AU2021233359A1

AU2021233359A1 - Composition for suppressing decrease in or improving skin barrier function, composition for suppressing decrease in or improving expression of type 4 collagen, and method for screening substances having action to suppress decrease in or improve skin barrier function

Info

Publication number: AU2021233359A1
Application number: AU2021233359A
Authority: AU
Inventors: Sota AKAZAWA
Original assignee: Suntory Holdings Ltd
Current assignee: Suntory Holdings Ltd
Priority date: 2020-03-10
Filing date: 2021-03-04
Publication date: 2022-11-03
Also published as: WO2021182283A1; TW202200185A; CN115243667A; KR20220151196A; JPWO2021182283A1

Abstract

The purpose of the present invention is to provide a composition for suppressing a decrease in or improving skin barrier function capable of suppressing a decrease in skin barrier function or improving said function, a composition for suppressing a decrease in or improving expression of type 4 collagen, a method for suppressing a decrease in skin barrier function or improving skin barrier function, a method for suppressing a decrease in expression of type 4 collagen or improving said expression, and a method for screening substances having an action to suppress a decrease in or improve skin barrier function. The present invention relates to a composition for suppressing a decrease in or improving skin barrier function via suppressing a decrease in or improving expression of type 4 collagen that includes as an active ingredient one or more extracts selected from the group consisting of extracts of plants of the genus Rosa (Rosaceae), extracts of grape seeds or pericarp, extracts of Yarrowia yeasts, extracts of plants of the genus Camellia (Theaceae), extracts of plants of the genus Filipendula (Rosaceae), and extract of plants of the genus Mentha (Labiatae).

Description

DESCRIPTION

TITLE OF INVENTION: COMPOSITION FOR SUPPRESSING DECREASE IN OR IMPROVING SKIN BARRIER FUNCTION, COMPOSITION FOR SUPPRESSING DECREASE IN OR IMPROVING EXPRESSION OF TYPE 4 COLLAGEN, AND METHOD FOR SCREENING SUBSTANCES HAVING ACTION TO SUPPRESS DECREASE IN OR IMPROVE SKIN BARRIER FUNCTION

TECHNICAL FIELD

[0001] The present invention relates to a composition for suppressing a decrease in or improving skin barrier function and a composition for suppressing a decrease in or improving expression of type IV collagen. The present invention also relates to a method of suppressing a decrease in skin barrier function or improving the skin barrier function and a method of suppressing a decrease in expression of type IV collagen or improving the expression. The present invention also relates to a method of screening for a substance having an effect of suppressing a decrease in or improving skin barrier function.

BACKGROUND ART

[0002] Skin is primarily composed of two layers including the epidermis and the dermis from the surface. The epidermis is composed of four layers including a basal layer, a spinous layer, a granular layer, and a cornified layer in ascending order. The basal layer adheres to the dermis via a basement membrane. The basement membrane is composed of components such as type IV collagen and type VII collagen and has functions such as a function of connecting the basal layer in the epidermis with the dermis. The functions unfortunately decrease with age.

[0003]

Skin has a barrier function that prevents entry of foreign substances into the living body from the outside and keeps water in the skin by preventing water in the body from excessively evaporating from the skin. A decrease in skin barrier function due to factors such as aging causes troubles such as dry skin. Patent Literature 1 discloses an agent for improving skin barrier function containing components such as an extract of Aspalathus linearis.

CITATION LIST - Patent Literature

[0004] Patent Literature 1: JP 2009-256244 A

SUMMARY OF INVENTION - Technical Problem

[0005] The present invention aims to provide a composition for suppressing a decrease in or improving skin barrier function capable of suppressing a decrease in skin barrier function or improving the function. The present invention also aims to provide a composition for suppressing a decrease in or improving expression of type IV collagen. The present invention also aims to provide a method of suppressing a decrease in skin barrier function or improving the function, and a method of suppressing a decrease in expression of type IV collagen or improving the expression. The present invention also aims to provide a method of screening for a substance having an effect of suppressing a decrease in or improving skin barrier function.

- Solution to problem

[0006] The present inventor made studies to solve the above problem, and found out that suppression of a decrease in expression of type IV collagen or improvement of the expression in skin cells suppresses a decrease in skin barrier function or improves the function. The present inventor made further studies based on the above finding, and completed the present invention.

[0007] The present invention encompasses but is not limited to the following composition for suppressing a decrease in or improving skin barrier function, the composition for suppressing a decrease in or improving expression of type IV collagen, and the like. (1) A composition for suppressing a decrease in or improving skin barrier function by suppressing a decrease in or improving expression of type IV collagen, the composition containing, as an active ingredient, at least one extract selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae. (2) The composition for suppressing a decrease in or improving skin barrier function according to (1) above, wherein the plant belonging to the genus Rosa of the family Rosaceae is a rose, the yeast belonging to the genus Yarrowia is Yarrowia lipolytica, the plant belonging to the genus Camellia of the family Theaceae is tea plant, the plant belonging to the genus Filipendula of the family Rosaceae is meadowsweet, and the plant belonging to the genus Mentha of the family Lamiaceae is peppermint. (3) The composition for suppressing a decrease in or improving skin barrier function according to (1) or (2) above, wherein the plant belonging to the genus Rosa of the family Rosaceae is a rose petal containing at least one compound selected from the group consisting of a delphinidin-type anthocyanin, a rosacyanin compound, and a rosadelphin compound, the plant belonging to the genus Camellia of the family Theaceae is a tea plant leaf, the plant belonging to the genus Filipendula of the family Rosaceae is a meadowsweet flower, and the plant belonging to the genus Mentha of the family Lamiaceae is a whole plant of peppermint. (4) A composition for suppressing a decrease in or improving skin barrier function, the composition containing, as an active ingredient, an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae and/or an extract of a plant belonging to the genus Plantago of the family Plantaginaceae. (5) The composition for suppressing a decrease in or improving skin barrier function according to (4) above, wherein the plant belonging to the genus Hamamelis of the family Hamamelidaceae is American witch-hazel and the plant belonging to the genus Plantago of the family Plantaginaceae is greater plantain. (6) The composition for suppressing a decrease in or improving skin barrier function according to (4) or (5) above, wherein the plant belonging to the genus Hamamelis of the family Hamamelidaceae is an American witch-hazel leaf and the plant belonging to the genus Plantago of the family Plantaginaceae is a greater plantain seed. (7) The composition for suppressing a decrease in or improving skin barrier function according to any one of (4) to (6) above, wherein the composition suppresses a decrease in or improves skin barrier function by suppressing a decrease in or improving basement membrane function. (8) The composition for suppressing a decrease in or improving skin barrier function according to (7) above, wherein the composition suppresses a decrease in or improving basement membrane function by suppressing a decrease in or improving expression of type IV collagen. (9) A composition for suppressing a decrease in or improving skin barrier function, the composition containing, as an active ingredient, an agent for suppressing a decrease in or improving basement membrane function. (10) The composition for suppressing a decrease in or improving skin barrier function according to (9) above, wherein the agent for suppressing a decrease in or improving basement membrane function suppresses a decrease in or improving basement membrane function by suppressing a decrease in or improving expression of type IV collagen. (11) A composition for suppressing a decrease in or improving expression of type IV collagen, the composition containing, as an active ingredient, at least one extract selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, an extract of a plant belonging to the genus Plantago of the family Plantaginaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae. (12) The composition for suppressing a decrease in or improving expression of type IV collagen according to (11) above, wherein the plant belonging to the genus Rosa of the family Rosaceae is a rose, the yeast belonging to the genus Yarrowia is Yarrowia lipolytica, the plant belonging to the genus Camellia of the family Theaceae is tea plant, the plant belonging to the genus Hamamelis of the family

