WO2020213743A1 - Méthode et dispositif de prévention et/ou d'atténuation du photovieillissement et/ou de la pigmentation dermique, agent inhibiteur de photovieillissement et/ou de pigmentation dermique, méthode de criblage d'un agent inhibiteur de photovieillissement et/ou de pigmentation dermique, méthode d'évaluation d'un traitement cosmétique inhibiteur de photovieillissement et/ou de pigmentation dermique, et méthode d'évaluation du photovieillissement et/ou de la pigmentation dermique - Google Patents

Méthode et dispositif de prévention et/ou d'atténuation du photovieillissement et/ou de la pigmentation dermique, agent inhibiteur de photovieillissement et/ou de pigmentation dermique, méthode de criblage d'un agent inhibiteur de photovieillissement et/ou de pigmentation dermique, méthode d'évaluation d'un traitement cosmétique inhibiteur de photovieillissement et/ou de pigmentation dermique, et méthode d'évaluation du photovieillissement et/ou de la pigmentation dermique Download PDF

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WO2020213743A1
WO2020213743A1 PCT/JP2020/017082 JP2020017082W WO2020213743A1 WO 2020213743 A1 WO2020213743 A1 WO 2020213743A1 JP 2020017082 W JP2020017082 W JP 2020017082W WO 2020213743 A1 WO2020213743 A1 WO 2020213743A1
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photoaging
skin
balance
pigmentation
dermal pigmentation
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English (en)
Japanese (ja)
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聡 堀場
純一 細井
雅哉 高木
有宇子 松浦
健太朗 加治屋
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株式会社 資生堂
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Priority to CN202410125329.4A priority Critical patent/CN117815216A/zh
Priority to CN202080029442.2A priority patent/CN113784753A/zh
Priority to JP2021514255A priority patent/JP7439059B2/ja
Publication of WO2020213743A1 publication Critical patent/WO2020213743A1/fr
Priority to JP2023212456A priority patent/JP2024037928A/ja

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61H—PHYSICAL THERAPY APPARATUS, e.g. DEVICES FOR LOCATING OR STIMULATING REFLEX POINTS IN THE BODY; ARTIFICIAL RESPIRATION; MASSAGE; BATHING DEVICES FOR SPECIAL THERAPEUTIC OR HYGIENIC PURPOSES OR SPECIFIC PARTS OF THE BODY
    • A61H23/00—Percussion or vibration massage, e.g. using supersonic vibration; Suction-vibration massage; Massage with moving diaphragms
    • A61H23/02—Percussion or vibration massage, e.g. using supersonic vibration; Suction-vibration massage; Massage with moving diaphragms with electric or magnetic drive
    • 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
    • A61K31/16—Amides, e.g. hydroxamic acids
    • 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)
    • A61K36/38—Clusiaceae, Hypericaceae or Guttiferae (Hypericum or Mangosteen family), e.g. common St. Johnswort
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • 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
    • A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
    • A61K8/42—Amides
    • 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
    • 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
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/16—Emollients or protectives, e.g. against radiation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • 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
    • A61Q19/08—Anti-ageing preparations
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/42—Apparatus for the treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C12Q1/06—Quantitative determination
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • 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
    • 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

Definitions

  • the present invention provides methods and devices for preventing or ameliorating photoaging and / or dermal pigmentation by adjusting the M1 / M2 balance, and anti-photoaging and / or dermal pigmentation inhibitors, and M1 / M2 balance.
  • the present invention relates to a method for screening an anti-photoaging and / or dermal pigmentation inhibitor as an index, a method for evaluating anti-photoaging and / or a cosmetic treatment for suppressing dermal pigmentation, and a method for evaluating the degree of photoaging and / or dermal pigmentation.
  • the aging phenomenon of human skin can be broadly divided into “natural aging” and “photoaging”.
  • Photoaging is a phenomenon that is observed specifically in the exposed area and is skin-specific.
  • the production of active oxygen and damage to cell DNA are induced by the influence of UV, etc., and it is thought that the skin fibrous tissue is damaged and phenotypes such as wrinkles and sagging appear.
  • skin photoaging causes a decrease in collagen fibers and degeneration of elastic fibers made of elastin.
  • melanocytes are also damaged, and a large amount of melanin pigment is produced, which causes stains and the like.
  • pigmentation such as stains and dullness is caused by the accumulation of melanin produced by melanocytes in the basal layer of the epidermis.
  • Melanin is usually present in the epidermis and basal layer, but since the epidermis turns over in a relatively fast cycle, such melanin is easily excreted.
  • melanin may be present in the dermis layer because melanin falls into the dermis through the gaps in the basement membrane. Since the turnover cycle of dermal cells is much slower than that of the epidermis, such melanin is often accumulated without being excreted. For this reason, it is very difficult to improve dermal pigmentation.
  • Patent Document 1 discloses a preventive or inhibitory agent for skin photoaging by preventing the inhibition of leukocyte elastase.
  • Patent Document 2 discloses a photoaging inhibitor composition containing a plant extract of the genus Amaranthaceae, which has a collagen synthesis promoting action, and an animal-derived collagen peptide.
  • Non-Patent Document 13 proposes to prevent melanin from falling into the dermis by strengthening the basement membrane.
  • Patent Document 3 discloses an anti-aging cosmetic composition containing a compound that induces autophagy activation as well as increased expression of adiponectin. It has also been suggested that the phagocytic action of macrophages is used to improve the pigmentation of the dermis.
  • Patent Document 15 states that dermal spots are caused by attracting and phagocytosing macrophages to fibroblasts that have taken up melanin that has fallen into the dermis. Disclose preventive / ameliorating agents.
  • Japanese Patent No. 5657723 JP-A-2017-203004 JP-A-2018-177805 Special Table 2014-504629 JP-A-2015-140334 Japanese Patent No. 6178088 Japanese Patent No. 6273304 Japanese Patent No. 5744409 Japanese Unexamined Patent Publication No. 2018-140953 JP-A-2019-043855 Japanese Unexamined Patent Publication No. 2013-053130 Japanese Unexamined Patent Publication No. 09-187248 Japanese Unexamined Patent Publication No. 2003-261455 Japanese Patent Application Laid-Open No. 09-227367 JP-A-2018-072098 Japanese Patent No. 6535146 JP-A-2019-031482 JP-A-2007-063192
  • PROBLEM TO BE SOLVED To prevent and / or improve photoaging and / or dermal pigmentation, and an anti-photoaging and / or dermal pigmentation inhibitor, anti-photoaging and / or dermal pigmentation inhibition.
  • an anti-photoaging and / or dermal pigmentation inhibitor for preventing and / or ameliorating photoaging and / or dermal pigmentation and an evaluation method for anti-photoaging and / or dermal pigmentation-suppressing cosmetic treatment. was established.
  • a novel cosmetological method and device having an anti-photoaging and / or dermal pigmentation inhibitory effect, and an anti-photoaging and / or dermal pigmentation inhibitor have also been developed.
  • a cosmetological method for preventing and / or ameliorating photoaging and / or dermal pigmentation by adjusting the M1 / M2 balance is to increase the ratio of M2 to M1, the method described in (1).
  • Adjusting the M1 / M2 balance includes the step of applying a weak physical stimulus to the skin, where the step is, for example, (a) stretching the target skin by 0.1% or more and 50.0% or less.
