WO2021182283A1 - 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 - Google Patents

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 Download PDF

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Publication number
WO2021182283A1
WO2021182283A1 PCT/JP2021/008390 JP2021008390W WO2021182283A1 WO 2021182283 A1 WO2021182283 A1 WO 2021182283A1 JP 2021008390 W JP2021008390 W JP 2021008390W WO 2021182283 A1 WO2021182283 A1 WO 2021182283A1
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Prior art keywords
extracts
rosaceae
plants
decrease
suppressing
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PCT/JP2021/008390
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French (fr)
Japanese (ja)
Inventor
壮太 赤澤
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サントリーホールディングス株式会社
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Priority to JP2022505997A priority Critical patent/JPWO2021182283A1/ja
Priority to KR1020227034866A priority patent/KR20220151196A/en
Priority to CN202180019249.5A priority patent/CN115243667A/en
Priority to AU2021233359A priority patent/AU2021233359A1/en
Publication of WO2021182283A1 publication Critical patent/WO2021182283A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A61K31/366—Lactones having six-membered rings, e.g. delta-lactones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06—Fungi, e.g. yeasts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
    • A61K36/534—Mentha (mint)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/68—Plantaginaceae (Plantain Family)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
    • A61K36/738—Rosa (rose)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/82—Theaceae (Tea family), e.g. camellia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/87—Vitaceae or Ampelidaceae (Vine or Grape family), e.g. wine grapes, muscadine or peppervine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/60—Sugars; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9728—Fungi, e.g. yeasts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/15—Medicinal preparations ; Physical properties thereof, e.g. dissolubility
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types

Definitions

  • the present invention relates to a composition for suppressing or improving a decrease in skin barrier function and a composition for suppressing or improving the expression of type 4 collagen.
  • the present invention also relates to a method for suppressing a decrease in the skin barrier function or improving the skin barrier function and a method for suppressing the decrease in the expression of type 4 collagen or improving the expression.
  • the present invention also relates to a method for screening a substance having an effect of suppressing or improving a decrease in skin barrier function.
  • the skin is roughly divided into two layers, the epidermis and the dermis, from the surface.
  • the epidermis is composed of four layers, from the deep layer to the basal layer, the stratum spinosum, the stratum granulosum, and the stratum granulosum, and the basal layer is adhered to the dermis via the basement membrane.
  • the basement membrane is composed of components such as type 4 collagen and type 7 collagen, and has a function of binding the basal layer of the epidermis to the dermis, but its function deteriorates with aging.
  • the skin has a barrier function of preventing foreign substances from entering the living body from the outside and maintaining the moisture of the skin by preventing excessive evaporation of water in the body from the skin.
  • Patent Document 1 describes a skin barrier function improving agent containing an extract of asparagus linearis and the like.
  • An object of the present invention is to provide a composition for suppressing or improving a decrease in skin barrier function, which can suppress or improve the decrease in skin barrier function. Another object of the present invention is to provide a composition for suppressing or improving the expression of type 4 collagen. Another object of the present invention is to provide a method for suppressing a decrease in skin barrier function or improving the function, and a method for suppressing a decrease in the expression of type 4 collagen or improving the expression. Furthermore, an object of the present invention is to provide a method for screening a substance having an effect of suppressing or improving a decrease in skin barrier function.
  • the present inventor suppresses the decrease in the expression of type 4 collagen in the skin cells or improves the expression, thereby suppressing the decrease in the skin barrier function or the function. Found to be improved.
  • the present inventor has conducted further research based on such findings and has completed the present invention.
  • the present invention includes, but is not limited to, the following compositions for suppressing or improving the decrease in skin barrier function, the compositions for suppressing or improving the expression of type 4 collagen, and the like.
  • Rosaceae Rosaceae plants include rose petals containing one or more compounds selected from the group consisting of delphinidin-type anthocyanin, Rosacyanin compounds and Rosadelfin compounds, Rosaceae Rosaceae plants are Rosaceae leaves, Rosaceae Labiatae
  • a composition for suppressing or improving a decrease in skin barrier function containing an extract of a witch hazel family plant and / or an extract of a plantain family plantago genus as an active ingredient.
  • composition for suppressing or improving the decrease in skin barrier function according to the above [7], wherein the suppression or improvement of the basement membrane function is mediated by the suppression or improvement of the expression of type 4 collagen.
  • a composition for suppressing or improving the deterioration of the skin barrier function which contains an agent for suppressing or improving the deterioration of the basement membrane function as an active ingredient.
  • the agent for suppressing or improving the decrease in basement membrane function suppresses or improves the decrease in basement membrane function through the suppression or improvement of the expression of type 4 collagen.
  • a composition for suppressing or improving the deterioration of the skin barrier function of the skin is mediated by the suppression or improvement of the expression of type 4 collagen.
  • Rosaceae plants of the genus Rosa, Yarrowia yeast are Yarrowia lipolytica, plants of the genus Rosaceae are Chanoki, plants of the genus Rosaceae are Hamamelis, and plants of the genus Filipendula are the plants of the genus Filipendula.
  • Rosaceae Rosaceae plants include rose petals containing one or more compounds selected from the group consisting of delphinidin-type anthocyanin, rosasianin compounds and rosadelphin compounds, Plantainaceae plants of the genus Plantain are leaves of Chanoki, genus Mansaku of the family Plantains.
  • the plant is a leaf of Hamamelis
  • the plant of the genus Rosaceae is the flower of Yukisou
  • the plant of the genus Plantain is the seed of the plantain
  • the plant of the genus Rosaceae is the whole plant of the genus Astragalus.
  • the composition for suppressing or improving the expression of type 4 collagen according to the above.
  • a method for suppressing deterioration of the skin barrier function or improving the skin barrier function by administering an extract of a witch hazel family plant and / or an extract of a plantain family plantago genus.
  • compositions for suppressing or improving a decrease in skin barrier function which can suppress or improve the decrease in skin barrier function.
  • a composition for suppressing or improving the expression of type 4 collagen it is possible to provide a method for suppressing a decrease in skin barrier function or improving the function, and a method for suppressing a decrease in the expression of type 4 collagen or improving the expression.
  • the first aspect of the present invention is an extract of Rosaceae plants, grape seeds and / or peels, an extract of Yarrowia yeast, an extract of Plantains, and a plantain.
  • the active ingredient is one or more extracts selected from the group consisting of extracts of genus plants, extracts of plants of the genus Rosaceae, extracts of plants of the genus Plantains, and extracts of plants of the genus Plantains. It is a composition for suppressing or improving the expression of type 4 collagen contained as.
  • the composition for suppressing or improving the expression of type 4 collagen according to the first aspect of the present invention is also referred to as the composition according to the first aspect of the present invention.
  • the above extract has an effect of suppressing a decrease in the expression of type 4 collagen or improving the expression.
  • Expression of type 4 collagen includes expression of a gene encoding type 4 collagen and expression of type 4 collagen at the protein level, which may be any or both.
  • the type 4 collagen is preferably human type 4 collagen, and more preferably human type 4 collagen ⁇ 1 chain.
  • Human type 4 collagen ⁇ 1 chain is a protein encoded by gene type 4 collagen A1 (COL4A1).
  • the composition for suppressing or improving the expression of type 4 collagen of the present invention suppresses or improves the expression of human type 4 collagen ⁇ 1 chain, thereby expressing the type 4 collagen in humans. It can be used to suppress the decrease in collagen or improve its expression.
  • type 4 collagen decreases (the expression level decreases) due to external factors such as ultraviolet rays and internal factors such as aging. As shown in the examples below, suppressing or improving the expression of type 4 collagen in epidermal keratinized cells suppressed or improved the decrease in transepithelial electrical resistance (TEER) of the skin. This indicates that the decrease in the skin barrier function was suppressed or the skin barrier function was improved.
  • TEER transepithelial electrical resistance
  • the above-mentioned plant or yeast extract has an excellent effect of suppressing or improving the expression of type 4 collagen, it can be used to suppress or improve the decrease of basement membrane function.
  • the above-mentioned extract is an extract of a Rosaceae plant, a grape seed and / or a fruit skin. It is preferable that the species is one or more selected from the group consisting of the plant and the extract of the plant belonging to the genus Filipendula of the Rosaceae family.
  • the skin barrier function refers to a skin barrier function that prevents foreign substances from entering the living body from the outside and maintains skin moisture by preventing excessive evaporation of moisture in the body from the skin. Suppression of deterioration of skin barrier function includes suppressing the progress of deterioration of skin barrier function, maintaining skin barrier function, reducing (alleviating) the degree of deterioration of skin barrier function, and the like. Improvement of the skin barrier function includes restoration of the skin barrier function and the like.
  • the decrease in the skin barrier function may be a decrease in the skin barrier function due to aging, a decrease in the skin barrier function due to exposure to ultraviolet rays, or the like.
  • a second aspect of the present invention is a composition for suppressing or improving the decrease in skin barrier function, which comprises an active ingredient for suppressing or improving the decrease in basement membrane function.
  • the composition for suppressing or improving the decrease in skin barrier function according to the second aspect of the present invention is also hereinafter referred to as the composition according to the second aspect of the present invention.
  • Known functions of the basement membrane include a mechanical support that connects the epidermis and the dermis, and a selective filter for substance exchange between the epidermis and the dermis.
  • Suppressing or improving the expression of basement membrane components such as type 4 collagen is effective in suppressing or improving the decrease in basement membrane function.
  • the agent for suppressing or improving the decrease in basement membrane function is not particularly limited as long as it has an effect of suppressing or improving the decrease in basement membrane function, for example, an effect of suppressing or improving the expression of components of the basement membrane.
  • the agent for suppressing or improving the decrease in basement membrane function is preferably one that suppresses or improves the decrease in basement membrane function through the suppression or improvement of the expression of type 4 collagen.
  • the agent for suppressing or improving the decrease in basement membrane function for example, the above-mentioned extract of a plant or yeast having an effect of suppressing or improving the expression of type 4 collagen can be used.
  • an extract of a Rosaceae plant, a grape seed and / or a peel extract an extract of a Yarrowia yeast, a Plantain plant.
  • One or more species selected from the group consisting of extracts, witch hazel plants, rosaceae plants, plantains, plantains, and plantains. Extracts can be used.
  • a group consisting of an extract of a Rosaceae plant, an extract of a grape seed and / or a fruit skin, and an extract of a Rosaceae Filipendula plant as an agent for suppressing or ameliorating a decrease in basal membrane function is preferable.
  • a third aspect of the present invention is an extract of Rosaceae plants, grape seeds and / or peels, an extract of Yarrowia yeast, an extract of Camellia plants, and Rosaceae.
  • a skin barrier through suppression or improvement of the expression of type 4 collagen which comprises, as an active ingredient, one or more extracts selected from the group consisting of extracts of genus plants and extracts of plants of the genus Camellia of the Rosaceae family. It is a composition for suppressing or improving the deterioration of function.
  • the composition for suppressing or improving the decrease in skin barrier function according to the third aspect of the present invention is also hereinafter referred to as the composition according to the third aspect of the present invention.
  • the above extracts are an extract of grape seed and / or peel, an extract of Yarrowia yeast, an extract of Theaceae Camellia plant, and a rose. It is preferably one or more species selected from the group consisting of extracts of plants belonging to the family Theaceae.
  • a fourth aspect of the present invention is a composition for suppressing or improving a decrease in skin barrier function, which comprises an extract of a witch hazel family plant and / or an extract of a plantain family plantago genus as an active ingredient.
  • the composition for suppressing or improving the decrease in skin barrier function according to the fourth aspect of the present invention is also hereinafter referred to as the composition according to the fourth aspect of the present invention.
  • the composition of the fourth aspect of the present invention preferably contains an extract of a witch hazel plant of the witch hazel family as an active ingredient.
  • composition of the third aspect of the present invention and the above-mentioned extract used in the composition of the fourth aspect have an action of suppressing or ameliorating a decrease in the expression of type 4 collagen. This makes it possible to suppress or improve the decrease in basement membrane function.
  • the composition for suppressing or improving the decrease in skin barrier function according to the third and fourth aspects of the present invention suppresses or improves the decrease in basement membrane function and suppresses or improves the decrease in skin barrier function. It can be used to improve the barrier function.
  • the composition for suppressing or improving the decrease in skin barrier function according to the fourth aspect of the present invention is a skin barrier through suppression or improvement of decrease in basement membrane function through suppression or improvement of expression of type 4 collagen.
  • composition for suppressing or improving the decrease in skin barrier function according to the fourth aspect of the present invention is used to suppress or improve the decrease in skin barrier function by suppressing or improving the expression of type 4 collagen. be able to.
  • compositions for suppressing or improving the expression of type 4 collagen according to the first aspect of the present invention a composition for suppressing or improving the decrease in skin barrier function of the second, third and fourth aspects.
  • the substances are also collectively referred to as the composition of the present invention.
  • two or more of the above extracts may be combined and used as an active ingredient.
  • the Rosaceae Rosa plant is a rose of the Rosaceae Rosa genus represented by Rosa hybrida (scientific name).
  • Rosa hybrida Rosa hybrida
  • a rose containing one or more compounds selected from the group consisting of delphinidin-type anthocyanins, rosasianin compounds and rosadelphin compounds is preferable, and a group consisting of delphinidin-type anthocyanins, rosasianin compounds and rosadelfin compounds on the petals. More preferably, roses containing one or more of the more selected compounds. Of these, roses containing delphinidin-type anthocyanins in the petals are preferable.
  • Delphinidin-type anthocyanins are pigments that are abundant in purple and blue petals.
  • Examples of delphinidin-type anthocyanins include delphinidin, malvidin, petunidin, and compounds in which they are modified with sugars, acyl groups, and the like.
  • Examples of roses containing delphinidin-type anthocyanins include roses having a bluish or wisteria color tone, for example, roses of varieties such as Suntory Blue Rose Applause (registered trademark).
  • the inclusion of delphinidin-type anthocyanins in roses can be confirmed by HPLC, for example, by the method described in Japanese Patent No. 5697040.
  • the rosasianin compound is one or more compounds selected from the group consisting of rosasianin A1, rosasianin A2 and rosasianin B.
  • the rose containing the rosasianin compound may be a rose containing at least one of the above compounds, but is preferably a rose containing rosa rosasianin A1, rosadelphin A2 and rosadelphin B.
  • Rosacyanin A1 is a compound in which R 1 is a hydrogen atom in the following general formula (I)
  • Rosacyanin A2 is a compound in which R 2 is a hydrogen atom in the following general formula (II)
  • Rosacyanin B is In the following general formula (III)
  • R 3 is a compound having a hydrogen atom.
  • R 1 represents a hydrogen atom or a hydroxyl group.
  • R 2 represents a hydrogen atom or a hydroxyl group.
  • R 3 represents a hydrogen atom or a hydroxyl group.
  • Roses containing rosasianin compounds that exhibit a bluish or mauve hue such as Madame Biore, Purple Rain, Labande, Manhattan Blue, Chantery Lace, Blue Moon, Tasogare, Charles de Goal, Violet Dolly, Blue Ribbon, Aozora. , Lady X, Blue Bayou, Sterling Silver and other varieties of roses. These roses are preferable because the petals contain a rosasianin compound.
  • the Rosa Delphin compound is one or more compounds selected from the group consisting of Rosa Delphin A1, Rosa Delphin A2 and Rosa Delphin B.
  • the rose containing the Rosa Delphin compound may be a rose containing at least one of the above compounds, but is preferably a rose containing Rosa Delphin A1, Rosa Delphin A2 and Rosa Delphin B.
  • Rosa Delphin A1 is a compound in which R 1 is a hydroxyl group in the above general formula (I)
  • Rosa Delphin A2 is a compound in which R 2 is a hydroxyl group in the above general formula (II)
  • Rosa Delphin B is in the above formula (III), a compound wherein R 3 is a hydroxyl group.
  • the above Rosadelphin compound does not exist in wild roses, and the gene encoding flavonoid 3', 5'-hydroxylase (hereinafter, also referred to as flavonoid 3', 5'-hydroxylase gene) is genetically modified.
  • Delfinidine which is a blue pigment, is synthesized in roses introduced by such methods and roses having flavonoids 3', 5'-hydroxylase genes obtained by using recombinant roses as parents, and gallates and terimaglandins are synthesized. Is a compound produced by binding.
  • roses containing the above Rosadelphin compound roses into which a flavonoid 3', 5'-hydroxylase gene has been introduced by a method such as gene recombination, and flavonoid 3'obtained from such a genetically modified rose as a parent.
  • a rose carrying the, 5'-hydroxylase gene can be used.
  • a variety of rose such as Suntory Blue Rose Applause (registered trademark) can be used. It is known that the petals of Suntory Blue Rose Applause (registered trademark) include Rosacyanin A1, Rosacyanin A2 and Rosacyanin B, and Rosadelfin A1, Rosadelfin A2 and Rosadelfin B (for example, WO2010 / 110382). ).
  • Suntory Blue Rose Applause (registered trademark) can be obtained from Suntory Flowers Limited (Tokyo, Japan). Roses having flavonoid 3', 5'-hydroxylase genes can also be prepared by the method described in WO2010 / 110382 and the like. Examples of the flavonoid 3', 5'-hydroxylase gene include plant-derived genes described in WO2004 / 02637, for example, derived from Violet pansy (Violet spp. Cv Black Pansy, etc.). Flavonoids 3', 5'-hydroxylase genes and the like are preferred. Roses carrying the flavonoid 3', 5'-hydroxylase gene are preferred because the petals contain the Rosadelfin compound. All academic and patent documents described herein are incorporated herein by reference.
  • the inclusion of the rosesianin compound and / or the rosadelphin compound in the roses can be confirmed by HPLC-TOF-MS by the method described in, for example, WO2010 / 110382, WO2015 / 178385 and the like.
  • the grape seeds and skins are the seeds and skins of grapes (scientific name: Vitis spp.) Of the genus Vitaceae Vitis.
  • the grape varieties are not particularly limited, and examples thereof include black grapes such as Cabernet Sauvignon, Concord, Merlot, and Muscat Berry A; and white grapes such as Chardonnay and Koshu. Among them, Chardonnay or black grapes are preferable, black grapes are more preferable, and Cabernet Sauvignon, Merlot and the like are further preferable.
  • the grapes in the grape seed and / or pericarp extract may be one or more.
  • the grape seed and / or pericarp extract is preferably a grape seed and pericarp extract.
  • Yarrowia lipolytica is preferable.
  • the extract of Yarrowia yeast is preferably an extract of Yarrowia lipolytica.
  • Plantago As a plant of the genus Plantago (Plantago plantago) of the Plantain family, plantain (scientific name Plantago major) is preferable. As a Lamiaceae Mentha plant of the Labiatae family, Mentha x mentha (scientific name: Mentha x piperita) is preferable.
  • any part of the above plant for example, root, rhizome, stem, leaf, branch, bark, flower, seed (including seed coat, germ, endosperm, hypocotyl, cotyledon, etc.), fruit (including seed coat, germ, endosperm, hypocotyl, cotyledon, etc.) (Including flesh, bark), or multiple combinations thereof can be used.
  • a plant extract can be produced by extracting any part of the plant with a solvent.
  • Preferred sites for producing a plant extract include, for example, the following sites.
  • an extract of the following part of the plant is preferable.
  • the Rosaceae plants of the genus Rosa are preferably flowers, especially petals.
  • seed and / or pericarp are used, preferably seeds and pericarp.
  • Leaves are preferable for the Theaceae Camellia plant and the Witch hazel plant. Flowers are preferred for plants of the genus Filipendula in the Rosaceae family. Seeds are preferred for Plantain plants of the Plantain family.
  • the Labiatae plant of the genus Mentha is preferably whole plant.
  • the plant parts may be subjected to the extraction step as they are, or may be subjected to the extraction step after being crushed, cut or dried.
  • the plant extract may be an extract obtained by extracting the above plant as it is, but may be an extract extracted from a crushed, cut or dried plant. Preferably, it is an extract from the crushed, cut or dried plant.
  • the method for extracting a plant for obtaining a plant extract is not particularly limited, and the plant can be obtained by a usual extraction method used for extracting plant components.
  • the extraction method can be set as appropriate, and the extraction conditions are not particularly limited.
  • the above-mentioned plant as a raw material may be subjected to the extraction step as it is, or may be subjected to the extraction step after being crushed, cut or dried.
  • the extraction solvent used for the preparation of the plant extract can be appropriately selected, and those usually used for the extraction of plant components can be used.
  • the extraction solvent for example, water; monohydric alcohol having 1 to 5 carbon atoms such as methanol, ethanol, propanol and butanol; ethylene glycol, propylene glycol, 1,2-butylene glycol, 1,3-butylene glycol, 1,4- Polyhydric alcohols having 2 to 5 carbon atoms such as butylene glycol and 2,3-butylene glycol; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; chain and cyclic ethers such as tetrahydrofuran and diethyl ether; polyethylene.
  • Polyethers such as glycol; squalane and the like can be mentioned. These may be used alone, or a mixed solvent (mixed solution) in which two or more kinds are combined may be used.
  • a mixed solvent mixed solution
  • the monohydric alcohol having 1 to 5 carbon atoms those having 1 to 4 carbon atoms are preferable, those having 2 to 4 carbon atoms are more preferable, and ethanol is further preferable.
  • the polyhydric alcohol having 2 to 5 carbon atoms a divalent or trivalent alcohol having 2 to 4 carbon atoms is preferable, a divalent alcohol is more preferable, and 1,3-butylene glycol is further preferable.
  • the extraction solvent water, a monohydric alcohol having 1 to 5 carbon atoms, a polyhydric alcohol having 2 to 5 carbon atoms, and a mixed solvent of two or more of these are preferable, and water and a monohydric alcohol having 2 to 4 carbon atoms are preferable.
  • an aqueous solution thereof, a dihydric alcohol having 2 to 4 carbon atoms or an aqueous solution thereof is more preferable, and water, ethanol, an aqueous ethanol solution, 1,3-butylene glycol, a 1,3-butylene glycol aqueous solution is more preferable, and water, an aqueous ethanol solution or an aqueous solution thereof.
  • an aqueous solution of 1,3-butylene glycol is particularly preferable.
  • the ethanol or 1,3-butylene glycol aqueous solution preferably has an ethanol or 1,3-butylene glycol concentration of 10 to 98% by volume, more preferably 30 to 90% by volume, and further 30 to 70% by volume. preferable.
  • the above solvent extract can be preferably used.
  • Acid or alkali may be added at the time of extraction to adjust the pH of the extraction solvent. After extraction, it is preferable to remove the plant residue (plant body after extraction or a portion thereof) from the extract.
  • the method for removing the plant residue from the extract is not particularly limited, and examples thereof include known separation means such as filtration and centrifugation.
  • the following methods can be used, for example.
  • the plant is left as it is or dried, crushed, and the extraction solvent is added in an amount of 0.1 to 30 times by weight, preferably under normal pressure at room temperature for 10 minutes to 15 days, more preferably 30 minutes to 10 days, still more preferably.
  • the obtained filtrate plant extract
  • the plant extract obtained by extraction can be used as it is as a plant extract. Further, as long as the effect of the present invention is not impaired, it may be diluted, concentrated or dried by a known method to form a diluted solution, a concentrate or a powder, or it may be prepared in the form of a paste and used. Examples of the drying method include freeze-drying and spray-drying. Further, the above-mentioned plant extract, its concentrate, dry powder and the like may be further purified by deodorization, decolorization and the like, if necessary, as long as the effects of the present invention are not impaired. Such a purification method may be carried out by arbitrarily selecting ordinary means.