Hamamelidaceae is American witch-hazel, the plant belonging to the genus Filipendula of the family Rosaceae is meadowsweet, the plant belonging to the genus Plantago of the family Plantaginaceae is greater plantain, and the plant belonging to the genus Mentha of the family Lamiaceae is peppermint. (13) The composition for suppressing a decrease in or improving expression of type IV collagen according to (11) or (12) above, wherein the plant belonging to the genus Rosa of the family Rosaceae is a rose petal containing at least one compound selected from the group consisting of a delphinidin-type anthocyanin, a rosacyanin compound, and a rosadelphin compound, the plant belonging to the genus Camellia of the family Theaceae is a tea plant leaf, the plant belonging to the genus Hamamelis of the family Hamamelidaceae is an American witch-hazel leaf, the plant belonging to the genus Filipendula of the family Rosaceae is a meadowsweet flower, the plant belonging to the genus Plantago of the family Plantaginaceae is a greater plantain seed, and the plant belonging to the genus Mentha of the family Lamiaceae is a whole plant of peppermint. (14) The composition according to any one of (1) to (13) above, wherein the composition is a topical agent for skin. (15) The composition according to any one of (1) to (14) above, wherein the composition is a cosmetic. (16) Use of an extract for suppressing a decrease in skin barrier function or improving the skin barrier function by suppressing a decrease in or improving expression of type IV collagen, the extract including at least one selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae. (17) Use of an extract for suppressing a decrease in skin barrier function or improving the skin barrier function, the extract including an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae and/or an extract of a plant belonging to the genus Plantago of the family Plantaginaceae. (18) Use of an extract for suppressing a decrease in expression of type IV collagen or improving the expression, the extract including at least one selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, an extract of a plant belonging to the genus Plantago of the family Plantaginaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae. (19) A method of suppressing a decrease in skin barrier function or improving the skin barrier function by suppressing a decrease in or improving expression of type IV collagen, the method including administering at least one extract selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae. (20) A method of suppressing a decrease in skin barrier function or improving the skin barrier function, the method including administering an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae and/or an extract of a plant belonging to the genus Plantago of the family Plantaginaceae. (21) A method of suppressing a decrease in expression of type IV collagen or improving the expression, the method including administering at least one extract selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, an extract of a plant belonging to the genus Plantago of the family Plantaginaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae. (22) A method of screening for a substance having an effect of suppressing a decrease in or improving skin barrier function, the method including: a step of culturing in vitro human epidermal keratinocytes in a medium containing a test substance and separately in a control medium not containing the test substance; and a step of comparing an expression level of type IV collagen between the keratinocytes cultured in the medium containing a test substance and the keratinocytes as a control cultured in the control medium, wherein the test substance is selected as a substance having an effect of suppressing a decrease in or improving skin barrier function when the expression level of type IV collagen in the keratinocytes cultured in the medium containing the test substance is higher than the expression level of type IV collagen in the keratinocytes as a control.

- Advantageous Effects of Invention

[00081 The present invention can provide a composition for suppressing a decrease in or improving skin barrier function capable of suppressing a decrease in skin barrier function or improving the function. The present invention can also provide a composition for suppressing a decrease in or improving expression of type IV collagen. The present invention can also provide a method of suppressing a decrease in skin barrier function or improving the function, and a method of suppressing a decrease in expression of type IV collagen or improving the expression. The present invention can also provide a method of screening for a substance having an effect of suppressing a decrease in or improving skin barrier function.

DESCRIPTION OF EMBODIMENTS

[00091 A first aspect of the present invention is a composition for suppressing a decrease in expression of type IV collagen, the composition containing, as an active ingredient, at least one extract selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, an extract of a plant belonging to the genus Plantago of the family

Plantaginaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae. The composition for suppressing a decrease in or improving expression of type IV collagen of the first aspect of the present invention is also referred to as a composition of the first aspect of the present invention. The at least one extract has an effect of suppressing a decrease in expression of type IV collagen or improving the expression. Suppressing a decrease in expression of type IV collagen includes suppression of progress of the decrease in expression of type IV collagen, maintenance of the expression of type IV collagen, reduction (ease) of the level of the decrease in expression of type IV collagen, and the like. Improving expression of type IV collagen includes recovery of expression of type IV collagen, and the like. Recovery includes at least partial recovery. The expression of type IV collagen includes expression of a gene encoding type IV collagen and expression of type IV collagen at the protein level, and herein may refer to one or both of them. Type IV collagen is preferably human type IV collagen, more preferably human collagen a-1(IV) chain. The human collagen a-1(IV) chain is a protein encoded by the COL4A1 gene. In one embodiment, the composition for suppressing a decrease in or improving expression of type IV collagen of the present invention can be used to suppress a decrease in expression of type IV collagen in a human or improving the expression by suppressing a decrease in expression of the human collagen a-1(IV) chain or improving the expression.

[0010] The expression of type IV collagen is decreased (the expression level is lowered) by external factors such as UV and internal factors such as aging. As described later in EXAMPLES, suppression of a decrease in expression of type IV collagen or improvement of the expression in epidermal keratinocytes suppressed a decrease in transepithelial electrical resistance (TEER) of the skin or improved the resistance. This demonstrates that a decrease in skin barrier function was suppressed or the skin barrier function was improved. A specific mechanism for suppression of a decrease in skin barrier function or improvement of the function by suppressing a decrease in or improving expression of type IV collagen is not clear but presumably as follows. Suppressing a decrease in or improving expression of type IV collagen which is a component of a basement membrane suppressed a decrease in basement membrane function or improved the function, which enabled maintenance of skin homeostasis and finally led to suppression of a decrease in skin barrier function or improvement of the function. Suppressing a decrease in basement membrane function or improving the function is therefore expected to be able to suppress a decrease in skin barrier function or improve the skin barrier function. The extract of any of the plants or the yeast have an excellent effect of suppressing a decrease in or improving expression of type IV collagen to be usable for suppressing a decrease in basement membrane function or improving the function. In terms of the effect of suppressing a decrease in or improving expression of type IV collagen, the at least one extract in the composition of the first aspect of the present invention preferably includes at least one selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, and an extract of a plant belonging to the genus Filipendula of the family Rosaceae.

[0011] The skin barrier function refers to a barrier function of the skin that prevents entry of foreign substances into the living body from the outside and keeps water in the skin by preventing water in the body from excessively evaporating from the skin. Suppressing a decrease in skin barrier function includes suppression of progress of the decrease in skin barrier function, maintenance of the skin barrier function, and reduction (ease) of the level of the decrease in skin barrier function. Improving the skin barrier function includes recovery of the skin barrier function. The decrease in skin barrier function may include a decrease in skin barrier function due to aging, a decrease in skin barrier function due to exposure to UV, and the like.

[0012] As described above, suppressing a decrease in or improving basement membrane function can suppress a decrease in skin barrier function or improve the function. A second aspect of the present invention is a composition for suppressing a decrease in or improving skin barrier function containing, as an active ingredient, an agent for suppressing a decrease in or improving basement membrane function. Hereafter, the composition for suppressing a decrease in or improving skin barrier function of the second aspect of the present invention may be also referred to as a composition of the second aspect of the present invention.

[0013] The basement membrane is known to function as a mechanical support connecting the epidermis and the dermis, a selective filter for substance exchange between the epidermis and the dermis, and the like. Suppressing a decrease in expression of a basement membrane component such as type IV collagen or improving the expression is effective for suppressing a decrease in or improving basement membrane function. The agent for suppressing a decrease in or improving basement membrane function may be any agent having an effect of suppressing a decrease in or improving basement membrane function, such as an agent having an effect of suppressing a decrease in or improving expression of a basement membrane component. The agent for suppressing a decrease in or improving basement membrane function preferably suppresses a decrease in basement membrane function or improves the basement membrane function by suppressing a decrease in or improving expression of type IV collagen. The agent for suppressing a decrease in or improving basement membrane function used may be, for example, the above-mentioned extract of any of the plants or the yeast having an effect of suppressing a decrease in or improving expression of type IV collagen. In one embodiment, the agent for suppressing a decrease in or improving basement membrane function used may include at least one extract selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, an extract of a plant belonging to the genus Plantago of the family Plantaginaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae. Among the above extracts, the agent for suppressing a decrease in or improving basement membrane function preferably includes at least one selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, and an extract of a plant belonging to the genus Filipendula of the family Rosaceae.

[0014]

A third aspect of the present invention is a composition for suppressing a decrease in or improving skin barrier function by suppressing a decrease in or improving expression of type IV collagen, the composition containing, as an active ingredient, at least one extract selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae. Hereafter, the composition for suppressing a decrease in or improving skin barrier function of the third aspect of the present invention is also referred to as a composition of the third aspect of the present invention. In the composition of the third aspect of the present invention, the extract preferably includes at least one selected from the group consisting of an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, and an extract of a plant belonging to the genus Filipendula of the family Rosaceae.

[0015] A fourth aspect of the present invention is a composition for suppressing a decrease in or improving skin barrier function, the composition containing, as an active ingredient, an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae and/or an extract of a plant belonging to the genus Plantago of the family Plantaginaceae. Hereafter, the composition for suppressing a decrease in or improving skin barrier function of the fourth aspect of the present invention is also referred to as a 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 belonging to the genus Hamamelis of the family Hamamelidaceae.

[00161 The extracts used in the composition of the third aspect and the composition of the fourth aspect of the present invention have an effect of suppressing a decrease in or improving expression of type IV collagen. Containing any of the extracts, the compositions can suppress a decrease in or improving basement membrane function. In one embodiment, the compositions for suppressing a decrease in or improving skin barrier function of the third and fourth aspects of the present invention can be used for suppressing a decrease in skin barrier function or improving the skin barrier function by suppressing a decrease in or improving basement membrane function. The composition for suppressing a decrease in or improving skin barrier function of the fourth aspect of the present invention can be used for suppressing a decrease in skin barrier function or improving the skin barrier function by suppressing a decrease in or improving basement membrane function which is achieved by suppressing a decrease in or improving expression of type IV collagen. In one embodiment, the composition for suppressing a decrease in or improving skin barrier function of the fourth aspect of the present invention can be used for suppressing a decrease in or improving skin barrier function by suppressing a decrease in or improving expression of type IV collagen.