  • the extension rate is (In the equation, the fixed points A and B are the epidermis or any position on the matrix to which the epidermis is adhered, where the straight line passing through the fixed points A and B is parallel to the extension direction).
  • a cosmetological device for preventing and / or ameliorating photoaging and / or dermal pigmentation by adjusting the M1 / M2 balance.
  • the device is The stimulus generator that causes physical stimulus and It is equipped with a stimulus-giving part that gives the physical stimulus generated in the stimulus-generating part to the skin.
  • the device is a device for performing a step of applying a weak physical stimulus to the skin, and the step is described here, for example.
  • Procedure for acquiring data on preset skin M1 / M2 balance reference values Procedure for acquiring data on M1 / M2 balance of the target skin; A procedure for calculating by referring to the reference value and comparing with the data on the M1 / M2 balance of the target skin; The method having a procedure for evaluating the degree of photoaging and / or dermal pigmentation inhibition of the skin based on the calculation result by the calculation procedure; and a procedure for displaying the evaluation result by the evaluation procedure.
  • a system for evaluating the degree of photoaging and / or dermal pigmentation A system for evaluating the degree of photoaging and / or dermal pigmentation.
  • Database section that stores data related to preset skin M1 / M2 balance reference values; Data input unit for inputting data related to M1 / M2 balance of the target skin; A calculation unit that refers to the reference value stored in the database unit and compares it with the data on the M1 / M2 balance of the target skin input by the data input unit; The system having an evaluation unit that evaluates the degree of photoaging and / or dermal pigmentation of the skin based on the calculation result by the calculation unit; and a display unit that displays the evaluation result by the evaluation unit. (9) Screening kit for anti-photoaging and / or dermal pigmentation inhibitor, which contains a drug for measuring M1 / M2 balance.
  • M1 / M2 balance adjuster / improver including Hypericum extract, Phellodendron amur extract, Phellodendron amur, and / or Eucalyptus extract.
  • the methods, devices, and agents of the present invention can prevent and / or improve photoaging and / or dermal pigmentation by adjusting the M1 / M2 balance.
  • M1 / M2 balance as an index, it becomes possible to objectively evaluate the degree of photoaging and / or dermal pigmentation, and photoaging and / or dermal pigmentation can be evaluated based on such a method.
  • Drugs and cosmetic treatments for prevention and / or improvement can be searched.
  • FIG. 1a shows the types of macrophage markers used in Experiment 1 as well as the subject's age, skin type, and gender.
  • FIG. 1b is a photomicrograph showing macrophages (CD11b: red) and M1 macrophages (CD86: green) in the skin tissue of young and elderly people.
  • FIG. 1c is a photomicrograph showing macrophages (CD11b: red) and M2 macrophages (CD206: green) in the skin tissue of young and elderly people.
  • Figure 1d is a graph showing the number of M1 and M2 macrophages and the total number of total macrophages in the skin tissues of young and elderly people (cells / mm 2 ).
  • Figure 1e Left is a photomicrograph showing co-staining of M1 macrophages (CD86: red) or M2 macrophages (CD206: red) and procollagen (green) in the skin tissue of young and elderly people.
  • the rectangles in each of the four panels in the figure on the left are an enlargement of the characteristic parts.
  • Figure 1e The figure on the right is a schematic diagram of the relationship between M1, M2, fibroblasts, and collagen production / destruction.
  • Figure 2a outlines the method of Experiment 2.
  • FIG. 2b is a photomicrograph of M0, M1 and M2 macrophages induced by Experiment 2. The upper figure of FIG.
  • FIG. 2c is a graph showing the gene expression levels of cytokines (IL-1beta, TNF-alpha, IL-10) produced by M0, M1 and M2 macrophages induced by Experiment 2.
  • the lower figure of FIG. 2c is a graph showing the expression level of mRNA of cell surface markers (M1: CD86, M2: CD206) of M1 and M2 macrophages. It is corrected by the mRNA expression level of GAPDH, and the case of M0 is shown as a relative value (%) as 100%.
  • Figure 3a outlines the method of Experiment 3.
  • FIG. 3a outlines the method of Experiment 3.
  • FIG. 3b shows the amount of procollagen ( ⁇ g / well) in fibroblasts to which the supernatants of M1 and M2 macrophages were added according to Experiment 3.
  • FIG. 3c is a photomicrograph showing collagen (red) and hyaluronic acid (green) in fibroblasts supplemented with supernatants of M1 and M2 macrophages according to Experiment 3.
  • FIG. 3d is a photomicrograph showing ⁇ -galactosidase ( ⁇ -Gal) in fibroblasts supplemented with supernatants of M1 and M2 macrophages according to Experiment 3.
  • Figure 3e shows the results of the aging-related ⁇ -galactosidase (SA ⁇ -Gal) assay in fibroblasts supplemented with the supernatants of M1 and M2 macrophages according to Experiment 3 (left figure: SA ⁇ -gal relative to the total number of DAPI-positive cells). It is a graph which shows the ratio (%) of positive cells, and the total number of DAPI positive cells per well (right figure).
  • Figure 3f shows the melanogenesis-enhancing factors (HGF, ET1, bFGF, IL-1alpha, SCF) and melanogenesis-suppressing factors (clusterin, DKK1) in the fibroblasts to which the supernatants of M1 and M2 macrophages were added according to Experiment 3. It is a graph which shows the mRNA expression level of.
  • FIG. 4a is a photomicrograph showing ⁇ -Gal in young and aged fibroblasts supplemented with supernatants of M1 and M2 macrophages according to Experiment 4.
  • Figure 4b shows the results of the SA ⁇ -Gal assay (SA ⁇ -gal relative to the total number of DAPI-positive cells) in young (top) and aged fibroblasts (bottom) to which the supernatants of M1 and M2 macrophages were added according to Experiment 4. It is a graph which shows the ratio (%) of positive cells) and the total number of DAPI positive cells per well.
  • FIG. 4c shows the amount of procollagen ( ⁇ g / well) in young and aged fibroblasts to which the supernatants of M1 and M2 macrophages were added according to Experiment 4.
  • FIG. 4d is a photomicrograph showing collagen (red) and DAPI-stained nuclei (blue) in young and aged fibroblasts supplemented with supernatants of M1 and M2 macrophages according to Experiment 3.
  • FIG. 5 is a photomicrograph showing type I collagen (red) and macrophages (CD68: green) in neonatal foreskin-derived fibroblasts to which supernatants of M1 and M2 macrophages were added according to Experiment 5. The upper left photo shows fibroblasts cultured without the addition of macrophage supernatant.
  • FIG. 5 is a photomicrograph showing collagen (red) and DAPI-stained nuclei (blue) in young and aged fibroblasts supplemented with supernatants of M1 and M2 macrophages according to Experiment 3.
  • FIG. 5 is a photomicrograph showing type I collagen (red) and macrophages (CD68: green) in neonatal foreskin-derived fibro
  • FIG. 6 is a photomicrograph showing macrophages (CD68: red) and M1 macrophages (CD86: green) or M2 macrophages (CD206: green) in a 3D model prepared according to Experiment 6.
  • the part indicated by the triangle indicates the part where CD68 and CD86 or CD68 and CD206 are double-stained.
  • Figure 7 is a photomicrograph showing p21 (red) and DAPI-stained nuclei (blue) in a 3D model created according to Experiment 6.
