Classifications

• • 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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• 2021
  • 2021-03-04 CN CN202180019249.5A patent/CN115243667A/zh active Pending
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