[0017] Hereinbelow, the composition for suppressing a decrease in or improving expression of type IV collagen of the first aspect of the present invention and the compositions for suppressing a decrease in or improving skin barrier function of the second, third, and fourth aspects of the present invention are collectively referred to as compositions of the present invention. In the compositions of the present invention, two or more of the extracts may be used in combination as active ingredients.

[0018] In the present invention, the plant belonging to the genus Rosa of the family Rosaceae is a rose belonging to the genus Rosa of the family Rosaceae, typified by Rosa hybrida (scientific name). The plant belonging to the genus Rosa of the family Rosaceae is preferably a rose containing at least one compound selected from the group consisting of a delphinidin-type anthocyanin, a rosacyanin compound, and a rosadelphin compound, more preferably a rose containing, in its petals, at least one compound selected from the group consisting of a delphinidin-type anthocyanin, a rosacyanin compound, and a rosadelphin compound. In particular, preferred is a rose containing a delphinidin-type anthocyanin in its petals.

[0019] The delphinidin-type anthocyanin is a pigment mostly contained in violet or blue petals. Examples of the delphinidin-type anthocyanin include delphinidin, malvidin, petunidin, and compounds obtained by modifying the foregoing pigments with a saccharide or an acyl group. Examples of a rose containing a delphinidin-type anthocyanin include roses showing blue to wisteria colors, such as SUNTORY blue rose APPLAUSE®. Whether or not the rose contains a delphinidin-type anthocyanin can be determined by HPLC by the method disclosed in JP 5697040 B2.

[0020] The rosacyanin compound includes at least one compound selected from the group consisting of rosacyanin

Al, rosacyanin A2, and rosacyanin B. A rose containing a rosacyanin compound may be any rose containing at least one of the above-mentioned compounds. The rose preferably contains rosacyanin Al, rosacyanin A2, and rosacyanin B. Rosacyanin Al is a compound represented by the following formula (I) wherein R1 is a hydrogen atom. Rosacyanin A2 is a compound represented by the following formula (II) 2 wherein R is a hydrogen atom. Rosacyanin B is a compound represented by the following formula (III) wherein R 3 is a hydrogen atom.

[0021]

[Chem. 1]

OH HO OH OH HO / OH

O O OH OH HO O OH O H HO O0 HO H

OH HO

[0022] In the formula, coordination (wave line) of a hydroxy group at position 1 of glucose indicates tautomerism between an a form and a P form, and R1 is a hydrogen atom or a hydrocxy group.

[0023]

[Chem. 2]

HO OH HO O HO OH

+ OHHO - OH HO O 2 O O OH O

OHOO OOH 0 H HO O0 HO 00 H HO

HO OH

[0024] In the formula, R 2 is a hydrogen atom or a hydroxy group.

[0025]

[Chem. 3]

OH OH

HO O R3

H0 0 (ill)

HO OH

[0026] In the formula, R 3 is a hydrogen atom or a hydroxy group.

[0027] Examples of the rose containing a rosacyanin compound include blue to wisteria-colored roses, such as Madame Violet, Purple Rain, Lavande, Manhattan Blue, Chantilly

Lace, Blue Moon, Tasogare, Charles de Gaulle, Violet Dolly, Blue Ribbon, Aozora, Lady X, Blue Bajou, and Sterling Silver. These roses are preferred as they contain a rosacyanin compound in their petals.

[0028] The rosadelphin compound includes at least one compound selected from the group consisting of rosadelphin Al, rosadelphin A2, and rosadelphin B. The rose containing a rosadelphin compound is a rose containing at least one of the above compounds. Preferably, the rose is a rose containing rosadelphin Al, rosadelphin A2, and rosadelphin B. Rosadelphin Al is a compound represented by the formula (I) wherein R' is a hydroxy group. Rosadelphin A2 is a 2 compound represented by the formula (II) wherein R is a hydroxy group. Rosadelphin B is a compound represented by the formula (III) wherein R 3 is a hydroxy group.

[0029] Wild roses do not contain the rosadelphin compound. In roses into which a gene encoding flavonoid 3',5' hydroxylase (hereafter, also referred to as a flavonoid 3',5'-hydroxylase gene) is introduced by a technique such as gene recombination or in roses carrying the flavonoid 3',5'-hydroxylase gene which are obtained from genetically modified roses as a parent, delphinidin which is a blue pigment is synthesized. To the synthesized delphinidin is bound a gallate or a tellimagrandin, thereby producing a rosadelphin compound.

[0030] Roses containing the rosadelphin compound used may be roses into which a flavonoid 3',5'-hydroxylase gene is introduced by a technique such as gene recombination or roses carrying a flavonoid 3',5'-hydroxylase gene which are obtained from genetically modified roses as a parent. Examples of the roses include roses such as SUNTORY blue rose APPLAUSE®. SUNTORY blue rose APPLAUSE® is known to have petals containing rosacyanin Al, rosacyanin A2, rosacyanin B, rosadelphin Al, rosadelphin A2, and rosadelphin B (e.g., WO 2010/110382). SUNTORY blue rose APPLAUSE® is available from Suntory Flowers Ltd. (Tokyo, Japan). Roses carrying a flavonoid 3', 5'-hydroxylase gene can be produced by the method disclosed in WO 2010/110382 or the like. Examples of the flavonoid 3',5'-hydroxylase gene include a plant-derived gene disclosed in WO 2004/020637. Preferred is a flavonoid 3',5'-hydroxylase gene derived from pansy belonging to the genus Viola (e.g., Viola spp. Cv. black Pansy). Roses containing a flavonoid 3',5'-hydroxylase gene are preferred as they contain a rosadelphin compound in their petals. All of the academic and patent literatures described herein are incorporated herein by reference.

[0031] The rosacyanin compound and/or the rosadelphin compound contained in roses can be confirmed, for example, by HPLC-TOF-MS by the method disclosed in WO 2010/110382, WO 2015/178385, or the like.

[0032] The grape seed and the grape skin refer to a seed and a skin of a grape (scientific name: Vitis spp.) which is a plant belonging to the genus Vitis of the family Vitaceae. The variety of a grape is not limited, and examples thereof include black grapes such as Cabernet Sauvignon, Concord, Merlot, Muscat Bailey A; and white grapes such as Chardonnay and Koshu. Among these, Chardonnay and black grapes are preferred, black grapes are more preferred, and Cabernet Sauvignon, Merlot, and the like are still more preferred. The grape for the extract of a grape seed and/or a grape skin may include one or more varieties of grapes. The extract of a grape seed and/or a grape skin is preferably an extract of a grape seed and a grape skin.

[0033]

The yeast belonging to the genus Yarrowia is preferably Yarrowia lipolytica. The extract of a yeast belonging to the genus Yarrowia is preferably an extract of Yarrowia lipolytica.

[00341 The plant belonging to the genus Camellia of the family Theaceae is preferably tea plant (scientific name: Camellia sinensis). The plant belonging to the genus Hamamelis of the family Hamamelidaceae is preferably American witch-hazel (scientific name: Hamamelis virginiana). The plant belonging to the genus Filipendula of the family Rosaceae is preferably meadowsweet (scientific name: Filipendula ulmaria)

[0035] The plant belonging to the genus Plantago of the family Plantaginaceae is preferably greater plantain (scientific name: Plantago major). The plant belonging to the genus Mentha of the family Lamiaceae is preferably peppermint (scientific name: Mentha x piperita).

[0036] The extract of the plant can be prepared using any of the following parts of the plants described above: root, rhizome, stem, leaf, branch, bark, flower, seed (including seed coat, germ, albumen, hypocotyl, cotyledon, and the like), fruit (including pulp and pericarp), or a combination of two or more of these parts. The extract of the plant can be produced by extracting any of these parts of the plants with a solvent. The following parts can be mentioned as examples of preferred parts for producing the extract of the plant. Preferred examples of the extract of the plant in the present invention include extracts of the following parts of the plant.