  • FIG. 9 is a graph showing the total number of macrophages, M1 and M2 macrophages in an ex vivo skin model irradiated with a solar simulator according to Experiment 7.
  • the number of cells of each macrophage without irradiation is set to 100% (left bar: irradiation (-)), and the relative value (%) of the number of cells of each macrophage irradiated to it is the right bar (irradiation (irradiation (-)). +)).
  • Figure 10 shows the setting of the extension stimulus performed in Experiment 8.
  • Figure 11 shows the equipment and skin sample used in Experiment 8.
  • FIG. 12 shows a comparison of the total number of M1 and M2 macrophages and the total number of macrophages when the ex vivo skin model was stretched and not stimulated (control) according to Experiment 8.
  • the figure on the left is a graph showing the number of macrophages (cells / 1 mm 2 ) per 1 mm 2 of skin sample.
  • the figure on the right is a graph showing the ratio (%) of each macrophage (M1, M2) to the total number of macrophages.
  • Figure 13 shows the results of the screening performed in Experiment 9.
  • the bar controls the CD86 expression level (CD86 / GAPDH value) when each concentration of Hypericum erectum extract (0.1%), thyme extract (0.1%), and tranexamic acid methylamide (0.03%, 0.06%) was added. It is a graph which shows a relative value (%) when cont) is 100%.
  • FIG. 14 shows an example of the apparatus of the present invention.
  • Figure 15 shows the results of the screening performed in Experiment 10.
  • the bar shows the relative value (CD86 / GAPDH value) of the CD86 expression level (CD86 / GAPDH value) when the Phellodendron amur extract (0.01%) and the eucalyptus extract (0.1%) of each concentration were added, when the control (cont) was 100%.
  • FIG. 16a shows the mRNA expression levels of each collagen-degrading enzyme (MMP-1, MMP-2) and inflammatory cytokine (IL-1 ⁇ ) in fibroblasts supplemented with the supernatants of M1 and M2 macrophages according to Experiment 11. It is a graph which shows a relative value (%) when the result in is taken as 100%.
  • Figure 16b shows the mRNA expression levels of each collagen production / maturation factor (COL1A1, COL1A2, HSP47, and ADAMTS-2) in fibroblasts supplemented with the supernatants of M1 and M2 macrophages according to Experiment 11 as a result of M1.
  • Figure 17a shows the age of each subject who took the sample in Experiment 12 for each young and elderly group.
  • Figure 17b is a graph showing the number of M1 and M2 macrophages in the skin tissues of young and elderly people (50 ⁇ m from just below the basement membrane) in Experiment 12 (cells / mm 2 ).
  • Figure 17c is a graph showing the number of M1 and M2 macrophages in the skin tissue of young and elderly people (200 ⁇ m from just below the basement membrane) in Experiment 12 (cells / mm 2 ).
  • Figure 17d is a graph showing the number of CD68-negative cells, M1 and M2 macrophages showing 3/4 collagen positive in the skin tissues of young and elderly people in Experiment 12 (cells / mm 2 ).
  • FIG. 18a shows the appearance of fibroblasts, M1 macrophages, and M2 macrophages 24 hours after the addition of melanin in Experiment 13. Cells that phagocytose melanin take a circular shape as shown in the figure on the right.
  • FIG. 18b shows the amount of melanin (melanin / almar blue) per cell taken up by fibroblasts, M1 macrophages, and M2 macrophages 24 hours after the addition of melanin in Experiment 13.
  • FIG. 18a shows the appearance of fibroblasts, M1 macrophages, and M2 macrophages 24 hours after the addition of melanin in Experiment 13.
  • FIG. 18b shows the amount of melanin (melanin / almar blue) per cell taken up
  • FIG. 19 shows the state of M1 macrophages and M2 macrophages 5 days after the addition of melanin in Experiment 13.
  • the upper part of FIG. 19 is a graph showing the number of melanin-phagocytic M1 macrophages and M2 macrophages in the dermis layer counted according to Experiment 14 for each age group.
  • the lower part of Fig. 19 shows the numbers of melanin-phagocytic M1 macrophages and M2 macrophages as an average in all subjects.
  • Macrophages are cells that are localized in various tissues in the body and elicit an immune response against foreign substances and pathogens, and are known to be involved in inflammation. Macrophages differentiate from undifferentiated M0 macrophages (hereinafter sometimes abbreviated as M0) into M1 type and M2 type. M1 macrophages (hereinafter sometimes abbreviated as M1) are known as inflammatory type, and M2 macrophages (hereinafter sometimes abbreviated as M2) are known as repair type (anti-inflammatory type). It has been reported that the imbalance between M1 macrophages and M2 macrophages is associated with diseases such as obesity, type 2 diabetes, and arteriosclerosis (Patent Documents 4 to 6, Non-Patent Documents 1 to 5). However, the relationship between skin photoaging and pigmentation and M1 / M2 balance was unclear.
  • the present inventors have discovered that the balance between these M1 macrophages and M2 macrophages (M1 / M2 balance) is specifically disturbed in the exposed skin and the site where pigmentation occurs, and adjust the M1 / M2 balance. However, it was found to be particularly important in the prevention and improvement of photoaging and pigmentation. Furthermore, the present inventors have also found that the M1 / M2 balance is used as an index of skin photoaging and pigmentation, and a favorable effect is obtained when a specific physical stimulus is applied to the target skin.
  • the present inventors used the M1 / M2 balance as an index of skin photoaging and pigmentation, and as a result of screening various substances, the compounds of the present invention such as otogirisou extract and tranexamic acid methylamide, thyme extract, We also found that tranexamic acid and eucalyptus extracts have the effect of adjusting / improving the M1 / M2 balance and function as anti-photoaging agents and anti-pigmenting agents. It has been reported that the Hypericum erectum extract has a type I collagen production promoting action, a hair growing action, and the like (Patent Documents 9 and 10).
  • the compounds of the present invention including tranexamic acid methylamide, have been reported to have a rough skin improving effect, a whitening effect, an antiallergic effect, and the like (Patent Documents 11, 14, etc.). Anti-allergic action, antibacterial action, etc. have been reported for thyme extract (Patent Documents 12 and 13). Phellodendron amur is also used as an antidiarrheal, a stomach medicine, an internal medicine for weak stomach and loss of appetite, or an external medicine for bruises and sprains, and has been reported to have antibacterial, antioxidant, skin barrier function improving effects, pigment cell activating effects, etc. ( Patent Documents 16, 17). Eucalyptus extract has been reported to have antibacterial and antioxidant effects (Patent Document 18). However, it has been discovered for the first time by the present inventors that these substances have an effect of adjusting / improving the M1 / M2 balance.
  • the present invention provides methods, devices, and anti-photoaging and / or pigmentation inhibitors for preventing and / or ameliorating photoaging and / or pigmentation by adjusting the M1 / M2 balance.
  • the method of the present invention is a method for the purpose of beauty and may not be treated by a doctor or a medical professional.
  • pigmentation refers to the deposition of pigments such as melanin in the dermis and epidermis.
  • the present invention is effective for both the dermis and the epidermis, it is highly expected as a countermeasure against pigmentation of the dermis in the sense that improvement methods other than phagocytosis by melanophages are limited.
  • stains, dullness, dark circles, etc. due to pigmentation are improved.
  • M1 macrophages can be measured with markers such as CD86, CD80, and iNOS.