  • the plant extract in the present invention includes various solvent extracts obtained by the above-mentioned extraction methods, diluted solutions thereof, concentrates thereof or dry powders thereof, and purified products thereof. Further, the extract can be used by diluting or dissolving it in a solvent different from the extraction solvent. In addition, the above plant extracts are commercially available, and commercially available products can also be used.
  • the extract of Yarrowia yeast is preferably an extract of bacterial cells or cell cultures of Yarrowia yeast.
  • the mycelium or the mycelium culture containing the mycelium preferably the mycelium culture solution
  • a mycelium crushing treatment such as autolysis treatment or enzymatic decomposition treatment.
  • yeast cells eluted in a culture medium crushed cells
  • cells that have undergone cell crushing treatment or cultures that have been subjected to cell crushing (crushed cells) from which bacterial residue has been removed.
  • the cells or a cell culture containing the cells is autolyzed.
  • sterilization treatment such as heat treatment may be performed on the crushed cell product and the product from which the bacterial cell residue has been removed.
  • the above-mentioned yeast extract may have undergone such a sterilization treatment or the like.
  • the form of the extract of Yarrowia yeast is not particularly limited, and examples thereof include paste, suspension, extract, liquid, and powder.
  • an ethanol extract of roses preferably petals
  • an aqueous ethanol extract of grape seed and / or pericarp
  • an ethanol extract or an aqueous ethanol extract of grape seed and / or pericarp is preferable.
  • an ethanol extract of tea plant preferably leaves
  • an aqueous ethanol extract is preferable.
  • the extract of the witch hazel genus plant the extract of Hamamelis (preferably leaves) 1,3-butylene glycol extract or the extract of 1,3-butylene glycol aqueous solution is preferable.
  • a 1,3-butylene glycol extract or a 1,3-butylene glycol aqueous solution extract of Meadowsweet is preferable.
  • an extract of a plantago genus Plantago of the Plantain family an ethanol extract of plantain (preferably seeds) or an aqueous ethanol extract is preferable.
  • an ethanol extract of mint preferably whole plant
  • an aqueous ethanol extract, or a hot water extract is preferable.
  • the form of the composition of the present invention is not particularly limited, and may be in the form of powder, granules, paste, solid, etc., or in liquid form, and can be appropriately selected.
  • other components can be added to the composition of the present invention as long as the effects of the present invention are not impaired.
  • examples of other ingredients include ingredients that can be used in cosmetics, foods and drinks, pharmaceuticals, quasi-drugs, etc., which will be described later.
  • the composition for suppressing or improving the expression of type 4 collagen of the present invention and the composition for suppressing or improving the decrease in skin barrier function can be provided, for example, in the form of an agent, but are limited to this form. It's not a thing.
  • the agent can be provided as it is as a composition or as a composition containing the agent.
  • the composition of the present invention can also be said to be an agent for suppressing or improving the expression of type 4 collagen, and an agent for suppressing or improving the decrease in skin barrier function.
  • the composition for suppressing or ameliorating the decrease in the expression of type 4 collagen according to the first aspect of the present invention is preferably type 4 in the target skin in order to suppress or improve the decrease in the expression of the target type 4 collagen. It is used to suppress the decrease in collagen expression or to improve the expression.
  • the composition for suppressing or improving the deterioration of the skin barrier function of the second aspect, the third aspect and the fourth aspect of the present invention is used for suppressing the deterioration of the skin barrier function of the subject or improving the function.
  • the composition of the present invention can be used, for example, for the prevention or improvement of defects caused by a decrease in skin barrier function, a decrease in basement membrane function, or a decrease in the expression of type 4 collagen.
  • the composition of the present invention suppresses or improves the decrease in skin barrier function, or suppresses or improves the decrease in the expression of type 4 collagen in the skin, thereby causing dryness, itchiness, dryness, sensitive skin, rough skin, etc. Can be used to prevent or improve.
  • Prevention of defects includes prevention of onset, delay of onset, reduction of onset rate, and the like. Improvement of defects includes improvement of symptoms, improvement of symptoms, suppression of progression of symptoms, and the like.
  • composition of the present invention can be used for various purposes such as cosmetics, foods and drinks, pharmaceuticals, and quasi-drugs.
  • the composition of the present invention itself is for suppressing or improving the decrease in the expression of type 4 collagen, or for suppressing or improving the decrease in the skin barrier function, such as cosmetics, foods and drinks, pharmaceuticals or quasi-drugs, etc. It may be a material or a preparation used by blending with the cosmetics, foods and drinks, pharmaceuticals, quasi-drugs, and the like.
  • the composition of the present invention is suitably used, for example, as an external preparation for skin.
  • the external preparation for skin includes cosmetics, pharmaceuticals, and quasi-drugs, and is preferably a cosmetic.
  • the composition of the present invention can also be used as a drug or quasi-drug other than an external preparation for skin.
  • the case where the composition of the present invention is used as a cosmetic, a pharmaceutical product, a skin external preparation such as a quasi-drug, a food or drink, etc. will be described below.
  • the dosage form and the like are not particularly limited, and may be in any form such as a solution, a milky lotion, a cream, a gel, a powder, an aerosol, a mist, a capsule and a sheet. ..
  • the external preparation for skin is preferably a cosmetic.
  • the product form of cosmetics is not particularly limited, and for example, skin care cosmetics such as facial cleansers, makeup removers, lotions, beauty liquids, packs, milky lotions, creams, and sunscreens; foundations, makeup bases, lipsticks, eye shadows, and eyes. Makeup cosmetics such as liner, mascara, eyebrow, cheek red, nail enamel; hair cosmetics such as hair shampoo, hair rinse, hair conditioner, hair dye, hair restorer; cleaning agents such as soap and body soap; bathing agents, etc. Be done.
  • the above-mentioned cosmetics include components such as carriers and additives that are acceptable for cosmetics, for example, water, alcohols, oils, surfactants, thickeners, metal salts, etc., as long as the effects of the present invention are not impaired.
  • One or two or more of extracts, keratolytic agents, enzymes, hormones, other vitamins, moisturizers, bactericidal agents, anti-inflammatory agents, fragrances and the like can be appropriately contained.
  • Cosmetics can be manufactured by a general manufacturing method. For example, it can be obtained by mixing the above-mentioned extract used as an active ingredient with one or more of the above-mentioned ingredients that can be used in cosmetics, and then processing the mixture into a desired form.
  • components such as carriers and additives permitted for the drug or quasi-drug may be used as long as the effects of the present invention are not impaired.
  • examples of such components include one or more of excipients, binders, disintegrants, lubricants, antioxidants, colorants and the like, which can be used as needed.
  • composition of the present invention can also be a drug or quasi-drug other than the above-mentioned external preparation for skin.
  • drugs or quasi-drugs may be administered orally or parenterally.
  • the drug or quasi-drug may be in the form of an orally-administered preparation (oral preparation) or a parenteral-administered preparation.
  • oral preparation oral preparation
  • parenteral-administered preparation examples include liquid preparations, tablets, powders, fine granules, granules, sugar-coated tablets, capsules, suspensions, emulsions, chewable agents and the like.
  • examples of the dosage form of the parenteral preparation include injections, infusions and the like.
  • components such as carriers and additives that are acceptable for the drugs or quasi-drugs may be used.
  • examples of such components include one or more of excipients, binders, disintegrants, lubricants, antioxidants, colorants, flavoring agents and the like, which can be used as needed.
  • Pharmaceuticals and quasi-drugs can be manufactured by a general manufacturing method. For example, it can be obtained by mixing the above extract used as an active ingredient with one or more of the ingredients that can be used in a drug or a quasi drug, and then processing the extract into a desired form.
  • the food or drink is not particularly limited, and examples thereof include general food and drink, health food, food with functional claims, food for specified health use and the like.
  • the form of food and drink is not particularly limited.
  • the above-mentioned health foods, foods with functional claims, and foods for specified health use include various preparations such as liquid preparations, tablets, powders, fine granules, granules, sugar-coated tablets, capsules, suspensions, emulsions, chewables, and liquid foods. It can also be in the form.
  • ingredients permitted in the food and drink for example, other food and drink materials, additives to be blended in the food and drink, and the like may be blended as long as the effects of the present invention are not impaired.
  • Such foods and drinks can be produced by a general production method.
  • the above-mentioned extract used as an active ingredient in the present invention may be blended with food and drink materials and the like.
  • the content of the extract used as the active ingredient is the composition.
  • the weight of the extract in terms of dry matter is preferably 0.000000001 to 100% by weight, more preferably 0.000001 to 100% by weight, still more preferably 0.00001 to 10% by weight.
  • the above-mentioned content is the total content thereof.
  • the amount of the composition of the present invention to be used is not particularly limited as long as it is an amount (effective amount) that can suppress or improve the decrease in skin barrier function and suppress or improve the expression of type 4 collagen. , Can be appropriately set according to the subject's condition, weight, gender, age or other factors.
  • the amount of the composition of the present invention used is, for example, as an external preparation for skin such as cosmetics, for example, as the above-mentioned extract (dry matter equivalent) used as an active ingredient per day per adult (60 kg). , 0.01-500 mg is preferable.
  • the dose is preferably, for example, 0.01 to 500 mg per day for an adult (60 kg) as the above extract (dry matter equivalent).
  • the intake thereof is preferably, for example, 0.01 to 500 mg per day for an adult (60 kg) as the above-mentioned extract (dry matter equivalent).
  • the above amount of extract can be ingested or administered in a single dose or in multiple doses.
  • the amount of the above extract is the total amount when two or more kinds of extracts are used.
  • the timing of applying the external preparation for skin containing the above extract to the skin, the timing of ingesting or administering foods and drinks, pharmaceuticals or quasi-drugs, and the like are not particularly limited.
  • the application target (which can also be said to be an administration target) of the composition of the present invention is not particularly limited and can be applied to humans, non-human mammals and the like, but humans are preferable.
  • Examples of the application target include a subject who desires or needs to suppress or need to suppress or improve the decrease in skin barrier function, and a subject who desires or needs to suppress or need to suppress or need to suppress or improve the expression of type 4 collagen.
  • Such targets include, for example, those who experience dry skin, itching, dryness, sensitive skin, and rough skin.
  • the composition of the present invention can be ingested or administered by various methods known per se, depending on its form, dosage form and the like. In one aspect, the compositions of the invention are preferably applied to the skin.
  • the above-mentioned extract used as the active ingredient of the composition of the present invention can be used to suppress a decrease in the expression of the target type 4 collagen or to improve the expression. Since the above extract suppresses the decrease in the skin barrier function or improves the skin barrier function of the subject, preferably, the decrease in the expression of type 4 collagen is suppressed or improved to suppress the decrease in the skin barrier function or the skin barrier function is improved. Can be used to
  • the present invention also includes the following uses and methods. Extracts of Rosaceae plants, grape seeds and / or peels, Yarrowia, for suppressing or improving the decrease in the expression of type 4 collagen to suppress the decrease in the skin barrier function or improve the skin barrier function. ) Use of one or more extracts selected from the group consisting of extracts of genus yeast, extracts of Camellia plants of the family Rosaceae, extracts of plants of the genus Rosaceae, and extracts of plants of the genus Camellia. ..
  • Extracts of Rosaceae plants, grape seeds and / or peels, extracts of Yarrowia yeast, extracts of Camellia plants, extracts of Rosaceae plants, and perilla A method of suppressing or improving the decrease in skin barrier function by suppressing or improving the expression of type 4 collagen by administering one or more extracts selected from the group consisting of extracts of Rosaceae plants. ..
  • witch hazel plant extract and / or plantain plantago extract to suppress deterioration of skin barrier function or improve skin barrier function.
  • Extracts of Rosaceae plants, grape seeds and / or peels, extracts of Yarrowia yeast, extracts of Rosaceae, and camellia for suppressing or improving the expression of type 4 collagen Selected from the group consisting of extracts of genus plants, extracts of plants of the genus Mansaku of the family Rosaceae, extracts of plants of the genus Rosaceae, extracts of plants of the genus Oobaco, and extracts of plants of the genus Rosaceae. Use of one or more extracts.
  • the above extract is preferably administered (applied) to the skin to suppress a decrease in the skin barrier function or improve the function, to suppress a decrease in the expression of type 4 collagen in the skin, or to improve the expression.
  • the extract can be used to prevent or ameliorate defects caused by decreased skin barrier function, decreased basement membrane function or decreased expression of type 4 collagen.
  • the above extract prevents or improves dryness, itchiness, dryness, sensitive skin, rough skin, etc. of the skin by, for example, suppressing or improving the decrease in skin barrier function, or suppressing or improving the decrease in the expression of type 4 collagen in the skin.
  • the above-mentioned extract, the subject to which the above-mentioned extract is administered, the dose of the extract, preferable embodiments thereof and the like are the same as those in the above-mentioned composition of the present invention.
  • the above extract may be used as it is or may be used in combination with other components.
  • the composition of the present invention may be used.
  • the above extract can be used in the form of the above-mentioned external preparation for skin, food and drink, and the like.
  • the above use may be therapeutic or non-therapeutic use.
  • the above method may be a therapeutic method or a non-therapeutic method.
  • Non-therapeutic is a concept that does not include medical practice, ie human surgery, treatment or diagnosis.
  • Another aspect of the present invention is the use of the above extract for producing a composition for suppressing or improving the decrease in skin barrier function; the composition for suppressing or improving the expression of type 4 collagen in the above extract. It is also used for manufacturing.
  • the extracts and preferred embodiments thereof are the same as those in the compositions of the present invention described above.
  • Yet another aspect of the present invention is the use of a basement membrane function decline suppressor or ameliorating agent to suppress or improve the skin barrier function decline; administer a basement membrane function decline suppressor or improve agent. It is also a method of suppressing a decrease in the skin barrier function or improving the skin barrier function.
  • the agent for suppressing or improving the decrease in basement membrane function and its preferred embodiment are the same as described above.
  • the present invention also includes a method for screening a substance having an effect of suppressing or improving a decrease in skin barrier function.
  • the screening method of the present invention will be described.
  • the method for screening a substance having an effect of suppressing or improving a decrease in skin barrier function of the present invention is a step of culturing human epidermal keratinized cells in a medium containing a test substance or a control medium not containing the above test substance in vitro. , Including the step of comparing the expression level of type 4 collagen in the skin cells cultured in the medium containing the above test substance and the control skin cells cultured in the control medium, and the expression of type 4 collagen in the skin cells cultured in the medium containing the above test substance.
  • the test substance is selected as a substance having an effect of suppressing or improving the decrease of the skin barrier function.
  • human epidermal keratinocytes are cultured in vitro in a medium containing a test substance or a control medium containing no test substance.
  • Human epidermal keratinocytes are commercially available, and commercially available products can be used.
  • the test substance is not particularly limited, and examples thereof include plant extracts, cell extracts, cell culture supernatants, fermentation products, proteins, peptides, vitamins, synthetic compounds and the like. These may be known substances or new substances.
  • the step of culturing human epidermal keratinocytes in vitro in a medium containing a test substance or a control medium not containing the test substance is also referred to as a culturing step.
  • the step of comparing the type 4 collagen expression level in the skin cells cultured in the medium containing the test substance (also referred to as the test skin cells) and the control skin cells cultured in the control medium is also referred to as a type 4 collagen expression level comparison step.
  • the conditions for culturing human epidermal keratinocytes are not particularly limited, and known conditions may be adopted.
  • the medium containing the test substance can be prepared by mixing the test substance with the medium.
  • the medium those used for culturing human epidermal keratinocytes can be used, and commercially available products may be used. Examples of the medium include HuMedia-KB2 medium (manufactured by Kurabo Industries Ltd.), DMEM medium and the like. Insulin, human EGF, hydrocortisone, serum, antibacterial agent and the like may be added to the medium.
  • the culturing time is not particularly limited, and may be appropriately set depending on the culturing conditions and the like.
  • control medium the same medium as the medium containing the test substance may be used except that the test substance is not contained.
  • human epidermal keratinocytes may be cultured in a medium containing a test substance or a control medium for preferably 1 to 72 hours, more preferably 3 to 48 hours.
  • stress loading such as ultraviolet irradiation and oxidative stress loading may be performed.
  • effective components can be screened by suppressing the decrease in skin barrier function due to the stress or improving the skin barrier function under stress.
  • a type 4 collagen expression level comparison step is performed. After the above culture, the expression level of type 4 collagen in the skin cells cultured in the medium containing the test substance and the control skin cells cultured in the control medium is compared. When the expression level of type 4 collagen in the skin cells cultured in the medium containing the test substance is higher than the expression level of type 4 collagen in the control skin cells, the test substance is a substance having an effect of suppressing or improving the decrease of the skin barrier function. Select as.
  • the expression level of type 4 collagen can be determined by measuring the amount of mRNA encoding type 4 collagen or the amount of protein of type 4 collagen.
  • the amount of mRNA or protein can be measured by a known method.
  • the amount of mRNA encoding type 4 collagen can be measured by quantitative PCR, Northern blotting, microarray, live imaging using a fluorescent probe, or the like.
  • the amount of protein of type 4 collagen can be measured by immunochemical assay, mass spectrometry and the like.
  • Immunochemical assays include Enzyme Linked Immunosorbent Assay (ELISA), Western blotting and the like.
  • ELISA Enzyme Linked Immunosorbent Assay
  • a commercially available ELISA kit manufactured by MyBioSource or the like can also be used for measuring the amount of protein.
  • the base sequence and amino acid sequence (ORF) of human type 4 collagen are known, and the base sequence (mRNA) is Accession No. It is registered as AH002741 in Genbank, which is a public database. Those skilled in the art can easily design and synthesize primers for measuring the amount of mRNA encoding type 4 collagen, amplify DNA, and the like.
  • the test substance is considered to be a substance having an effect of suppressing or improving the decrease of the skin barrier function. It can be determined. According to the above screening method, it is possible to screen for a substance having an effect of suppressing or improving the decrease of the skin barrier function.
  • the crushed products of the plants (1) to (7) above were immersed in a solvent and extracted. More specifically, with respect to the above (1), (2), (3), (6) and (7), the crushed plant is immersed in a 30-90 vol% ethanol aqueous solution in an amount 10 times the weight, and at room temperature. The mixture was stirred once a day and extracted for 10 minutes to 7 days to obtain an extract. Regarding (4) and (5) above, an aqueous solution of 1,3-butylene glycol was used instead of the aqueous ethanol solution, and the mixture was extracted by the above method.
  • the plant residue was removed from the obtained extract by filtration, and the filtrate was obtained as a plant extract.
  • the obtained plant extract was concentrated with an evaporator and then freeze-dried to obtain an extract of each plant as a powder.
  • the powdered plant extract (rose petal extract, grape seed and peel extract, chanoki leaf extract, hamamelis leaf extract, sardine flower extract, sardine seed extract, sardine sardine extract) It was used as an extract sample for the evaluation described later.
  • Yarrowia lipolytica extract was obtained by culturing Yarrowia lipolytica, autolyzing it, and then performing a heat sterilization sterilization operation to obtain a powdery Yarrowia lipolytica extract.
  • the above powder was used as an extract sample for the Yarrowia lipolytica extract.
  • Example 1 Effect of overexpression of type 4 collagen on skin barrier during inflammation stimulation
  • a plasmid (hereinafter referred to as COL4A1 plasmid) in which COL4A1 (RefSeq ID: NM_001845.6) was incorporated downstream of the CMV promoter was prepared.
  • Opti-MEM solutions Two kinds of solutions having different plasmid concentrations (hereinafter referred to as Opti-MEM solutions) were prepared (plasmid concentration: 5 ⁇ g / mL or 20 ⁇ g / mL). As a control, a solution in which Opti-MEM was added instead of the plasmid was also prepared.
  • HuMedia-KG2 HuMedia-KG2 (hereinafter referred to as recommended medium) (manufactured by Keratinocyte)
  • HuMedia-KG2 normal human adult epidermal keratinized cells
  • Cell Culture Inserts manufactured by Millipore
  • 10 ⁇ L of any of the above Opti-MEM solutions was immediately added.
  • 900 ⁇ L of the recommended medium was added to the 24-well plate (manufactured by AGC Technograss Co., Ltd.).
  • Transepithelial electrical resistance (TEER ( ⁇ ⁇ cm 2 ) was evaluated using Millicel ERS-2 (manufactured by Merck) 96 hours, 120 hours, and 144 hours after sowing. In addition, 144 hours after seeding, cells were lysed using isogen (manufactured by Nippon Gene Co., Ltd.), and then mRNA was extracted. After that, cDNA was synthesized using High-Capacity cDNA Reverse Transition Kit and RNase Inhibitor, and TaqMan Fast Universal PCR Master Mix (both Thermo Fisher Scientific 4) The expression level of (COL4A1) was evaluated. Eukaryotic 18S rRNA was used for endogenous control.
  • Tables 1 and 2 The results are shown in Tables 1 and 2.
  • Table 1 plasmid-added and H 2 O 2 untreated group (H 2 O 2 (-) plasmid (-)) showing a gene expression amount of COL4A1 when was the 1 COL4A1 expression level of.
  • H 2 O 2 (+) is a group subjected to H 2 O 2 treatment.
  • the plasmid (-) indicates that the COL4A1 plasmid was not added, and the plasmid (+) indicates that the COL4A1 plasmid was added.
  • * Indicates that there was a significant difference (p ⁇ 0.05) (Dunnett's test (vs plasmid-added and H 2 O 2 treatment group)).
  • Table 2 shows the value of TEER after 144 hours of sowing. * Indicates that there was a significant difference (p ⁇ 0.05) (Dunnett's test (vs plasmid-added and H 2 O 2 treatment group)). As the H 2 O 2 stimulation, but significant decrease in TEER was seen, the introduction of COL4A1 plasmid, relative to H 2 O 2 stimulation group, dose-dependent and significant TEER improvement in the high dose group The action was confirmed (*: p ⁇ 0.05).
  • Examples 2 to 9 Evaluation of inhibitory or ameliorating effect of reducing or ameliorating type 4 collagen gene expression in an extract sample Using a recommended medium, normal human adult epidermal keratinized cells (manufactured by Kurabou) were placed on a 12-well plate (AGC technograss). The cells were seeded at a concentration of 5 ⁇ 10 4 cells / mL / well and cultured for 48 hours. After exchanging with the recommended medium containing the extract sample and starting the sample treatment, hydrogen peroxide solution (manufactured by Nacalai Tesque Co., Ltd.) was added 30 minutes after the start of the sample treatment so that the final concentration became 100 ⁇ M.
  • hydrogen peroxide solution manufactured by Nacalai Tesque Co., Ltd.
  • DMSO dimethyl sulfoxide
  • the extract samples used in Examples 2 to 9 were rose petal extract, grape seed and peel extract, Chanoki leaf extract, Hamamelis leaf extract, and Yukisou flower extract prepared in Preparation Examples 1 and 2. It is a sardine seed extract, a Yarowia lipolytica extract or a sardine extract.
  • the extract sample was dissolved in ultrapure water or DMSO and added to the medium. The concentration of the extract sample in the medium was 5 to 20 ⁇ g / mL. When dissolved in ultrapure water, DMSO was added separately so that the final concentration of DMSO in the recommended medium containing any of the extract samples was 0.1%.
  • the results shown in Table 3 are relative values (%) of the expression level of COL4A1 in the cells treated with the extract sample when the expression level of COL4A1 of the control is 100%. * Indicates that there was a significant difference (p ⁇ 0.05) with respect to the control (Student's t-test).
  • Table 3 shows the results when the concentration of the extract sample (test substance) was in the range of 5 to 20 ⁇ g / mL and the expression of COL4A1 was the highest.