In the case of the plant belonging to the genus Rosa of the family Rosaceae, a flower, in particular, a petal is preferred. In the case of a grape, a seed and/or a skin is/are used, and a seed and a skin are preferably used. In the cases of the plant belonging to the genus Camellia of the family Theaceae and the plant belonging to the genus Hamamelis of the family Hamamelidaceae, a leaf is preferred. In the case of the plant belonging to the genus Filipendula of the family Rosaceae, a flower is preferred. In the case of the plant belonging to the genus Plantago of the family Plantaginaceae, a seed is preferred. In the case of the plant belonging to the genus Mentha of the family Lamiaceae, a whole plant is preferred. Any of the above-mentioned parts of the plant may be directly extracted, or may be crushed, cut, or dried before extraction. In the present invention, a plant extract may be directly extracted from any of the above plants, or may be extracted from a crushed, cut, or dried product of any of the above plants. An extract from a crushed, cut, or dried product of any of the above plants is preferred.

[0037] Any plant extraction method may be used to obtain a plant extract. An extraction method commonly used to extract plant components can be used. The extraction method can be suitably selected, and extraction conditions are also not limited. For example, preferably, a plant extract is extracted from any of the above plants with a solvent (extraction solvent) at room temperature or under heating. In preparation of a plant extract, any of the above plants as raw materials may be directly extracted, or may be crushed, cut, or dried before extraction.

[0038] The extraction solvent used to prepare a plant extract may be suitably selected, and one commonly used to extract plant components can be used. Examples of the extraction solvent include water; C1-C5 monohydric alcohols such as methanol, ethanol, propanol, and butanol; C2-C5 polyhydric alcohols 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; open-chain or cyclic ethers such as tetrahydrofuran and diethyl ether; polyethers such as polyethylene glycol; and squalane. Any of these may be used alone, or a mixed solvent (mixture) of two or more thereof may be used. Among the C1-C5 monohydric alcohols, C1-C4 monohydric alcohols are preferred; C2-C4 monohydric alcohols are more preferred; and ethanol is still more preferred. Among the C2-C5 polyhydric alcohols, C2-C4 di or trihydric alcohols are preferred; dihydric alcohols are more preferred; and 1,3-butylene glycol is still more preferred. Of these, the extraction solvent is preferably water, a C1-C5 monohydric alcohol, a C2-C5 polyhydric alcohol, or a mixed solvent of two or more thereof; more preferably water, a C2-C4 monohydric alcohol or an aqueous solution thereof, or a C2-C4 dihydric alcohol or an aqueous solution thereof; still more preferably water, ethanol, an aqueous solution of ethanol, 1,3-butylene glycol, or an aqueous solution of 1,3-butylene glycol; particularly preferably water, an aqueous solution of ethanol, or an aqueous solution of 1,3-butylene glycol. In one embodiment, the concentration of ethanol or 1,3-butylene glycol in an aqueous solution of ethanol or an aqueous solution of 1,3 butylene glycol, respectively, is preferably 10 to 98 vol%, more preferably 30 to 90 vol%, still more preferably 30 to 70 vol%. The solvent extracts can be suitably used as the plant extracts in the present invention.

[00391 An acid or alkali may be added during extraction to adjust the pH of the extraction solvent. The extraction is preferably followed by removal of plant residues (the plant or its parts after extraction) from the resulting liquid extract. The method of removing the plant residues from the liquid extract is not limited. For example, a known separation means such as filtration or centrifugation can be used.

[0040] For example, the following method can be used as an example of the plant extraction method. A plant is directly finely crushed, or a dried product of a plant is finely crushed. Then, an extraction solvent in an amount 0.1 to 30 times that of the crushed product is added for extraction at normal pressure and room temperature preferably for 10 minutes to 15 days, more preferably 30 minutes to 10 days, still more preferably 1 hour to 7 days, or at a temperature near the boiling point of the extraction solvent preferably for about 10 minutes to 1 day (more preferably for 10 minutes to 2 hours), followed by filtration to obtain a filtrate. The extraction may be performed under standing still or suitable stirring. The resulting liquid filtrate (liquid plant extract) may be directly used as a plant extract, or may be diluted, concentrated, dried, or the like, if necessary.

[0041] In the present invention, a liquid plant extract obtained by extraction can be directly used as a plant extract. The liquid plant extract may be diluted, concentrated, or dried by a known method to provide a dilute solution, concentrate, or powder, or may be prepared in the form of a paste, as long as the effects of the present invention are not impaired. Examples of the drying method include freeze drying and spray drying. In addition, the liquid plant extract or its concentrate, dried powder, or the like may be further purified by deodorizing, decolorizing, or the like, if necessary, without impairing the effects of the present invention. Such a purification method may be any usual means. The plant extracts in the present invention encompass liquid extracts extracted with various solvents which were obtained by the extraction method as described above, and diluted solutions, concentrates, and dried powders of such liquid extracts; and purified products thereof. The extracts may be diluted or dissolved in a solvent different from the extraction solvent. The plant extracts are also commercially available, and commercial products can be used.

[0042] The extract of a yeast belonging to the genus Yarrowia is preferably an extract of a cell or a cell culture of a yeast belonging to the genus Yarrowia. Examples of the extract of a cell or a cell culture include those obtained by subjecting a cell or a cell culture (preferably a cell culture liquid) containing a cell to a fungal disruption treatment such as autolysis or enzymatic decomposition to dissolve a yeast content into the culture medium or the like (cell disruption product), and those obtained by removing cell residues from the cell or the cell culture (cell disruption product) after the cell disruption treatment. Preferred is an extract obtained by subjecting a cell or a cell culture (preferably a cell culture liquid) containing a cell to autolysis. The cell disruption product or a product obtained by removing cell residues from the cell disruption product may be subjected to sterilization such as heat treatment, if necessary. The yeast extract may be one subjected to such sterilization. The extract of a yeast belonging to the genus Yarrowia may be in any form such as a paste, a suspension, a concentrate, a liquid, or a powder.

[0043] In one embodiment, the extract of a plant belonging to the genus Rosa of the family Rosaceae is preferably an extract of a rose (preferably its petals) in ethanol or in an aqueous solution of ethanol. The extract of a grape seed and/or a grape skin is preferably an extract of a grape seed and/or a grape skin in ethanol or in an aqueous solution of ethanol. The extract of a plant belonging to the genus Camellia of the family Theaceae is preferably an extract of tea plant (preferably its leaves) in ethanol or in an aqueous solution of ethanol. The extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae is preferably an extract of American witch-hazel (preferably its leaves) in 1,3-butylene glycol or in an aqueous solution of 1,3 butylene glycol. The extract of a plant belonging to the genus Filipendula of the family Rosaceae is preferably an extract of meadowsweet (preferably its flowers) in 1,3-butylene glycol or in an aqueous solution of 1,3-butylene glycol. The extract of a plant belonging to the genus Plantago of the family Plantaginaceae is preferably an extract of greater plantain (preferably its seeds) in ethanol or in an aqueous solution of ethanol. The extract of a plant belonging to the genus Mentha of the family Lamiaceae is preferably an extract of peppermint (preferably a whole plant thereof) in ethanol, in an aqueous solution of ethanol, or in hot water.

[0044] The compositions of the present invention may have any form. Examples include powders, granules, pastes, solids, and liquids, and any of which can be suitably selected. As long as the effects of the present invention are not impaired, other components may be added, in addition to the extract, to the compositions of the present invention. The other components may be components that can be used in products such as cosmetics, foods and beverages, and pharmaceutical and quasi-pharmaceutical products, which are described later. The composition for suppressing a decrease in or improving expression of type IV collagen and the composition for suppressing a decrease in or improving skin barrier function of the present invention may be provided as an agent, for example, but it is not limited thereto. The agent may be provided as a composition by itself or as a composition containing the agent. In one embodiment, the compositions of the present invention may be referred to as agents for suppressing a decrease in or improving expression of type IV collagen or agents for suppressing a decrease in or improving skin barrier function.

[0045] The composition for suppressing a decrease in or improving expression of type IV collagen of the first aspect of the present invention is used to suppress a decrease in expression of type IV collagen or improve the expression in a subject, preferably to suppress a decrease in expression of type IV collagen or improve the expression in the skin of the subject. The compositions for suppressing a decrease in or improving skin barrier function of the second, third, and forth aspects of the present invention are used to suppress a decrease in skin barrier function or improve the function. The compositions of the present invention can be used to prevent or ameliorate troubles caused by a decrease in skin barrier function, a decrease in basement membrane function, or a decrease in expression of type IV collagen, for example. Examples of the troubles include dry skin, itchy skin, rough skin, sensitive skin, and damaged skin. In one embodiment, the compositions of the present invention can be used to prevent or ameliorate dry skin, itchy skin, rough skin, sensitive skin, damaged skin, or the like by suppressing a decrease in or improving skin barrier function or by suppressing a decrease in or improving expression of type IV collagen. Prevention of troubles encompasses prevention of onset, delay of onset, and lower incidence. Amelioration of troubles encompasses alleviation of symptoms, good reversal of symptoms, and suppression of progression of symptoms.