  • M2 macrophages can be measured with markers such as CD206, CD163, Agr1, etc. Examples of markers for all macrophages including M1 and M2 include CD11b and CD68. Additional or alternative, it may be measured by quantifying M1-specific cytokines such as IL-1beta, TNF-alpha and M2-specific cytokines such as IL-10. However, any marker that can measure the M1 / M2 balance is not limited to the above markers.
  • the M1 / M2 balance may refer to the ratio of the number of M1 macrophages to the number of M2 macrophages, and may refer to the amount of mRNA of a marker of M1 macrophages (for example, CD86, CD80, iNOS, etc.). It may refer to the ratio of the amount of mRNA of M2 macrophage markers (for example, CD206, CD163, Agr1, etc.).
  • the ratio of M1 is high and the ratio of M2 is low, and it is M2 that has a high melanin phagocytosis effect. Therefore, the adjustment / improvement of the M1 / M2 balance is the ratio of M2 to M1 (M2).
  • the number / number of M1 and / or the amount of mRNA of the M2 marker / the amount of mRNA of the M1 marker may be increased.
  • the increase may be, for example, an increase with a statistically significant difference (eg, Student's t-test) with a significance level of 5%, and / or, for example, 1% or more, 5% or more, 10% or more,
  • the increase may be 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more.
  • adjusting / improving the M1 / M2 balance can be performed by keeping the ratio of M2 to M1 (the number of M2 / the number of M1 and / or the amount of mRNA of the M2 marker / the amount of mRNA of the M1 marker) within a certain range, for example, about. It may be 4/6 to about 9/1, about 5/5 to about 8/2, about 5/5 to about 7/3, etc., or it may be close to the above range, and the above It may be maintained within the range, or homeostasis may be maintained around the above range.
  • Adjustment / improvement of M1 / M2 balance may be achieved by applying physical stimuli such as extension stimulus, pressing stimulus, and massage to the skin.
  • the adjustment / improvement of the M1 / M2 balance includes a step of applying a weak physical stimulus to the skin, and the device of the present invention is a device for performing a step of applying a weak physical stimulus to the skin.
  • the cycle including (a) stretching the skin to a stretch rate of 0.1% or more and 50.0% or less; and (b) recovering from the stretched state; is performed at a frequency of 60 Hz or less. There may be.
  • the extension rate is calculated by the above formula 1.
  • Physical stimulation may be performed at an extension rate of 0.001% or more and 80.0 or less, 0.01% or more and 60.0% or less, 0.1% or more and 50.0% or less, preferably 0.1% or more and 50.0% or less.
  • Extension speed refers to the speed (% / s) until the maximum extension rate (%) is reached in one cycle.
  • the recovery rate refers to the rate (% / s) of returning from the maximum extension rate to the non-extension state.
  • Extension speed / recovery speed is 0.010% / s-40% / s, 0.05% / s-30% / s, 0.10% / s-20% / s, 0.2% / s-15% / s, 0.3% / It may be carried out at any speed such as s to 10% / s.
  • the extension rate and recovery rate may be the same or different.
  • the adjustment / improvement of the M1 / M2 balance includes a step of applying a weak physical stimulus to the skin
  • the device of the present invention is a device for performing a step of applying a weak physical stimulus to the skin.
  • the step includes, for example, (a-1) pressing the subject skin by 1 ⁇ m to 1000 ⁇ m; and (b-1) recovering the subject skin from the pressed state; 60 Hz or less. It may be performed at the frequency of.
  • Pressing the skin from 1 ⁇ m to 1000 ⁇ m means pressing the skin to a depth of 1 ⁇ m to 1000 ⁇ m from the outermost surface of the skin.
  • the depth of pressing can be arbitrarily set from 1 ⁇ m to 1000 ⁇ m, 10 ⁇ m to 1000 ⁇ m, 10 ⁇ m to 300 ⁇ m, 10 ⁇ m to 100 ⁇ m, etc. from the outermost surface of the skin.
  • the frequency refers to the number of cycles per second when one cycle is the cycle of recovery from the start of extension or pressing to the non-extension or non-pressing state.
  • One cycle may include maintaining a stretched or pressed state for a period of time and / or resting in a non-extended or non-pressed state. For example, during one cycle, after (a') (a) and before (b), or after (a-1) and before (b-1), 0 seconds to 30 minutes, 1 second.
  • the frequency may be, for example, 0.0000001 Hz or more and 10 kHz or less, 0.000001 Hz or more and 1 kHz or less, 0.00001 Hz or more and 100 Hz or less, preferably 0.0001 Hz or more and 60 Hz or less, 0.0001 Hz or more and 10 Hz or less.
  • facial equipment, massage equipment, etc. include those that use electromagnetic waves with a frequency of about 0.3 to 300 MHz, such as RF waves, and those that use ultrasonic waves with a frequency of about 1 MHz to 7 MHz. Compared with these frequencies / frequencies, the frequencies adopted by the method / apparatus of the present invention are extremely low. If a strong frequency is applied to the skin like a conventional facial device, there is a risk of adverse effects such as redness, pressure marks, scratches, pain, and inflammation on the skin, but if a frequency like the present invention is adopted, the above risks Is low and allows non-invasive physical stimulation. This is because the present inventors have discovered that if the extension rate and the frequency are too high, the stimulation is too strong, and it is preferable to gently stimulate the skin by adjusting these values to appropriate values.
  • low- and medium-frequency devices such as EMS devices are commercially available, they are designed to act especially in deep layers such as muscle and subcutaneous fat, and the effect on the skin surface layer as in the present invention is unknown. .. Further, such a device may be accompanied by a tingling stimulus when an electric current is passed even if the frequency is low, which is different from the present invention that gives a gentle stimulus to the skin.
  • the present invention enables a cosmetological method by a simple method of directly giving a stretch stimulus to the skin without applying energy such as ultrasonic waves, electric current, or magnetic field. Further, the extension stimulus having such a frequency is a gentle stimulus, but has an effect of adjusting / improving the M1 / M2 balance as described in the examples. Therefore, when the method / apparatus of the present invention is used, it is expected to prevent and / or improve photoaging and / or pigmentation without adversely affecting the skin.
  • the physical stimulus may be by an instrument such as a facial device, an experimental device, a massage using human hands or instruments, a facial exercise, or may be achieved by contact or non-contact. ..
  • a mechanically generated physical stimulus is applied to the skin by using a device having a stimulus generating part that generates a physical stimulus and a stimulating part that applies the physical stimulus by contact or non-contact.
  • Physical irritation may be achieved, for example, by contact such as pulling, pressing, tapping, kneading, aspirating, and / or applying a shock wave to the skin, for example, with something like ultrasonic waves, air pressure or water pressure. It may be achieved by non-contact such as displacement with.
  • Facial exercises include swelling the cheeks and opening the eyes. Examples of the massage include a massage using an instrument such as a hand or a roller by the subject to be treated or a practitioner such as a cosmetologist. However, it is not limited as long as it is within the range of the physical stimulation of the present invention.
  • Examples of the device of the present invention include a beauty device provided with a skin contact portion that contacts the skin of the user and gives the physical stimulus of the present invention.
  • a beauty device provided with a skin contact portion that contacts the skin of the user and gives the physical stimulus of the present invention.
  • it may be composed of a grip portion and a skin extension portion or a skin pressing portion.
