in petals are preferred.

Delphinidin-type anthocyanin is a pigment that is contained in a lot of purple or blue petals. Delphinidin-type anthocyanins include delphinidin, malvidin, petunidin, or compounds modified with sugars, acyl groups, or the like. Roses containing delphinidin-type anthocyanins include roses exhibiting a blue or light purple color tone, for example, roses of varieties such as Suntory Blue Rose Applause (registered trademark). The inclusion of delphinidin-type anthocyanins in roses can be confirmed by HPLC, for example, by a method described in Japanese Patent No. 5697040 and the like.

The rosacyanin compound is at least one compound selected from the group consisting of rosacyanin A1, rosacyanin A2 and rosacyanin B. Roses containing rosacyanin compounds may be roses containing one or more of the above compounds, but are preferably roses containing rosaceanin A1, rosadelphin A2 and rosadelphin B. Rosacyanin A1 is a compound in which R ¹ is a hydrogen atom in the following general formula (I), and rosacyanin A2 is a compound in which R ² is a hydrogen atom in the following general formula (II), B, which is not, is a compound in which R ³ is a hydrogen atom in the following general formula (III).

(In the above formula, the configuration (broken line) of the hydroxyl group at the 1st position of glucose indicates that the α-form and the β-form are tautomers. R ¹ represents a hydrogen atom or a hydroxyl group.)

(In the above formula, R ² represents a hydrogen atom or a hydroxyl group.)

(In the above formula, R ³ represents a hydrogen atom or a hydroxyl group.)

A rose containing a rosacyanin compound, which exhibits blue or light purple tones, such as Madame Biore, Purple Rain, Ravende, Manhattan Blue, Chantilly Lace, Blue Moon, Tassogare, Charles de Gaulle, and Violet Dolly , Blue Ribbon, Aozora, Lady X, Blue Bajo and Sterling Silver roses. These roses are preferable because petals contain a rosacyanin compound.

Rosadelphin compound is at least one compound selected from the group consisting of rosadelphin A1, rosadelphin A2 and rosadelphin B. A rose containing a rosadelphin compound may be a rose containing at least one of the above compounds, but is preferably a rose containing rosadelphin A1, rosadelphin A2 and rosadelphin B. Rosadelphin A1 is a compound in which R ¹ is a hydroxyl group in the general formula (I), Rosadelphin A2 is a compound in which R ² is a hydroxyl group in the general formula (II), and rosadelphin B is a compound in which R 1 is a hydroxyl group, In the general formula (III), R ³ is a hydroxyl group.

The rosardelphin compound does not exist in wild roses, and a gene encoding flavonoid 3',5'-hydroxylase (hereinafter also referred to as a flavonoid 3',5'-hydroxylase gene) is introduced by methods such as genetic recombination. Delphinidin, a blue pigment, is synthesized in a rose or a rose having a flavonoid 3',5'-hydroxylase gene obtained from a genetically modified rose as a parent, and it is a compound produced by combining gallate or telimagrandins. .

Roses containing the rosadelphin compound, roses in which the flavonoid 3',5'-hydroxylase gene was introduced by a method such as genetic recombination, and flavonoid 3',5'-hydroxylase obtained from such a genetically modified rose as a parent Roses with enzyme genes can be used. As such a rose, roses of varieties such as Suntory Blue Rose Applause (registered trademark) can be used, for example. It is known that the petals of Suntory Blue Rose Applause (registered trademark) contain rosacyanin A1, rosacyanin A2, rosacyanin B, and rosacyanin A1, rosadelphin A2, and rosacyanin B (e.g., WO2010 /110382). Suntory Blue Rose Applause (registered trademark) can be obtained from Suntory Flowers Co., Ltd. (Tokyo, Japan). A rose having a flavonoid 3',5'-hydroxylase gene can also be produced by the method described in WO2010/110382 and the like. Examples of flavonoid 3',5'-hydroxylase genes include plant-derived genes described in WO2004/020637, such as those derived from Viola spp. cv Black Pansy, etc. Flavonoid 3',5'-hydroxylase genes and the like are preferred. Roses having the flavonoid 3',5'-hydroxylase gene are preferred because they contain the rosadelphin compound in their petals. On the other hand, all of the academic literature and patent literature described in this specification are incorporated into this specification as a reference.

The inclusion of a rosacyanin compound and/or a rosadelphin compound in a rose can be confirmed by HPLC-TOF-MS by methods described in, for example, WO2010/110382, WO2015/178385, and the like.

Grape seeds and skins are seeds and skins of grapes (scientific name Vitis spp. ) of plants belonging to the genus Vitaceae Vitis. Although the grape variety is not particularly limited, examples include black grapes such as Cabernet Sauvignon, Concord, Merlot, and Muscat Bailey A; and white grapes such as Chardonnay and Kosch. Among them, Chardonnay or black grapes are preferable, black grapes are more preferable, and Cabernet Sauvignon, Merlot and the like are still more preferable. The number of grapes in the grape seed and/or skin extract may be 1 type or 2 or more types. The extract of grape seeds and/or pericarp is preferably an extract of grape seeds and pericarp.

As yeast of the genus Yarrowia , Yarrowia lipolytica is preferred. The yeast extract of the genus Yarrowia is preferably an extract of Yarrowia lipolytica .

As a plant of the genus Theaceae Camellia, the tea tree (scientific name Camellia sinensis ) is preferred.

As a Hamamelidaceae Hamamelis plant, Hamamelis (scientific name: Hamamelis virginiana ) is preferable.

As a Rosaceae Filipendula plant, meadowsweet (scientific name Filipendula ulmaria ) is preferred.

As a plant of the genus Plantainaceae Plantago (Plantaginaceae Plantago), royal plantain (scientific name Plantago major ) is preferred.

As a plant of the genus Lamiaceae Mentha, peppermint (scientific name Mentha x piperita ) is preferred.

In the preparation of plant extracts, any part of the plant, such as roots, rhizomes, stems, leaves, branches, bark, flowers, seeds (including seed coat, embryo, endosperm, hypocotyl, lateral side, etc.), fruit (fruit flesh) , skin), or a combination of a plurality of these may be used. A plant extract can be prepared by extracting any part of the plant with a solvent.