[0046] The compositions of the present invention can be used in various applications such as cosmetics, foods and beverages, and pharmaceutical and quasi-pharmaceutical products. The compositions of the present invention each may be a cosmetic, a food or beverage, or a pharmaceutical or quasi-pharmaceutical product by itself for suppressing a decrease in or improving expression of type IV collagen, or for suppressing a decrease in or improving skin barrier function; or may be a material, a preparation, or the like that is added to such a cosmetic, food or beverage, pharmaceutical or quasi-pharmaceutical product, or the like.

[0047] The compositions of the present invention are each suitably used as a topical agent for skin. The topical agent for skin encompasses cosmetics and pharmaceutical and quasi-pharmaceutical products. Preferably, the topical agent for skin is a cosmetic. The compositions of the present invention each can also be used as a pharmaceutical or quasi-pharmaceutical product, other than the topical agent for skin. In another embodiment, the compositions of the present invention each can also be provided as a food or beverage. The following describes a case where the compositions of the present invention each are provided as a topical agent for skin such as a cosmetic or a pharmaceutical or quasi-pharmaceutical product, a food or beverage, or the like.

[0048] When the compositions or the like of the present invention are each provided as a topical agent for skin, the dosage form and the like are not limited, and may be provided in any form. Examples include solutions, emulsions, creams, gels, powders, aerosols, mists, capsules, and sheets. Preferably, the topical agent for skin is a cosmetic. The product form of the cosmetic is also not limited. Examples include skin care cosmetics such as face wash, makeup remover, skin lotion, essence lotion, face pack, emulsion, cream, and sunscreen lotion; makeup cosmetics such as foundation, makeup base, lipstick, eyeshadow, eyeliner, mascara, eyebrow pencil, brush, and nail enamel; hair cosmetics such as shampoo, hair conditioner, hair styling products, hair dyes, and hair growth products; cleansing agents such as soap and body wash; and bath salts.

[0049] The above cosmetics may suitably contain one or more cosmetically acceptable carriers, additives, and like other components, without impairing the effects of the present invention. Examples include water, alcohols, oils, surfactants, thickeners, metal soaps, gelling agents, powders, chelating agents, water-soluble polymers, film forming agents, resins, inclusion compounds, antimicrobial agents, deodorants, salts, pH adjusting agents, ultraviolet light absorbers, extracts from microorganisms/plants/animals other than those mentioned above, keratolytic agents, enzymes, hormones, other vitamins, humectants, antiseptics, anti-inflammatory agents, and perfumes. The cosmetics can be produced by a usual production method in which, for example, any of the extracts used as an active ingredient is mixed with one or more cosmetically acceptable components described above, and the mixture is then processed into a desired form.

[0050] When the topical agent for skin is provided as a pharmaceutical or quasi-pharmaceutical product, components such as pharmaceutically or quasi-pharmaceutically acceptable carriers and additives may be used, without impairing the effects of the present invention. Examples of such components include excipients, binders, disintegrants, lubricants, antioxidants, and colorants. One or more of these components can be used, if necessary.

[0051] The compositions of the present invention each can also be provided as a pharmaceutical or quasi pharmaceutical product, other than the topical agent for skin described above. Such a pharmaceutical or quasi pharmaceutical product may be administered orally or parenterally. The pharmaceutical or quasi-pharmaceutical product can be provided in the form of a formulation for oral administration (agent for internal use) or a formulation for parenteral administration. Examples of the dosage form of the formulation for oral administration include liquids, tablets, powders, fine granules, granules, sugar-coated tablets, capsules, suspensions, emulsions, and chewable tablets. Examples of the dosage form of the formulation for parenteral administration include injections and infusions. In such a pharmaceutical or quasi-pharmaceutical product, components such as pharmaceutically or quasi-pharmaceutically acceptable carriers and additives may be used. Examples of such components include excipients, binders, disintegrants, lubricants, antioxidants, colorants, and taste masking agents. One or more of these components can be used, if necessary. The pharmaceutical or quasi-pharmaceutical product can be produced by a usual production method in which, for example, any of the extracts used as an active ingredient is mixed with one or more pharmaceutically or quasi-pharmaceutically acceptable components, and the mixture is then processed into a desired form.

[0052] When the compositions of the present invention each are provided as a food or beverage, the food or beverage is not limited, and examples include general foods and beverages, health foods, foods with function claims, and foods for specified health uses. The form of the food or beverage is also not limited. The health foods, foods with function claims, and foods for specified health uses can be provided in the form of various formulations such as liquids, tablets, powders, fine granules, granules, sugar coated tablets, capsules, suspensions, emulsions, chewable tablets, and liquid foods.

[0053] These foods and beverages may contain food- or beverage acceptable ingredients, such as other food or beverage materials and additives for foods and beverages, without impairing the effects of the present invention. Such foods and beverages can be produced by a usual production method. For example, the method may only require adding any of the extracts used as an active ingredient in the present invention to food or beverage materials or the like in the production of foods and beverages.

[0054] When the compositions of the present invention each are a topical agent for skin, a pharmaceutical or quasi pharmaceutical product other than the topical agent for skin, or a food or beverage, the amount of the extract used as an active ingredient (in terms of dry matter) in the composition is preferably 0.0000001 to 100 wt%, more preferably 0.000001 to 100 wt%, still more preferably 0.00001 to 10 wt%. The amount is the total amount when the composition contains two or more extracts.

[0055] The amount of each of the compositions of the present invention to be used is not limited, as long as the amount achieves the effect of suppressing a decrease in or improving skin barrier function or the effect of suppressing a decrease in or improving expression of type IV collagen (effective dose), and can be suitably set according to the subject's conditions, weight, sex, age, or other factors. Preferably, the amount of each of the compositions of the present invention is determined such that the amount of the extract (in terms of dry matter) as an active ingredient in a topical agent for skin such as a cosmetic is, for example 0.01 to 500 mg per adult (60 kg) per day. Preferably, the dose of the extract (in terms of dry matter) when orally administered as a pharmaceutical or quasi-pharmaceutical product is, for example, 0.01 to 500 mg per adult (60 kg) per day. Preferably, the intake of the extract (in terms of dry matter) as a food or beverage is, for example, 0.01 to 500 mg per adult (60 kg) per day. The above amount of the extract can be applied, fed, or administered at once or in several portions. The amount is the total amount when the composition contains two or more extracts. The timing at which a topical agent for skin containing the extract is applied to the skin and the timing at which a food, beverage, pharmaceutical product, or quasi-pharmaceutical product containing the plant extract is fed (administered) are not limited.

[00561 Subjects to which the compositions of the present invention are applicable (subject for administration) are not limited. For example, the compositions are applicable to a human or non-human mammal, preferably a human. Examples of subjects include those who want or need to suppress a decrease in or improve skin barrier function and those who want or need to suppress a decrease in or improve expression of type IV collagen. Examples of such subjects include those who realize their skin troubles such as dry skin, itchy skin, rough skin, sensitive skin, or damaged skin. The compositions of the present invention each can be fed or administered by various methods known per se, in accordance with the form, dosage form, or the like thereof. In one embodiment, the compositions of the present invention each are preferably applied to the skin.

[0057] Any of the extracts used as an active ingredient of the compositions of the present invention can be used for suppressing a decrease in expression of type IV collagen of a subject or improving the expression. Any of the extracts can be used for suppressing a decrease in skin barrier function of a subject or improving the skin barrier function, preferably for suppressing a decrease in skin barrier function or improving the skin barrier function by suppressing a decrease in or improving expression of type IV collagen.

[0058] The present invention also encompasses the following uses and methods. Use of an extract for suppressing a decrease in skin barrier function or improving the skin barrier function by suppressing a decrease in or improving expression of type IV collagen, the extract including at least one selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae. A method of suppressing a decrease in skin barrier function or improving skin barrier function by suppressing a decrease in or improving expression of type IV collagen, the method including administering at least one extract selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae.

[00591 Use of an extract for suppressing a decrease in skin barrier function or improving skin barrier function, the extract including an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae and/or an extract of a plant belonging to the genus Plantago of the family Plantaginaceae. A method of suppressing a decrease in skin barrier function or improving skin barrier function, the method including administering an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae and/or an extract of a plant belonging to the genus Plantago of the family Plantaginaceae.

[00601 Use of an extract for suppressing a decrease in expression of type IV collagen or improving the expression, the extract including at least one selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, an extract of a plant belonging to the genus Plantago of the family Plantaginaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae. A method of suppressing expression of type IV collagen or improving the expression, the method including administering the at least one extract.