  • the device shown on the left in FIG. 14 is designed to stretch the skin at a specific frequency and extension rate when the skin contact part touches the skin.
  • the device of the present invention includes a power supply, a stimulus generating unit, and a skin stimulating unit, the power supply generates an electric signal, and the stimulus generating unit converts the electric signal from the power source into a physical stimulus.
  • the skin stimulus portion may be one that receives the physical stimulus generated by the stimulus generator and gives the physical stimulus to the user's skin.
  • the device shown on the left in FIG. 14 includes a grip portion, a power source, a control unit that controls physical stimulation, a stimulation generation unit, and a skin contact portion including a skin stimulation portion and a skin fixing portion.
  • the user holds the grip part and puts the skin contact part on the skin, fixes the skin with the skin fixing part, and operates the control part so that the electrical signal from the power supply is physically stimulated by the stimulation generating part.
  • the stimulus generator may be driven by a motor or the like to convert an electrical signal into a physical stimulus.
  • the skin stimulating portion shown on the left side of FIG. 14 gives a stretching stimulus to the skin, but may be, for example, a pressing stimulus to the skin.
  • the device of the present invention may be a beauty device including a power supply, a control unit for controlling physical stimulation, a stimulation generating unit, and a skin contact portion including a skin contact surface made of a sheet-like material.
  • a beauty device including a power supply, a control unit for controlling physical stimulation, a stimulation generating unit, and a skin contact portion including a skin contact surface made of a sheet-like material.
  • the sheet-like material can carry an electric current and may convert an electrical signal from a power source into a physical stimulus. Examples of such sheet-like materials include dielectric elastomer actuators (DEA), conductive polymers, IPMC, PVC gel, Mckinnen type and the like.
  • DEA dielectric elastomer actuators
  • the power source of the device of the present invention may be an internal power source or an external power source, or may be a rechargeable type. Further, the device of the present invention may be one that uses data stored in a mobile phone, a cloud, or the like, or one that is remotely operated wirelessly.
  • the physical stimulus may be the one that achieves the extension of the skin by giving a physical stimulus by contact or non-contact as in the case of upper surgery.
  • the physical stimulus may be applied in the direction parallel to the skin surface, that is, in the lateral direction, in the direction perpendicular to the skin surface, that is, in the vertical direction, or in any direction such as an oblique direction or a twisting direction. it can.
  • the number of cycles to perform physical stimulation is not limited. For example, any number of cycles such as 10 to 500 cycles, 20 to 400 cycles, 30 to 300 cycles, 40 to 200 cycles, and 50 to 100 cycles may be performed. For example, as described in the examples, 27 cycles may be sufficient.
  • any number of these cycles may be regarded as one set, and this set may be performed with an arbitrary number of sets, for example, 1 to 100 sets, 2 to 50 sets, or 3 to 10 sets.
  • the time to perform physical stimulation is not limited.
  • the cycle may be repeated with or without a pause time for a certain period of time such as 5 minutes to 3 hours, 10 minutes to 2 hours, and 30 minutes to 1 hour.
  • the time interval between cycles or sets is not limited.
  • the extension or pressing stimulus may be one set or multiple sets alone, with one or more sets daily, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days. It may be performed continuously or intermittently, periodically or irregularly, such as once, once every 1, 2, 3, or 4 weeks.
  • the frequency, extension rate, number of cycles, and frequency as described above are not limited.
  • the waveform for physical stimulation can be set arbitrarily, such as a square wave, a sine wave, a triangular wave, and a sawtooth wave.
  • the adjustment / improvement of the M1 / M2 balance may be the administration of an M1 / M2 balance adjustment / improvement agent or a composition containing the same.
  • the present inventors have discovered that the compounds of the present invention, otogirisou extract, thyme extract, aubergine, and / or eucalyptus extract function as such M1 / M2 balance adjusting / improving agents.
  • the present invention comprises the compound of the present invention, otogirisou extract, thyme extract, aubergine, and / or eucalyptus extract, or the compound of the present invention, otogirisou extract, thyme extract, aubergine, and / or eucalyptus.
  • M1 / M2 balance adjusting / improving agents, anti-photoaging agents, pigmentation inhibitors, and compositions containing them, which contain an extract as an active ingredient, are also provided.
  • the compound of the present invention is N-methyl-trans-4- (aminomethyl) cyclohexanecarboxamide (methylamide tranexamic acid (C 9 H 19 C 1 N 2 O)) or a salt thereof.
  • the above-mentioned compound may be synthesized, for example, by a method as described in Patent Document 14, or a commercially available product may be used.
  • Hypericum erectum is a perennial plant belonging to the genus Hypericum in the family Hypericum.
  • the extract of Hypericum erectum used in the present invention the extract of the above-ground part of Hypericum erectum is preferable, but since the seeds, roots, etc. also contain the active ingredient, one or more of these extracts. Can also be used. Commercially available products can also be used as the extract of Hypericum erectum.
  • Thymus is a perennial plant belonging to the genus Thymus of the Labiatae family.
  • various times such as common time (T. vulgaris), citrus time (T. x citriodorus), and wild time (T. serpyllum) can be used.
  • T. vulgaris common time
  • citrus time T. x citriodorus
  • wild time T. serpyllum
  • an extract of whole thyme is preferable, but since seeds, flowers, roots and the like also contain active ingredients, one or more of these are extracted. You can also use things.
  • Commercially available products can also be used as the thyme extract.
  • Oubaku is a crude drug obtained by drying the bark of Phellodendron amurense or Phellodendron chinense of the Rutaceae family.
  • the Phellodendron amurensis or its extract may be produced by a conventional method, or a commercially available product may be used.
  • Eucalyptus is a tree of the genus Eucalyptus in the family Myrtaceae.
  • an extract of eucalyptus leaves is preferable, but since the seeds, flowers, bark, roots, etc. also contain active ingredients, any one or more of these are contained. Extracts can also be used.
  • the eucalyptus extract may be produced by a conventional method, or a commercially available product may be used.
  • the extraction method is not particularly limited, but an extraction method using a solvent is preferable.
  • the plant can be used as it is, but it is better to crush it into granules or powder and use it for extraction in a short time with high extraction efficiency under mild conditions. It can be carried out.
  • the extraction temperature is not particularly limited, and may be appropriately set according to the particle size of the pulverized product, the type of solvent, and the like. Usually, it is set in the range from room temperature to the boiling point of the solvent.
  • the extraction time is not particularly limited, and may be appropriately set according to the particle size of the pulverized product, the type of solvent, the extraction temperature, and the like. Further, at the time of extraction, stirring may be performed, the mixture may be allowed to stand without stirring, or ultrasonic waves may be applied.
  • the type of solvent is not particularly limited, but water, lower alcohols such as hydrous ethanol and ethanol, organic solvents such as hexane, and mixed solvents such as hexane / ethanol are preferable. Extraction may be carried out at room temperature, but may be carried out under heating (for example, using a heated solvent such as hot water or hot water). Further, the extraction treatment may be carried out by adding an enzyme to the solvent. By adding the enzyme, the cell tissue of the plant can be disrupted, which can further improve the extraction efficiency. As the enzyme, it is preferable to use a cell tissue disrupting enzyme.
  • enzymes examples include pectinase, cellulase, hemicellulase, ⁇ -amylase, and phytase. One of these enzymes may be used alone, or two or more of these enzymes may be mixed and used.