Preferred sites for producing plant extracts include, for example, the following sites. As the plant extract in the present invention, extracts of the following parts of plants are preferable.

A rose plant of the genus Rosaceae is preferably a flower, particularly a petal. For grapes, seeds and/or skins are used, and seeds and skins are preferably used. Camellia and Camellia genus plants, Pungnyeonhwa and Pungnyeonhwa genus plants preferably have leaves. The plant of the genus Rosaceae has a preferable flower. Seeds are preferred for plants of the genus Plantain and Plantain. Perilla and mint plants are preferably outposts.

The plant parts may be used in the extraction step as is, or may be used in the extraction step after being pulverized, cut or dried. In the present invention, the plant extract may be an extract obtained by extracting the plant as it is, or may be an extract obtained by crushing, cutting or drying the plant. Preferably, it is an extract from grinding, cutting or drying the plant.

The plant extraction method for obtaining the plant extract is not particularly limited, and can be obtained by a conventional extraction method used for extraction of plant components. The extraction method can be set appropriately, and the extraction conditions are not particularly limited either. For example, it is preferable to obtain a plant extract by extracting the plant with a solvent (extraction solvent) at room temperature or at elevated temperature. In preparation of the plant extract, the plant as a raw material may be used in the extraction step as it is or may be used in the extraction step after being pulverized, cut or dried.

The extraction solvent used for preparation of the plant extract can be selected appropriately, and those commonly used for extraction of plant components can be used. As the extraction solvent, for example, water; monohydric alcohols having 1 to 5 carbon atoms, such as methanol, ethanol, propanol and butanol; polyhydric alcohols having 2 to 5 carbon atoms such as ethylene glycol, propylene glycol, 1,2-butylene glycol, 1,3-butylene glycol, 1,4-butylene glycol, and 2,3-butylene glycol; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; chain and cyclic ethers such as tetrahydrofuran and diethyl ether; polyethers such as polyethylene glycol; Squalene etc. are mentioned. These may be used independently or may be used as a mixed solvent (mixture) in combination of two or more. As the monohydric alcohol having 1 to 5 carbon atoms, those having 1 to 4 carbon atoms are preferable, those having 2 to 4 carbon atoms are more preferable, and ethanol is still more preferable. As the polyhydric alcohol having 2 to 5 carbon atoms, a dihydric or trihydric alcohol having 2 to 4 carbon atoms is preferable, a dihydric alcohol is more preferable, and 1,3-butylene glycol is still more preferable. Especially, as the extraction solvent, water, a monohydric alcohol having 1 to 5 carbon atoms, a polyhydric alcohol having 2 to 5 carbon atoms, and a mixed solvent of two or more of these are preferable, and water, a monohydric alcohol having 2 to 4 carbon atoms, or an aqueous solution thereof. , A dihydric alcohol having 2 to 4 carbon atoms or an aqueous solution thereof is more preferable, and water, ethanol, aqueous ethanol solution, 1,3-butylene glycol, and 1,3-butylene glycol aqueous solution are more preferable, and water, ethanol aqueous solution or A 1,3-butylene glycol aqueous solution is particularly preferred. In one aspect, the aqueous ethanol solution and the 1,3-butylene glycol aqueous solution have an ethanol or 1,3-butylene glycol concentration of preferably 10 to 98 vol%, more preferably 30 to 90 vol%, and 30 to 90 vol%. 70% by volume is more preferred. As the plant extract in the present invention, the solvent extract can be suitably used.

During extraction, acid or alkali may be added to adjust the pH of the extraction solvent. After extraction, it is preferable to remove plant residues (plant bodies or parts thereof after extraction) from the extract. The method for removing plant residues from the extract is not particularly limited, and known separation means such as filtration and centrifugation can be used.

As an example of a plant extraction method, for example, the following method can be used. Plants are ground as they are or after drying, 0.1 to 30 times the amount of extraction solvent is added by weight, and at room temperature under atmospheric pressure, preferably for 10 minutes to 15 days, more preferably for 30 minutes to 10 days, still more preferably for 1 hour to After extracting for 7 days or preferably near the boiling point of the extraction solvent for about 10 minutes to 1 day (more preferably 10 minutes to 2 hours), it is filtered to obtain a filtrate. At the time of extraction, you may leave still and may stir suitably. In addition, the obtained filtrate (plant extract) may be used as a plant extract as it is, or diluted, concentrated, or dried as necessary.

In the present invention, a plant extract obtained by extraction can be used as a plant extract as it is. In addition, as long as the effect of the present invention is not impaired, it may be diluted, concentrated, or dried by a known method to obtain a diluted solution, concentrate, or powder, or may be prepared and used in a paste form. As a drying method, freeze drying, spray drying, etc. are mentioned, for example. Further, the above-mentioned plant extracts, concentrates and dried powders thereof may be further purified, such as deodorization and discoloration, as necessary within a range not impairing the effects of the present invention. Such a purification method may be performed by arbitrarily selecting conventional means.

Plant extracts in the present invention include various solvent extracts obtained by the above extraction methods, diluted solutions thereof, concentrates thereof or dried powders thereof, and purified products thereof. In addition, the extract may be used after being diluted or dissolved in a solvent different from the extraction solvent.

In addition, the plant extract is commercially available, and commercially available products may be used.

It is preferable that the Yarrowia genus yeast extract is a Yarrowia genus yeast cell body or an extract of a cell culture. As an extract of cells or cell cultures, cells or cell cultures containing cells (preferably cell cultures) are subjected to cell disruption treatment such as auto-digestion treatment or enzymatic degradation treatment to elute the yeast cell contents into the culture solution, etc. (cell lysate), and microbial cell residues removed from cells or cell cultures subjected to cell lysis treatment (cell lysate). Preferably, cells or a cell culture containing cells (preferably a cell culture solution) are subjected to self-digestion treatment. Sterilization treatment, such as heat treatment, etc. may be performed to the cell lysate or the thing from which microbial cell residues were removed from it, if necessary. The yeast extract may be one subjected to such a sterilization treatment or the like. The form of the yeast extract of the genus Yarrowia is not particularly limited, but examples thereof include paste form, suspension form, extract form, liquid form and powder form.