  • Examples 10 to 15 Evaluation of suppression or ameliorating effect on skin barrier function deterioration of extract sample Using the recommended medium containing the extract sample, normal human adult epidermal keratinized cells were subjected to Millicell Cell Culture Inserts in 3 ⁇ 10 4 cells. Seeded at a concentration of / 200 ⁇ L / well. Thirty minutes after sowing, hydrogen peroxide solution was added so that the final concentration was 100 ⁇ M. After 4 hours from seeding, the medium was replaced with the recommended medium containing no extract sample. Twenty-four hours after sowing, the medium was replaced with an irregular recommended medium. Then, the transepithelial electrical resistance value (TEER) was evaluated by the same method as in Example 1.
  • TEER transepithelial electrical resistance value
  • cells were cultured in the same manner as above, except that the recommended medium containing the extract sample was replaced with the recommended medium in which DMSO was added to a final concentration of 0.1%, and hydrogen peroxide was used. Water was added and the transepithelial electrical resistance was evaluated.
  • the extract samples used in Examples 10 to 15 are the extracts shown in Table 4 prepared in Preparation Examples 1 and 2.
  • the extract sample was dissolved in ultrapure water or DMSO and added to the medium.
  • the concentration of the extract sample in the medium was 5 to 20 ⁇ g / mL.
  • DMSO was added separately so that the final concentration of DMSO in the recommended medium containing any of the extract samples was 0.1%.
  • the results are shown in Table 4.
  • the results shown in Table 4 are the relative values (%) of the TEER in the cells treated with the extract sample, where the control TEER is 100%.
  • Table 4 shows the results when the concentration of the extract sample (test substance) was 5 to 20 ⁇ g / mL and the measurement time was in the range of 96 to 144 hours after sowing, and the TER value was the highest. It was confirmed that 6 kinds of extracts have a high effect of suppressing or improving the decrease of TEER, and further, grape seed and peel extract, Chanoki leaf extract, Hamamelis leaf extract, Seiyounatsuyukisou flower extract, and Yarowia lipolytica extract. For the substance, a significant change was confirmed with respect to the control (*: p ⁇ 0.05) (Student's t-test).

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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

皮膚バリア機能の低下抑制又は改善用組成物、４型コラーゲンの発現の低下抑制又は改善用組成物及び皮膚バリア機能の低下抑制又は改善作用を有する物質のスクリーニング方法A composition for suppressing or improving the decrease in skin barrier function, a composition for suppressing or improving the expression of type 4 collagen, and a method for screening a substance having an effect of suppressing or improving the decrease in skin barrier function.
本発明は皮膚バリア機能の低下抑制又は改善用組成物及び４型コラーゲンの発現の低下抑制又は改善用組成物に関する。また、本発明は、皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善する方法及び４型コラーゲンの発現の低下を抑制又は当該発現を改善する方法に関する。本発明はまた、皮膚バリア機能の低下抑制又は改善作用を有する物質のスクリーニング方法等に関する。 The present invention relates to a composition for suppressing or improving a decrease in skin barrier function and a composition for suppressing or improving the expression of type 4 collagen. The present invention also relates to a method for suppressing a decrease in the skin barrier function or improving the skin barrier function and a method for suppressing the decrease in the expression of type 4 collagen or improving the expression. The present invention also relates to a method for screening a substance having an effect of suppressing or improving a decrease in skin barrier function.
皮膚は、大きく分けて表面から表皮、真皮の２層から構成されている。表皮は、深層から基底層、有棘層、顆粒層、角層の４つの層から構成されており、基底層は基底膜を介して真皮と接着している。基底膜は、４型コラーゲン、７型コラーゲン等の成分で構成されており、表皮の基底層と真皮とを結合させる等の働きがあるが、加齢に伴いその機能が低下する。 The skin is roughly divided into two layers, the epidermis and the dermis, from the surface. The epidermis is composed of four layers, from the deep layer to the basal layer, the stratum spinosum, the stratum granulosum, and the stratum granulosum, and the basal layer is adhered to the dermis via the basement membrane. The basement membrane is composed of components such as type 4 collagen and type 7 collagen, and has a function of binding the basal layer of the epidermis to the dermis, but its function deteriorates with aging.
皮膚には、外部からの生体への異物の侵入を防いだり、体内の水分が皮膚から過剰に蒸発することを防ぐことで皮膚の水分を維持したりするバリア機能がある。加齢等の原因で皮膚バリア機能が低下すると、乾燥肌等の原因となる。特許文献１には、アスパラサスリネアリスの抽出物等を含む皮膚バリア機能改善剤が記載されている。 The skin has a barrier function of preventing foreign substances from entering the living body from the outside and maintaining the moisture of the skin by preventing excessive evaporation of water in the body from the skin. When the skin barrier function deteriorates due to aging or the like, it causes dry skin or the like. Patent Document 1 describes a skin barrier function improving agent containing an extract of asparagus linearis and the like.
特開２００９－２５６２４４号公報JP-A-2009-256244
本発明は、皮膚バリア機能の低下を抑制又は当該機能を改善することができる皮膚バリア機能の低下抑制又は改善用組成物を提供することを目的とする。本発明はまた、４型コラーゲンの発現の低下抑制又は改善用組成物を提供することを目的とする。また、本発明は、皮膚バリア機能の低下を抑制又は当該機能を改善する方法、及び、４型コラーゲンの発現の低下を抑制又は当該発現を改善する方法を提供することを目的とする。さらに、本発明は、皮膚バリア機能の低下抑制又は改善作用を有する物質のスクリーニング方法を提供することを目的とする。 An object of the present invention is to provide a composition for suppressing or improving a decrease in skin barrier function, which can suppress or improve the decrease in skin barrier function. Another object of the present invention is to provide a composition for suppressing or improving the expression of type 4 collagen. Another object of the present invention is to provide a method for suppressing a decrease in skin barrier function or improving the function, and a method for suppressing a decrease in the expression of type 4 collagen or improving the expression. Furthermore, an object of the present invention is to provide a method for screening a substance having an effect of suppressing or improving a decrease in skin barrier function.
本発明者は、上記課題を解決するために研究を行った結果、皮膚細胞において４型コラーゲンの発現の低下を抑制又は当該発現を改善することによって、皮膚バリア機能の低下が抑制又は当該機能が改善されることを見出した。本発明者は、かかる知見に基づきさらに研究を行い、本発明を完成するに至った。 As a result of conducting research to solve the above problems, the present inventor suppresses the decrease in the expression of type 4 collagen in the skin cells or improves the expression, thereby suppressing the decrease in the skin barrier function or the function. Found to be improved. The present inventor has conducted further research based on such findings and has completed the present invention.
本発明は、これに限定されるものではないが、以下の皮膚バリア機能の低下抑制又は改善用組成物、４型コラーゲンの発現の低下抑制又は改善用組成物等を包含する。
〔１〕バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、バラ科シモツケソウ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物を有効成分として含む、４型コラーゲンの発現の低下抑制又は改善を介した皮膚バリア機能の低下抑制又は改善用組成物。
〔２〕バラ科バラ属植物がバラ、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母がヤロウィア・リポリティカ（Ｙａｒｒｏｗｉａ　ｌｉｐｏｌｙｔｉｃａ）、ツバキ科ツバキ属植物がチャノキ、バラ科シモツケソウ属植物がセイヨウナツユキソウ、シソ科ハッカ属植物がセイヨウハッカである、上記〔１〕に記載の皮膚バリア機能の低下抑制又は改善用組成物。
〔３〕バラ科バラ属植物がデルフィニジン型アントシアニン、ロザシアニン化合物及びロザデルフィン化合物からなる群より選択される１種以上の化合物を含むバラ花弁、ツバキ科ツバキ属植物がチャノキの葉、バラ科シモツケソウ属植物がセイヨウナツユキソウの花、シソ科ハッカ属植物がセイヨウハッカ全草である、上記〔１〕又は〔２〕に記載の皮膚バリア機能の低下抑制又は改善用組成物。
〔４〕マンサク科マンサク属植物の抽出物及び／又はオオバコ科オオバコ属植物の抽出物を有効成分として含む皮膚バリア機能の低下抑制又は改善用組成物。
〔５〕マンサク科マンサク属植物がハマメリス、オオバコ科オオバコ属植物がセイヨウオオバコである上記〔４〕に記載の皮膚バリア機能の低下抑制又は改善用組成物。
〔６〕マンサク科マンサク属植物がハマメリスの葉、オオバコ科オオバコ属植物がセイヨウオオバコ種子である上記〔４〕又は〔５〕に記載の皮膚バリア機能の低下抑制又は改善用組成物。
〔７〕基底膜機能の低下抑制又は改善を介して皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善する、上記〔４〕～〔６〕のいずれかに記載の皮膚バリア機能の低下抑制又は改善用組成物。
〔８〕基底膜機能の低下抑制又は改善が４型コラーゲンの発現の低下抑制又は改善を介したものである、上記〔７〕に記載の皮膚バリア機能の低下抑制又は改善用組成物。
〔９〕基底膜機能の低下抑制又は改善剤を有効成分として含む皮膚バリア機能の低下抑制又は改善用組成物。
〔１０〕基底膜機能の低下抑制又は改善剤が４型コラーゲンの発現の低下抑制又は改善を介して基底膜機能の低下を抑制又は基底膜機能を改善するものである、上記〔９〕に記載の皮膚バリア機能の低下抑制又は改善用組成物。
〔１１〕バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、マンサク科マンサク属植物の抽出物、バラ科シモツケソウ属植物の抽出物、オオバコ科オオバコ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物を有効成分として含む４型コラーゲンの発現の低下抑制又は改善用組成物。
〔１２〕バラ科バラ属植物がバラ、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母がヤロウィア・リポリティカ（Ｙａｒｒｏｗｉａ　ｌｉｐｏｌｙｔｉｃａ）、ツバキ科ツバキ属植物がチャノキ、マンサク科マンサク属植物がハマメリス、バラ科シモツケソウ属植物がセイヨウナツユキソウ、オオバコ科オオバコ属植物がセイヨウオオバコ、シソ科ハッカ属植物がセイヨウハッカである、上記〔１１〕に記載の４型コラーゲンの発現の低下抑制又は改善用組成物。
〔１３〕バラ科バラ属植物がデルフィニジン型アントシアニン、ロザシアニン化合物及びロザデルフィン化合物からなる群より選択される１種以上の化合物を含むバラ花弁、ツバキ科ツバキ属植物がチャノキの葉、マンサク科マンサク属植物がハマメリスの葉、バラ科シモツケソウ属植物がセイヨウナツユキソウの花、オオバコ科オオバコ属植物がセイヨウオオバコ種子、シソ科ハッカ属植物がセイヨウハッカ全草である、上記〔１１〕又は〔１２〕に記載の４型コラーゲンの発現の低下抑制又は改善用組成物。
〔１４〕皮膚外用剤である上記〔１〕～〔１３〕のいずれかに記載の組成物。
〔１５〕化粧料である上記〔１〕～〔１４〕のいずれかに記載の組成物。
〔１６〕４型コラーゲンの発現の低下抑制又は改善により皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善するための、バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、バラ科シモツケソウ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物の使用。
〔１７〕皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善するための、マンサク科マンサク属植物の抽出物及び／又はオオバコ科オオバコ属植物の抽出物の使用。
〔１８〕４型コラーゲンの発現の低下を抑制又は前記発現を改善するための、バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、マンサク科マンサク属植物の抽出物、バラ科シモツケソウ属植物の抽出物、オオバコ科オオバコ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物の使用。
〔１９〕バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、バラ科シモツケソウ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物を投与する、４型コラーゲンの発現の低下抑制又は改善により皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善する方法。
〔２０〕マンサク科マンサク属植物の抽出物及び／又はオオバコ科オオバコ属植物の抽出物を投与する、皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善する方法。
〔２１〕バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、マンサク科マンサク属植物の抽出物、バラ科シモツケソウ属植物の抽出物、オオバコ科オオバコ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物を投与する４型コラーゲンの発現の低下を抑制又は前記発現を改善する方法。
〔２２〕インビトロで、ヒト表皮角化細胞を、被験物質を含む培地又は上記被験物質を含まないコントロール培地で培養する工程、及び、上記被験物質を含む培地で培養した皮膚細胞及びコントロール培地で培養したコントロール皮膚細胞における４型コラーゲン発現量を比較する工程を含み、上記被験物質を含む培地で培養した皮膚細胞における４型コラーゲン発現量が、上記コントロール皮膚細胞における４型コラーゲン発現量より多い場合に、上記被験物質を皮膚バリア機能の低下抑制又は改善作用を有する物質として選択する、皮膚バリア機能の低下抑制又は改善作用を有する物質のスクリーニング方法。 The present invention includes, but is not limited to, the following compositions for suppressing or improving the decrease in skin barrier function, the compositions for suppressing or improving the expression of type 4 collagen, and the like.
[1] Extracts of Rosaceae plants, grape seeds and / or peels, extracts of Yarrowia yeast, extracts of Camellia plants, extracts of Rosaceae plants, In addition, suppression or improvement of skin barrier function through suppression or improvement of expression of type 4 collagen containing at least one extract selected from the group consisting of extracts of Rosaceae plants as an active ingredient. Composition for.
[2] Rosaceae, Rosaceae, Rosaceae, Yarrowia yeast, Yarrowia lipolytica, Theaceae, Camellia, Chanoki, Rosaceae, Filipendula, Labiatae, Lamiaceae The composition for suppressing or improving the deterioration of the skin barrier function according to the above [1], which is a mint.
[3] Rosaceae Rosaceae plants include rose petals containing one or more compounds selected from the group consisting of delphinidin-type anthocyanin, Rosacyanin compounds and Rosadelfin compounds, Rosaceae Rosaceae plants are Rosaceae leaves, Rosaceae Labiatae The composition for suppressing or improving the deterioration of the skin barrier function according to the above [1] or [2], wherein the plant is a flower of Filipendula and the plant of the genus Rosaceae is a whole plant of Rosaceae.
[4] A composition for suppressing or improving a decrease in skin barrier function containing an extract of a witch hazel family plant and / or an extract of a plantain family plantago genus as an active ingredient.
[5] The composition for suppressing or improving the deterioration of the skin barrier function according to the above [4], wherein the witch hazel family witch hazel plant is Hamamelis and the plantain family plantago genus is Plantain.
[6] The composition for suppressing or improving the deterioration of the skin barrier function according to the above [4] or [5], wherein the witch hazel family witch hazel is a leaf of Hamamelis and the plantain family is a plantain seed.
[7] Suppression of deterioration of skin barrier function or improvement of skin barrier function according to any one of the above [4] to [6], which suppresses or improves reduction of skin barrier function through suppression or improvement of basement membrane function. Composition for improvement.
[8] The composition for suppressing or improving the decrease in skin barrier function according to the above [7], wherein the suppression or improvement of the basement membrane function is mediated by the suppression or improvement of the expression of type 4 collagen.
[9] A composition for suppressing or improving the deterioration of the skin barrier function, which contains an agent for suppressing or improving the deterioration of the basement membrane function as an active ingredient.
[10] The above-mentioned [9], wherein the agent for suppressing or improving the decrease in basement membrane function suppresses or improves the decrease in basement membrane function through the suppression or improvement of the expression of type 4 collagen. A composition for suppressing or improving the deterioration of the skin barrier function of the skin.
[11] Extracts of Rosaceae plants, grape seeds and / or peels, extracts of Yarrowia yeast, extracts of Plantains, extracts of Plantains, extracts of Plantains, Type 4 collagen containing at least one extract selected from the group consisting of extracts of Rosaceae plants of the genus Rosaceae, extracts of plants of the genus Plantains, and extracts of plants of the genus Witch-hazelaceae as active ingredients. A composition for suppressing or improving the decrease in expression.
[12] Rosaceae plants of the genus Rosa, Yarrowia yeast are Yarrowia lipolytica, plants of the genus Rosaceae are Chanoki, plants of the genus Rosaceae are Hamamelis, and plants of the genus Filipendula are the plants of the genus Filipendula. The composition for suppressing or ameliorating the decrease in the expression of type 4 collagen according to the above [11], wherein the plant belonging to the genus Rosaceae of the Rosaceae family is Filipendula and the plant belonging to the genus Rosaceae is Filipendula.
[13] Rosaceae Rosaceae plants include rose petals containing one or more compounds selected from the group consisting of delphinidin-type anthocyanin, rosasianin compounds and rosadelphin compounds, Plantainaceae plants of the genus Plantain are leaves of Chanoki, genus Mansaku of the family Plantains. In [11] or [12] above, the plant is a leaf of Hamamelis, the plant of the genus Rosaceae is the flower of Yukisou, the plant of the genus Plantain is the seed of the plantain, and the plant of the genus Rosaceae is the whole plant of the genus Astragalus. The composition for suppressing or improving the expression of type 4 collagen according to the above.
[14] The composition according to any one of the above [1] to [13], which is an external preparation for skin.
[15] The composition according to any one of the above [1] to [14], which is a cosmetic.
[16] Extracts of Rosaceae plants, grape seeds and / or peels, for suppressing or improving the decrease in the expression of type 4 collagen to suppress the decrease in the skin barrier function or improve the skin barrier function. One or more extracts selected from the group consisting of extracts of Yarrowia yeast, extracts of Camellia plants of the family Rosaceae, extracts of plants of the genus Rosaceae, and extracts of plants of the genus Camellia. Use of things.
[17] Use of an extract of a witch hazel family plant and / or an extract of a plantain family plantago genus to suppress a decrease in the skin barrier function or improve the skin barrier function.
[18] An extract of a Rosaceae plant, an extract of grape seeds and / or a peel, an extract of Yarrowia yeast, for suppressing a decrease in the expression of type 4 collagen or improving the expression. From a group consisting of extracts of plants of the genus Tsubaki of the family Rosaceae, extracts of plants of the genus Mansaku of the family Rosaceae, extracts of plants of the genus Rosaceae, extracts of plants of the genus Oobaco of the family Rosaceae, and extracts of plants of the genus Hakka of the family Rosaceae. Use of one or more selected extracts.
[19] Extracts of Rosaceae plants, grape seeds and / or peels, extracts of Yarrowia yeast, extracts of Camellia plants, extracts of Rosaceae plants, In addition, by administering one or more extracts selected from the group consisting of extracts of Rosaceae plants, the decrease in the expression of type 4 collagen is suppressed or improved to suppress the decrease in the skin barrier function or the skin barrier function. How to improve.
[20] A method for suppressing deterioration of the skin barrier function or improving the skin barrier function by administering an extract of a witch hazel family plant and / or an extract of a plantain family plantago genus.
[21] Extracts of Rosaceae plants, grape seeds and / or peels, extracts of Yarrowia yeast, extracts of Plantains, extracts of Plantains, extracts of Plantains, Expression of type 4 collagen administered by one or more extracts selected from the group consisting of extracts of Rosaceae plants of the genus Rosaceae, extracts of plants of the genus Plantains, and extracts of plants of the genus Witch-hazelaceae. A method for suppressing a decrease or improving the expression.
[22] In vitro, a step of culturing human epidermal keratinized cells in a medium containing the test substance or a control medium not containing the test substance, and culturing the skin cells and the control medium cultured in the medium containing the test substance. When the expression level of type 4 collagen in the skin cells cultured in the medium containing the test substance is higher than the expression level of type 4 collagen in the control skin cells, which includes a step of comparing the expression level of type 4 collagen in the control skin cells. , A method for screening a substance having an effect of suppressing or improving the decrease in skin barrier function, which selects the above-mentioned test substance as a substance having an effect of suppressing or improving the decrease of skin barrier function.
本発明により、皮膚バリア機能の低下を抑制又は当該機能を改善することができる皮膚バリア機能の低下抑制又は改善用組成物を提供することができる。また、本発明によれば、４型コラーゲンの発現の低下抑制又は改善用組成物を提供することができる。また、本発明によれば、皮膚バリア機能の低下を抑制又は当該機能を改善する方法、及び、４型コラーゲンの発現の低下を抑制又は当該発現を改善する方法を提供することができる。本発明によれば、皮膚バリア機能の低下抑制又は改善作用を有する物質のスクリーニング方法を提供することができる。 INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a composition for suppressing or improving a decrease in skin barrier function, which can suppress or improve the decrease in skin barrier function. Further, according to the present invention, it is possible to provide a composition for suppressing or improving the expression of type 4 collagen. Further, according to the present invention, it is possible to provide a method for suppressing a decrease in skin barrier function or improving the function, and a method for suppressing a decrease in the expression of type 4 collagen or improving the expression. According to the present invention, it is possible to provide a method for screening a substance having an effect of suppressing or improving a decrease in skin barrier function.
本発明の第一の態様は、バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、マンサク科マンサク属植物の抽出物、バラ科シモツケソウ属植物の抽出物、オオバコ科オオバコ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物を有効成分として含む４型コラーゲンの発現の低下抑制又は改善用組成物である。本発明の第一の態様の４型コラーゲンの発現の低下抑制又は改善用組成物を、本発明の第一の態様の組成物ともいう。
上記の抽出物は、４型コラーゲンの発現の低下を抑制又は当該発現を改善する作用を有する。４型コラーゲンの発現の低下抑制は、４型コラーゲンの発現低下の進行を抑えること、４型コラーゲンの発現を維持すること、４型コラーゲンの発現低下の程度を軽減（緩和）すること等を含む。４型コラーゲンの発現の改善は、４型コラーゲンの発現を回復させること等を含む。回復は、少なくとも部分的に回復させることを含む。４型コラーゲンの発現には、４型コラーゲンをコードする遺伝子の発現及び４型コラーゲンのタンパク質レベルでの発現が含まれ、これらのいずれか又は両方であってよい。４型コラーゲンは、好ましくはヒトの４型コラーゲンであり、より好ましくはヒト４型コラーゲンα１鎖である。ヒト４型コラーゲンα１鎖は、遺伝子４型コラーゲンＡ１（ＣＯＬ４Ａ１）にコードされるタンパク質である。一態様において、本発明の４型コラーゲンの発現の低下抑制又は改善用組成物は、ヒト４型コラーゲンα１鎖の発現の低下を抑制又は当該発現を改善することによって、ヒトにおける４型コラーゲンの発現の低下を抑制又は当該発現を改善するために使用することができる。 The first aspect of the present invention is an extract of Rosaceae plants, grape seeds and / or peels, an extract of Yarrowia yeast, an extract of Plantains, and a plantain. The active ingredient is one or more extracts selected from the group consisting of extracts of genus plants, extracts of plants of the genus Rosaceae, extracts of plants of the genus Plantains, and extracts of plants of the genus Plantains. It is a composition for suppressing or improving the expression of type 4 collagen contained as. The composition for suppressing or improving the expression of type 4 collagen according to the first aspect of the present invention is also referred to as the composition according to the first aspect of the present invention.
The above extract has an effect of suppressing a decrease in the expression of type 4 collagen or improving the expression. Suppression of decreased expression of type 4 collagen includes suppressing the progression of decreased expression of type 4 collagen, maintaining the expression of type 4 collagen, reducing (alleviating) the degree of decreased expression of type 4 collagen, and the like. .. Improving the expression of type 4 collagen includes restoring the expression of type 4 collagen and the like. Recovery involves at least partial recovery. Expression of type 4 collagen includes expression of a gene encoding type 4 collagen and expression of type 4 collagen at the protein level, which may be any or both. The type 4 collagen is preferably human type 4 collagen, and more preferably human type 4 collagen α1 chain. Human type 4 collagen α1 chain is a protein encoded by gene type 4 collagen A1 (COL4A1). In one aspect, the composition for suppressing or improving the expression of type 4 collagen of the present invention suppresses or improves the expression of human type 4 collagen α1 chain, thereby expressing the type 4 collagen in humans. It can be used to suppress the decrease in collagen or improve its expression.