  • the active ingredient is extracted and dissolved in the solvent.
  • the solvent containing the extract may be used as it is, or may be used after undergoing conventional purification treatments such as sterilization, washing, filtration, decolorization, and deodorization. Further, it may be used after being concentrated or diluted if necessary. Further, the solvent may be completely volatilized to form a solid (dried product) before use, or the dried product may be redissolved in an arbitrary solvent before use.
  • the squeezed liquid obtained by squeezing the raw material plant contains the same active ingredient as the extract, the squeezed liquid can be used instead of the extract.
  • the M1 / M2 balance adjusting / improving agent and the composition are not limited to the above substances, and it is known that any substance, for example, the substance described in Patent Documents 4 to 7 and the like, adjusts / improves the M1 / M2 balance.
  • Known substances may be used.
  • the administration route can be arbitrarily selected, for example, transdermal administration, oral administration, subcutaneous administration, transmucosal administration, intramuscular administration, etc., but photoaging is a phenomenon specifically observed in the skin in the exposed part. Therefore, transdermal administration, which can be administered to a specific part of the skin, may be preferable.
  • transdermal administration may be preferable so that the pigmentation can reach the epidermis and dermis from the skin.
  • the adjustment / improvement of the M1 / M2 balance may be, for example, induction of differentiation into M2 macrophages, or may be performed by any other M1 / M2 balance adjustment / improvement method.
  • the agent or composition of the present invention regulates / improves the M1 / M2 balance and, as a result, suppresses photoaging and / or pigmentation of the skin.
  • the M1 / M2 balance adjusting / improving agent, anti-photoaging agent, and pigmentation inhibitor of the present invention are any one of the above active ingredients.
  • the seeds may be contained alone, or two or more kinds may be contained in any combination and ratio.
  • the agent of the present invention may be a composition in which the above active ingredient is combined with one or more other ingredients such as excipients, carriers and / or diluents.
  • the composition and form of the composition are arbitrary, and may be appropriately selected according to the conditions such as the active ingredient and the intended use.
  • the composition can be produced by a conventional method with a formulation appropriately combined with an excipient, a carrier and / or a diluent and the like and other components according to the dosage form.
  • the agent of the present invention can be blended in cosmetics or the like and used for humans and animals, or can be administered to humans and animals as a pharmaceutical preparation. In addition, it may be mixed with various foods and drinks and feeds and ingested by humans and animals.
  • the blending amount (dry mass) of the plant or its extract depends on the type, purpose, form, usage, etc. , Can be decided as appropriate. For example, 0.00001% to 50% (in the case of an extract or crude drug, converted to dry mass) of the compound of the present invention, Hypericum erectum extract, Phellodendron amur extract, Phellodendron amur extract, and / or Eucalyptus extract, respectively, in the total amount of cosmetics. Can be mixed.
  • ingredients usually used for external skin preparations such as cosmetics, pharmaceuticals, quasi-drugs, etc., such as antioxidants, oils, UV protection agents, within the range that does not impair the effects of the present invention.
  • ingredients usually used for external skin preparations such as cosmetics, pharmaceuticals, quasi-drugs, etc., such as antioxidants, oils, UV protection agents, within the range that does not impair the effects of the present invention.
  • Surfactants, thickeners, alcohols, powder components, coloring materials, aqueous components, water, various skin nutrients, etc. can be appropriately blended as needed.
  • the external preparation for skin of the present invention can be applied as a cosmetic, a non-medicinal product, etc. applied to the outer skin, particularly preferably as a cosmetic, and the dosage form is not limited as long as it can be applied to the skin.
  • Any dosage form such as solution system, solubilization system, emulsification system, powder dispersion system, water-oil two-layer system, water-oil-powder three-layer system, ointment, lotion, gel, aerosol, etc. is applied.
  • the agent of the present invention when used as cosmetics, it may be used in lotions, emulsions, foundations, lipsticks, lip balms, cleansing creams, massage creams, facial masks, hand creams, hand powders, body shampoos, body lotions, body creams, bath cosmetics, etc. It may be used as a form.
  • agent and composition of the present invention are not limited to the above-mentioned dosage forms and forms.
  • agent or composition of the present invention may be used in combination with the device or method of the present invention or other device or method.
  • the subject to which the method, apparatus, agent and composition of the present invention is applied is skin photoaging and / or skin photoaging even if pigmentation (for example, dermal pigmentation) is objectively or subjectively observed. And / or may be the subject wishing to prevent pigmentation. For example, it may be an object that is judged to be out of M1 / M2 balance. In one embodiment, the subject may be a subject judged to have a high degree of skin photoaging and / or degree of pigmentation (for example, degree of dermal pigmentation) using the M1 / M2 balance in the skin as an index. Alternatively, the phenotype specific to photoaging of the skin, for example, spots, wrinkles, sagging, etc.
  • epidermis and pigmentation for example, spots, dullness, bruises, tattoo marks, etc. May be the subject of concern. Spots, wrinkles, sagging, dullness, bruises, tattoo marks, etc. can be determined by visual judgment or using known indexes.
  • the candidate drug is applied to the biological sample; the M1 / M2 balance in the biological sample before and after the candidate drug is applied; the M1 / M2 balance in the biological sample to which the candidate drug is applied is the drug.
  • methods of screening for anti-photoaging agents including assessing that the agent has anti-photoaging and / or anti-pigmentation activity if it improves as compared to before the application. Applying a cosmetic treatment to a skin sample; measuring the M1 / M2 balance in the skin sample before and after the cosmetic treatment; comparing the M1 / M2 balance in the skin sample with the cosmetic treatment with that before the treatment.
  • the method of the present invention enables screening as to whether or not a candidate drug or cosmetic treatment has an anti-photoaging and / or pigmentation-suppressing effect, and enables product development and proposal of new skin care. That is, according to the present invention, an anti-photoaging agent and / or a pigmentation inhibitor and an anti-photoaging and / or pigment for preventing and / or ameliorating photoaging and / or pigmentation by adjusting the M1 / M2 balance. Deposition control cosmetic treatments are provided.
  • the pigmentation may be dermal pigmentation.
  • the skin sample may be a skin sample after collection, for example, a skin sample in an ex vivo state after being collected from an animal such as a human, or cultured skin cells, for example, monolayer or multilayer cultured cells, cultured. It may be in an in vitro state such as keratinocytes or cultured fibroblasts. Alternatively, it may be an artificial skin sample such as a 3D skin model.
  • the immune cell sample may be an immune cell present in the skin after collection, an immune cell in the blood after collecting blood infiltrating the skin, a cultured immune cell, or the like.
  • the 3D skin model is not limited, it may be created by the methods described in, for example, Patent Document 8 and Non-Patent Documents 10 to 12, or may be created by modifying such a method so that the M1 / M2 balance can be easily measured. .. Biological samples are not limited as long as the M1 / M2 balance can be measured.
  • immune cells such as THP-1 are differentiated into M0, the culture supernatant is collected after adding the drug and culturing for a certain period of time, and ELISA is used. It may be achieved by quantifying the amount of M2-specific cytokines such as IL-10 by a measurement method such as IL-10.
  • a screening method cells differentiated into M2 by adding IL-4, IL-13, etc. may be used as a positive control, and further, a candidate drug is added to IL-4, IL-13. May be compared.