In one aspect, as the extract of a plant of the genus Rosaceae, an ethanol extract or aqueous ethanol extract of a rose (preferably petals) is preferred.

As the extract of grape seeds and/or skins, ethanol extracts or aqueous ethanol extracts of grape seeds and/or skins are preferred.

As the extract of Camellia and Camellia genus plants, an ethanol extract or an aqueous ethanol extract of tea plant (preferably leaves) is preferred.

As an extract of a plant of the Pomponaceae family Pomponaceae, a 1,3-butylene glycol extract or an aqueous 1,3-butylene glycol extract of Hamamelis (preferably leaves) is preferred.

As an extract of a plant of the genus Rosaceae, a 1,3-butylene glycol extract or an aqueous 1,3-butylene glycol extract of meadowsweet (preferably flowers) is preferred. As an extract of a plant of the genus Plantain and Plantago, preferred is an ethanol extract or an aqueous ethanol extract of Pyophyllum (preferably seeds).

As an extract of a plant of the genus Perilla and Mint, an ethanol extract, an aqueous ethanol extract or a hot water extract of peppermint (preferably a whole plant) is preferable.

The form of the composition of the present invention is not particularly limited, and may be in powder form, granular form, paste form, solid form, or the like, or may be in liquid form, and may be appropriately selected. In the composition of the present invention, other components may be incorporated in addition to the above extract as long as the effect of the present invention is not impaired. Examples of other components include components that can be used in cosmetics, food and beverages, pharmaceuticals, quasi-drugs, and the like, which will be described later.

The composition for suppressing or improving the expression of type 4 collagen and the composition for suppressing or improving the skin barrier function of the present invention can be provided in the form of a formulation as an example, but are not limited thereto. The agent may be provided as a composition as it is or as a composition containing the agent. In one embodiment, the composition of the present invention can also be referred to as an agent for suppressing or improving the expression of type 4 collagen and an agent for suppressing or improving the reduction of skin barrier function.

The composition for suppressing or improving the decrease in the expression of type 4 collagen of the first aspect of the present invention is intended to suppress or improve the decrease in the expression of type 4 collagen in a subject, preferably for type 4 collagen expression in the skin of a subject. It is used to inhibit a decrease in the expression of collagen or to improve the expression. The composition for suppressing or improving the decrease in skin barrier function of the second, third and fourth aspects of the present invention is used to suppress the decrease in the skin barrier function of a subject or to improve the function.

The composition of the present invention can be used, for example, to prevent or improve problems caused by a decrease in skin barrier function, a decrease in basement membrane function, or a decrease in the expression of type 4 collagen. As such problems, for example, skin dryness, itchiness, roughness, sensitive skin, rough skin, and the like can be cited. In one embodiment, the composition of the present invention inhibits or improves the decrease in skin barrier function, or suppresses or improves the decrease in the expression of type 4 collagen in the skin, thereby reducing dryness, itching, roughness, sensitive skin, It can be used to prevent or improve rough skin and the like. Prevention of the problem includes preventing the onset, delaying the onset, reducing the rate of onset, and the like. Amelioration of a problem includes relief of symptoms, improvement of symptoms, suppression of progression of symptoms, and the like.

The composition of the present invention can be used in various applications such as cosmetics, food and beverages, pharmaceuticals and quasi-drugs. The composition of the present invention itself may be a cosmetic, food or drink, pharmaceutical or quasi-drug for suppressing or improving the expression of type 4 collagen or suppressing or improving the skin barrier function, or It may be a material or formulation used in combination with the cosmetics, food and beverages, pharmaceuticals, quasi-drugs, and the like.

The composition of the present invention is suitably used, for example, as an external preparation for skin. External skin preparations include cosmetics, pharmaceuticals, and quasi-drugs, and are preferably cosmetics.

The composition of the present invention can also be used as a pharmaceutical or quasi-drug other than an external skin preparation.

In another embodiment, it is also preferable to use the composition of the present invention as food or drink. The case where the composition of the present invention is used as external skin preparations such as cosmetics, pharmaceuticals and quasi-drugs, food and beverages, etc. will be described below.

When the composition of the present invention is used as an external skin preparation, the dosage form is not particularly limited, and may be in any form, such as a solution, emulsion, cream, gel, powder, aerosol, mist, capsule, or sheet. The skin external preparation is preferably a cosmetic. The product form of cosmetics is not particularly limited either, and examples thereof include skin care cosmetics such as face wash, makeup remover, toner, serum, pack, lotion, cream, and sunblock; makeup cosmetics such as foundation, makeup base, lipstick, eye shadow, eye liner, mascara, eyebrow, blush, and nail enamel; hair cosmetics such as hair shampoo, hair conditioner, hairdressing agent, hair dye agent, and hair restorer; detergents such as soap and body soap; A bath salt etc. are mentioned.

The cosmetics include components such as carriers and additives acceptable for cosmetics, such as water, alcohols, oils, surfactants, thickeners, metal soaps, gelling agents, etc. Powders, chelating agents, water-soluble polymers, film formers, resins, inclusion compounds, antibacterial agents, deodorants, salts, pH adjusters, ultraviolet absorbers, extracts derived from animals, plants and microorganisms other than the above, keratolytic agents, enzymes, hormones, and other vitamins , a moisturizing agent, a bactericide, an anti-inflammatory agent, and a flavoring agent, or one or two or more thereof may be incorporated as appropriate. Cosmetics can be manufactured by a general manufacturing method. For example, it can be obtained by mixing the extract used for the active ingredient with one or two or more ingredients that can be used for the cosmetic, and then processing it into a desired form.

When the external skin preparation is a pharmaceutical or quasi-drug, components such as carriers and additives acceptable for pharmaceuticals or quasi-drugs may be used as long as the effect of the present invention is not impaired. Examples of such components include one or two or more of excipients, binders, disintegrants, lubricants, antioxidants, colorants, and the like, and these may be used as necessary.