紫外線等の外的要因や、加齢等の内的要因によって、４型コラーゲンの発現は低下（発現量が減少）する。
後記の実施例に示されるように、表皮角化細胞において４型コラーゲンの発現低下を抑制又は発現を改善すると、皮膚の経上皮電気抵抗値（ＴＥＥＲ）の低下が抑制又は改善された。これは、皮膚バリア機能の低下が抑制又は皮膚バリア機能が改善されたことを示す。４型コラーゲンの発現の低下抑制又は改善によって皮膚バリア機能の低下が抑制又は当該機能が改善された詳細なメカニズムは定かではないが、４型コラーゲンは基底膜の成分であることから、４型コラーゲンの発現の低下を抑制又は改善することによって、基底膜の機能の低下が抑制又は当該機能が改善され、皮膚の恒常性が維持された結果皮膚バリア機能の低下が抑制又は当該機能が改善されたと推定される。従って、基底膜機能の低下を抑制又は当該機能を改善することによって、皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善することができると期待される。上記の植物又は酵母の抽出物は、優れた４型コラーゲンの発現の低下抑制又は改善作用を有することから、基底膜機能の低下を抑制又は当該機能を改善するために使用することができる。
４型コラーゲンの発現の低下抑制又は改善効果の観点から、本発明の第一の態様の組成物において、上記の抽出物は、バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、並びに、バラ科シモツケソウ属植物の抽出物からなる群より選択される１種以上であることが好ましい。 The expression of type 4 collagen decreases (the expression level decreases) due to external factors such as ultraviolet rays and internal factors such as aging.
As shown in the examples below, suppressing or improving the expression of type 4 collagen in epidermal keratinized cells suppressed or improved the decrease in transepithelial electrical resistance (TEER) of the skin. This indicates that the decrease in the skin barrier function was suppressed or the skin barrier function was improved. The detailed mechanism by which the decrease in skin barrier function was suppressed or improved by suppressing or improving the expression of type 4 collagen is not clear, but since type 4 collagen is a component of the basement membrane, type 4 collagen By suppressing or ameliorating the decrease in the expression of the skin, the decrease in the function of the basement membrane is suppressed or the function is improved, and as a result of maintaining the homeostasis of the skin, the decrease in the skin barrier function is suppressed or the function is improved. Presumed. Therefore, it is expected that the decrease in the skin barrier function can be suppressed or the skin barrier function can be improved by suppressing the decrease in the basement membrane function or improving the function. Since the above-mentioned plant or yeast extract has an excellent effect of suppressing or improving the expression of type 4 collagen, it can be used to suppress or improve the decrease of basement membrane function.
From the viewpoint of suppressing or improving the expression of type 4 collagen, in the composition of the first aspect of the present invention, the above-mentioned extract is an extract of a Rosaceae plant, a grape seed and / or a fruit skin. It is preferable that the species is one or more selected from the group consisting of the plant and the extract of the plant belonging to the genus Filipendula of the Rosaceae family.
皮膚バリア機能とは、外部からの生体への異物の侵入を防いだり、体内の水分が皮膚から過剰に蒸発することを防ぐことで皮膚の水分を維持したりする、皮膚のバリア機能をいう。皮膚バリア機能の低下抑制は、皮膚バリア機能の低下の進行を抑えること、皮膚バリア機能を維持すること、皮膚バリア機能の低下の程度の軽減（緩和）すること等を含む。皮膚バリア機能の改善は、皮膚バリア機能を回復させること等を含む。皮膚バリア機能の低下は、加齢による皮膚バリア機能の低下、紫外線曝露による皮膚バリア機能の低下等であってもよい。 The skin barrier function refers to a skin barrier function that prevents foreign substances from entering the living body from the outside and maintains skin moisture by preventing excessive evaporation of moisture in the body from the skin. Suppression of deterioration of skin barrier function includes suppressing the progress of deterioration of skin barrier function, maintaining skin barrier function, reducing (alleviating) the degree of deterioration of skin barrier function, and the like. Improvement of the skin barrier function includes restoration of the skin barrier function and the like. The decrease in the skin barrier function may be a decrease in the skin barrier function due to aging, a decrease in the skin barrier function due to exposure to ultraviolet rays, or the like.
上記のように、基底膜機能の低下抑制又は改善により、皮膚バリア機能の低下を抑制又は当該機能を改善することが可能となる。
本発明の第二の態様は、基底膜機能の低下抑制又は改善剤を有効成分として含む皮膚バリア機能の低下抑制又は改善用組成物である。本発明の第二の態様の皮膚バリア機能の低下抑制又は改善用組成物を、以下では本発明の第二の態様の組成物ともいう。 As described above, by suppressing or improving the decrease in basement membrane function, it is possible to suppress or improve the decrease in skin barrier function.
A second aspect of the present invention is a composition for suppressing or improving the decrease in skin barrier function, which comprises an active ingredient for suppressing or improving the decrease in basement membrane function. The composition for suppressing or improving the decrease in skin barrier function according to the second aspect of the present invention is also hereinafter referred to as the composition according to the second aspect of the present invention.
基底膜の機能として、表皮と真皮をつなぐ機械的支えや、表皮と真皮の間の物質交換の選択的フィルターなどの機能が知られている。４型コラーゲン等の基底膜成分の発現の低下を抑制又は当該発現を改善することは、基底膜機能の低下抑制又は改善に有効である。
基底膜機能の低下抑制又は改善剤は、基底膜機能の低下抑制又は改善作用、例えば、基底膜の構成成分の発現の低下抑制又は改善作用を有するものであればよく、特に限定されない。基底膜機能の低下抑制又は改善剤は、好ましくは、４型コラーゲンの発現の低下抑制又は改善を介して基底膜機能の低下を抑制又は基底膜機能を改善するものである。基底膜機能の低下抑制又は改善剤には、例えば、４型コラーゲンの発現の低下抑制又は改善作用を有する上記の植物又は酵母の抽出物を使用することができる。一態様において、基底膜機能の低下抑制又は改善剤として、バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、マンサク科マンサク属植物の抽出物、バラ科シモツケソウ属植物の抽出物、オオバコ科オオバコ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物を使用することができる。
上記の抽出物の中でも、基底膜機能の低下抑制又は改善剤として、バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、並びに、バラ科シモツケソウ属植物の抽出物からなる群より選択される１種以上が好ましい。 Known functions of the basement membrane include a mechanical support that connects the epidermis and the dermis, and a selective filter for substance exchange between the epidermis and the dermis. Suppressing or improving the expression of basement membrane components such as type 4 collagen is effective in suppressing or improving the decrease in basement membrane function.
The agent for suppressing or improving the decrease in basement membrane function is not particularly limited as long as it has an effect of suppressing or improving the decrease in basement membrane function, for example, an effect of suppressing or improving the expression of components of the basement membrane. The agent for suppressing or improving the decrease in basement membrane function is preferably one that suppresses or improves the decrease in basement membrane function through the suppression or improvement of the expression of type 4 collagen. As the agent for suppressing or improving the decrease in basement membrane function, for example, the above-mentioned extract of a plant or yeast having an effect of suppressing or improving the expression of type 4 collagen can be used. In one embodiment, as an agent for suppressing or ameliorating a decrease in basal membrane function, an extract of a Rosaceae plant, a grape seed and / or a peel extract, an extract of a Yarrowia yeast, a Plantain plant. One or more species selected from the group consisting of extracts, witch hazel plants, rosaceae plants, plantains, plantains, and plantains. Extracts can be used.
Among the above extracts, a group consisting of an extract of a Rosaceae plant, an extract of a grape seed and / or a fruit skin, and an extract of a Rosaceae Filipendula plant as an agent for suppressing or ameliorating a decrease in basal membrane function. One or more selected from the above is preferable.
本発明の第三の態様は、バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、バラ科シモツケソウ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物を有効成分として含む、４型コラーゲンの発現の低下抑制又は改善を介した皮膚バリア機能の低下抑制又は改善用組成物である。本発明の第三の態様の皮膚バリア機能の低下抑制又は改善用組成物を、以下では本発明の第三の態様の組成物ともいう。
本発明の第三の態様の組成物において、上記の抽出物は、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、並びに、バラ科シモツケソウ属植物の抽出物からなる群より選択される１種以上であることが好ましい。 A third aspect of the present invention is an extract of Rosaceae plants, grape seeds and / or peels, an extract of Yarrowia yeast, an extract of Camellia plants, and Rosaceae. A skin barrier through suppression or improvement of the expression of type 4 collagen, which comprises, as an active ingredient, one or more extracts selected from the group consisting of extracts of genus plants and extracts of plants of the genus Camellia of the Rosaceae family. It is a composition for suppressing or improving the deterioration of function. The composition for suppressing or improving the decrease in skin barrier function according to the third aspect of the present invention is also hereinafter referred to as the composition according to the third aspect of the present invention.
In the composition of the third aspect of the present invention, the above extracts are an extract of grape seed and / or peel, an extract of Yarrowia yeast, an extract of Theaceae Camellia plant, and a rose. It is preferably one or more species selected from the group consisting of extracts of plants belonging to the family Theaceae.
本発明の第四の態様は、マンサク科マンサク属植物の抽出物及び／又はオオバコ科オオバコ属植物の抽出物を有効成分として含む皮膚バリア機能の低下抑制又は改善用組成物である。本発明の第四の態様の皮膚バリア機能の低下抑制又は改善用組成物を、以下では本発明の第四の態様の組成物ともいう。本発明の第四の態様の組成物は、好ましくは、マンサク科マンサク属植物の抽出物を有効成分として含む。 A fourth aspect of the present invention is a composition for suppressing or improving a decrease in skin barrier function, which comprises an extract of a witch hazel family plant and / or an extract of a plantain family plantago genus as an active ingredient. The composition for suppressing or improving the decrease in skin barrier function according to the fourth aspect of the present invention is also hereinafter referred to as the composition according to the fourth aspect of the present invention. The composition of the fourth aspect of the present invention preferably contains an extract of a witch hazel plant of the witch hazel family as an active ingredient.
本発明の第三の態様の組成物及び第四の態様の組成物で使用される上記の抽出物は、４型コラーゲンの発現の低下抑制又は改善作用を有する。これにより、基底膜機能の低下抑制又は改善が可能となる。一態様において、本発明の第三の態様及び第四の態様の皮膚バリア機能の低下抑制又は改善用組成物は、基底膜機能の低下抑制又は改善を介して皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善するために使用することができる。また、本発明の第四の態様の皮膚バリア機能の低下抑制又は改善用組成物は、４型コラーゲンの発現の低下抑制又は改善を介した基底膜機能の低下抑制又は改善を介して、皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善するために使用することができる。一態様において、本発明の第四の態様の皮膚バリア機能の低下抑制又は改善用組成物は、４型コラーゲンの発現の低下抑制又は改善により皮膚バリア機能の低下を抑制又は改善するために使用することができる。 The composition of the third aspect of the present invention and the above-mentioned extract used in the composition of the fourth aspect have an action of suppressing or ameliorating a decrease in the expression of type 4 collagen. This makes it possible to suppress or improve the decrease in basement membrane function. In one aspect, the composition for suppressing or improving the decrease in skin barrier function according to the third and fourth aspects of the present invention suppresses or improves the decrease in basement membrane function and suppresses or improves the decrease in skin barrier function. It can be used to improve the barrier function. In addition, the composition for suppressing or improving the decrease in skin barrier function according to the fourth aspect of the present invention is a skin barrier through suppression or improvement of decrease in basement membrane function through suppression or improvement of expression of type 4 collagen. It can be used to suppress deterioration of function or improve skin barrier function. In one aspect, the composition for suppressing or improving the decrease in skin barrier function according to the fourth aspect of the present invention is used to suppress or improve the decrease in skin barrier function by suppressing or improving the expression of type 4 collagen. be able to.
以下では、本発明の第一の態様の４型コラーゲンの発現の低下抑制又は改善用組成物、第二の態様、第三の態様及び第四の態様の皮膚バリア機能の低下抑制又は改善用組成物を、まとめて本発明の組成物ともいう。本発明の組成物においては、上記の抽出物を２種以上組み合わせて有効成分として使用してもよい。 In the following, a composition for suppressing or improving the expression of type 4 collagen according to the first aspect of the present invention, a composition for suppressing or improving the decrease in skin barrier function of the second, third and fourth aspects. The substances are also collectively referred to as the composition of the present invention. In the composition of the present invention, two or more of the above extracts may be combined and used as an active ingredient.
本発明において、バラ科バラ属（Ｒｏｓａｃｅａｅ　Ｒｏｓａ）植物は、Ｒｏｓａ　ｈｙｂｒｉｄａ（学名）に代表されるバラ科バラ属のバラである。
バラ科バラ属植物として、デルフィニジン型アントシアニン、ロザシアニン化合物及びロザデルフィン化合物からなる群より選択される１種以上の化合物を含むバラが好ましく、花弁にデルフィニジン型アントシアニン、ロザシアニン化合物及びロザデルフィン化合物からなる群より選択される１種以上の化合物を含むバラがより好ましい。中でも、花弁にデルフィニジン型アントシアニンを含むバラが好ましい。 In the present invention, the Rosaceae Rosa plant is a rose of the Rosaceae Rosa genus represented by Rosa hybrida (scientific name).
As a plant belonging to the genus Rose of the family Rose, a rose containing one or more compounds selected from the group consisting of delphinidin-type anthocyanins, rosasianin compounds and rosadelphin compounds is preferable, and a group consisting of delphinidin-type anthocyanins, rosasianin compounds and rosadelfin compounds on the petals. More preferably, roses containing one or more of the more selected compounds. Of these, roses containing delphinidin-type anthocyanins in the petals are preferable.
デルフィニジン型アントシアニンは紫色や青色を呈する花弁に多く含まれている色素である。デルフィニジン型アントシアニンとして、デルフィニジン、マルビジン、ペチュニジン、又はそれらが糖、アシル基などで修飾された化合物が挙げられる。デルフィニジン型アントシアニンを含むバラとして、青系又は藤色系の色調を呈するバラ、例えば、サントリーブルーローズアプローズ（登録商標）等の品種のバラが挙げられる。バラにデルフィニジン型アントシアニンが含まれることは、例えば、特許５６９７０４０号公報等に記載の方法で、ＨＰＬＣで確認することができる。 Delphinidin-type anthocyanins are pigments that are abundant in purple and blue petals. Examples of delphinidin-type anthocyanins include delphinidin, malvidin, petunidin, and compounds in which they are modified with sugars, acyl groups, and the like. Examples of roses containing delphinidin-type anthocyanins include roses having a bluish or wisteria color tone, for example, roses of varieties such as Suntory Blue Rose Applause (registered trademark). The inclusion of delphinidin-type anthocyanins in roses can be confirmed by HPLC, for example, by the method described in Japanese Patent No. 5697040.
ロザシアニン化合物は、ロザシアニンＡ１、ロザシアニンＡ２及びロザシアニンＢからなる群より選択される１以上の化合物である。ロザシアニン化合物を含むバラは、上記の化合物を１種以上含むバラであればよいが、好ましくは、ロザロザシアニンＡ１、ロザデルフィンＡ２及びロザデルフィンＢを含むバラである。ロザシアニンＡ１は、下記一般式（Ｉ）において、Ｒ１が水素原子である化合物であり、ロザシアニンＡ２は、下記一般式（ＩＩ）において、Ｒ２が水素原子である化合物であり、ロザシアニンＢは、下記一般式（ＩＩＩ）において、Ｒ３が水素原子である化合物である。 The rosasianin compound is one or more compounds selected from the group consisting of rosasianin A1, rosasianin A2 and rosasianin B. The rose containing the rosasianin compound may be a rose containing at least one of the above compounds, but is preferably a rose containing rosa rosasianin A1, rosadelphin A2 and rosadelphin B. Rosacyanin A1 is a compound in which R 1 is a hydrogen atom in the following general formula (I) , Rosacyanin A2 is a compound in which R 2 is a hydrogen atom in the following general formula (II), and Rosacyanin B is In the following general formula (III), R 3 is a compound having a hydrogen atom.
Figure JPOXMLDOC01-appb-C000001
Figure JPOXMLDOC01-appb-C000001
（上記式中、グルコースの１位の水酸基の配位（波線）はα体とβ体の互変異性であることを示す。Ｒ１は、水素原子又は水酸基を表す。） (In the above formula, the coordination (wavy line) of the hydroxyl group at the 1-position of glucose indicates that the α-form and the β-form are tautomerized. R 1 represents a hydrogen atom or a hydroxyl group.)
Figure JPOXMLDOC01-appb-C000002
Figure JPOXMLDOC01-appb-C000002
（上記式中、Ｒ２は、水素原子又は水酸基を表す。） (In the above formula, R 2 represents a hydrogen atom or a hydroxyl group.)
Figure JPOXMLDOC01-appb-C000003
Figure JPOXMLDOC01-appb-C000003
（上記式中、Ｒ３は、水素原子又は水酸基を表す。） (In the above formula, R 3 represents a hydrogen atom or a hydroxyl group.)
ロザシアニン化合物を含むバラとして、青系又は藤色系の色調を呈するバラ、例えば、マダムビオレ、パープルレイン、ラバンデ、マンハッタンブルー、シャンテリーレース、ブルームーン、タソガレ、シャルルドゴール、バイオレットドリー、ブルーリボン、アオゾラ、レディエックス、ブルーバユー、スターリングシルバー等の品種のバラが挙げられる。これらのバラは、花弁にロザシアニン化合物を含むため好ましい。 Roses containing rosasianin compounds that exhibit a bluish or mauve hue, such as Madame Biore, Purple Rain, Labande, Manhattan Blue, Chantery Lace, Blue Moon, Tasogare, Charles de Goal, Violet Dolly, Blue Ribbon, Aozora. , Lady X, Blue Bayou, Sterling Silver and other varieties of roses. These roses are preferable because the petals contain a rosasianin compound.
ロザデルフィン化合物は、ロザデルフィンＡ１、ロザデルフィンＡ２及びロザデルフィンＢからなる群より選択される１以上の化合物である。ロザデルフィン化合物を含むバラは、上記の化合物を１種以上含むバラであればよいが、好ましくは、ロザデルフィンＡ１、ロザデルフィンＡ２及びロザデルフィンＢを含むバラである。ロザデルフィンＡ１は、上記一般式（Ｉ）において、Ｒ１が水酸基である化合物であり、ロザデルフィンＡ２は、上記一般式（ＩＩ）において、Ｒ２が水酸基である化合物であり、ロザデルフィンＢは、上記一般式（ＩＩＩ）において、Ｒ３が水酸基である化合物である。 The Rosa Delphin compound is one or more compounds selected from the group consisting of Rosa Delphin A1, Rosa Delphin A2 and Rosa Delphin B. The rose containing the Rosa Delphin compound may be a rose containing at least one of the above compounds, but is preferably a rose containing Rosa Delphin A1, Rosa Delphin A2 and Rosa Delphin B. Rosa Delphin A1 is a compound in which R 1 is a hydroxyl group in the above general formula (I) , Rosa Delphin A2 is a compound in which R 2 is a hydroxyl group in the above general formula (II), and Rosa Delphin B is in the above formula (III), a compound wherein R 3 is a hydroxyl group.
上記ロザデルフィン化合物は野生種のバラには存在せず、フラボノイド３’，５’－水酸化酵素をコードする遺伝子（以下、フラボノイド３’，５’－水酸化酵素遺伝子ともいう）を遺伝子組換え等の手法で導入したバラや、遺伝子組換えバラを親として得られるフラボノイド３’，５’－水酸化酵素遺伝子を有するバラにおいて青色色素であるデルフィニジンが合成され、これにガレートやテリマグランジン類が結合して生成する化合物である。 The above Rosadelphin compound does not exist in wild roses, and the gene encoding flavonoid 3', 5'-hydroxylase (hereinafter, also referred to as flavonoid 3', 5'-hydroxylase gene) is genetically modified. Delfinidine, which is a blue pigment, is synthesized in roses introduced by such methods and roses having flavonoids 3', 5'-hydroxylase genes obtained by using recombinant roses as parents, and gallates and terimaglandins are synthesized. Is a compound produced by binding.
上記ロザデルフィン化合物を含むバラとして、フラボノイド３’，５’－水酸化酵素遺伝子を、遺伝子組換え等の手法で導入したバラ、及び、このような遺伝子組換えバラを親として得られるフラボノイド３’，５’－水酸化酵素遺伝子を有するバラを使用することができる。このようなバラとして、例えば、サントリーブルーローズアプローズ（登録商標）等の品種のバラを使用することができる。サントリーブルーローズアプローズ（登録商標）の花弁には、ロザシアニンＡ１、ロザシアニンＡ２及びロザシアニンＢ並びにロザデルフィンＡ１、ロザデルフィンＡ２及びロザデルフィンＢが含まれていることが知られている（例えば、ＷＯ２０１０／１１０３８２）。サントリーブルーローズアプローズ（登録商標）は、サントリーフラワーズ株式会社（日本国、東京都）より入手することができる。フラボノイド３’，５’－水酸化酵素遺伝子を有するバラは、ＷＯ２０１０／１１０３８２等に記載の方法で作製することもできる。フラボノイド３’，５’－水酸化酵素遺伝子としては、ＷＯ２００４／０２０６３７に記載されている植物由来の遺伝子が挙げられ、例えばスミレ属パンジー（スミレ属ブラックパンジー（Ｖｉｏｌａ　ｓｐｐ．　ｃｖ　Ｂｌａｃｋ　Ｐａｎｓｙ）等）由来のフラボノイド３’，５’－水酸化酵素遺伝子等が好ましい。フラボノイド３’，５’－水酸化酵素遺伝子を有するバラは、花弁にロザデルフィン化合物を含むため好ましい。なお、本明細書中に記載された学術文献及び特許文献の全ては、参照として本明細書に組み入れられる。 As roses containing the above Rosadelphin compound, roses into which a flavonoid 3', 5'-hydroxylase gene has been introduced by a method such as gene recombination, and flavonoid 3'obtained from such a genetically modified rose as a parent. A rose carrying the, 5'-hydroxylase gene can be used. As such a rose, for example, a variety of rose such as Suntory Blue Rose Applause (registered trademark) can be used. It is known that the petals of Suntory Blue Rose Applause (registered trademark) include Rosacyanin A1, Rosacyanin A2 and Rosacyanin B, and Rosadelfin A1, Rosadelfin A2 and Rosadelfin B (for example, WO2010 / 110382). ). Suntory Blue Rose Applause (registered trademark) can be obtained from Suntory Flowers Limited (Tokyo, Japan). Roses having flavonoid 3', 5'-hydroxylase genes can also be prepared by the method described in WO2010 / 110382 and the like. Examples of the flavonoid 3', 5'-hydroxylase gene include plant-derived genes described in WO2004 / 02637, for example, derived from Violet pansy (Violet spp. Cv Black Pansy, etc.). Flavonoids 3', 5'-hydroxylase genes and the like are preferred. Roses carrying the flavonoid 3', 5'-hydroxylase gene are preferred because the petals contain the Rosadelfin compound. All academic and patent documents described herein are incorporated herein by reference.
バラにロザシアニン化合物及び／又はロザデルフィン化合物が含まれることは、例えば、ＷＯ２０１０／１１０３８２、ＷＯ２０１５／１７８３８５等に記載の方法で、ＨＰＬＣ－ＴＯＦ－ＭＳにより確認することができる。 The inclusion of the rosesianin compound and / or the rosadelphin compound in the roses can be confirmed by HPLC-TOF-MS by the method described in, for example, WO2010 / 110382, WO2015 / 178385 and the like.