  • M2 since M2 has a high IL-10-producing ability, if IL-10 in the supernatant is increased after M0 differentiation compared to the control cultured only in the medium, it can be said that it has differentiated into M2.
  • a drug may be used as an "improving agent", and here, even if the presence or absence of the drug is compared in the presence of IL-4, IL-13, etc. to evaluate whether the amount of IL-10 is different. Good.
  • the candidate drug means a drug whose anti-photoaging and / or pigmentation-suppressing effect is investigated, and includes drugs already sold as products, drugs in the development stage, and cosmetics. It may be the drug selected for cosmetics in the development of the drug.
  • the cosmetological treatment is not particularly limited, but includes any treatment considered to be effective in suppressing photoaging and / or pigmentation, such as application of a cosmetic containing the agent of the present invention or other ingredients.
  • Cosmetics refer to cosmetics applied to the skin, such as lotions, milky lotions, beauty essences, creams, foundations, etc., but are not limited to these, and are not intended to directly improve the skin condition. However, it is intended to include everything that is applied to the skin, including, for example, sunscreens.
  • the cosmetological treatment may be to give the skin physical stimuli such as extension stimulus, pressing stimulus, and massage.
  • the cosmetological procedure may be a single procedure or a continuous procedure performed over days to weeks.
  • the beauty treatment may be performed individually, or may be performed at a beauty salon, a cosmetics store, an esthetic salon, or the like.
  • the present invention measures the M1 / M2 balance, for example, an antibody that detects arbitrary markers such as CD86, CD206, and CD163 that can measure the number of M1 and M2, and a drug that measures the amount of mRNA of those markers. Also provided are screening kits for anti-photoaging and / or anti-pigmentation agents, including agents for this. Furthermore, the present invention also provides the following methods, systems, and devices.
  • a method of assessing the degree of photoaging and / or pigmentation of the subject's skin performed by one or more computers with the procedure;
  • the analysis department of the server performed machine learning using a neural network method using teacher data based on the M1 / M2 balance of the skin and the photoaging and / or the degree of pigmentation stored in the storage unit.
  • the procedure for analyzing the relationship between / M2 balance and photoaging and / or the degree of pigmentation; the procedure for the receiving unit of the server to receive the data of the M1 / M2 balance of the target skin; and the evaluation unit of the server. The procedure for evaluating and outputting the photoaging and / or the degree of pigmentation of the target skin by inputting the M1 / M2 balance of the received target skin based on the analyzed relationship; A method for evaluating the degree of photoaging and / or pigmentation.
  • a database unit that stores data related to the preset M1 / M2 balance of the skin; a data input unit that inputs data related to the M1 / M2 balance of the target skin; refers to the reference value stored in the database unit. , Calculated by comparing with the data on the M1 / M2 balance of the target skin input by the data input unit; Calculation unit; Photoaging and / or pigmentation degree of the skin based on the calculation result by the calculation unit.
  • a system for evaluating the degree of photoaging and / or pigmentation of the target skin which has an evaluation unit for evaluating the skin; and a display unit for displaying the evaluation result by the evaluation unit.
  • a photoaging and / or pigmentation degree calculation device that calculates the photoaging and / or pigmentation degree of the target skin using data on the M1 / M2 balance of the target skin, and the device is the target skin.
  • the calculation unit is provided to calculate the photoaging and / or the degree of pigmentation of the target, and the calculation unit is when the data on the M1 / M2 balance of the skin is input.
  • a photoaging and / or pigmentation degree calculator having a trained neural network that has undergone machine learning processing using teacher data to calculate the estimated photoaging and / or pigmentation degree.
  • the method, system, and apparatus of the present invention enable objective measurement of photoaging and / or pigmentation based on M / M2 balance.
  • Experiment 1 Tissue staining The skin of the outer corners of the eyes of white-derived young people (20s to 30s) and elderly people (60s to 70s) with photoaging shown in Fig. 1a was made into frozen sections and thinned. After cutting, the cells were stained with the macrophage markers shown below.
  • Staining of M1 macrophages Double staining was performed with an anti-human CD86 antibody (R & D) derived from goats and an anti-human CD11b antibody (abcam) derived from rabbits (Fig. 1b).
  • FIGS. 1b and 1c The tissue staining of (1) to (3) is shown in FIGS. 1b and 1c, the graph of the counting result is shown in FIG.
  • M1 macrophages were more common in the elderly with photoaging, while M2 macrophages were decreased. More specifically, as shown in Fig. 1b, M1 macrophages were present only around blood vessels in young people, while they were scattered throughout the tissue in elderly people. In addition, as shown in Fig. 1c, M2 macrophages were present throughout the tissue in young people, while their number decreased in elderly people. Furthermore, as shown in Fig.
  • the total number of macrophages (the number of M1 + the number of M2) did not change between the young and the elderly, only the M1 / M2 balance changed.
  • the ratio of M2 to M1 (number of M2 / number of M1) is in the range of about 5/5 to about 7/3, while in elderly people, the ratio of M2 to M1 decreases sharply and M1 / The M2 balance was greatly disturbed, which was significantly different from that of the young.
  • THP-1 M1 and M2 differentiation stimulation experiment of THP-1 Differentiation was induced into M1 and M2 macrophages using THP-1, which is a human-derived cell, according to the method described in Non-Patent Document 1. Specifically, THP-1 was cultured by adding 1 mM Na Pyruvate (Nakalai), 2 mM L Glutamine (Nakalai), and 10% FBS to RPMI1640 (Nakalai) by the method shown in Fig. 2a. Then, 100 nM PMA (abcam) was further added and stimulated for 24 hours to differentiate into macrophages.
  • Nakalai Na Pyruvate
  • Nakalai 2 mM L Glutamine
  • FBS RPMI1640
  • RNA of each differentiated or undifferentiated cell is extracted, and real-time PCR is performed by TaqMan Gene expression assay using IL-1beta, TNF-alpha, and IL-10 probes (Applied Biosystems). , The expression level was quantified (Fig. 2c, upper figure). Furthermore, using the same CD86 antibody (R & D) and CD206 antibody (BD) as in Experiment 1, PCR was performed in the same manner to quantify the expression level (Fig. 2c, lower figure). Each value was corrected by the mRNA expression level of GAPDH.
  • the macrophages differentiated by the method described in Experiment 2 have inflammatory cytokines (IL-1beta, TNF-alpha) characteristic of M1 and anti-inflammatory cytokines (IL) characteristic of M2, respectively. It was shown to produce -10). Furthermore, these differentiation-inducing macrophages showed increased expression of CD86 and CD206, which are surface markers of M1 and M2 macrophages, respectively. From these results, it was confirmed that the differentiation induction was successful. Therefore, the M1 and M2 macrophages differentiated by the method of Experiment 2 and the undifferentiated M0 macrophages were used in Experiments 3 and 4 below.
  • IL-1beta inflammatory cytokines
  • IL anti-inflammatory cytokines
  • Experiment 3 Addition of M1 and M2 macrophage supernatants to fibroblasts As shown in Fig. 3a, THP-1 was differentiated into M1 and M2 in the same manner as in Experiment 2, or M0 remained undifferentiated. After removing the supernatant and washing once with PBS, the medium was added and the cells were cultured for 48 hours. Their supernatants containing M1 and M2 secretions such as inflammatory and anti-inflammatory cytokines were added to neonatal fibroblasts. Cells supplemented with RPMI1640 (Nakalai) were used as a control.