The composition of the present invention can also be used as a pharmaceutical or quasi-drug other than the above-mentioned external skin preparation. These drugs or quasi-drugs may be administered orally or parenterally. Drugs or quasi-drugs may be in the form of oral administration preparations (internal administration) or parenteral administration preparations. Examples of dosage forms of preparations for oral administration include solutions, tablets, powders, fine granules, granules, dragees, capsules, suspensions, emulsions, chewables, and the like. Formulations of parenteral administration preparations include injections, infusions, and the like. For such drugs or quasi-drugs, components such as carriers and additives that are acceptable for drugs or quasi-drugs may be used. Examples of such components include one or two or more of excipients, binders, disintegrants, lubricants, antioxidants, colorants, corrigents, and the like, and these may be used as necessary. Pharmaceuticals and quasi-drugs can be produced by general manufacturing methods. For example, it can be obtained by mixing the extract used as an active ingredient with one or two or more ingredients that can be used in pharmaceuticals or quasi-drugs, and then processing them into a desired form.

When the composition of the present invention is used as a food or drink, the food or drink is not particularly limited, and examples thereof include general food and drink, health food, functional display food, and specific health use food. The shape of the food or drink is not particularly limited either. The above health foods, foods with functional claims, and foods for specified health use may be in the form of various formulations, such as solutions, tablets, powders, fine granules, granules, dragees, capsules, suspensions, emulsions, chewables, and liquid formulas.

In the food-drinks described above, ingredients acceptable for food-drinks, for example, other food-drinks ingredients, additives incorporated in food-drinks, and the like may be blended within a range that does not impair the effects of the present invention. These foods and beverages can be manufactured by a general manufacturing method. For example, what is necessary is just to mix the said extract used as an active ingredient in this invention with food-drinks material etc. in manufacture of food-drinks.

Even when the composition of the present invention is in any form such as an external preparation for skin, a pharmaceutical or quasi-drug other than external preparation for skin, food or drink, the content of the extract used as an active ingredient in the composition, for example, in terms of dry matter of the extract As weight, 0.0000001 to 100% by weight is preferable, 0.000001 to 100% by weight is more preferable, and 0.00001 to 10% by weight is still more preferable. The said content is content of the sum total, when two or more types of extracts are included.

The amount of the composition of the present invention used is not particularly limited, and may be an amount (effective amount) capable of suppressing or improving a decrease in the skin barrier function and suppressing or improving the expression of type 4 collagen, and the subject's condition, weight, and gender , age or other factors can be appropriately set. The amount of the composition of the present invention used for external skin preparations such as cosmetics is preferably 0.01 to 500 mg, for example, as the extract used for the active ingredient (in terms of dry matter) per adult (60 kg) per day. The dose in the case of oral administration of a drug or quasi-drug is preferably 0.01 to 500 mg, for example, as the extract (in terms of dry matter) per adult (60 kg) per day. If it is a food or drink, the intake amount is preferably, for example, 0.01 to 500 mg as the extract (in terms of dry matter) per adult (60 kg) per person per day. The above amount of the extract can be ingested or administered once or divided into multiple times. The quantity of the said extract is the total amount, when using 2 or more types of extracts. The timing of applying the external skin preparation containing the extract to the skin, the timing of ingesting or administering food, drink, medicine, or quasi-drug is not particularly limited.

The target of application of the composition of the present invention (can also be referred to as the target of administration) is not particularly limited, and can be applied to humans or non-human mammals, but humans are preferred. Applicable targets include, for example, targets in which suppression or improvement in skin barrier function is desired or required, and targets in which suppression or improvement in type 4 collagen expression is desired or required. Examples of such subjects include people who experience dryness, itchiness, roughness, sensitive skin, and rough skin of the skin. The composition of the present invention can be ingested or administered by various methods known per se, depending on its form, dosage form, and the like. In one aspect, the composition of the present invention is preferably applied to the skin.

The extract used in the active ingredient of the composition of the present invention can be used to suppress the decrease in the expression of type 4 collagen or to improve the expression. The extract suppresses the decrease in the skin barrier function of the subject or improves the skin barrier function, preferably by suppressing or improving the decrease in the expression of type 4 collagen, or improving the skin barrier function. can be used to do

The present invention also includes the following uses and methods.

An extract of a plant of the genus Rosaceae, an extract of grape seeds and/or peel, Yarrowia genus for suppressing or improving the decrease in skin barrier function by inhibiting or improving the expression of type 4 collagen Use of at least one extract selected from the group consisting of a yeast extract, an extract of a plant of the genus Camellia and Camellia, an extract of a plant of the genus Rosaceae, and an extract of a plant of the genus Perilla and Mint.

An extract of a plant of the genus Rosaceae, an extract of grape seeds and/or skin, an extract of yeast of the genus Yarrowia , an extract of a plant of the genus Camellia and the genus Camellia, an extract of a plant of the genus Rosaceae, and an extract of a plant of the genus Perilla and Mint A method for suppressing a decrease in skin barrier function or improving a skin barrier function by suppressing or improving a decrease in the expression of type 4 collagen, by administering at least one extract selected from the group consisting of:

A use of an extract of a plant belonging to the genus Pomponaceae and/or an extract of a plant belonging to the genus Plantain and plantain for suppressing a decrease in skin barrier function or improving the skin barrier function.

A method for suppressing a decrease in skin barrier function or improving a skin barrier function, comprising administering an extract of a plant of the genus Pomponaceae and/or a plant of the genus Plantain and/or the genus Plantain.

An extract of a plant of the genus Rosaceae, an extract of grape seeds and/or skin, an extract of yeast of the genus Yarrowia , an extract of a plant of the genus Camellia and Camellia, for inhibiting or improving the expression of type 4 collagen, , Use of at least one extract selected from the group consisting of extracts of plants of the genus Pompanoaceae, extracts of plants of the genus Rosaceae, plant extracts of the genus Plantain and plantain, and extracts of plants of the genus Perilla and Mint.

A method for suppressing the decrease in the expression of type 4 collagen or improving the expression by administering the one or more extracts.