ブドウ種子及び果皮は、ブドウ科ブドウ属（Ｖｉｔａｃｅａｅ　Ｖｉｔｉｓ）植物のブドウ（学名Ｖｉｔｉｓ　ｓｐｐ．）の種子及び果皮である。ブドウの品種は特に限定されないが、例えば、カベルネ・ソーヴィニヨン、コンコード、メルロー、マスカットベリーＡ等の黒ブドウ；シャルドネ、甲州等の白ブドウ等が挙げられる。中でも、シャルドネ又は黒ブドウが好ましく、黒ブドウがより好ましく、カベルネ・ソーヴィニヨン、メルロー等がさらに好ましい。ブドウ種子及び／又は果皮の抽出物におけるブドウは、１種であってもよく、２種以上であってもよい。ブドウ種子及び／又は果皮の抽出物は、好ましくはブドウ種子及び果皮の抽出物である。 The grape seeds and skins are the seeds and skins of grapes (scientific name: Vitis spp.) Of the genus Vitaceae Vitis. The grape varieties are not particularly limited, and examples thereof include black grapes such as Cabernet Sauvignon, Concord, Merlot, and Muscat Berry A; and white grapes such as Chardonnay and Koshu. Among them, Chardonnay or black grapes are preferable, black grapes are more preferable, and Cabernet Sauvignon, Merlot and the like are further preferable. The grapes in the grape seed and / or pericarp extract may be one or more. The grape seed and / or pericarp extract is preferably a grape seed and pericarp extract.
ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母として、ヤロウィア・リポリティカ（Ｙａｒｒｏｗｉａ　ｌｉｐｏｌｙｔｉｃａ）が好ましい。ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物は、好ましくは、ヤロウィア・リポリティカ（Ｙａｒｒｏｗｉａ　ｌｉｐｏｌｙｔｉｃａ）の抽出物である。 As the yeast of the genus Yarrowia, Yarrowia lipolytica is preferable. The extract of Yarrowia yeast is preferably an extract of Yarrowia lipolytica.
ツバキ科ツバキ属（Ｔｈｅａｃｅａｅ　Ｃａｍｅｌｌｉａ）植物として、チャノキ（学名Ｃａｍｅｌｌｉａ　ｓｉｎｅｎｓｉｓ）が好ましい。
マンサク科マンサク属（Ｈａｍａｍｅｌｉｄａｃｅａｅ　Ｈａｍａｍｅｌｉｓ）植物として、ハマメリス（学名Ｈａｍａｍｅｌｉｓ　ｖｉｒｇｉｎｉａｎａ）が好ましい。
バラ科シモツケソウ属（Ｒｏｓａｃｅａｅ　Ｆｉｌｉｐｅｎｄｕｌａ）植物として、セイヨウナツユキソウ（学名Ｆｉｌｉｐｅｎｄｕｌａ　ｕｌｍａｒｉａ）が好ましい。 As a plant of the genus Camellia (Theaceae Camellia) of Theaceae, tea plant (scientific name Camellia sinensis) is preferable.
As a plant of the genus Witch hazel (Hamamelidaceae Hamamelis), Hamamelis (scientific name: Hamamelis virginiana) is preferable.
As a plant belonging to the genus Filipendula of the Rosaceae family, Meadowsweet (scientific name: Filipendula ulmaria) is preferable.
オオバコ科オオバコ属（Ｐｌａｎｔａｇｉｎａｃｅａｅ　Ｐｌａｎｔａｇｏ）植物として、セイヨウオオバコ（学名Ｐｌａｎｔａｇｏ　ｍａｊｏｒ）が好ましい。
シソ科ハッカ属（Ｌａｍｉａｃｅａｅ　Ｍｅｎｔｈａ）植物として、セイヨウハッカ（学名Ｍｅｎｔｈａ　ｘ　ｐｉｐｅｒｉｔａ）が好ましい。 As a plant of the genus Plantago (Plantago plantago) of the Plantain family, plantain (scientific name Plantago major) is preferable.
As a Lamiaceae Mentha plant of the Labiatae family, Mentha x mentha (scientific name: Mentha x piperita) is preferable.
植物抽出物の調製には、上記植物の任意の部位、例えば、根、根茎、茎、葉、枝、樹皮、花、種子（種皮、胚芽、胚乳、胚軸、子葉等を含む）、果実（果肉、果皮を含む）、又はこれらの複数の組み合わせを使用することができる。この植物の任意の部位を溶媒で抽出することにより植物抽出物を製造することができる。
植物抽出物を作製するのに好ましい部位として、例えば、以下の部位が挙げられる。本発明における植物抽出物としては、植物の下記部位の抽出物が好ましい。
バラ科バラ属植物は、花、特に花弁が好ましい。ブドウは、種子及び／又は果皮を使用し、好ましくは種子及び果皮を使用する。ツバキ科ツバキ属植物、マンサク科マンサク属植物は、葉が好ましい。バラ科シモツケソウ属植物は、花が好ましい。オオバコ科オオバコ属植物は、種子が好ましい。シソ科ハッカ属植物は全草が好ましい。
上記植物の部位は、そのまま抽出工程に付されてもよく、粉砕、切断又は乾燥された後に抽出工程に付されてもよい。本発明において植物抽出物は、上記植物をそのまま抽出した抽出物であってよいが、該植物を粉砕、切断又は乾燥したものから抽出した抽出物であってよい。好ましくは、上記植物を粉砕、切断又は乾燥したものからの抽出物である。 For the preparation of plant extracts, any part of the above plant, for example, root, rhizome, stem, leaf, branch, bark, flower, seed (including seed coat, germ, endosperm, hypocotyl, cotyledon, etc.), fruit (including seed coat, germ, endosperm, hypocotyl, cotyledon, etc.) (Including flesh, bark), or multiple combinations thereof can be used. A plant extract can be produced by extracting any part of the plant with a solvent.
Preferred sites for producing a plant extract include, for example, the following sites. As the plant extract in the present invention, an extract of the following part of the plant is preferable.
The Rosaceae plants of the genus Rosa are preferably flowers, especially petals. For grapes, seeds and / or pericarp are used, preferably seeds and pericarp. Leaves are preferable for the Theaceae Camellia plant and the Witch hazel plant. Flowers are preferred for plants of the genus Filipendula in the Rosaceae family. Seeds are preferred for Plantain plants of the Plantain family. The Labiatae plant of the genus Mentha is preferably whole plant.
The plant parts may be subjected to the extraction step as they are, or may be subjected to the extraction step after being crushed, cut or dried. In the present invention, the plant extract may be an extract obtained by extracting the above plant as it is, but may be an extract extracted from a crushed, cut or dried plant. Preferably, it is an extract from the crushed, cut or dried plant.
植物抽出物を得るための植物の抽出方法は特に限定されず、植物成分の抽出に用いられる通常の抽出方法により得ることができる。抽出方法は適宜設定することができ、抽出条件も特に限定されない。例えば、上記植物を常温又は加温下にて溶媒（抽出溶媒）で抽出することにより植物抽出物を得ることが好ましい。植物抽出物の調製において、原料である上記植物はそのまま抽出工程に付されてもよく、粉砕、切断又は乾燥された後に抽出工程に付されてもよい。 The method for extracting a plant for obtaining a plant extract is not particularly limited, and the plant can be obtained by a usual extraction method used for extracting plant components. The extraction method can be set as appropriate, and the extraction conditions are not particularly limited. For example, it is preferable to obtain a plant extract by extracting the above-mentioned plant with a solvent (extraction solvent) at room temperature or heating. In the preparation of the plant extract, the above-mentioned plant as a raw material may be subjected to the extraction step as it is, or may be subjected to the extraction step after being crushed, cut or dried.
植物抽出物の調製に用いる抽出溶媒は適宜選択することができ、植物成分の抽出に通常用いられるものを使用することができる。抽出溶媒として、例えば水；メタノール、エタノール、プロパノール、ブタノール等の炭素数１～５の１価アルコール；エチレングリコール、プロピレングリコール、１，２－ブチレングリコール、１，３－ブチレングリコール、１，４－ブチレングリコール、２，３－ブチレングリコール等の炭素数２～５の多価アルコール；アセトン、メチルエチルケトン等のケトン；酢酸メチル、酢酸エチル等のエステル；テトラヒドロフラン、ジエチルエーテル等の鎖状及び環状エーテル；ポリエチレングリコール等のポリエーテル；スクワラン等が挙げられる。これらは単独で用いてもよいし、２種以上を組み合わせた混合溶媒（混合液）を用いてもよい。炭素数１～５の１価アルコールとして、炭素数１～４のものが好ましく、炭素数２～４のものがより好ましく、エタノールがさらに好ましい。炭素数２～５の多価アルコールとして、炭素数２～４の２価又は３価のアルコールが好ましく、２価のアルコールがより好ましく、１，３－ブチレングリコールがさらに好ましい。中でも、抽出溶媒として、水、炭素数１～５の１価アルコール、炭素数２～５の多価アルコール、これらの２種以上の混合溶媒が好ましく、水、炭素数２～４の１価アルコール又はその水溶液、炭素数２～４の２価アルコール又はその水溶液がより好ましく、水、エタノール、エタノール水溶液、１，３－ブチレングリコール、１，３－ブチレングリコール水溶液がさらに好ましく、水、エタノール水溶液又は１，３－ブチレングリコール水溶液が特に好ましい。一態様において、エタノール水溶液及び１，３－ブチレングリコール水溶液は、エタノール又は１，３－ブチレングリコール濃度が１０～９８体積％が好ましく、３０～９０体積％がより好ましく、３０～７０体積％がさらに好ましい。本発明における植物抽出物として、上記の溶媒抽出物を好適に使用することができる。 The extraction solvent used for the preparation of the plant extract can be appropriately selected, and those usually used for the extraction of plant components can be used. As the extraction solvent, for example, water; monohydric alcohol having 1 to 5 carbon atoms such as methanol, ethanol, propanol and butanol; ethylene glycol, propylene glycol, 1,2-butylene glycol, 1,3-butylene glycol, 1,4- Polyhydric alcohols having 2 to 5 carbon atoms such as butylene glycol and 2,3-butylene glycol; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; chain and cyclic ethers such as tetrahydrofuran and diethyl ether; polyethylene. Polyethers such as glycol; squalane and the like can be mentioned. These may be used alone, or a mixed solvent (mixed solution) in which two or more kinds are combined may be used. As the monohydric alcohol having 1 to 5 carbon atoms, those having 1 to 4 carbon atoms are preferable, those having 2 to 4 carbon atoms are more preferable, and ethanol is further preferable. As the polyhydric alcohol having 2 to 5 carbon atoms, a divalent or trivalent alcohol having 2 to 4 carbon atoms is preferable, a divalent alcohol is more preferable, and 1,3-butylene glycol is further preferable. Among them, as the extraction solvent, water, a monohydric alcohol having 1 to 5 carbon atoms, a polyhydric alcohol having 2 to 5 carbon atoms, and a mixed solvent of two or more of these are preferable, and water and a monohydric alcohol having 2 to 4 carbon atoms are preferable. Alternatively, an aqueous solution thereof, a dihydric alcohol having 2 to 4 carbon atoms or an aqueous solution thereof is more preferable, and water, ethanol, an aqueous ethanol solution, 1,3-butylene glycol, a 1,3-butylene glycol aqueous solution is more preferable, and water, an aqueous ethanol solution or an aqueous solution thereof. An aqueous solution of 1,3-butylene glycol is particularly preferable. In one embodiment, the ethanol or 1,3-butylene glycol aqueous solution preferably has an ethanol or 1,3-butylene glycol concentration of 10 to 98% by volume, more preferably 30 to 90% by volume, and further 30 to 70% by volume. preferable. As the plant extract in the present invention, the above solvent extract can be preferably used.
抽出に際して酸又はアルカリを添加し、抽出溶媒のｐＨを調整してもよい。抽出後には、植物残渣（抽出後の植物体又はその部分）を抽出液から除去することが好ましい。植物残渣を抽出液から除去する方法は特に限定されず、ろ過、遠心分離等の公知の分離手段が挙げられる。 Acid or alkali may be added at the time of extraction to adjust the pH of the extraction solvent. After extraction, it is preferable to remove the plant residue (plant body after extraction or a portion thereof) from the extract. The method for removing the plant residue from the extract is not particularly limited, and examples thereof include known separation means such as filtration and centrifugation.
植物の抽出方法の一例として、例えば以下の方法が使用できる。植物をそのまま又は乾燥させて、細砕し、抽出溶媒を重量で０．１～３０倍量加え、常圧下、室温で好ましくは１０分～１５日、より好ましくは３０分～１０日、さらに好ましくは１時間～７日抽出するか、又は、抽出溶媒の沸点付近で好ましくは１０分～１日（より好ましくは１０分～２時間）程抽出してからろ過してろ液を得る。抽出の際は、静置してもよく、適宜攪拌を行ってもよい。また、得られたろ液（植物抽出液）をそのまま植物抽出物としてもよく、必要に応じ希釈、濃縮又は乾燥等してもよい。 As an example of the plant extraction method, the following methods can be used, for example. The plant is left as it is or dried, crushed, and the extraction solvent is added in an amount of 0.1 to 30 times by weight, preferably under normal pressure at room temperature for 10 minutes to 15 days, more preferably 30 minutes to 10 days, still more preferably. Is extracted for 1 hour to 7 days, or is extracted near the boiling point of the extraction solvent for preferably about 10 minutes to 1 day (more preferably 10 minutes to 2 hours) and then filtered to obtain a filtrate. At the time of extraction, it may be allowed to stand or stirred as appropriate. Further, the obtained filtrate (plant extract) may be used as it is as a plant extract, or may be diluted, concentrated or dried as necessary.
本発明においては、抽出により得られた植物抽出液をそのまま植物抽出物として用いることができる。また、本発明の効果を損なわない限り、公知の方法により希釈、濃縮又は乾燥して、希釈液、濃縮物や粉末としてもよく、ペースト状に調製して用いることもできる。乾燥方法として、例えば、凍結乾燥、噴霧乾燥等が挙げられる。さらに、上記の植物抽出液、その濃縮物や乾燥粉末等について、本発明の効果を損なわない範囲で、必要に応じて、さらに脱臭、脱色等の精製を行ってもよい。このような精製方法は、通常の手段を任意に選択して行えばよい。
本発明における植物抽出物は、上記のような抽出方法で得られた各種溶媒抽出液、その希釈液、その濃縮物又はその乾燥粉末、これらの精製物を含む。また、抽出物は、抽出溶媒とは異なる溶媒で希釈又は溶解させて用いることもできる。
また、上記の植物抽出物は市販されており、市販品を使用することもできる。 In the present invention, the plant extract obtained by extraction can be used as it is as a plant extract. Further, as long as the effect of the present invention is not impaired, it may be diluted, concentrated or dried by a known method to form a diluted solution, a concentrate or a powder, or it may be prepared in the form of a paste and used. Examples of the drying method include freeze-drying and spray-drying. Further, the above-mentioned plant extract, its concentrate, dry powder and the like may be further purified by deodorization, decolorization and the like, if necessary, as long as the effects of the present invention are not impaired. Such a purification method may be carried out by arbitrarily selecting ordinary means.
The plant extract in the present invention includes various solvent extracts obtained by the above-mentioned extraction methods, diluted solutions thereof, concentrates thereof or dry powders thereof, and purified products thereof. Further, the extract can be used by diluting or dissolving it in a solvent different from the extraction solvent.
In addition, the above plant extracts are commercially available, and commercially available products can also be used.
ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物は、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の菌体又は菌体培養物の抽出物であることが好ましい。菌体又は菌体培養物の抽出物として、菌体又は菌体を含む菌体培養物（好ましくは菌体培養液）に、自己消化処理や酵素分解処理等の菌体破砕処理を行って、酵母菌体内容物を培養液中等に溶出したもの（菌体破砕物）、菌体破砕処理を行った菌体又は菌体培養物（菌体破砕物）から菌体残渣を除去したものが挙げられる。好ましくは、菌体又は菌体を含む菌体培養物（好ましくは菌体培養液）を自己消化処理したものである。菌体破砕物やこれから菌体残渣を除去したものには、必要に応じて加熱処理等の殺菌処理等を行ってもよい。上記の酵母の抽出物は、このような殺菌処理等が行われたものであってよい。ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物の形態は特に限定されないが、例えば、ペースト状、懸濁状、エキス状、液状、粉末状等が挙げられる。 The extract of Yarrowia yeast is preferably an extract of bacterial cells or cell cultures of Yarrowia yeast. As an extract of the mycelium or the mycelium culture, the mycelium or the mycelium culture containing the mycelium (preferably the mycelium culture solution) is subjected to a mycelium crushing treatment such as autolysis treatment or enzymatic decomposition treatment. Examples include yeast cells eluted in a culture medium (crushed cells), and cells that have undergone cell crushing treatment or cultures that have been subjected to cell crushing (crushed cells) from which bacterial residue has been removed. Be done. Preferably, the cells or a cell culture containing the cells (preferably a cell culture solution) is autolyzed. If necessary, sterilization treatment such as heat treatment may be performed on the crushed cell product and the product from which the bacterial cell residue has been removed. The above-mentioned yeast extract may have undergone such a sterilization treatment or the like. The form of the extract of Yarrowia yeast is not particularly limited, and examples thereof include paste, suspension, extract, liquid, and powder.
一態様において、バラ科バラ属植物の抽出物として、バラ（好ましくは花弁）のエタノール抽出物又はエタノール水溶液抽出物が好ましい。
ブドウ種子及び／又は果皮の抽出物として、ブドウ種子及び／又は果皮のエタノール抽出物又はエタノール水溶液抽出物が好ましい。
ツバキ科ツバキ属植物の抽出物として、チャノキ（好ましくは葉）のエタノール抽出物又はエタノール水溶液抽出物が好ましい。
マンサク科マンサク属植物の抽出物として、ハマメリス（好ましくは葉）の１，３－ブチレングリコール抽出物又は１，３－ブチレングリコール水溶液抽出物が好ましい。
バラ科シモツケソウ属植物の抽出物として、セイヨウナツユキソウ（好ましくは花）の１，３－ブチレングリコール抽出物又は１，３－ブチレングリコール水溶液抽出物が好ましい。オオバコ科オオバコ属植物の抽出物として、セイヨウオオバコ（好ましくは種子）のエタノール抽出物又はエタノール水溶液抽出物が好ましい。
シソ科ハッカ属植物の抽出物として、セイヨウハッカ（好ましくは全草）のエタノール抽出物、エタノール水溶液抽出物、又は熱水抽出物が好ましい。 In one embodiment, as the extract of the Rosaceae plant belonging to the genus Rosa, an ethanol extract of roses (preferably petals) or an aqueous ethanol extract is preferable.
As the grape seed and / or pericarp extract, an ethanol extract or an aqueous ethanol extract of grape seed and / or pericarp is preferable.
As the extract of the Theaceae plant of the genus Camellia, an ethanol extract of tea plant (preferably leaves) or an aqueous ethanol extract is preferable.
As the extract of the witch hazel genus plant, the extract of Hamamelis (preferably leaves) 1,3-butylene glycol extract or the extract of 1,3-butylene glycol aqueous solution is preferable.
As an extract of a plant belonging to the genus Filipendula of the Rosaceae family, a 1,3-butylene glycol extract or a 1,3-butylene glycol aqueous solution extract of Meadowsweet (preferably a flower) is preferable. As an extract of a plantago genus Plantago of the Plantain family, an ethanol extract of plantain (preferably seeds) or an aqueous ethanol extract is preferable.
As the extract of the Labiatae plant of the genus Peppermint, an ethanol extract of mint (preferably whole plant), an aqueous ethanol extract, or a hot water extract is preferable.
本発明の組成物の形態は特に限定されず、粉末状、顆粒状、ペースト状、固形状等であってもよく、液状であってもよく、適宜選択することができる。本発明の組成物には、本発明の効果を損なわない限り、上記の抽出物に加えて、その他の成分を配合することができる。その他の成分としては、例えば、後述する化粧料、飲食品、医薬品、医薬部外品等に使用し得る成分が挙げられる。
本発明の４型コラーゲンの発現の低下抑制又は改善用組成物、皮膚バリア機能の低下抑制又は改善用組成物は、一例として、剤の形態で提供することができるが、本形態に限定されるものではない。当該剤をそのまま組成物として、又は、当該剤を含む組成物として提供することもできる。一態様において、本発明の組成物は、４型コラーゲンの発現の低下抑制又は改善剤、皮膚バリア機能の低下抑制又は改善剤ということもできる。 The form of the composition of the present invention is not particularly limited, and may be in the form of powder, granules, paste, solid, etc., or in liquid form, and can be appropriately selected. In addition to the above-mentioned extracts, other components can be added to the composition of the present invention as long as the effects of the present invention are not impaired. Examples of other ingredients include ingredients that can be used in cosmetics, foods and drinks, pharmaceuticals, quasi-drugs, etc., which will be described later.
The composition for suppressing or improving the expression of type 4 collagen of the present invention and the composition for suppressing or improving the decrease in skin barrier function can be provided, for example, in the form of an agent, but are limited to this form. It's not a thing. The agent can be provided as it is as a composition or as a composition containing the agent. In one aspect, the composition of the present invention can also be said to be an agent for suppressing or improving the expression of type 4 collagen, and an agent for suppressing or improving the decrease in skin barrier function.
本発明の第一の態様の４型コラーゲンの発現の低下抑制又は改善用組成物は、対象の４型コラーゲンの発現の低下を抑制又は当該発現を改善するため、好ましくは対象の皮膚における４型コラーゲンの発現の低下を抑制又は当該発現を改善するために使用される。本発明の第二の態様、第三の態様及び第四の態様の皮膚バリア機能の低下抑制又は改善用組成物は、対象の皮膚バリア機能の低下を抑制又は当該機能を改善するために使用される。
本発明の組成物は、例えば、皮膚バリア機能の低下、基底膜の機能低下又は４型コラーゲンの発現低下に起因する不具合の予防又は改善のために使用することができる。このような不具合として、例えば、肌の乾燥、かゆみ、かさつき、敏感肌、肌荒れ等が挙げられる。一態様において、本発明の組成物は、皮膚バリア機能の低下抑制若しくは改善、又は、皮膚における４型コラーゲンの発現の低下抑制若しくは改善により、肌の乾燥、かゆみ、かさつき、敏感肌、肌荒れ等を予防又は改善するために使用され得る。不具合の予防は、発症の防止、発症の遅延、発症率の低下等を包含する。不具合の改善は、症状の軽快、症状の好転、症状の進行抑制等を包含する。 The composition for suppressing or ameliorating the decrease in the expression of type 4 collagen according to the first aspect of the present invention is preferably type 4 in the target skin in order to suppress or improve the decrease in the expression of the target type 4 collagen. It is used to suppress the decrease in collagen expression or to improve the expression. The composition for suppressing or improving the deterioration of the skin barrier function of the second aspect, the third aspect and the fourth aspect of the present invention is used for suppressing the deterioration of the skin barrier function of the subject or improving the function. NS.
The composition of the present invention can be used, for example, for the prevention or improvement of defects caused by a decrease in skin barrier function, a decrease in basement membrane function, or a decrease in the expression of type 4 collagen. Examples of such defects include dry skin, itchiness, dryness, sensitive skin, and rough skin. In one aspect, the composition of the present invention suppresses or improves the decrease in skin barrier function, or suppresses or improves the decrease in the expression of type 4 collagen in the skin, thereby causing dryness, itchiness, dryness, sensitive skin, rough skin, etc. Can be used to prevent or improve. Prevention of defects includes prevention of onset, delay of onset, reduction of onset rate, and the like. Improvement of defects includes improvement of symptoms, improvement of symptoms, suppression of progression of symptoms, and the like.