[0061] The extract is administered (applied) preferably to the skin and can be used for suppressing a decrease in skin barrier function or improving the function, for suppressing a decrease in expression of type IV collagen in the skin or improving the expression, or for suppressing a decrease in basement membrane function or improving the function. In one embodiment, the extract can be used to prevent or ameliorate troubles caused by a decrease in skin barrier function, a decrease in basement membrane function, or a decrease in expression of type IV collagen. The extract can be used for preventing or ameliorating troubles such as dry skin, itchy skin, rough skin, sensitive skin, or damaged skin by suppressing a decrease in or improving skin barrier function, or by suppressing a decrease in or improving expression of type IV collagen in the skin.

[0062] In the uses and methods, the extracts, subjects for administration, the amount of the extract, preferred embodiments thereof, and the like are the same as described above for the compositions of the present invention. Each extract may be directly used or may be used in combination with other components. The compositions of the present invention may be used. Each extract can be used in the form of the topical agent for skin, food or beverage, or the like described above. Each use may be therapeutic or non-therapeutic. Each method may be therapeutic or non-therapeutic. The "non- therapeutic" is a concept that does not include medical activities, i.e., a concept that does not include methods of surgery, therapy, or diagnosis of humans.

[00631 In other aspects, the present invention provides use of any of the extracts to produce a composition for suppressing a decrease in or improving skin barrier function; and use of any of the extracts to produce a composition for suppressing a decrease in or improving expression of type IV collagen. The extracts, preferred embodiments thereof, and the like are the same as described above for the compositions of the present invention. In still other aspects, the present invention provides use of an agent for suppressing a decrease in or improving basement membrane function for suppressing a decrease in skin barrier function or improving the skin barrier function; and a method of suppressing a decrease in skin barrier function or improving the skin barrier function, including administering an agent for suppressing a decrease in or improving basement membrane function. The agent for suppressing a decrease in or improving basement membrane function, preferred embodiments thereof, and the like are the same as described above.

[0064] The present invention also encompasses a method of screening for a substance having an effect of suppressing a decrease in or improving skin barrier function. Next, the screening method of the present invention is described. The method of screening for a substance having an effect of suppressing a decrease in or improving skin barrier function of the present invention includes: a step of culturing in vitro human epidermal keratinocytes in a medium containing a test substance and separately in a control medium not containing the test substance; and a step of comparing the expression level of type IV collagen between the keratinocytes cultured in the medium containing a test substance and the keratinocytes as a control cultured in the control medium, wherein the test substance is selected as a substance having an effect of suppressing a decrease in or improving skin barrier function when the expression level of type IV collagen in the keratinocytes cultured in the medium containing the test substance is higher than the expression level of type IV collagen in the keratinocytes as a control.

[00651 The screening method of the present invention includes culturing in vitro human epidermal keratinocytes in a medium containing a test substance and separately in a control medium not containing the test substance. The human epidermal keratinocytes are commercially available, and a commercial product can be used. The test substance is not limited. Examples thereof include liquid plant extracts, cell extracts, supernatants of cell cultures, fermentation products, proteins, peptides, vitamins, and synthetic compounds. These may be known substances or novel substances.

[00661 The step of culturing in vitro human epidermal keratinocytes in a medium containing a test substance and separately in a control medium not containing the test substance is also referred to as a culturing step. The step of comparing the expression level of type IV collagen between the keratinocytes (also referred to as test keratinocytes) cultured in the medium containing the test substance and the keratinocytes as a control cultured in the control medium is also referred to as a type IV collagen expression level comparing step.

[0067] The conditions for culturing human epidermal keratinocytes in the culturing step are not limited, and known conditions may be employed. The medium containing a test substance can be prepared by mixing a test substance in a medium. The medium used may be one used for culturing of human epidermal keratinocytes, and a commercial product may be used. Examples of the medium include HuMedia-KB2 (available from Kurabo Industries ltd.) and DMEM. To the medium may be added additives such as insulin, human EGF, hydrocortisone, a blood serum, and an antibacterial agent. The culture time is not limited, and may be suitably set according to culture conditions and the like. The control medium used may be the same as the medium containing a test substance except that the test substance is not contained. For example, human epidermal keratinocytes may be cultured in the medium containing a test substance and separately in the control medium preferably for 1 to 72 hours, more preferably 3 to 48 hours. In the culturing step, a stress may be imposed such as irradiation with UV or an oxidative stress. For example, culturing under stress such as irradiation with UV or an oxidative stress enables screening for a substance which is more effective for suppressing a decrease in skin barrier function due to the stress or improving the skin barrier function under stress.

[00681 The culturing step is followed by the type IV collagen expression level comparing step. After the culturing, the expression level of type IV collagen is compared between the keratinocytes cultured in the medium containing a test substance and the keratinocytes as a control cultured in the control medium. When the expression level of type IV collagen in the keratinocytes cultured in the medium containing a test substance is higher than the expression level of type IV collagen in the keratinocytes as a control, the test substance is selected as a substance having an effect of suppressing a decrease in or improving skin barrier function.

[00691 The expression level of type IV collagen can be determined by measuring the amount of mRNA encoding type IV collagen or the amount of type IV collagen protein. The amount of mRNA or protein can be measured by a known method. The amount of mRNA encoding type IV collagen can be measured by a technique such as quantitative PCR, Northern blotting, microarray analysis, or live imaging using a fluorescent probe. The amount of type IV collagen protein can be measured by a technique such as immunochemical assay or mass analysis. Examples of the immunochemical assay include Enzyme Linked Immunosorbent Assay (ELISA) and Western blotting. The amount of protein can be also measured using a commercial ELISA kit (available from MyBioSource, Inc.). The base sequence and amino acid sequence (ORF) of human type IV collagen are known. The base sequence (mRNA) is registered with Accession No. AH002741 in Genbank (public database). A person skilled in the art can easily design and synthesize a primer for measuring the amount of mRNA encoding type IV collagen, as well as amplification of DNA.

[0070] As described above, suppression of a decrease in expression of type IV collagen or improvement of the expression can suppress a decrease in or improve skin barrier function. Therefore, when the expression level of type IV collagen in the test keratinocytes is higher than the expression level of type IV collagen in the keratinocytes as a control, the test substance can be determined as a substance having an effect of suppressing a decrease in or improving skin barrier function. The screening method enables screening for a substance having an effect of suppressing a decrease in or improving skin barrier function.

EXAMPLES

[00711 The following describes the present invention with reference to examples, but the present invention is not limited to these examples.

[0072] <Preparation Example 1> Plant-derived extracts were prepared. The plants and their parts used in the extraction are listed below. (1) Rose (Rosa hybrida, "SUNTORY blue rose APPLAUSE®", Suntory Flowers Ltd.): petal (2) Grape (Vitis spp.) (Cabernet Sauvignon): seed and skin (3) Tea plant (Camellia sinensis): leaf (4) American witch-hazel (Hamamelis virginiana): leaf (5) Meadowsweet (Filipendula ulmaria): flower (6) Greater plantain (Plantago major): seed (7) Peppermint (Mentha x piperita): whole plant The rose ("SUNTORY blue rose APPLAUSE") is a blue colored rose containing a delphinidin-type anthocyanin, rosacyanin Al, rosacyanin A2, and rosacyanin B, as well as rosadelphin Al, rosadelphin A2, and rosadelphin B, in its petals.

[0073] Crushed products of the plants (1) to (7) were each immersed in a solvent for extraction. More specifically, crushed products of the plants (1), (2), (3), (6), and (7) were each immersed in a 30 to 90 vol% aqueous solution of ethanol in an amount of 10 times the amount of the crushed product, and extracted over 10 minutes to 7 days with stirring once a day at room temperature. Thus, liquid extracts were obtained. The plants (4) and (5) were each extracted by the above method except that the aqueous solution of ethanol was changed to an aqueous solution of 1,3-butylene glycol.

[0074] The obtained liquid extracts were subjected to filtering for removal of plant residues, and the resulting filtrates were obtained as liquid plant extracts. The obtained liquid plant extracts were each concentrated in an evaporator, and then freeze-dried. Thus, the extract of each plant was obtained in the form of powder. The powdery plant extracts (rose petal extract, grape seed and skin extract, tea plant leaf extract, American witch-hazel leaf extract, meadowsweet flower extract, greater plantain seed extract, and peppermint extract) were used as extract samples in the following evaluation.

[0075] <Preparation Example 2> Yarrowia lipolytica was cultured, followed by autolysis and subsequent heating for disinfection and sterilization. Thus, a powdery Yarrowia lipolytyca extract was obtained. In the following examples, the above powder was used as an extract sample of Yarrowia lipolytyca.

[0076] <Example 1> Influence of overexpression of type IV collagen on skin barrier under inflammatory stimulus A plasmid incorporating COL4A1 (RefSeq ID: NM_001845.6) downstream of a CMV promoter was created (hereafter, referred to as COL4A1 plasmid). According to the recommended protocol, the COL4A1 plasmid and Lipofectamine 3000 Reagent (available from Thermo Fisher Scientific) were diluted with Opti-MEM@ I Reduced Serum Media (available from Thermo Fisher Scientific) (hereafter, referred to as Opti-MEM) to give two solutions (hereafter, referred to as Opti-MEM solutions) different in plasmid concentration (plasmid concentration: 5 pg/mL or 20 pg/mL). As a control, a solution containing Opti-MEM instead of the plasmid was also produced.