The extract is preferably administered (applied) to the skin to suppress or improve the decrease in skin barrier function, to suppress the decrease in the expression of type 4 collagen in the skin or to improve the expression, Alternatively, it can be used to suppress the deterioration of the basement membrane function or to improve the function. In one embodiment, the extract can be used to prevent or improve problems caused by a decrease in skin barrier function, a decrease in basement membrane function, or a decrease in the expression of type 4 collagen. The extract prevents, for example, dryness, itchiness, roughness, sensitive skin, rough skin, etc. of the skin by inhibiting or improving the decrease in the skin barrier function or by inhibiting or improving the expression of type 4 collagen in the skin. Or it can be used to improve.

In the above uses and methods, the extract, the subject to which it is administered, the dosage of the extract and their preferred embodiments, etc. are the same as those in the above-described composition of the present invention. The extract may be used as it is or may be used in combination with other components. You may use the composition of this invention. The extract can be used in the form of the above-mentioned external preparation for skin, food or drink.

The use may be a therapeutic use or a non-therapeutic use. The method may be a therapeutic method or a non-therapeutic method. Non-therapeutic is a concept that does not include medical practice, that is, surgery, treatment or diagnosis of humans.

A separate aspect of the present invention, use of the extract, for preparing a composition for inhibiting or improving the decrease in skin barrier function; It is also a use for preparing a composition for suppressing or improving the decrease in the expression of type 4 collagen of the extract. The extracts and preferred embodiments thereof are the same as those in the composition of the present invention described above.

Another aspect of the present invention, use of an agent for inhibiting or improving a decrease in basement membrane function, for inhibiting a decrease in skin barrier function or improving a skin barrier function; It is also a method of suppressing the decline of the skin barrier function or improving the skin barrier function by administering an agent for suppressing or improving the basement membrane function.

The agent for suppressing or improving a decrease in basement membrane function and preferred embodiments thereof are the same as described above.

The present invention also includes a screening method for a substance having an effect of suppressing or improving a decrease in skin barrier function. The screening method of the present invention is described.

The screening method for a substance having an action of inhibiting or improving skin barrier function of the present invention is a step of culturing human epidermal keratinocytes in vitro in a medium containing a test substance or a control medium not containing the test substance. and, comparing the expression level of type 4 collagen in skin cells cultured in a medium containing the test substance and in control skin cells cultured in a control medium, the skin cultured in the medium containing the test substance When the expression level of type 4 collagen in the cells is higher than the expression level of type 4 collagen in the control skin cells, the test substance is selected as a substance having an effect of suppressing or improving the decrease in skin barrier function.

In the screening method of the present invention, human epidermal keratinocytes are cultured in vitro in a medium containing a test substance or a control medium not containing the test substance. Human epidermal keratinocytes are commercially available, and commercial products can be used.

The test substance is not particularly limited, and examples thereof include plant extracts, cell extracts, cell culture supernatants, fermentation products, proteins, peptides, vitamins, and synthetic compounds. These may be known substances or may be new substances.

A step of culturing human epidermal keratinocytes in vitro in a medium containing a test substance or a control medium not containing the test substance is also referred to as a culture step. The step of comparing the type 4 collagen expression level in skin cells (also referred to as test skin cells) cultured in a medium containing the test substance and the control skin cells cultured in a control medium is also referred to as the type 4 collagen expression level comparison step. .

In the culturing step, conditions for culturing human epidermal keratinocytes are not particularly limited, and known conditions may be employed. A medium containing a test substance can be prepared by mixing the test substance into a medium. As the medium, those used for culturing human epidermal keratinocytes can be used, and commercially available products may be used. Examples of the medium include HuMedia-KB2 medium (manufactured by Kurabo Industries), DMEM medium, and the like. Insulin, human EGF, hydrocortisone, serum, antibacterial agent, etc. may be added to the medium. The culturing time is not particularly limited, and may be appropriately set depending on the culturing conditions and the like. As the control medium, the same medium as the medium containing the test substance may be used except that the test substance is not contained. For example, human epidermal keratinocytes may be cultured in a medium containing a test substance or a control medium for preferably 1 to 72 hours, more preferably 3 to 48 hours.

In the culturing step, a stress load such as ultraviolet irradiation or oxidative stress load may be applied. For example, when culture is performed under stress load conditions such as ultraviolet irradiation or oxidative stress load, effective components can be screened for suppressing the decrease in skin barrier function due to the stress or improving the skin barrier function under stress.

After the culturing step, a type 4 collagen expression level comparison step is performed. After the culturing, the expression levels of type 4 collagen in skin cells cultured in a medium containing the test substance and in control skin cells cultured in a control medium are compared. When the expression level of type 4 collagen in skin cells cultured in a medium containing the test substance is greater than the expression level of type 4 collagen in the control skin cells, the test substance exhibits an effect of suppressing or improving a decrease in skin barrier function. Choose as a material with

The amount of type 4 collagen expression can be measured by measuring the amount of mRNA encoding type 4 collagen or the amount of protein of type 4 collagen. The amount of mRNA or protein can be measured by a known method.

The amount of mRNA encoding type 4 collagen can be measured by quantitative PCR, northern blotting, microarray, live imaging using a fluorescent probe, or the like. The amount of protein in type 4 collagen can be measured by immunochemical assay, mass spectrometry or the like. Examples of immunochemical assays include Enzyme Linked Immunosorbent Assay (ELISA), Western blotting, and the like. For the measurement of the amount of protein, a commercially available ELISA kit (manufactured by MyBioSource) or the like can also be used. The nucleotide sequence and amino acid sequence (ORF) of human type 4 collagen are known, and the nucleotide sequence (mRNA) is registered in Genbank, a public database, as Accession No.AH002741. A person skilled in the art can easily design and synthesize primers for measuring the amount of mRNA encoding type 4 collagen, amplify DNA, and the like.

As described above, by suppressing the decrease in the expression of type 4 collagen or improving the expression, it is possible to suppress or improve the decrease in the skin barrier function. For this reason, when the expression level of type 4 collagen in the test skin cells is greater than the expression level of type 4 collagen in the control skin cells, the test substance can be judged as a substance having an effect of suppressing or improving the decrease in skin barrier function. have. According to the above screening method, it is possible to screen for a substance having an effect of suppressing or improving a decrease in skin barrier function.

Example

Hereinafter, the present invention will be described in more detail by examples, but the scope of the present invention is not limited thereby.

<Preparation Example 1>

A plant-derived extract was prepared. Plants used for extraction and their parts are as follows.