本発明の組成物は、化粧料、飲食品、医薬品、医薬部外品等様々な用途に使用することができる。本発明の組成物は、それ自体が、４型コラーゲンの発現の低下抑制又は改善のため、又は、皮膚バリア機能の低下抑制又は改善のための化粧料、飲食品、医薬品又は医薬部外品等であってもよく、又は、該化粧料、飲食品、医薬品又は医薬部外品等に配合して使用される素材又は製剤等であってもよい。 The composition of the present invention can be used for various purposes such as cosmetics, foods and drinks, pharmaceuticals, and quasi-drugs. The composition of the present invention itself is for suppressing or improving the decrease in the expression of type 4 collagen, or for suppressing or improving the decrease in the skin barrier function, such as cosmetics, foods and drinks, pharmaceuticals or quasi-drugs, etc. It may be a material or a preparation used by blending with the cosmetics, foods and drinks, pharmaceuticals, quasi-drugs, and the like.
本発明の組成物は、例えば、皮膚外用剤として好適に使用される。皮膚外用剤は、化粧料、医薬品、医薬部外品を含み、好ましくは化粧料である。
本発明の組成物は、皮膚外用剤以外の医薬品又は医薬部外品として使用することもできる。
別の実施態様においては、本発明の組成物を飲食品とすることも好ましい。本発明の組成物を化粧料、医薬品、医薬部外品等の皮膚外用剤、飲食品等とする場合について以下に説明する。 The composition of the present invention is suitably used, for example, as an external preparation for skin. The external preparation for skin includes cosmetics, pharmaceuticals, and quasi-drugs, and is preferably a cosmetic.
The composition of the present invention can also be used as a drug or quasi-drug other than an external preparation for skin.
In another embodiment, it is also preferable to use the composition of the present invention as a food or drink. The case where the composition of the present invention is used as a cosmetic, a pharmaceutical product, a skin external preparation such as a quasi-drug, a food or drink, etc. will be described below.
本発明の組成物等を皮膚外用剤とする場合、剤形等は特に限定されず、例えば溶液、乳液、クリーム、ゲル、粉末、エアゾール、ミスト、カプセル及びシート等任意の形態とすることができる。皮膚外用剤は、好ましくは化粧料である。化粧料の製品形態も特に限定されず、例えば、洗顔料、メーク落とし、化粧水、美容液、パック、乳液、クリーム、日焼け止め等のスキンケア化粧料；ファンデーション、化粧下地、口紅、アイシャドー、アイライナー、マスカラ、アイブロー、頬紅、ネイルエナメル等のメイクアップ化粧料；ヘアシャンプー、ヘアリンス、整髪料、染毛料、育毛剤等の毛髪化粧料；石けん、ボディソープ等の洗浄料；入浴剤等が挙げられる。 When the composition of the present invention is used as an external preparation for skin, the dosage form and the like are not particularly limited, and may be in any form such as a solution, a milky lotion, a cream, a gel, a powder, an aerosol, a mist, a capsule and a sheet. .. The external preparation for skin is preferably a cosmetic. The product form of cosmetics is not particularly limited, and for example, skin care cosmetics such as facial cleansers, makeup removers, lotions, beauty liquids, packs, milky lotions, creams, and sunscreens; foundations, makeup bases, lipsticks, eye shadows, and eyes. Makeup cosmetics such as liner, mascara, eyebrow, cheek red, nail enamel; hair cosmetics such as hair shampoo, hair rinse, hair conditioner, hair dye, hair restorer; cleaning agents such as soap and body soap; bathing agents, etc. Be done.
上記化粧料には、本発明の効果を損なわない範囲で、化粧料に許容される担体、添加剤等の成分、例えば、水、アルコール類、油剤、界面活性剤、増粘剤、金属セッケン、ゲル化剤、粉体、キレート剤、水溶性高分子、皮膜形成剤、樹脂、包接化合物、抗菌剤、消臭剤、塩類、ｐＨ調整剤、紫外線吸収剤、上記以外の動植物・微生物由来の抽出物、角質溶解剤、酵素、ホルモン類、他のビタミン類、保湿剤、殺菌剤、抗炎症剤、香料等の１又は２以上を適宜含有させることができる。化粧料は、一般的な製造法により製造することができる。例えば、有効成分に使用される上記の抽出物と、上記の化粧料に使用し得る成分１又は２以上とを混合した後、所望の形態に加工することによって得ることができる。 The above-mentioned cosmetics include components such as carriers and additives that are acceptable for cosmetics, for example, water, alcohols, oils, surfactants, thickeners, metal salts, etc., as long as the effects of the present invention are not impaired. Gelling agents, powders, chelating agents, water-soluble polymers, film-forming agents, resins, inclusion compounds, antibacterial agents, deodorants, salts, pH adjusters, UV absorbers, derived from animals, plants and microorganisms other than the above One or two or more of extracts, keratolytic agents, enzymes, hormones, other vitamins, moisturizers, bactericidal agents, anti-inflammatory agents, fragrances and the like can be appropriately contained. Cosmetics can be manufactured by a general manufacturing method. For example, it can be obtained by mixing the above-mentioned extract used as an active ingredient with one or more of the above-mentioned ingredients that can be used in cosmetics, and then processing the mixture into a desired form.
皮膚外用剤を医薬品又は医薬部外品とする場合、本発明の効果を損なわない範囲で、医薬品又は医薬部外品に許容される担体、添加剤等の成分を使用してもよい。そのような成分として、例えば、賦形剤、結合剤、崩壊剤、滑沢剤、抗酸化剤、着色剤等の１又は２以上が挙げられ、これらは必要に応じて使用できる。 When the external preparation for skin is a drug or a quasi-drug, components such as carriers and additives permitted for the drug or quasi-drug may be used as long as the effects of the present invention are not impaired. Examples of such components include one or more of excipients, binders, disintegrants, lubricants, antioxidants, colorants and the like, which can be used as needed.
本発明の組成物は、上述した皮膚外用剤以外の医薬品、医薬部外品とすることもできる。このような医薬品又は医薬部外品は、経口投与されてもよく、非経口投与されてもよい。医薬品又は医薬部外品は経口投与製剤（内服剤）又は非経口投与製剤の形態とすることができる。経口投与製剤の剤形としては、液剤、錠剤、散剤、細粒剤、顆粒剤、糖衣錠、カプセル剤、懸濁液、乳剤、チュアブル剤等が挙げられる。非経口投与製剤の剤形としては、注射剤、輸液剤等が挙げられる。このような医薬品又は医薬部外品には、医薬品又は医薬部外品に許容される担体、添加剤等の成分を使用してもよい。そのような成分として、例えば、賦形剤、結合剤、崩壊剤、滑沢剤、抗酸化剤、着色剤、矯味剤等の１又は２以上が挙げられ、これらは必要に応じて使用できる。医薬品、医薬部外品は、一般的な製造法により製造することができる。例えば、有効成分に使用される上記の抽出物と、医薬品又は医薬部外品において使用し得る成分の１又は２以上とを混合した後、所望の形態に加工することによって得ることができる。 The composition of the present invention can also be a drug or quasi-drug other than the above-mentioned external preparation for skin. Such drugs or quasi-drugs may be administered orally or parenterally. The drug or quasi-drug may be in the form of an orally-administered preparation (oral preparation) or a parenteral-administered preparation. Examples of the dosage form of the orally administered preparation include liquid preparations, tablets, powders, fine granules, granules, sugar-coated tablets, capsules, suspensions, emulsions, chewable agents and the like. Examples of the dosage form of the parenteral preparation include injections, infusions and the like. For such drugs or quasi-drugs, components such as carriers and additives that are acceptable for the drugs or quasi-drugs may be used. Examples of such components include one or more of excipients, binders, disintegrants, lubricants, antioxidants, colorants, flavoring agents and the like, which can be used as needed. Pharmaceuticals and quasi-drugs can be manufactured by a general manufacturing method. For example, it can be obtained by mixing the above extract used as an active ingredient with one or more of the ingredients that can be used in a drug or a quasi drug, and then processing the extract into a desired form.
本発明の組成物を飲食品とする場合、飲食品は特に限定されず、例えば、一般的な飲食品、健康食品、機能性表示食品、特定保健用食品等が挙げられる。飲食品の形態も特に限定されない。上記健康食品、機能性表示食品、特定保健用食品は、例えば、液剤、錠剤、散剤、細粒剤、顆粒剤、糖衣錠、カプセル剤、懸濁液、乳剤、チュアブル剤、流動食等の各種製剤形態とすることもできる。 When the composition of the present invention is used as a food or drink, the food or drink is not particularly limited, and examples thereof include general food and drink, health food, food with functional claims, food for specified health use and the like. The form of food and drink is not particularly limited. The above-mentioned health foods, foods with functional claims, and foods for specified health use include various preparations such as liquid preparations, tablets, powders, fine granules, granules, sugar-coated tablets, capsules, suspensions, emulsions, chewables, and liquid foods. It can also be in the form.
上記飲食品には、本発明の効果を損なわない範囲で、飲食品に許容される成分、例えば、他の飲食品材料、飲食品に配合される添加剤等を配合してもよい。このような飲食品は、一般的な製造法により製造することができる。例えば、飲食品の製造において、飲食品材料等に、本発明において有効成分に使用される上記の抽出物を配合すればよい。 In the above-mentioned food and drink, ingredients permitted in the food and drink, for example, other food and drink materials, additives to be blended in the food and drink, and the like may be blended as long as the effects of the present invention are not impaired. Such foods and drinks can be produced by a general production method. For example, in the production of foods and drinks, the above-mentioned extract used as an active ingredient in the present invention may be blended with food and drink materials and the like.
本発明の組成物が、皮膚外用剤、皮膚外用剤以外の医薬品又は医薬部外品、飲食品等のいずれの形態の場合も、有効成分に使用される上記抽出物の含有量は、該組成物中に、例えば、該抽出物の乾燥物換算の重量として、０．００００００１～１００重量％が好ましく、０．０００００１～１００重量％がより好ましく、０．００００１～１０重量％がさらに好ましい。上記含有量は、２種以上の抽出物を含む場合は、その合計の含有量である。 When the composition of the present invention is in any form such as a skin external preparation, a pharmaceutical product other than the skin external preparation, a quasi drug, a food or drink, etc., the content of the extract used as the active ingredient is the composition. In the substance, for example, the weight of the extract in terms of dry matter is preferably 0.000000001 to 100% by weight, more preferably 0.000001 to 100% by weight, still more preferably 0.00001 to 10% by weight. When two or more kinds of extracts are contained, the above-mentioned content is the total content thereof.
本発明の組成物の使用量は、特に限定されず、皮膚バリア機能の低下抑制又は改善効果、４型コラーゲンの発現の低下抑制又は改善効果が得られるような量（有効量）であればよく、対象者の状態、体重、性別、年齢又はその他の要因に従って適宜設定することができる。本発明の組成物の使用量は、化粧料等の皮膚外用剤であれば、例えば、成人（６０ｋｇ）１人当たり１日当たり、有効成分に使用される上記の抽出物（乾燥物換算）として、例えば、０．０１～５００ｍｇが好ましい。医薬品又は医薬部外品を経口投与する場合の投与量は、例えば、成人（６０ｋｇ）１人当たり１日当たり、上記の抽出物（乾燥物換算）として、例えば、０．０１～５００ｍｇが好ましい。飲食品であれば、その摂取量は、例えば、成人（６０ｋｇ）１人当たり１日当たり、上記の抽出物（乾燥物換算）として、例えば、０．０１～５００ｍｇが好ましい。上記量の抽出物を、単回又は複数回に分けて摂取又は投与することができる。上記の抽出物の量は、２種以上の抽出物を使用する場合は、その合計量である。上記抽出物を含む皮膚外用剤を皮膚に適用するタイミング、飲食品、医薬品又は医薬部外品を摂取又は投与するタイミング等は特に限定されない。 The amount of the composition of the present invention to be used is not particularly limited as long as it is an amount (effective amount) that can suppress or improve the decrease in skin barrier function and suppress or improve the expression of type 4 collagen. , Can be appropriately set according to the subject's condition, weight, gender, age or other factors. The amount of the composition of the present invention used is, for example, as an external preparation for skin such as cosmetics, for example, as the above-mentioned extract (dry matter equivalent) used as an active ingredient per day per adult (60 kg). , 0.01-500 mg is preferable. When the drug or quasi-drug is orally administered, the dose is preferably, for example, 0.01 to 500 mg per day for an adult (60 kg) as the above extract (dry matter equivalent). In the case of food and drink, the intake thereof is preferably, for example, 0.01 to 500 mg per day for an adult (60 kg) as the above-mentioned extract (dry matter equivalent). The above amount of extract can be ingested or administered in a single dose or in multiple doses. The amount of the above extract is the total amount when two or more kinds of extracts are used. The timing of applying the external preparation for skin containing the above extract to the skin, the timing of ingesting or administering foods and drinks, pharmaceuticals or quasi-drugs, and the like are not particularly limited.
本発明の組成物の適用対象（投与対象ということもできる）としては、特に限定されず、ヒト又は非ヒト哺乳動物等に適用することができるが、ヒトが好ましい。適用対象としては、例えば、皮膚バリア機能の低下抑制又は改善を希望する又は必要とする対象、４型コラーゲンの発現の低下抑制又は改善を希望する又は必要とする対象等が挙げられる。このような対象として、例えば、肌の乾燥、かゆみ、かさつき、敏感肌、肌荒れを実感する人が挙げられる。本発明の組成物は、その形態、剤形等に応じて自体公知の種々の方法で摂取又は投与することができる。一態様において、本発明の組成物は、皮膚に適用することが好ましい。 The application target (which can also be said to be an administration target) of the composition of the present invention is not particularly limited and can be applied to humans, non-human mammals and the like, but humans are preferable. Examples of the application target include a subject who desires or needs to suppress or need to suppress or improve the decrease in skin barrier function, and a subject who desires or needs to suppress or need to suppress or need to suppress or improve the expression of type 4 collagen. Such targets include, for example, those who experience dry skin, itching, dryness, sensitive skin, and rough skin. The composition of the present invention can be ingested or administered by various methods known per se, depending on its form, dosage form and the like. In one aspect, the compositions of the invention are preferably applied to the skin.
本発明の組成物の有効成分に使用される上記の抽出物は、対象の４型コラーゲンの発現の低下を抑制又は当該発現を改善するために使用することができる。上記の抽出物は、対象の皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善するため、好ましくは４型コラーゲンの発現の低下抑制又は改善により皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善するために使用することができる。 The above-mentioned extract used as the active ingredient of the composition of the present invention can be used to suppress a decrease in the expression of the target type 4 collagen or to improve the expression. Since the above extract suppresses the decrease in the skin barrier function or improves the skin barrier function of the subject, preferably, the decrease in the expression of type 4 collagen is suppressed or improved to suppress the decrease in the skin barrier function or the skin barrier function is improved. Can be used to
本発明は、以下の使用及び方法も包含する。
４型コラーゲンの発現の低下抑制又は改善により皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善するための、バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、バラ科シモツケソウ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物の使用。
バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、バラ科シモツケソウ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物を投与する、４型コラーゲンの発現の低下抑制又は改善により皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善する方法。 The present invention also includes the following uses and methods.
Extracts of Rosaceae plants, grape seeds and / or peels, Yarrowia, for suppressing or improving the decrease in the expression of type 4 collagen to suppress the decrease in the skin barrier function or improve the skin barrier function. ) Use of one or more extracts selected from the group consisting of extracts of genus yeast, extracts of Camellia plants of the family Rosaceae, extracts of plants of the genus Rosaceae, and extracts of plants of the genus Camellia. ..
Extracts of Rosaceae plants, grape seeds and / or peels, extracts of Yarrowia yeast, extracts of Camellia plants, extracts of Rosaceae plants, and perilla A method of suppressing or improving the decrease in skin barrier function by suppressing or improving the expression of type 4 collagen by administering one or more extracts selected from the group consisting of extracts of Rosaceae plants. ..
皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善するための、マンサク科マンサク属植物の抽出物及び／又はオオバコ科オオバコ属植物の抽出物の使用。
マンサク科マンサク属植物の抽出物及び／又はオオバコ科オオバコ属植物の抽出物を投与する、皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善する方法。 Use of witch hazel plant extract and / or plantain plantago extract to suppress deterioration of skin barrier function or improve skin barrier function.
A method of administering an extract of a witch hazel plant and / or an extract of a plantain plant, which suppresses a decrease in the skin barrier function or improves the skin barrier function.
４型コラーゲンの発現の低下を抑制又は当該発現を改善するための、バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、マンサク科マンサク属植物の抽出物、バラ科シモツケソウ属植物の抽出物、オオバコ科オオバコ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物の使用。
上記１種以上の抽出物を投与する４型コラーゲンの発現の低下を抑制又は当該発現を改善する方法。 Extracts of Rosaceae plants, grape seeds and / or peels, extracts of Yarrowia yeast, extracts of Rosaceae, and camellia for suppressing or improving the expression of type 4 collagen. Selected from the group consisting of extracts of genus plants, extracts of plants of the genus Mansaku of the family Rosaceae, extracts of plants of the genus Rosaceae, extracts of plants of the genus Oobaco, and extracts of plants of the genus Rosaceae. Use of one or more extracts.
A method for suppressing a decrease in the expression of type 4 collagen or improving the expression by administering one or more of the above extracts.
上記抽出物は、好ましくは皮膚に投与（適用）され、皮膚バリア機能の低下を抑制又は当該機能を改善するため、皮膚における４型コラーゲンの発現の低下を抑制又は当該発現を改善するため、又は、基底膜機能の低下を抑制又は当該機能を改善するために使用することができる。一態様において、上記抽出物は、皮膚バリア機能の低下、基底膜の機能低下又は４型コラーゲンの発現低下に起因する不具合の予防又は改善のために使用することができる。上記抽出物は、例えば、皮膚バリア機能の低下抑制若しくは改善、又は、皮膚における４型コラーゲンの発現の低下抑制若しくは改善により、肌の乾燥、かゆみ、かさつき、敏感肌、肌荒れ等を予防又は改善するために使用され得る。 The above extract is preferably administered (applied) to the skin to suppress a decrease in the skin barrier function or improve the function, to suppress a decrease in the expression of type 4 collagen in the skin, or to improve the expression. , Can be used to suppress deterioration of basement membrane function or improve the function. In one aspect, the extract can be used to prevent or ameliorate defects caused by decreased skin barrier function, decreased basement membrane function or decreased expression of type 4 collagen. The above extract prevents or improves dryness, itchiness, dryness, sensitive skin, rough skin, etc. of the skin by, for example, suppressing or improving the decrease in skin barrier function, or suppressing or improving the decrease in the expression of type 4 collagen in the skin. Can be used to
上記使用及び方法において、上記の抽出物、これを投与する対象、抽出物の投与量及びこれらの好ましい態様等は、上述した本発明の組成物におけるものと同じである。上記抽出物は、そのまま使用してもよく、その他の成分と組み合わせて使用してもよい。本発明の組成物を使用してもよい。上記の抽出物は上述した皮膚外用剤、飲食品等の形態で使用することができる。
上記使用は、治療的使用であってもよく、非治療的使用であってもよい。上記方法は、治療的な方法であってもよく、非治療的な方法であってもよい。非治療的とは、医療行為、すなわち人間の手術、治療又は診断を含まない概念である。 In the above-mentioned uses and methods, the above-mentioned extract, the subject to which the above-mentioned extract is administered, the dose of the extract, preferable embodiments thereof and the like are the same as those in the above-mentioned composition of the present invention. The above extract may be used as it is or may be used in combination with other components. The composition of the present invention may be used. The above extract can be used in the form of the above-mentioned external preparation for skin, food and drink, and the like.
The above use may be therapeutic or non-therapeutic use. The above method may be a therapeutic method or a non-therapeutic method. Non-therapeutic is a concept that does not include medical practice, ie human surgery, treatment or diagnosis.
本発明の別の態様は、上記抽出物の、皮膚バリア機能の低下抑制又は改善用組成物を製造するための使用；上記抽出物の、４型コラーゲンの発現の低下抑制又は改善用組成物を製造するための使用でもある。抽出物及びこれらの好ましい態様等は、上述した本発明の組成物におけるものと同じである。
本発明のさらに別の態様は、皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善するための、基底膜機能の低下抑制又は改善剤の使用；基底膜機能の低下抑制又は改善剤を投与する、皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善する方法でもある。
基底膜機能の低下抑制又は改善剤及びその好ましい態様は、上記と同じである。 Another aspect of the present invention is the use of the above extract for producing a composition for suppressing or improving the decrease in skin barrier function; the composition for suppressing or improving the expression of type 4 collagen in the above extract. It is also used for manufacturing. The extracts and preferred embodiments thereof are the same as those in the compositions of the present invention described above.
Yet another aspect of the present invention is the use of a basement membrane function decline suppressor or ameliorating agent to suppress or improve the skin barrier function decline; administer a basement membrane function decline suppressor or improve agent. It is also a method of suppressing a decrease in the skin barrier function or improving the skin barrier function.
The agent for suppressing or improving the decrease in basement membrane function and its preferred embodiment are the same as described above.
本発明は、皮膚バリア機能の低下抑制又は改善作用を有する物質のスクリーニング方法も包含する。本発明のスクリーニング方法について説明する。
本発明の皮膚バリア機能の低下抑制又は改善作用を有する物質のスクリーニング方法は、インビトロで、ヒト表皮角化細胞を、被験物質を含む培地又は上記被験物質を含まないコントロール培地で培養する工程、及び、上記被験物質を含む培地で培養した皮膚細胞及びコントロール培地で培養したコントロール皮膚細胞における４型コラーゲン発現量を比較する工程を含み、上記被験物質を含む培地で培養した皮膚細胞における４型コラーゲン発現量が、上記コントロール皮膚細胞における４型コラーゲン発現量より多い場合に、上記被験物質を皮膚バリア機能の低下抑制又は改善作用を有する物質として選択する。 The present invention also includes a method for screening a substance having an effect of suppressing or improving a decrease in skin barrier function. The screening method of the present invention will be described.
The method for screening a substance having an effect of suppressing or improving a decrease in skin barrier function of the present invention is a step of culturing human epidermal keratinized cells in a medium containing a test substance or a control medium not containing the above test substance in vitro. , Including the step of comparing the expression level of type 4 collagen in the skin cells cultured in the medium containing the above test substance and the control skin cells cultured in the control medium, and the expression of type 4 collagen in the skin cells cultured in the medium containing the above test substance. When the amount is larger than the expression level of type 4 collagen in the control skin cells, the test substance is selected as a substance having an effect of suppressing or improving the decrease of the skin barrier function.
本発明のスクリーニング方法では、インビトロで、ヒト表皮角化細胞を、被験物質を含む培地又は上記被験物質を含まないコントロール培地で培養する。ヒト表皮角化細胞は市販されており、市販品を使用することができる。
被験物質は特に限定されず、例えば、植物抽出液、細胞抽出液、細胞培養上清、発酵生産物、タンパク質、ペプチド、ビタミン類、合成化合物等が挙げられる。これらは既知物質であってもよく、新規物質であってもよい。 In the screening method of the present invention, human epidermal keratinocytes are cultured in vitro in a medium containing a test substance or a control medium containing no test substance. Human epidermal keratinocytes are commercially available, and commercially available products can be used.