[0077] Using HuMedia-KG2 (hereafter, referred to as a recommended medium) (available from Kurabo Industries ltd.), normal adult human epidermal keratinocytes (available from Kurabo Industries ltd.) were seeded to Cell Culture Insert for 24-well plates (available from Millipore Corporation) at a concentration of 3 x 104 cells/200 pL/well. Immediately after the seeding, 10 pL of either of the Opti MEM solutions were added. To the 24-well plate (available from AGC Techno Glass Co., Ltd.) was added 900 pL of the recommended medium. After 30 minutes, a hydrogen peroxide solution (Nacalai Tesque, Inc.) was added to some of the groups to a final concentration of 100 pM (H 2 0 2 treatment) After four hours from seeding, the medium was changed with the recommended medium. After 24 hours from seeding, the medium was changed with the recommended medium having a Ca 2 + concentration increased to 1.41 mM by addition of CaCl 2

(hereafter, referred to as an irregular recommended medium). The medium was changed with the irregular recommended medium every several days.

[0078] The transepithelial electrical resistance (TEER 2 (Q-cm )) was evaluated using Millicell ERS-2 (available from Merck KGaA) after 96, 120, and 144 hours from seeding. The cells were lysed using isogen (available from Nippon Gene Co., Ltd.) after 144 hours from seeding, and then mRNA was extracted. Subsequently, cDNA was synthesized using High-Capacity cDNA Reverse Transcription Kit and RNase Inhibitor (both available from Thermo Fisher Scientific), followed by quantitative PCR using TaqMan Fast Universal PCR Master Mix (available from Thermo Fisher Scientific) and a primer. Thus, the expression level of human type IV collagen Al (COL4A1) was evaluated. As an endogenous control, Eukaryotic 18S rRNA was used.

[0079] The primer (COL4Al) for measuring the COL4A1 expression level and the primer (Eukaryotic 18S rRNA) for quantifying the endogenous control were #Hs00266237_ml and #Hs99999901_s1 (both available from Thermo Fisher Scientific), respectively.

[0080] Tables 1 and 2 show the results. Table 1 shows the COL4A1 gene expression level relative to the COL4A1 expression level of a plasmid-free and H 20 2 -untreated group (H 2 0 2 (-), plasmid (-) ) which is set to 1. In Tables 1 and 2, "H 20 2 (+)" refers to a H 2 0 2 -treated group. "Plasmid (-)" means that the COL4A1 plasmid was not added and "Plasmid (+)" means that the COL4A1 plasmid was added. The symbol "*" indicates a significant difference (p < 0.05) (Dunnett' s test (vs a plasmid-free and H 2 0 2 -treated group)) In response to H 2 0 2 stimulation, the COL4A1 expression level was lowered. Introduction of the COL4A1 plasmid was confirmed to have a significant and dose-dependent effect of suppressing a decrease in or improving the expression on the H 20 2 -stimulated groups (*: p < 0.05).

[0081]

[Table 1] H 20 2 (-), Plasmid (-) H 20 2 (+), Plasmid (-) H 20 2 (+), Plasmid (+) H 20 2 (+), Plasmid(+) H2 0 2 - H 20 2 100 pM H 2 0 2 100 pM H2 0 2 100 pM Plasmid - - Plasmid 0.05 pg Plasmid 0.20 pg COL4A1 expression level 1.00 0.61 25.70* 86.39*

[0082] Table 2 shows the TEER values after 144 hours from seeding. The symbol "*" indicates a significant difference (p < 0.05) (Dunnett's test (vs a plasmid-free and H 2 0 2 treated group)). In response to H 2 0 2 stimulation, the TEER was significantly lowered. Introduction of the COL4A1 plasmid was confirmed to have a dose-dependent and significant (in the high-dose group) effect of improving TEER on the H 2 0 2 -stimulated groups (*: p < 0.05) .

[0083]

[Table 2] H2 0 2 (-), Plasmid (-) H2 0 2 (+), Plasmid (-) H 2 0 2 (+), Plasmid (+) H2 0 2 (+), Plasmid (+) H2 0 2 - H2 0 2 100pM H2 0 2 100pM H20 2 100pM Plasmid - - Plasmid 0.05 pg Plasmid 0.20 pg TEER (cm 2 ) 51.00* 38.33 44.19 50.06*

[0084] <Examples 2 to 9> Evaluation on effect of suppressing a decrease in or improving type IV collagen gene expression in extract sample Using the recommended medium, a normal adult human epidermal keratinocyte (available from Kurabo Industries ltd.) was seeded to a 12-well plate (available from AGC Techno Glass Co., Ltd.) at a concentration of 5 x 104 cells/mL/well and cultured for 48 hours. The medium was changed with the recommended medium containing an extract sample to start sample treatment. After 30 minutes from the start of sample treatment, a hydrogen peroxide solution (available from Nacalai Tesque, Inc.) was added to a final concentration of 100 pM. After three hours from the start of hydrogen peroxide solution treatment, the medium was changed with the recommended medium not containing the extract sample. After 48 hours from the start of sample treatment, the cells were lysed in Buffer RLT (containing 1% 2-mercapto ethanol, available from Qiagen N. V.) and collected, and mRNA was extracted using RNeasy Mini Kit and RNase-Free DNase Set (available from Qiagen N. V.). Then, synthesis of cDNA and quantitative PCR were performed and the COL4A1 expression level was evaluated by the same methods as in Example 1.

[0085] As the control, cells were cultured in the same manner except that dimethyl sulfoxide (DMSO) was added, instead of the extract sample, to a final concentration of 0.1%. To the cultured cells was added a hydrogen peroxide solution, and the COL4A1 expression level was evaluated by the same methods.

[00861 The extract sample used in each of Examples 2 to 9 was the rose petal extract, the grape seed and skin extract, the tea plant leaf extract, the American witch-hazel leaf extract, the meadowsweet flower extract, the greater plantain seed extract, the Yarrowia lipolytica extract, or the peppermint extract prepared in Preparation Example 1 or 2. The extract sample was dissolved in ultra-pure water or DMSO before addition to the medium. The concentration of the extract sample in the medium was set to 5 to 20 pg/mL. In the case where the extract sample was dissolved in ultra-pure water, DMSO was separately added. Thus, the recommended medium containing any of the extract samples was set to have a final concentration of DMSO of 0.1%.

[0087] Table 3 shows the results of Examples 2 to 9 (average of N = 3 to 4). The results in Table 3 are relative values (%) of the COL4A1 expression level in the cells treated with the extract sample to the COL4A1 expression level of the control which is set to 100%. The symbol "*" indicates a significant difference (p < 0.05) to the control (Student's t-test). Table 3 shows the results of the cases where the COL4A1 expression level was highest within the ranges of the extract sample (test substance) concentration of 5 to 20 pg/mL. The rose petal extract, the grape seed and skin extract, the tea plant leaf extract, the American witch-hazel leaf extract, the meadowsweet flower extract, the greater plantain seed extract, the Yarrowia lipolytica extract, and the peppermint extract were each confirmed to have a significant effect of suppressing a decrease in COL4A1 expression or improving the expression relative to the control (*: p < 0.05).

[00881

[Table 3]

Sample COL4A1 expression level Significant difference Example 2 Rose petal extract 277%

* Example 3 Grape seed and skin extract 248%

* Example 4 Tea plant leaf extract 155%

* Example 5 American witch-hazel leaf extract 151%

* Example 6 Meadowsweet flower extract 271%

* Example 7 Greater plantain seed extract 142%

* Example 8 Yarroia lipolytica extract 147%

* Example 9 Peppermint extract 178%

*

[0089] <Examples 10 to 15> Evaluation on effect of suppressing a decrease in or improving skin barrier function of extract sample Using the recommended medium containing an extract sample, a normal adult human epidermal keratinocyte was seeded to Millicell Cell Culture Insert at a concentration of 3 x 104 cells/200 pL/well. After 30 minutes from seeding, a hydrogen peroxide solution was added to a final concentration of 100 pM. After four hours from seeding, the medium was changed with the recommended medium not containing the extract sample. After 24 hours from seeding, the medium was changed with the irregular recommended medium. Then, the transepithelial electrical resistance (TEER) was evaluated by the same method as in Example 1.

[0090] As the control, cells were cultured by the same method except that the recommended medium to which DMSO was added to a final concentration of 0.1% was used instead of the recommended medium containing the extract sample. To the cells was added a hydrogen peroxide solution, and the transepithelial electrical resistance was evaluated by the same methods.