(1) Rose ( Rosa hybrida , cultivar "Suntory Blue Rose Applause (registered trademark), Suntory Flowers Co., Ltd.): petals

(2) Grapes ( Vitis spp. ) (Cabernet Sauvignon): seeds and skins

(3) Tea tree ( Camellia sinensis ): leaves

(4) Hamamelis ( Hamamelis virginiana ): leaves

(5) Meadowsweet ( Filipendula ulmaria ): Flower

(6) Plantago major : Seed

(7) Peppermint ( Mentha x piperita ): Outpost

The rose (variety "Suntory Blue Rose Applause") is a blue system containing delphinidin-type anthocyanins, rosacyanin A1, rosacyanin A2 and rosacyanin B, and rosadelphin A1, rosadelphin A2 and rosacyanin B in petals is the rose of

The plant debris of the above (1) to (7) was immersed in a solvent and extracted. More specifically, regarding the above (1), (2), (3), (6), and (7), the plant debris is immersed in a 30 to 90 vol% ethanol aqueous solution in an amount 10 times the weight, Then, stirring operation was added once a day to extract for 10 minutes to 7 days to obtain an extract. Regarding the above (4) and (5), an aqueous solution of 1,3-butylene glycol was used instead of the aqueous ethanol solution, and extraction was performed by the above method.

Plant residues were removed from the obtained extract by filtration, and a filtrate was obtained as a plant extract.

After concentrating the obtained plant extract with an evaporator, the extract of each plant was obtained as a powder by freeze-drying. The powdered plant extracts (rose petal extract, grape seed and pericarp extract, tea tree leaf extract, hamamelis leaf extract, meadowsweet flower extract, psyllium seed extract, peppermint extract) were used as extract samples for the following evaluation.

<Preparation Example 2>

Yarrowia lipolytica extract was obtained by culturing Yarrowia lipolytica, self-digesting, and subjecting to heat sterilization and disinfection operation to obtain a powdery Yarrowia lipolytica extract. In the following examples, the powder was used as an extract sample for Yarrowia lipolytica extract.

<Example 1> Effect of overexpression of type 4 collagen on skin barrier during inflammatory stimulation

A plasmid (hereinafter referred to as COL4A1 plasmid) was prepared by inserting COL4A1 (RefSeq ID: NM_001845.6) downstream of the CMV promoter. According to the recommended protocol, the COL4A1 plasmid and Lipofectamine 3000 Reagent (manufactured by Thermo Fisher Scientific) were diluted using Opti-MEM (registered trademark) I Reduced Serum Media (manufactured by Thermo Fisher Scientific) (hereinafter referred to as Opti-MEM). , Two types of solutions (hereinafter referred to as Opti-MEM solutions) having different plasmid concentrations were prepared (plasmid concentration: 5 µg/mL or 20 µg/mL). On the other hand, as a control, a solution in which Opti-MEM was added instead of the plasmid was also prepared.

Using HuMedia-KG2 (hereinafter referred to as the recommended medium) (manufactured by Kurabo Industries), normal adult epidermal keratinocytes (manufactured by Kurabo Industries) were added to Cell Culture Inserts (manufactured by Millipore) for 24 wells at 3×10 ⁴ cells/200 After seeding at a concentration of μL/well, 10 μL of either Opti-MEM solution was quickly added. On the other hand, 900 µL of the recommended medium was added to a 24-well plate (manufactured by AGC Technoglass Co., Ltd.). Then, after 30 minutes, hydrogen peroxide solution (manufactured by Nacalai Tesque Co., Ltd.) was added (H ₂ O ₂ treatment) to a final concentration of 100 µM for only some of the groups. 4 hours after sowing, the medium was replaced with the recommended medium. Twenty-four hours after seeding, the medium was replaced with a recommended medium in which the concentration of Ca ²⁺ in the medium was raised to 1.41 mM by the addition of CaCl ₂ (hereinafter referred to as an irregular recommended medium). The medium was exchanged every 2 to 3 days using an irregular recommendation medium.

After 96 hours, 120 hours, and 144 hours after sowing, the transepithelial electrical resistance value (TEER (Ω·cm 2 )) was evaluated using Millicel ERS-2 (manufactured by Merck). In addition, 144 hours after seeding, cells were lysed using isogen (manufactured by Nippon Gene), and then mRNA was extracted. Then, cDNA was synthesized using the High-Capacity cDNA Reverse Transcription Kit and RNase Inhibitor, and quantitative PCR was performed using TaqMan Fast Universal PCR Master Mix (both manufactured by Thermo Fisher Scientific) and primers to obtain human type 4 collagen A1 ( The expression level of COL4A1) was evaluated. Meanwhile, Eukaryotic 18S rRNA was used for endogenous control.

Primers for measuring COL4A1 expression level (COL4A1) and primers for quantifying endogenous control (Eukaryotic 18S rRNA) were respectively #Hs00266237_m1 and #Hs99999901_s1 (both manufactured by Thermo Fisher Scientific).

The results are shown in Table 1 and Table 2. Table 1 shows the COL4A1 gene expression level when the COL4A1 expression level of the plasmid-free and H ₂ O ₂ non-treated group (H ₂ O ₂ (-), plasmid (-)) was set to 1. In Tables 1 and 2, H ₂ O ₂ (+) is a group subjected to H ₂ O ₂ treatment. Plasmid (-) indicates no addition of COL4A1 plasmid, and plasmid (+) indicates addition of COL4A1 plasmid. * indicates that there is a significant difference (p<0.05) (Dunnett's test (vs plasmid not added and H ₂ O ₂ treated group)). Although a decrease in COL4A1 expression level was observed with H ₂ O ₂ stimulation, introduction of the COL4A1 plasmid resulted in a significant and dose-dependent suppression of expression decrease or improvement in the H ₂ O ₂ stimulation group (*: p< 0.05).

Table 2 shows the TEER values 144 hours after seeding. * indicates that there is a significant difference (p<0.05) (Dunnett's test (vs plasmid not added and H ₂ O ₂ treated group)). A significant decrease in TEER was observed with H ₂ O ₂ stimulation, but by introducing the COL4A1 plasmid, a dose-dependent and significant TEER improving effect was confirmed in the high-dose group with respect to the H ₂ O ₂ stimulation group (*: p <0.05).