The test substance is not particularly limited, and examples thereof include plant extracts, cell extracts, cell culture supernatants, fermentation products, proteins, peptides, vitamins, synthetic compounds and the like. These may be known substances or new substances.
インビトロで、ヒト表皮角化細胞を、被験物質を含む培地又は上記被験物質を含まないコントロール培地で培養する工程を、培養工程ともいう。上記被験物質を含む培地で培養した皮膚細胞（被験皮膚細胞ともいう）及びコントロール培地で培養したコントロール皮膚細胞における４型コラーゲン発現量を比較する工程を、４型コラーゲン発現量比較工程ともいう。 The step of culturing human epidermal keratinocytes in vitro in a medium containing a test substance or a control medium not containing the test substance is also referred to as a culturing step. The step of comparing the type 4 collagen expression level in the skin cells cultured in the medium containing the test substance (also referred to as the test skin cells) and the control skin cells cultured in the control medium is also referred to as a type 4 collagen expression level comparison step.
培養工程において、ヒト表皮角化細胞を培養する条件は特に限定されず、公知の条件を採用すればよい。被験物質を含む培地は、被験物質を培地に混合することにより調製することができる。培地は、ヒト表皮角化細胞の培養に使用されるものを使用することができ、市販品を使用してもよい。培地の一例として、ＨｕＭｅｄｉａ－ＫＢ２培地（クラボウ社製）、ＤＭＥＭ培地等が挙げられる。培地には、インスリン、ヒトＥＧＦ、ハイドロコーチゾン、血清、抗菌剤等を添加してもよい。培養時間は特に限定されず、培養条件等により適宜設定すればよい。コントロール培地には、被験物質を含有しない以外は、被験物質を含有する培地と同じ培地を使用すればよい。例えば、被験物質を含む培地又はコントロール培地中で、ヒト表皮角化細胞を好ましくは１～７２時間、より好ましくは３～４８時間培養すればよい。
培養工程において、紫外線照射、酸化ストレス負荷等のストレス負荷を行ってもよい。例えば、紫外線照射、酸化ストレス負荷等のストレス負荷条件で培養を行うと、当該ストレスによる皮膚バリア機能の低下抑制又はストレス下での皮膚バリア機能の改善により有効な成分をスクリーニングすることができる。 In the culturing step, the conditions for culturing human epidermal keratinocytes are not particularly limited, and known conditions may be adopted. The medium containing the test substance can be prepared by mixing the test substance with the medium. As the medium, those used for culturing human epidermal keratinocytes can be used, and commercially available products may be used. Examples of the medium include HuMedia-KB2 medium (manufactured by Kurabo Industries Ltd.), DMEM medium and the like. Insulin, human EGF, hydrocortisone, serum, antibacterial agent and the like may be added to the medium. The culturing time is not particularly limited, and may be appropriately set depending on the culturing conditions and the like. As the control medium, the same medium as the medium containing the test substance may be used except that the test substance is not contained. For example, human epidermal keratinocytes may be cultured in a medium containing a test substance or a control medium for preferably 1 to 72 hours, more preferably 3 to 48 hours.
In the culturing step, stress loading such as ultraviolet irradiation and oxidative stress loading may be performed. For example, when culturing under stress loading conditions such as ultraviolet irradiation and oxidative stress loading, effective components can be screened by suppressing the decrease in skin barrier function due to the stress or improving the skin barrier function under stress.
培養工程の後に、４型コラーゲン発現量比較工程を行う。上記の培養後に、上記被験物質を含む培地で培養した皮膚細胞及びコントロール培地で培養したコントロール皮膚細胞における４型コラーゲン発現量を比較する。上記被験物質を含む培地で培養した皮膚細胞における４型コラーゲン発現量が、上記コントロール皮膚細胞における４型コラーゲン発現量より多い場合に、上記被験物質を皮膚バリア機能の低下抑制又は改善作用を有する物質として選択する。 After the culturing step, a type 4 collagen expression level comparison step is performed. After the above culture, the expression level of type 4 collagen in the skin cells cultured in the medium containing the test substance and the control skin cells cultured in the control medium is compared. When the expression level of type 4 collagen in the skin cells cultured in the medium containing the test substance is higher than the expression level of type 4 collagen in the control skin cells, the test substance is a substance having an effect of suppressing or improving the decrease of the skin barrier function. Select as.
４型コラーゲン発現量は、４型コラーゲンをコードするｍＲＮＡ量又は４型コラーゲンのタンパク質量を測定することにより行うことができる。ｍＲＮＡ又はタンパク質量の測定は、公知の方法で行うことができる。
４型コラーゲンをコードするｍＲＮＡ量は、定量ＰＣＲ、ノーザンブロッティング、マイクロアレイ、蛍光プローブを用いたライブイメージング等により測定することが可能である。４型コラーゲンのタンパク質量は、免疫化学アッセイ、質量分析等により測定することが可能である。免疫化学アッセイとして、Ｅｎｚｙｍｅ　Ｌｉｎｋｅｄ　Ｉｍｍｕｎｏｓｏｒｂｅｎｔ　Ａｓｓａｙ（ＥＬＩＳＡ）、ウエスタンブロッティング等が挙げられる。タンパク質量の測定には、市販のＥＬＩＳＡキット（ＭｙＢｉｏＳｏｕｒｃｅ社製）等を使用することもできる。ヒトの４型コラーゲンの塩基配列やアミノ酸配列（ＯＲＦ）は公知であり、塩基配列（ｍＲＮＡ）はＡｃｃｅｓｓｉｏｎ　Ｎｏ．ＡＨ００２７４１として、公共のデータベースであるＧｅｎｂａｎｋに登録されている。当業者であれば、４型コラーゲンをコードするｍＲＮＡ量を測定するためのプライマーの設計や合成、ＤＮＡの増幅等を容易に行うことができる。 The expression level of type 4 collagen can be determined by measuring the amount of mRNA encoding type 4 collagen or the amount of protein of type 4 collagen. The amount of mRNA or protein can be measured by a known method.
The amount of mRNA encoding type 4 collagen can be measured by quantitative PCR, Northern blotting, microarray, live imaging using a fluorescent probe, or the like. The amount of protein of type 4 collagen can be measured by immunochemical assay, mass spectrometry and the like. Immunochemical assays include Enzyme Linked Immunosorbent Assay (ELISA), Western blotting and the like. A commercially available ELISA kit (manufactured by MyBioSource) or the like can also be used for measuring the amount of protein. The base sequence and amino acid sequence (ORF) of human type 4 collagen are known, and the base sequence (mRNA) is Accession No. It is registered as AH002741 in Genbank, which is a public database. Those skilled in the art can easily design and synthesize primers for measuring the amount of mRNA encoding type 4 collagen, amplify DNA, and the like.
上記のように、４型コラーゲンの発現の低下を抑制又は当該発現を改善することによって、皮膚バリア機能の低下抑制又は改善が可能となる。このため、コントロール皮膚細胞における４型コラーゲン発現量よりも、被験皮膚細胞における４型コラーゲンの発現量が多い場合には、当該被験物質を皮膚バリア機能の低下抑制又は改善作用を有する物質であると判定することができる。上記スクリーニング方法によれば、皮膚バリア機能の低下抑制又は改善作用を有する物質のスクリーニングを行うことができる。 As described above, by suppressing the decrease in the expression of type 4 collagen or improving the expression, it is possible to suppress or improve the decrease in the skin barrier function. Therefore, when the expression level of type 4 collagen in the test skin cells is higher than the expression level of type 4 collagen in the control skin cells, the test substance is considered to be a substance having an effect of suppressing or improving the decrease of the skin barrier function. It can be determined. According to the above screening method, it is possible to screen for a substance having an effect of suppressing or improving the decrease of the skin barrier function.
以下、本発明を実施例によりさらに詳しく説明するが、これにより本発明の範囲を限定するものではない。 Hereinafter, the present invention will be described in more detail with reference to Examples, but this does not limit the scope of the present invention.
＜調製例１＞
植物由来の抽出物を調製した。抽出に用いた植物及びその部位は以下の通りである。
（１）バラ（Ｒｏｓａ　ｈｙｂｒｉｄａ、品種「サントリーブルーローズアプローズ」（登録商標）、サントリーフラワーズ（株））：花弁
（２）ブドウ（Ｖｉｔｉｓ　ｓｐｐ．）（カベルネ・ソーヴィニヨン）：種子及び果皮
（３）チャノキ（Ｃａｍｅｌｌｉａ　ｓｉｎｅｎｓｉｓ）：葉
（４）ハマメリス（Ｈａｍａｍｅｌｉｓ　ｖｉｒｇｉｎｉａｎａ）：葉
（５）セイヨウナツユキソウ（Ｆｉｌｉｐｅｎｄｕｌａ　ｕｌｍａｒｉａ）：花
（６）セイヨウオオバコ（Ｐｌａｎｔａｇｏ　ｍａｊｏｒ）：種子
（７）セイヨウハッカ（Ｍｅｎｔｈａ　ｘ　ｐｉｐｅｒｉｔａ）：全草
上記のバラ（品種「サントリーブルーローズアプローズ」）は、花弁にデルフィニジン型アントシアニン、ロザシアニンＡ１、ロザシアニンＡ２及びロザシアニンＢ並びにロザデルフィンＡ１、ロザデルフィンＡ２及びロザデルフィンＢを含む青系のバラである。 <Preparation Example 1>
Plant-derived extracts were prepared. The plants used for extraction and their parts are as follows.
(1) Rose (Rosa hybrida, variety "Suntory Blue Rose Applause" (registered trademark), Suntory Flowers Limited): Petals (2) Grape (Vitis spp.) (Caberne sauvignon): Seeds and peels (3) Chanoki (Camellia sinensis): Leaves (4) Hamamelis virginiana: Leaves (5) Filipendula ulmaria: Flowers (6) Blue roses (Plantago maga): Seeds Whole plant The above rose (variety "Suntory Blue Rose Applause") is a bluish rose containing Delphinidin type anthocyanin, Rosacyanin A1, Rosacyanin A2 and Rosacyanin B and Rosa Delphin A1, Rosa Delphin A2 and Rosa Delphin B in the petals. ..
上記（１）～（７）の植物の破砕物を、溶媒に浸し抽出した。より詳細には、上記（１）、（２）、（３）、（６）、（７）については、植物の破砕物を重量の１０倍量の３０～９０ｖｏｌ％エタノール水溶液に浸し、室温下、１日１回攪拌操作を加えて１０分～７日間抽出して、抽出液を得た。上記（４）、（５）については、エタノール水溶液の代わりに１，３－ブチレングリコール水溶液を使用し、上記方法で抽出した。 The crushed products of the plants (1) to (7) above were immersed in a solvent and extracted. More specifically, with respect to the above (1), (2), (3), (6) and (7), the crushed plant is immersed in a 30-90 vol% ethanol aqueous solution in an amount 10 times the weight, and at room temperature. The mixture was stirred once a day and extracted for 10 minutes to 7 days to obtain an extract. Regarding (4) and (5) above, an aqueous solution of 1,3-butylene glycol was used instead of the aqueous ethanol solution, and the mixture was extracted by the above method.
得られた抽出液から、ろ過により植物残渣を除去し、ろ液を植物抽出液として得た。
得られた植物抽出液をエバポレーターで濃縮後、凍結乾燥することにより各植物の抽出物を粉末として得た。当該粉末状の植物抽出物（バラ花弁抽出物、ブドウ種子及び果皮抽出物、チャノキ葉抽出物、ハマメリス葉抽出物、セイヨウナツユキソウ花抽出物、セイヨウオオバコ種子抽出物、セイヨウハッカ抽出物）を、抽出物サンプルとして後記の評価に用いた。 The plant residue was removed from the obtained extract by filtration, and the filtrate was obtained as a plant extract.
The obtained plant extract was concentrated with an evaporator and then freeze-dried to obtain an extract of each plant as a powder. The powdered plant extract (rose petal extract, grape seed and peel extract, chanoki leaf extract, hamamelis leaf extract, sardine flower extract, sardine seed extract, sardine sardine extract) It was used as an extract sample for the evaluation described later.
＜調製例２＞
ヤロウィア・リポリティカ（Ｙａｒｒｏｗｉａ　ｌｉｐｏｌｙｔｉｃａ）抽出物は、ヤロウィア・リポリティカを培養し、自己消化させた後、加熱殺菌除菌操作を行うことで、粉末状のヤロウィア・リポリティカ抽出物を得た。
後記の実施例では、ヤロウィア・リポリティカ抽出物には、上記粉末を抽出物サンプルとして用いた。 <Preparation example 2>
The Yarrowia lipolytica extract was obtained by culturing Yarrowia lipolytica, autolyzing it, and then performing a heat sterilization sterilization operation to obtain a powdery Yarrowia lipolytica extract.
In the examples described below, the above powder was used as an extract sample for the Yarrowia lipolytica extract.
＜実施例１＞炎症刺激時における、４型コラーゲンの過剰発現が皮膚バリアに及ぼす影響
ＣＭＶプロモータの下流にＣＯＬ４Ａ１（ＲｅｆＳｅｑ　ＩＤ：ＮＭ＿００１８４５．６）を組み込んだプラスミド（以下、ＣＯＬ４Ａ１プラスミド）を作製した。推奨プロトコールに従い、ＣＯＬ４Ａ１プラスミド、Ｌｉｐｏｆｅｃｔａｍｉｎｅ　３０００　Ｒｅａｇｅｎｔ（Ｔｈｅｒｍｏ　Ｆｉｓｈｅｒ　Ｓｃｉｅｎｔｉｆｉｃ社製）を、Ｏｐｔｉ－ＭＥＭ（登録商標）　Ｉ　Ｒｅｄｕｃｅｄ　Ｓｅｒｕｍ　Ｍｅｄｉａ（Ｔｈｅｒｍｏ　Ｆｉｓｈｅｒ　Ｓｃｉｅｎｔｉｆｉｃ社製）（以下、Ｏｐｔｉ－ＭＥＭ）を用いて希釈し、プラスミド濃度が異なる２種の溶液（以下、Ｏｐｔｉ－ＭＥＭ溶液）を作製した（プラスミド濃度：５μｇ／ｍＬ又は２０μｇ／ｍＬ）。なお対照として、プラスミドの代わりにＯｐｔｉ－ＭＥＭを添加した溶液も作製した。 <Example 1> Effect of overexpression of type 4 collagen on skin barrier during inflammation stimulation A plasmid (hereinafter referred to as COL4A1 plasmid) in which COL4A1 (RefSeq ID: NM_001845.6) was incorporated downstream of the CMV promoter was prepared. According to the recommended protocol, COL4A1 plasmid, Lipofectamine 3000 Reagent (manufactured by Thermo Fisher Scientific), Opti-MEM (registered trademark) I Reduced Serum Media (manufactured by Thermo Fisher), using Diluted Serum Media (Thermo Fisher) Two kinds of solutions having different plasmid concentrations (hereinafter referred to as Opti-MEM solutions) were prepared (plasmid concentration: 5 μg / mL or 20 μg / mL). As a control, a solution in which Opti-MEM was added instead of the plasmid was also prepared.
ＨｕＭｅｄｉａ－ＫＧ２（以下、推奨培地）（クラボウ社製）を用いて、正常ヒト成人表皮角化細胞（クラボウ社製）を２４ｗｅｌｌ用Ｃｅｌｌ　Ｃｕｌｔｕｒｅ　Ｉｎｓｅｒｔｓ（Ｍｉｌｌｉｐｏｒｅ社製）に３×１０４細胞／２００μＬ／ウェルの濃度で播種した後、速やかに１０μＬの上記いずれかのＯｐｔｉ－ＭＥＭ溶液を添加した。なお、２４穴プレート（ＡＧＣテクノグラス（株）製）内には９００μＬの推奨培地を添加した。続いて、３０分後に、一部の群に対してのみ、終濃度が１００μＭとなるよう過酸化水素水（ナカライテスク（株）製）を添加した（Ｈ２Ｏ２処理）。播種４時間後、推奨培地に交換した。播種２４時間後にＣａＣｌ２添加により培地中Ｃａ２＋濃度を１．４１ｍＭまで高めた推奨培地（以下、変則推奨培地）に交換した。２～３日おきに変則推奨培地を用いて培地交換を行った。 Using HuMedia-KG2 (hereinafter referred to as recommended medium) (manufactured by Keratinocyte), normal human adult epidermal keratinized cells (manufactured by Keratinocyte) were added to Cell Culture Inserts (manufactured by Millipore) for 24 wells in 3 × 10 4 cells / 200 μL / After seeding at a well concentration, 10 μL of any of the above Opti-MEM solutions was immediately added. In addition, 900 μL of the recommended medium was added to the 24-well plate (manufactured by AGC Technograss Co., Ltd.). Subsequently, after 30 minutes, hydrogen peroxide solution (manufactured by Nacalai Tesque Co., Ltd.) was added to a final concentration of 100 μM only for a part of the groups (H 2 O 2 treatment). After 4 hours of sowing, the medium was replaced with the recommended medium. Twenty-four hours after sowing, the medium was replaced with a recommended medium (hereinafter referred to as an irregular recommended medium) in which the Ca 2+ concentration in the medium was increased to 1.41 mM by adding CaCl 2. Medium exchange was performed using an irregular recommended medium every 2 to 3 days.
播種９６時間後、１２０時間後、１４４時間後にミリセルＥＲＳ－２（Ｍｅｒｃｋ社製）を用いて経上皮電気抵抗値（ＴＥＥＲ（Ω・ｃｍ２））を評価した。また播種１４４時間後にｉｓｏｇｅｎ（（株）ニッポンジーン製）を用いて細胞を溶解した後、ｍＲＮＡを抽出した。その後、Ｈｉｇｈ－Ｃａｐａｃｉｔｙ　ｃＤＮＡ　Ｒｅｖｅｒｓｅ　Ｔｒａｎｓｃｒｉｐｔｉｏｎ　Ｋｉｔ及び、ＲＮａｓｅ　Ｉｎｈｉｂｉｔｏｒを用いてｃＤＮＡを合成、ＴａｑＭａｎ　Ｆａｓｔ　Ｕｎｉｖｅｒｓａｌ　ＰＣＲ　Ｍａｓｔｅｒ　Ｍｉｘ（いずれもＴｈｅｒｍｏ　Ｆｉｓｈｅｒ　Ｓｃｉｅｎｔｉｆｉｃ社製）、プライマーを用いて定量ＰＣＲを行い、ヒト４型コラーゲンＡ１（ＣＯＬ４Ａ１）の発現量を評価した。なお、内因性のコントロールにはＥｕｋａｒｙｏｔｉｃ　１８Ｓ　ｒＲＮＡを用いた。 Transepithelial electrical resistance (TEER (Ω · cm 2 )) was evaluated using Millicel ERS-2 (manufactured by Merck) 96 hours, 120 hours, and 144 hours after sowing. In addition, 144 hours after seeding, cells were lysed using isogen (manufactured by Nippon Gene Co., Ltd.), and then mRNA was extracted. After that, cDNA was synthesized using High-Capacity cDNA Reverse Transition Kit and RNase Inhibitor, and TaqMan Fast Universal PCR Master Mix (both Thermo Fisher Scientific 4) The expression level of (COL4A1) was evaluated. Eukaryotic 18S rRNA was used for endogenous control.
ＣＯＬ４Ａ１発現量を測定するためのプライマー（ＣＯＬ４Ａ１）、内因性のコントロールを定量するためのプライマー（Ｅｕｋａｒｙｏｔｉｃ　１８Ｓ　ｒＲＮＡ）には、それぞれ＃Ｈｓ００２６６２３７＿ｍ１、＃Ｈｓ９９９９９９０１＿ｓ１（いずれもＴｈｅｒｍｏ　Ｆｉｓｈｅｒ　Ｓｃｉｅｎｔｉｆｉｃ社製）を使用した。 As the primer for measuring the expression level of COL4A1 (COL4A1) and the primer for quantifying the endogenous control (Eukaryotic 18S rRNA), # Hs00266237_m1 and # Hs99999901_s1 (both manufactured by Thermo Fisher Scientific) were used.
結果を表１及び表２に示す。表１に、プラスミド非添加かつＨ２Ｏ２非処理群（Ｈ２Ｏ２（－）、プラスミド（－））のＣＯＬ４Ａ１発現量を１としたときのＣＯＬ４Ａ１の遺伝子発現量を示す。表１及び２において、Ｈ２Ｏ２（＋）は、Ｈ２Ｏ２処理を行った群である。プラスミド（－）はＣＯＬ４Ａ１プラスミド非添加、プラスミド（＋）はＣＯＬ４Ａ１プラスミド添加を示す。＊は、有意差（ｐ＜０．０５）があったことを示す（Ｄｕｎｎｅｔｔ’ｓ　ｔｅｓｔ（ｖｓプラスミド非添加かつＨ２Ｏ２処理群））。Ｈ２Ｏ２刺激に伴い、ＣＯＬ４Ａ１発現量の低下が見られたが、ＣＯＬ４Ａ１プラスミドを導入することで、Ｈ２Ｏ２刺激群に対して、有意かつ用量依存的な発現の低下抑制又は改善が確認された（＊：ｐ＜０．０５）。 The results are shown in Tables 1 and 2. Table 1, plasmid-added and H 2 O 2 untreated group (H 2 O 2 (-) plasmid (-)) showing a gene expression amount of COL4A1 when was the 1 COL4A1 expression level of. In Tables 1 and 2, H 2 O 2 (+) is a group subjected to H 2 O 2 treatment. The plasmid (-) indicates that the COL4A1 plasmid was not added, and the plasmid (+) indicates that the COL4A1 plasmid was added. * Indicates that there was a significant difference (p <0.05) (Dunnett's test (vs plasmid-added and H 2 O 2 treatment group)). As the H 2 O 2 stimulation was observed lowering of COL4A1 expression level, by introducing the COL4A1 plasmid, relative to H 2 O 2 stimulation group, decrease suppression or amelioration of the significant and dose-dependent expression Confirmed (*: p <0.05).
Figure JPOXMLDOC01-appb-T000004
Figure JPOXMLDOC01-appb-T000004
表２に、播種１４４時間後のＴＥＥＲの値を示す。＊は、有意差（ｐ＜０．０５）があったことを示す（Ｄｕｎｎｅｔｔ’ｓ　ｔｅｓｔ（ｖｓプラスミド非添加かつＨ２Ｏ２処理群））。Ｈ２Ｏ２刺激に伴い、ＴＥＥＲの有意な低下が見られたが、ＣＯＬ４Ａ１プラスミドを導入することで、Ｈ２Ｏ２刺激群に対して、用量依存的かつ高用量群においては有意なＴＥＥＲ改善作用が確認された（＊：ｐ＜０．０５）。 Table 2 shows the value of TEER after 144 hours of sowing. * Indicates that there was a significant difference (p <0.05) (Dunnett's test (vs plasmid-added and H 2 O 2 treatment group)). As the H 2 O 2 stimulation, but significant decrease in TEER was seen, the introduction of COL4A1 plasmid, relative to H 2 O 2 stimulation group, dose-dependent and significant TEER improvement in the high dose group The action was confirmed (*: p <0.05).