[0091] The extract sample used in each of Examples 10 to 15 was the extract shown in Table 4 prepared in Preparation Example 1 or 2. The extract sample was dissolved in ultra pure water or DMSO before addition to the medium. The concentration of the extract sample in the medium was set to 5 to 20 pg/mL. In the case where the extract sample was dissolved in ultra-pure water, DMSO was separately added. Thus, the recommended medium containing any of the extract samples was set to have a final concentration of DMSO of 0.1%.

[0092] Table 4 shows the results (average of N = 4 to 6). The results in Table 4 are relative values (%) of TEER of the cells treated with the extract sample to the TEER of the control which is set to 100%. Table 4 shows the results of the cases where the TEER value was highest within the ranges of the concentration of the extract sample (test substance) of 5 to 20 pg/mL and the measuring time of 96 to 144 hours after seeding. Six extracts were confirmed to have an effect of suppressing a decrease in or improving TEER. The grape seed and skin extract, the tea plant leaf extract, the American witch-hazel leaf extract, the meadowsweet flower extract, and the Yarrowia lipolytica extract were confirmed to have a significant difference relative to the control (*: p < 0.05) (Student's t-test).

[0093]

[Table 4]

Sample TEER Significant difference Example 10 Rose petal extract 117% Example 11 Grape seed and skin extract 119% *

Example 12 Tea plant leaf extract 114% *

Example 13 American witch-hazel leaf extract 123% *

Example 14 Meadowsweet flower extract 135% *

Example 15 YarroWialipolytica extract 117% *

[0094] The above plant extracts and the Yarrowia lipolytica extract were found to have an effect of suppressing a decrease in skin barrier function or improving the function.

Claims

Claim 1. A composition for suppressing a decrease in or improving skin barrier function by suppressing a decrease in or improving expression of type IV collagen, the composition comprising, as an active ingredient: at least one extract selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae.
Claim 2. The composition for suppressing a decrease in or improving skin barrier function according to claim 1, wherein the plant belonging to the genus Rosa of the family Rosaceae is a rose, the yeast belonging to the genus Yarrowia is Yarrowia lipolytica, the plant belonging to the genus Camellia of the family Theaceae is tea plant, the plant belonging to the genus Filipendula of the family Rosaceae is meadowsweet, and the plant belonging to the genus Mentha of the family Lamiaceae is peppermint.
Claim 3. The composition for suppressing a decrease in or improving skin barrier function according to claim 1 or 2, wherein the plant belonging to the genus Rosa of the family Rosaceae is a rose petal containing at least one compound selected from the group consisting of a delphinidin-type anthocyanin, a rosacyanin compound, and a rosadelphin compound, the plant belonging to the genus Camellia of the family Theaceae is a tea plant leaf, the plant belonging to the genus Filipendula of the family Rosaceae is a meadowsweet flower, and the plant belonging to the genus Mentha of the family Lamiaceae is a whole plant of peppermint.
Claim 4. A composition for suppressing a decrease in or improving skin barrier function, the composition comprising, as an active ingredient: an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae and/or an extract of a plant belonging to the genus Plantago of the family Plantaginaceae.
Claim 5. The composition for suppressing a decrease in or improving skin barrier function according to claim 4, wherein the plant belonging to the genus Hamamelis of the family Hamamelidaceae is American witch-hazel and the plant belonging to the genus Plantago of the family Plantaginaceae is greater plantain.
Claim 6. The composition for suppressing a decrease in or improving skin barrier function according to claim 4 or 5, wherein the plant belonging to the genus Hamamelis of the family Hamamelidaceae is an American witch-hazel leaf and the plant belonging to the genus Plantago of the family Plantaginaceae is a greater plantain seed.
Claim 7. The composition for suppressing a decrease in or improving skin barrier function according to any one of claims 4 to 6, wherein the composition suppresses a decrease in skin barrier function or improves skin barrier function by suppressing a decrease in or improving basement membrane function.
Claim 8. The composition for suppressing a decrease in or improving skin barrier function according to claim 7, wherein the composition suppresses a decrease in or improving basement membrane function by suppressing a decrease in or improving expression of type IV collagen.
Claim 9. A composition for suppressing a decrease in or improving skin barrier function, the composition comprising, as an active ingredient: an agent for suppressing a decrease in or improving basement membrane function.
Claim 10. The composition for suppressing a decrease in or improving skin barrier function according to claim 9, wherein the agent for suppressing a decrease in or improving basement membrane function suppresses a decrease in or improving basement membrane function by suppressing a decrease in or improving expression of type IV collagen.
Claim 11. A composition for suppressing a decrease in or improving expression of type IV collagen, the composition comprising, as an active ingredient: at least one extract selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Hamamelis of the family

Hamamelidaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, an extract of a plant belonging to the genus Plantago of the family Plantaginaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae.
Claim 12. The composition for suppressing a decrease in or improving expression of type IV collagen according to claim 11, wherein the plant belonging to the genus Rosa of the family Rosaceae is a rose, the yeast belonging to the genus Yarrowia is Yarrowia lipolytica, the plant belonging to the genus Camellia of the family Theaceae is tea plant, the plant belonging to the genus Hamamelis of the family Hamamelidaceae is American witch-hazel, the plant belonging to the genus Filipendula of the family Rosaceae is meadowsweet, the plant belonging to the genus Plantago of the family Plantaginaceae is greater plantain, and the plant belonging to the genus Mentha of the family Lamiaceae is peppermint.
Claim 13. The composition for suppressing a decrease in or improving expression of type IV collagen according to claim 11 or 12, wherein the plant belonging to the genus Rosa of the family Rosaceae is a rose petal containing at least one compound selected from the group consisting of a delphinidin-type anthocyanin, a rosacyanin compound, and a rosadelphin compound, the plant belonging to the genus Camellia of the family Theaceae is a tea plant leaf, the plant belonging to the genus Hamamelis of the family Hamamelidaceae is an American witch-hazel leaf, the plant belonging to the genus Filipendula of the family Rosaceae is a meadowsweet flower, the plant belonging to the genus Plantago of the family Plantaginaceae is a greater plantain seed, and the plant belonging to the genus Mentha of the family Lamiaceae is a whole plant of peppermint.
Claim 14. The composition according to any one of claims 1 to 13, wherein the composition is a topical agent for skin.
Claim 15. The composition according to any one of claims 1 to 14, wherein the composition is a cosmetic.
Claim 16. Use of an extract for suppressing a decrease in skin barrier function or improving the skin barrier function by suppressing a decrease in or improving expression of type IV collagen, the extract comprising at least one selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae.
Claim 17. Use of an extract for suppressing a decrease in skin barrier function or improving the skin barrier function, the extract comprising an extract of a plant belonging to the genus Hamamelis of the family

Hamamelidaceae and/or an extract of a plant belonging to the genus Plantago of the family Plantaginaceae.
Claim 18. Use of an extract for suppressing a decrease in expression of type IV collagen or improving the expression, the extract comprising at least one selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, an extract of a plant belonging to the genus Plantago of the family Plantaginaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae.
Claim 19. A method of suppressing a decrease in skin barrier function or improving skin barrier function by suppressing a decrease in or improving expression of type IV collagen, the method comprising administering at least one extract selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae.
Claim 20. A method of suppressing a decrease in skin barrier function or improving skin barrier function, the method comprising administering an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae and/or an extract of a plant belonging to the genus Plantago of the family Plantaginaceae.
Claim 21. A method of suppressing a decrease in expression of type IV collagen or improving the expression, the method comprising administering at least one extract selected from the group consisting of an extract of a plant belonging to the genus Rosa of the family Rosaceae, an extract of a grape seed and/or a grape skin, an extract of a yeast belonging to the genus Yarrowia, an extract of a plant belonging to the genus Camellia of the family Theaceae, an extract of a plant belonging to the genus Hamamelis of the family Hamamelidaceae, an extract of a plant belonging to the genus Filipendula of the family Rosaceae, an extract of a plant belonging to the genus Plantago of the family Plantaginaceae, and an extract of a plant belonging to the genus Mentha of the family Lamiaceae.
Claim 22. A method of screening for a substance having an effect of suppressing a decrease in or improving skin barrier function, the method comprising: a step of culturing in vitro human epidermal keratinocytes in a medium containing a test substance and separately in a control medium not containing the test substance ; and a step of comparing an expression level of type IV collagen between the keratinocytes cultured in the medium containing a test substance and the keratinocytes as a control cultured in the control medium, wherein the test substance is selected as a substance having an effect of suppressing a decrease in or improving skin barrier function when the expression level of type IV collagen in the keratinocytes cultured in the medium containing the test substance is higher than the expression level of type IV collagen in the keratinocytes as a control.