<Examples 2 to 9> Evaluation of extract samples for inhibiting or improving the decrease in type 4 collagen gene expression

Using the recommended medium, normal adult epidermal keratinocytes (manufactured by Kurabo Industries) were seeded in a 12-well plate (manufactured by AGC Technoglass) at a concentration of 5×10 ⁴ cells/mL/well, and cultured for 48 hours. After replacing the sample with the recommended medium containing the extract sample and starting the sample treatment, 30 minutes after the start of the sample treatment, hydrogen peroxide solution (manufactured by Nacalai Tesque Co., Ltd.) was added to a final concentration of 100 µM. After 3 hours from the start of the hydrogen peroxide treatment, the medium was replaced with a recommended medium containing no extract sample. After 48 hours from the start of sample treatment, cells were lysed and collected using RLT Buffer (containing 1% 2-mercaptoethanol), and then mRNA was analyzed using RNeasy Mini Kit and RNase-Free DNase Set (both manufactured by Qiagen). was extracted. Thereafter, cDNA synthesis and quantitative PCR were performed in the same manner as in Example 1 to evaluate the expression level of COL4A1.

Regarding the control, the cells were cultured in the same manner as above, except that instead of the extract sample, dimethyl sulfoxide (DMSO) was treated with the recommended medium added so that the final concentration was 0.1%, hydrogen peroxide was added, The expression level of COL4A1 was evaluated.

The extract samples used in Examples 2 to 9 were the rose petal extract, grape seed and pericarp extract, tea tree leaf extract, hamamelis leaf extract, meadowsweet flower extract, king plantain seed extract, and Yarrowia Lee prepared in Preparation Examples 1 and 2. Polytica extract or peppermint extract. Extract samples were dissolved in ultrapure water or DMSO and added to the medium. The concentration of the extract sample in the medium was 5 to 20 µg/mL. On the other hand, when dissolved in ultrapure water, DMSO was separately added so that the final concentration of DMSO was 0.1% in all recommended media containing extract samples.

Table 3 shows the results of Examples 2 to 9 (average values of N = 3 to 4). The results shown in Table 3 are the relative values (%) of the expression level of COL4A1 in cells treated with the extract sample when the expression level of COL4A1 in the control was taken as 100%. * indicates that there is a significant difference (p<0.05) with respect to the control (Student's t-test). On the other hand, Table 3 shows the results when the expression of COL4A1 was the highest in the concentration range of the extract sample (test substance) of 5 to 20 μg/mL. Rose petal extract, grape seed and pericarp extract, tea tree leaf extract, hamamelis leaf extract, meadowsweet flower extract, king plantain seed extract, Yarrowia lipolytica extract, peppermint extract, decrease in COL4A1 expression with respect to control A significant effect of inhibiting or improving the expression was confirmed (*: p<0.05).

<Examples 10 to 15> Evaluation of skin barrier function deterioration inhibition or improvement action of extract samples

Using the recommended medium containing the extract sample, normal adult epidermal keratinocytes were seeded on Millicell Cell Culture Inserts at a concentration of 3×10 ⁴ cells/200 μL/well. After 30 minutes from seeding, hydrogen peroxide solution was added to a final concentration of 100 µM. Four hours after seeding, the medium was replaced with the recommended medium not containing the extract sample. After 24 hours from sowing, the culture medium was replaced with the recommended anomalous medium. Thereafter, transepithelial electrical resistance (TEER) was evaluated by the same method as in Example 1.

Regarding control, cells were cultured in the same manner as above, except for using the recommended medium in which DMSO was added to a final concentration of 0.1% instead of the recommended medium containing the extract sample, hydrogen peroxide solution was added, and Blood electrical resistance was evaluated.

The extract samples used in Examples 10 to 15 are extracts shown in Table 4 prepared in Preparation Examples 1 and 2. Extract samples were dissolved in ultrapure water or DMSO and added to the medium. The concentration of the extract sample in the medium was 5 to 20 µg/mL. On the other hand, when dissolved in ultrapure water, DMSO was separately added so that the final concentration of DMSO was 0.1% in all recommended media containing extract samples.

The results (average values of N = 4 to 6) are shown in Table 4. The results shown in Table 4 are the relative values (%) of TEER in the cells treated with the extract sample when the TEER of the control was taken as 100%. On the other hand, Table 4 shows the results when the concentration of the extract sample (test substance) was 5 to 20 µg/mL and the measurement time was the highest in the range of 96 to 144 hours after sowing. A high effect of suppressing or improving the decrease in TEER was confirmed in six extracts, and furthermore, grape seed and pericarp extract, tea tree leaf extract, hamamelis leaf extract, meadowsweet flower extract, and Yarrowia lipolytica extract were confirmed. , a significant change was confirmed with respect to the control (*: p<0.05) (Student's t-test).

It has been clarified that the extract of the above plant and the extract of Yarrowia lipolytica have an action of suppressing a decrease in the skin barrier function or improving the function.

Claims (22)