Figure JPOXMLDOC01-appb-T000005
Figure JPOXMLDOC01-appb-T000005
＜実施例２～９＞抽出物サンプルの４型コラーゲン遺伝子発現の低下抑制又は改善作用の評価
推奨培地を用いて、正常ヒト成人表皮角化細胞（クラボウ社製）を１２穴プレート（ＡＧＣテクノグラス社製）に５×１０４細胞／ｍＬ／ウェルの濃度で播種し、４８時間培養した。抽出物サンプルを含む推奨培地に交換してサンプル処理を開始した後、サンプル処理開始から３０分後に、終濃度が１００μＭとなるよう過酸化水素水（ナカライテスク（株）製）を添加した。過酸化水素水処理開始から３時間後に、抽出物サンプルを含まない推奨培地に交換した。サンプル処理開始時より４８時間後にＲＬＴ　Ｂｕｆｆｅｒ（１％　２－メルカプトエタノール含有）を用いて細胞を溶解及び回収した後、ＲＮｅａｓｙ　Ｍｉｎｉ　Ｋｉｔ及びＲＮａｓｅ－Ｆｒｅｅ　ＤＮａｓｅ　Ｓｅｔ（いずれもＱｉａｇｅｎ社製）を用いてｍＲＮＡを抽出した。その後、実施例１と同じ方法により、ｃＤＮＡ合成、定量ＰＣＲを行い、ＣＯＬ４Ａ１の発現量を評価した。 <Examples 2 to 9> Evaluation of inhibitory or ameliorating effect of reducing or ameliorating type 4 collagen gene expression in an extract sample Using a recommended medium, normal human adult epidermal keratinized cells (manufactured by Kurabou) were placed on a 12-well plate (AGC technograss). The cells were seeded at a concentration of 5 × 10 4 cells / mL / well and cultured for 48 hours. After exchanging with the recommended medium containing the extract sample and starting the sample treatment, hydrogen peroxide solution (manufactured by Nacalai Tesque Co., Ltd.) was added 30 minutes after the start of the sample treatment so that the final concentration became 100 μM. Three hours after the start of the hydrogen peroxide solution treatment, the medium was replaced with a recommended medium containing no extract sample. Forty-eight hours after the start of sample treatment, cells were lysed and recovered using RLT Buffer (containing 1% 2-mercaptoethanol), and then mRNA was used using RNeasy Mini Kit and RNase-Free DNase Set (both manufactured by Qiagen). Was extracted. Then, cDNA synthesis and quantitative PCR were carried out by the same method as in Example 1 to evaluate the expression level of COL4A1.
コントロールについては、抽出物サンプルの代わりに、ジメチルスルホキシド（ＤＭＳＯ）を終濃度が０．１％となるよう添加した推奨培地で処理した以外は、上記と同じ方法で細胞を培養し、過酸化水素水を添加し、ＣＯＬ４Ａ１の発現量を評価した。 For controls, cells were cultured in the same manner as above and hydrogen peroxide, except that instead of the extract sample, cells were treated with recommended medium supplemented with dimethyl sulfoxide (DMSO) to a final concentration of 0.1%. Water was added and the expression level of COL4A1 was evaluated.
実施例２～９で使用した抽出物サンプルは、調製例１及び２で調製したバラ花弁抽出物、ブドウ種子及び果皮抽出物、チャノキ葉抽出物、ハマメリス葉抽出物、セイヨウナツユキソウ花抽出物、セイヨウオオバコ種子抽出物、ヤロウィア・リポリティカ抽出物又はセイヨウハッカ抽出物である。抽出物サンプルは、超純水又はＤＭＳＯに溶解させて培地に添加した。培地中の抽出物サンプルの濃度は、５～２０μｇ／ｍＬとした。なお、超純水に溶解させた場合は、別途ＤＭＳＯを添加し、いずれの抽出物サンプルを含む推奨培地もＤＭＳＯ終濃度が０．１％となるようにした。 The extract samples used in Examples 2 to 9 were rose petal extract, grape seed and peel extract, Chanoki leaf extract, Hamamelis leaf extract, and Yukisou flower extract prepared in Preparation Examples 1 and 2. It is a sardine seed extract, a Yarowia lipolytica extract or a sardine extract. The extract sample was dissolved in ultrapure water or DMSO and added to the medium. The concentration of the extract sample in the medium was 5 to 20 μg / mL. When dissolved in ultrapure water, DMSO was added separately so that the final concentration of DMSO in the recommended medium containing any of the extract samples was 0.1%.
実施例２～９の結果（Ｎ＝３～４の平均値）を表３に示す。表３に示す結果は、コントロールのＣＯＬ４Ａ１の発現量を１００％とした場合の、抽出物サンプルで処理した細胞におけるＣＯＬ４Ａ１の発現量の相対値（％）である。＊は、コントロールに対して有意差（ｐ＜０．０５）があったことを示す（Ｓｔｕｄｅｎｔ’ｓ　ｔ－ｔｅｓｔ）。なお、抽出物サンプル（被験物質）の濃度が５～２０μｇ／ｍＬの範囲で、ＣＯＬ４Ａ１の発現が最も高かった場合の結果を表３に示した。バラ花弁抽出物、ブドウ種子及び果皮抽出物、チャノキ葉抽出物、ハマメリス葉抽出物、セイヨウナツユキソウ花抽出物、セイヨウオオバコ種子抽出物、ヤロウィア・リポリティカ抽出物、セイヨウハッカ抽出物について、コントロールに対してＣＯＬ４Ａ１の発現の低下を抑制又は当該発現を改善する有意な効果が確認された（＊：ｐ＜０．０５）。 The results of Examples 2 to 9 (the average value of N = 3 to 4) are shown in Table 3. The results shown in Table 3 are relative values (%) of the expression level of COL4A1 in the cells treated with the extract sample when the expression level of COL4A1 of the control is 100%. * Indicates that there was a significant difference (p <0.05) with respect to the control (Student's t-test). Table 3 shows the results when the concentration of the extract sample (test substance) was in the range of 5 to 20 μg / mL and the expression of COL4A1 was the highest. For controls, rose petal extract, grape seed and peel extract, chanoki leaf extract, hamamelis leaf extract, blue-green algae flower extract, blue-green algae seed extract, yarowia lipolytica extract, and blue-green algae extract. It was confirmed that there is a significant effect of suppressing the decrease in the expression of COL4A1 or improving the expression (*: p <0.05).
Figure JPOXMLDOC01-appb-T000006
Figure JPOXMLDOC01-appb-T000006
＜実施例１０～１５＞抽出物サンプルの皮膚バリア機能低下抑制又は改善作用の評価
抽出物サンプルを含む推奨培地を用いて、正常ヒト成人表皮角化細胞をＭｉｌｌｉｃｅｌｌ　Ｃｅｌｌ　Ｃｕｌｔｕｒｅ　Ｉｎｓｅｒｔｓに３×１０４細胞／２００μＬ／ウェルの濃度で播種した。播種より３０分後に、終濃度が１００μＭとなるよう過酸化水素水を添加した。播種から４時間後、抽出物サンプルを含まない推奨培地に交換した。播種より２４時間後に変則推奨培地に交換した。その後、実施例１と同じ方法により、経上皮電気抵抗値（ＴＥＥＲ）を評価した。 <Examples 10 to 15> Evaluation of suppression or ameliorating effect on skin barrier function deterioration of extract sample Using the recommended medium containing the extract sample, normal human adult epidermal keratinized cells were subjected to Millicell Cell Culture Inserts in 3 × 10 4 cells. Seeded at a concentration of / 200 μL / well. Thirty minutes after sowing, hydrogen peroxide solution was added so that the final concentration was 100 μM. After 4 hours from seeding, the medium was replaced with the recommended medium containing no extract sample. Twenty-four hours after sowing, the medium was replaced with an irregular recommended medium. Then, the transepithelial electrical resistance value (TEER) was evaluated by the same method as in Example 1.
コントロールについては、抽出物サンプルを含む推奨培地の代わりに、ＤＭＳＯを終濃度が０．１％となるよう添加した推奨培地を用いた以外は、上記と同じ方法で細胞を培養し、過酸化水素水を添加し、経上皮電気抵抗値を評価した。 For control, cells were cultured in the same manner as above, except that the recommended medium containing the extract sample was replaced with the recommended medium in which DMSO was added to a final concentration of 0.1%, and hydrogen peroxide was used. Water was added and the transepithelial electrical resistance was evaluated.
実施例１０～１５で使用した抽出物サンプルは、調製例１及び２で調製した表４に示す抽出物である。抽出物サンプルは、超純水又はＤＭＳＯに溶解させて培地に添加した。培地中の抽出物サンプルの濃度は、５～２０μｇ／ｍＬとした。なお、超純水に溶解させた場合は、別途ＤＭＳＯを添加し、いずれの抽出物サンプルを含む推奨培地もＤＭＳＯ終濃度が０．１％となるようにした。 The extract samples used in Examples 10 to 15 are the extracts shown in Table 4 prepared in Preparation Examples 1 and 2. The extract sample was dissolved in ultrapure water or DMSO and added to the medium. The concentration of the extract sample in the medium was 5 to 20 μg / mL. When dissolved in ultrapure water, DMSO was added separately so that the final concentration of DMSO in the recommended medium containing any of the extract samples was 0.1%.
結果（Ｎ＝４～６の平均値）を表４に示す。表４に示す結果は、コントロールのＴＥＥＲを１００％とした場合の、抽出物サンプルで処理した細胞におけるＴＥＥＲの相対値（％）である。なお、抽出物サンプル（被験物質）の濃度が５～２０μｇ／ｍＬ、及び測定時間が播種後９６～１４４時間の範囲で、ＴＥＥＲの値が最も高かった場合の結果を表４に示した。６種の抽出物でＴＥＥＲの低下を抑制又は改善する高い効果が確認され、さらに、ブドウ種子及び果皮抽出物、チャノキ葉抽出物、ハマメリス葉抽出物、セイヨウナツユキソウ花抽出物、ヤロウィア・リポリティカ抽出物については、コントロールに対して有意な変化が確認された（＊：ｐ＜０．０５）（Ｓｔｕｄｅｎｔ’ｓ　ｔ－ｔｅｓｔ）。 The results (average value of N = 4 to 6) are shown in Table 4. The results shown in Table 4 are the relative values (%) of the TEER in the cells treated with the extract sample, where the control TEER is 100%. Table 4 shows the results when the concentration of the extract sample (test substance) was 5 to 20 μg / mL and the measurement time was in the range of 96 to 144 hours after sowing, and the TER value was the highest. It was confirmed that 6 kinds of extracts have a high effect of suppressing or improving the decrease of TEER, and further, grape seed and peel extract, Chanoki leaf extract, Hamamelis leaf extract, Seiyounatsuyukisou flower extract, and Yarowia lipolytica extract. For the substance, a significant change was confirmed with respect to the control (*: p <0.05) (Student's t-test).
Figure JPOXMLDOC01-appb-T000007
Figure JPOXMLDOC01-appb-T000007
上記の植物の抽出物及びヤロウィア・リポリティカの抽出物は、皮膚バリア機能の低下を抑制又は当該機能を改善する作用を有することが明らかとなった。
  It has been clarified that the above-mentioned plant extract and Yarrowia lipolytica extract have an effect of suppressing a decrease in skin barrier function or improving the function.

Claims (22)

  1. バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、バラ科シモツケソウ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物を有効成分として含む、４型コラーゲンの発現の低下抑制又は改善を介した皮膚バリア機能の低下抑制又は改善用組成物。 Extracts of Rosaceae plants, grape seeds and / or peels, extracts of Yarrowia yeast, extracts of Camellia plants, extracts of Rosaceae plants, and perilla A composition for suppressing or improving a decrease in skin barrier function through suppression or improvement of expression of type 4 collagen, which comprises at least one extract selected from the group consisting of extracts of Rosaceae plants as an active ingredient. ..
  2. バラ科バラ属植物がバラ、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母がヤロウィア・リポリティカ（Ｙａｒｒｏｗｉａ　ｌｉｐｏｌｙｔｉｃａ）、ツバキ科ツバキ属植物がチャノキ、バラ科シモツケソウ属植物がセイヨウナツユキソウ、シソ科ハッカ属植物がセイヨウハッカである、請求項１に記載の皮膚バリア機能の低下抑制又は改善用組成物。 Rosaceae Rosaceae is rose, Yarrowia yeast is Yarrowia lipolytica, Theaceae Camellia is Chanoki, Rosaceae Filipendula is Lamiaceae, and Labiatae is Lamiaceae. A composition for suppressing or improving a decrease in skin barrier function according to claim 1.
  3. バラ科バラ属植物がデルフィニジン型アントシアニン、ロザシアニン化合物及びロザデルフィン化合物からなる群より選択される１種以上の化合物を含むバラ花弁、ツバキ科ツバキ属植物がチャノキの葉、バラ科シモツケソウ属植物がセイヨウナツユキソウの花、シソ科ハッカ属植物がセイヨウハッカ全草である、請求項１又は２に記載の皮膚バリア機能の低下抑制又は改善用組成物。 Rosaceae Rosaceae plants include rose petals containing one or more compounds selected from the group consisting of delphinidin-type anthocyanin, Rosacyanin compounds and Rosadelfin compounds, Theaceae Camellia plants are tea leaves, and Rosaceae Labiatae plants are Labiatae. The composition for suppressing or improving the deterioration of the skin barrier function according to claim 1 or 2, wherein the flower of Theaceae and the plant of the genus Camellia of the Rosaceae family are whole plants of Camellia.
  4. マンサク科マンサク属植物の抽出物及び／又はオオバコ科オオバコ属植物の抽出物を有効成分として含む皮膚バリア機能の低下抑制又は改善用組成物。 A composition for suppressing or improving a decrease in skin barrier function containing an extract of a witch hazel family plant and / or an extract of a plantain family plantago genus as an active ingredient.
  5. マンサク科マンサク属植物がハマメリス、オオバコ科オオバコ属植物がセイヨウオオバコである請求項４に記載の皮膚バリア機能の低下抑制又は改善用組成物。 The composition for suppressing or improving the deterioration of the skin barrier function according to claim 4, wherein the witch hazel family witch hazel genus plant is Hamamelis and the plantain family plantain genus plant is broadleaf plantain.
  6. マンサク科マンサク属植物がハマメリスの葉、オオバコ科オオバコ属植物がセイヨウオオバコ種子である請求項４又は５に記載の皮膚バリア機能の低下抑制又は改善用組成物。 The composition for suppressing or improving the deterioration of the skin barrier function according to claim 4 or 5, wherein the witch hazel family witch hazel plant is a leaf of Hamamelis, and the plantain family plantago genus plant is a broadleaf plantain seed.
  7. 基底膜機能の低下抑制又は改善を介して皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善する、請求項４～６のいずれか一項に記載の皮膚バリア機能の低下抑制又は改善用組成物。 The composition for suppressing or improving the decrease in skin barrier function according to any one of claims 4 to 6, which suppresses or improves the decrease in skin barrier function through suppression or improvement of basement membrane function. ..
  8. 基底膜機能の低下抑制又は改善が４型コラーゲンの発現の低下抑制又は改善を介したものである、請求項７に記載の皮膚バリア機能の低下抑制又は改善用組成物。 The composition for suppressing or improving the decrease in skin barrier function according to claim 7, wherein the suppression or improvement of the basement membrane function is mediated by the suppression or improvement of the expression of type 4 collagen.
  9. 基底膜機能の低下抑制又は改善剤を有効成分として含む皮膚バリア機能の低下抑制又は改善用組成物。 A composition for suppressing or improving the deterioration of skin barrier function, which comprises an active ingredient for suppressing or improving the deterioration of basement membrane function.
  10. 基底膜機能の低下抑制又は改善剤が４型コラーゲンの発現の低下抑制又は改善を介して基底膜機能の低下を抑制又は基底膜機能を改善するものである、請求項９に記載の皮膚バリア機能の低下抑制又は改善用組成物。 The skin barrier function according to claim 9, wherein the agent for suppressing or improving the decrease in basement membrane function suppresses or improves the decrease in basement membrane function through the suppression or improvement of the expression of type 4 collagen. Composition for suppressing or improving the decrease in collagen.
  11. バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、マンサク科マンサク属植物の抽出物、バラ科シモツケソウ属植物の抽出物、オオバコ科オオバコ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物を有効成分として含む４型コラーゲンの発現の低下抑制又は改善用組成物。 Extracts of Rosaceae plants, grape seeds and / or peels, extracts of Yarrowia yeast, extracts of Camellia plants, extracts of plants of the family Rosaceae, extracts of Rosaceae, Rosaceae Decreased expression of type 4 collagen containing at least one extract selected from the group consisting of extracts of genus plants, extracts of plants of the genus Camellia of the family Rosaceae, and extracts of plants of the genus Camellia of the family Rosaceae as active ingredients Composition for suppression or improvement.
  12. バラ科バラ属植物がバラ、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母がヤロウィア・リポリティカ（Ｙａｒｒｏｗｉａ　ｌｉｐｏｌｙｔｉｃａ）、ツバキ科ツバキ属植物がチャノキ、マンサク科マンサク属植物がハマメリス、バラ科シモツケソウ属植物がセイヨウナツユキソウ、オオバコ科オオバコ属植物がセイヨウオオバコ、シソ科ハッカ属植物がセイヨウハッカである、請求項１１に記載の４型コラーゲンの発現の低下抑制又は改善用組成物。 Rosaceae Rosaceae plants are roses, Yarrowia yeast is Yarrowia lipolytica, Tsubaki family Tsubaki plants are Chanoki, Mansaku family Mansaku plants are Hamamelis, Rosaceae Filipendula plants are Seiyounatsukiso. The composition for suppressing or improving the expression of type 4 collagen according to claim 11, wherein the plant belonging to the genus Rosaceae is a plant belonging to the genus Rosaceae and the plant belonging to the genus Rosaceae is a plant belonging to the genus Rosaceae.
  13. バラ科バラ属植物がデルフィニジン型アントシアニン、ロザシアニン化合物及びロザデルフィン化合物からなる群より選択される１種以上の化合物を含むバラ花弁、ツバキ科ツバキ属植物がチャノキの葉、マンサク科マンサク属植物がハマメリスの葉、バラ科シモツケソウ属植物がセイヨウナツユキソウの花、オオバコ科オオバコ属植物がセイヨウオオバコ種子、シソ科ハッカ属植物がセイヨウハッカ全草である、請求項１１又は１２に記載の４型コラーゲンの発現の低下抑制又は改善用組成物。 Rosaceae Rosaceae plants include rose petals containing one or more compounds selected from the group consisting of delphinidin-type anthocyanins, rosasianin compounds and rosadelphin compounds, Plantains Rosaceae plants are Chanoki leaves, and Plantains Mansaku plants are Hamamelis. The type 4 collagen according to claim 11 or 12, wherein the leaves, Rosaceae plants of the genus Rosaceae are flowers of Yukisou, the plants of the genus Plantains are seeds of Plantains, and the plants of the genus Rosaceae are whole plants of the genus Pseudomonas aeruginosa. A composition for suppressing or improving the decrease in expression.
  14. 皮膚外用剤である請求項１～１３のいずれか一項に記載の組成物。 The composition according to any one of claims 1 to 13, which is an external preparation for skin.
  15. 化粧料である請求項１～１４のいずれか一項に記載の組成物。 The composition according to any one of claims 1 to 14, which is a cosmetic.
  16. ４型コラーゲンの発現の低下抑制又は改善により皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善するための、バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、バラ科シモツケソウ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物の使用。 Extracts of Rosaceae plants, grape seeds and / or peels, Yarrowia, for suppressing or improving the decrease in the expression of type 4 collagen to suppress the decrease in the skin barrier function or improve the skin barrier function. ) Use of one or more extracts selected from the group consisting of extracts of genus yeast, extracts of Camellia plants of the family Rosaceae, extracts of plants of the genus Rosaceae, and extracts of plants of the genus Camellia. ..
  17. 皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善するための、マンサク科マンサク属植物の抽出物及び／又はオオバコ科オオバコ属植物の抽出物の使用。 Use of witch hazel plant extract and / or plantain plantago extract to suppress deterioration of skin barrier function or improve skin barrier function.
  18. ４型コラーゲンの発現の低下を抑制又は前記発現を改善するための、バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、マンサク科マンサク属植物の抽出物、バラ科シモツケソウ属植物の抽出物、オオバコ科オオバコ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物の使用。 Extracts of Rosaceae plants, grape seeds and / or peels, extracts of Yarrowia yeast, extracts of Rosaceae, and camellia for suppressing the decrease in expression of type 4 collagen or improving the expression. Selected from the group consisting of extracts of genus plants, extracts of plants of the genus Mansaku of the family Rosaceae, extracts of plants of the genus Rosaceae, extracts of plants of the genus Oobaco, and extracts of plants of the genus Rosaceae. Use of one or more extracts.
  19. バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、バラ科シモツケソウ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物を投与する、４型コラーゲンの発現の低下抑制又は改善により皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善する方法。 Extracts of Rosaceae plants, grape seeds and / or peels, extracts of Yarrowia yeast, extracts of Camellia plants, extracts of Rosaceae plants, and perilla A method of suppressing or improving the decrease in skin barrier function by suppressing or improving the expression of type 4 collagen by administering one or more extracts selected from the group consisting of extracts of Rosaceae plants. ..
  20. マンサク科マンサク属植物の抽出物及び／又はオオバコ科オオバコ属植物の抽出物を投与する、皮膚バリア機能の低下を抑制又は皮膚バリア機能を改善する方法。 A method of administering an extract of a witch hazel plant and / or an extract of a plantain plant, which suppresses a decrease in the skin barrier function or improves the skin barrier function.
  21. バラ科バラ属植物の抽出物、ブドウ種子及び／又は果皮の抽出物、ヤロウィア（Ｙａｒｒｏｗｉａ）属酵母の抽出物、ツバキ科ツバキ属植物の抽出物、マンサク科マンサク属植物の抽出物、バラ科シモツケソウ属植物の抽出物、オオバコ科オオバコ属植物の抽出物、並びに、シソ科ハッカ属植物の抽出物からなる群より選択される１種以上の抽出物を投与する４型コラーゲンの発現の低下を抑制又は前記発現を改善する方法。 Extracts of Rosaceae plants, grape seeds and / or peels, extracts of Yarrowia yeast, extracts of Camellia plants, extracts of plants of the family Rosaceae, extracts of Rosaceae, Rosaceae Suppresses the decrease in the expression of type 4 collagen by administering one or more extracts selected from the group consisting of extracts of genus plants, extracts of plants of the genus Camellia of the family Rosaceae, and extracts of plants of the genus Camellia of the family Rosaceae. Or a method for improving the expression.
  22. インビトロで、ヒト表皮角化細胞を、被験物質を含む培地又は前記被験物質を含まないコントロール培地で培養する工程、及び、
    前記被験物質を含む培地で培養した皮膚細胞及びコントロール培地で培養したコントロール皮膚細胞における４型コラーゲン発現量を比較する工程を含み、
    前記被験物質を含む培地で培養した皮膚細胞における４型コラーゲン発現量が、前記コントロール皮膚細胞における４型コラーゲン発現量より多い場合に、前記被験物質を皮膚バリア機能の低下抑制又は改善作用を有する物質として選択する、皮膚バリア機能の低下抑制又は改善作用を有する物質のスクリーニング方法。
          In vitro, a step of culturing human epidermal keratinocytes in a medium containing a test substance or a control medium not containing the test substance, and
    Including a step of comparing the expression level of type 4 collagen in the skin cells cultured in the medium containing the test substance and the control skin cells cultured in the control medium.
    When the expression level of type 4 collagen in the skin cells cultured in the medium containing the test substance is higher than the expression level of type 4 collagen in the control skin cells, the test substance is a substance having an effect of suppressing or improving the decrease of the skin barrier function. A method for screening a substance having an effect of suppressing or improving a decrease in skin barrier function, which is selected as.
PCT/JP2021/008390 2020-03-10 2021-03-04 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 WO2021182283A1 (en)

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