An extract of a plant of the genus Rosaceae, an extract of grape seeds and/or skin, an extract of yeast of the genus Yarrowia , an extract of a plant of the genus Camellia and the genus Camellia, an extract of a plant of the genus Rosaceae, and an extract of a plant of the genus Perilla and Mint A composition for inhibiting or improving a decrease in skin barrier function through inhibition or improvement in the expression of type 4 collagen, comprising as an active ingredient one or more extracts selected from the group consisting of. The method of claim 1, wherein the Rosaceae Rosaceae plant is rose, Yarrowia (Yarrowia ) yeast is Yarrowia lipolytica ( Yarrowia lipolytica ), Camellia and Camellia plants are tea trees, Rosaceae plants of the genus Meadowsweet, Perilla and Mint A composition for suppressing or improving a decrease in skin barrier function, wherein the plant is peppermint. The method according to claim 1 or 2, wherein the rose petals containing at least one compound selected from the group consisting of delphinidin-type anthocyanins, rosacyanin compounds, and rosadelphin compounds, the rose petals of the rose family, and the camellia plants of the genus Camellia are tea trees A composition for inhibiting or improving skin barrier function deterioration, wherein the leaves of the genus Rosaceae are flowers of meadowsweet, and the plants of the genus Perilla and Mint are peppermint sheaths. A composition for inhibiting or improving a decrease in skin barrier function, comprising an extract of a plant belonging to the genus Cornucopia and/or a plant belonging to the genus Plantain and/or the genus Plantain as an active ingredient. The composition for inhibiting or improving a decrease in skin barrier function according to claim 4, wherein the plant of the genus Pomponaceae is Hamamelis, and the plant of the genus Plantain and Plantain is royal plantain. The composition for suppressing or improving a decrease in skin barrier function according to claim 4 or 5, wherein the Pomponaceae Pomonias plant is a leaf of Hamamelis, and the Plantain family Plantain plant is a Pyramid seed. The composition for inhibiting or improving a decrease in skin barrier function according to any one of claims 4 to 6, wherein the decrease in skin barrier function is suppressed or the skin barrier function is improved by suppressing or improving a decrease in basement membrane function. [Claim 8] The composition for inhibiting or improving the deterioration of the skin barrier function according to claim 7, wherein the inhibition or improvement of the basement membrane function is through the inhibition or improvement of the expression of type 4 collagen. A composition for inhibiting or improving a decrease in skin barrier function, comprising an agent for inhibiting or improving a basement membrane function as an active ingredient. The method of claim 9, wherein the agent for inhibiting or improving the deterioration of the basement membrane function inhibits or improves the deterioration of the basement membrane function by inhibiting or improving the expression of type 4 collagen, for inhibiting or improving the deterioration of the skin barrier function composition. Rosaceae Rosaceae Plant Extract, Grape Seed and/or Peel Extract, Yarrowia Genus Yeast Extract, Camellia and Camellia Plant Extract, Pomponaceae Polliniaceae Plant Extract, Rosaceae Plant Extract, Plantain and Plantain A composition for inhibiting or improving the decrease in the expression of type 4 collagen, comprising as an active ingredient at least one extract selected from the group consisting of extracts of plants of the genus Perilla and extracts of the genus Perilla and Mint. According to claim 11, the Rosaceae Rosaceae plant is rose, Yarrowia ( Yarrowia ) yeast is Yarrowia lipolytica ( Yarrowia lipolytica ), Camellia and Camellia plant is tea tree, Pomponaceae Pomponaceae plant is Hamamelis, Rosaceae A composition for suppressing or improving the decrease in the expression of type 4 collagen, wherein the plant of the genus Meadowsweet, the plant of the genus Plantain and plantain is royal plantain, and the plant of the genus Perilla and mint is peppermint. The method according to claim 11 or 12, wherein the rose petals containing at least one compound selected from the group consisting of delphinidin-type anthocyanins, rosacyanin compounds, and rosadelphin compounds, the rose petals of the rose family, and the camellia plants of the genus Camellia are tea trees Inhibiting the decrease in the expression of type 4 collagen, which is the leaf of Hamamelis, the leaf of Hamamelis, the leaf of Hamamelis, the flower of Meadowsweet, the flower of Meadowsweet, the flower of Meadowsweet, the plant of Plantago and Plantain, and the outpost of peppermint. Or a composition for improvement. The composition according to any one of claims 1 to 13, which is an external preparation for skin. The composition according to any one of claims 1 to 14, which is a cosmetic. An extract of a plant of the genus Rosaceae, an extract of grape seeds and/or peel, Yarrowia genus for suppressing or improving the decrease in skin barrier function by inhibiting or improving the expression of type 4 collagen Use of at least one extract selected from the group consisting of a yeast extract, an extract of a plant of the genus Camellia and Camellia, an extract of a plant of the genus Rosaceae, and an extract of a plant of the genus Perilla and Mint. A use of an extract of a plant belonging to the genus Pomponaceae and/or an extract of a plant belonging to the genus Plantain and plantain for suppressing a decrease in skin barrier function or improving the skin barrier function. An extract of a plant of the genus Rosaceae, an extract of grape seeds and/or skin, an extract of yeast of the genus Yarrowia , an extract of a plant of the genus Camellia and Camellia, for inhibiting or improving the expression of type 4 collagen, , Use of at least one extract selected from the group consisting of extracts of plants of the genus Pompanoaceae, extracts of plants of the genus Rosaceae, plant extracts of the genus Plantain and plantain, and extracts of plants of the genus Perilla and Mint. An extract of a plant of the genus Rosaceae, an extract of grape seeds and/or skin, an extract of yeast of the genus Yarrowia , an extract of a plant of the genus Camellia and the genus Camellia, an extract of a plant of the genus Rosaceae, and an extract of a plant of the genus Perilla and Mint A method for suppressing a decrease in skin barrier function or improving a skin barrier function by suppressing or improving a decrease in the expression of type 4 collagen, by administering at least one extract selected from the group consisting of: A method for suppressing a decrease in skin barrier function or improving a skin barrier function, comprising administering an extract of a plant of the genus Pomponaceae and/or a plant of the genus Plantain and/or the genus Plantain. Rosaceae Rosaceae Plant Extract, Grape Seed and/or Peel Extract, Yarrowia Genus Yeast Extract, Camellia and Camellia Plant Extract, Pomponaceae Polliniaceae Plant Extract, Rosaceae Plant Extract, Plantain and Plantain A method for suppressing a decrease in the expression of type 4 collagen or improving the expression by administering at least one extract selected from the group consisting of an extract of a plant of the genus Perilla and an extract of a plant of the genus Perilla and Mint. In vitro, a step of culturing human epidermal keratinocytes in a medium containing a test substance or a control medium not containing the test substance; and
Comparing the expression level of type 4 collagen in skin cells cultured in a medium containing the test substance and in control skin cells cultured in a control medium;
When the expression level of type 4 collagen in skin cells cultured in a medium containing the test substance is greater than the expression level of type 4 collagen in the control skin cells, the test substance exhibits an effect of suppressing or improving a decrease in skin barrier function. A screening method for a substance having an effect of suppressing or improving a decrease in skin barrier function, selected as a substance having an effect.