  • RPMI1640 Nakalai
  • fibroblasts were cultured for 72 hours, and the amount of procollagen in the supernatant was quantified using a PIP ELISA kit (TAKARA) (Fig. 3b).
  • Cell fractions were stained with rabbit-derived anti-human collagen antibody (CEDERLANE) and biotinylated hyaluronic acid-binding protein (HOKUDO) (Fig. 3c).
  • ⁇ -gal was used as an index of senescence, the nucleus of the cell was stained with DAPI, and then ⁇ -gal in the cell was stained with the Senescence Detection Kit (abcam) (Fig. 3d), and ⁇ -gal positive cells and ⁇ -gal positive cells and The number of DAPI-positive cells was counted (Fig. 3e).
  • Figure 3b shows the amount of procollagen after adding the supernatant of each macrophage (M1, M2) to fibroblasts and culturing for 72 hours.
  • Figure 3c shows the localization of collagen and hyaluronic acid in fibroblasts after adding the supernatant of each macrophage (M1, M2) and culturing for 72 hours. From Figures 3b and c, it can be seen that M1 remarkably suppresses collagen production.
  • Figure 3d shows intracellular ⁇ -gal in fibroblasts after adding the supernatant of each macrophage (M1, M2) and culturing for 72 hours.
  • Figure 3d suggests that M1 increases ⁇ -gal-positive cells and promotes aging, while M2 suppresses aging.
  • Figure 3e shows a graph counting the number of ⁇ -gal-positive cells and DAPI-positive cells. The figure on the right shows the total number of DAPI-positive cells per well. The right figure in Fig. 3e suggests that M2 not only suppresses cell aging but also promotes cell proliferation.
  • Figure 3e Left shows the ratio (%) of the number of ⁇ -gal-positive cells to the total number of DAPI-positive cells. M1 has aging and cell death promoting effects, while M2 has aging suppressing and cell proliferation effects. Is suggested. Furthermore, as shown in Fig.
  • fibroblasts are known to secrete factors such as SCF and HGF by photostimulation such as UV and cause cell death.
  • fibroblasts act directly or indirectly on melanocytes by secreting factors such as HGF, ET1, bFGF, SCF, and clusterin by photostimulation. Produces melanin. Therefore, it is suggested that cell death and melanin production by light stimulation are due to the imbalance of M1 / M2.
  • each macrophage supernatant collected by the same method as in Experiment 3 was collected from neonatal human capsule-derived fibroblasts (young-derived fibroblasts) and 68-year-old human-derived fibroblasts (old-age-derived fibroblasts). was added to and cultured for 72 hours. After that, ⁇ -gal and DAPI staining were performed in the same manner as in Experiment 3, and the number of ⁇ -gal-positive cells and DAPI-positive cells was counted (Figs. 4a and 4b).
  • FIG. 4a shows a ⁇ -gal staining diagram.
  • FIG. 4b shows a graph of the results of the number of ⁇ -gal-positive cells and the total number of cells per well of young-derived fibroblasts and old-age-derived fibroblasts. From the figure below in Fig. 4a, it was found that the addition of M2 supernatant significantly reduced ⁇ -gal-positive cells and suppressed aging even in aged cells. It was also found from Fig. 4b that M1 promotes aging and M2 suppresses aging regardless of age. This is supported by the result that, as shown in FIGS. 4c and 4d, the amount of collagen produced in the cells to which the M1 supernatant was added was significantly lower than that in the M2 supernatant, regardless of age.
  • Experiment 5 Co-culture experiment of M1 macrophages and M2 macrophages with neonatal sac-derived fibroblasts
  • RPMI1640 Nakalai
  • the amount of collagen produced when neonatal foramen-derived fibroblasts and half the number of M1 or M2 macrophages were cultured in RPMI was stained with rabbit-derived anti-human collagen antibody (CEDERLANE) and compared.
  • macrophages were visualized by staining with mouse-derived anti-human CD68 antibody (abcam).
  • the 3D skin model was created as follows. Human dermal fibroblasts (0.2 x 10 6 cells) were seeded in cell culture inserts ( ⁇ 12 mm, average pore size of porous membrane: 0.4 ⁇ m), 200 ⁇ M magnesium ascorbic acid-2-phosphate (APM), 10% FBS-DMEM. The medium was changed once every two days and cultured for one week.
  • the control model is 0.5% type I collagen-10% FBS-DMEM solution containing human dermal fibroblasts
  • the M1 model, and the M2 model are M1 macrophages and M2 macrophages differentiated by the method of Experiment 2.
  • Collagen gels were prepared on human dermal fibroblasts and cultured for 1 to 5 days.
  • epidermal keratinocytes dispersed in Humedia-KG2 (Kurabou) medium were seeded on collagen gel so as to be 5 ⁇ 10 5 cells / well, and Humedia-KG2 and 10% FBS-DMEM were placed outside the insert.
  • Medium mixed at 1: 1 and added with 200 ⁇ M APM was added to the same level as the inside and cultured for 3 days.
  • the cultured skin model was fixed with 4% PFA / PBS (Nakarai), and then stained with anti-CD206 antibody (abcam), anti-CD68 antibody (abcam), and anti-CD86 antibody (abcam) in the same manner as in Experiment 1.
  • the presence of M1 and M2 macrophages was confirmed (Fig. 6).
  • the cells were stained with anti-p21 antibody (abcam) and DAPI (VECTOR), and the total number of cells stained with DAPI in each of the epidermal cell layer and the fibroblast layer was taken as the total number of cells, and the anti-p21 antibody was also stained.
  • the number of cells was counted as the number of p21 positive cells.
  • the ratio of the number of p21-positive cells to the total number of cells was calculated by the following formula.
  • Experiment 7 Sunlight irradiation experiment on an ex vivo model using human skin Native Skin (skin collected from a 38-year-old woman) manufactured by Genoskin was cultured for 1 day in the attached culture medium after arrival. The next day, an optical filter was placed in an Oriel 1000W solar simulator, and only UVA and UVB were irradiated at 11.5 J / cm 2 , and the culture was continued with the attached culture solution (irradiation (+)). A non-irradiated sample was used as a control (irradiation (-)). Samples were collected 5 days after irradiation, and M1 macrophages, M2 macrophages, and total macrophages were stained in the same manner as in Experiment 1, and the numbers were counted and graphed.
  • Experiment 8 Extension stimulation experiment on an ex vivo model using human skin Next, we examined a method for adjusting or improving the M1 / M2 balance.
  • Sample Native Skin manufactured by Genoskin (skin collected from a 38-year-old woman) (6 well size, diameter about 2 to 2.5 cm) was used.
  • Extension conditions An extension device was created that has grips that grip both ends of the skin in the well as shown in Fig. 11 and stretches the skin by pulling the grips. The well containing the above tissue pieces was placed horizontally, and the grip was operated to extend the skin from both ends, and as shown in Fig. 10, extension with a 10% extension rate was performed at a rate of 10% / s.
  • the sample was returned to its original non-extended state at a recovery rate of% / s. This was regarded as one cycle, and a total of 90 cycles were performed over 30 minutes. These 90 cycles were regarded as one set, and a total of 3 sets (total of 270 cycles) were performed over 3 hours with a rest period of 30 minutes to 1 hour between sets.
  • a sample without extension stimulation was used (control).
  • M1 macrophages and M2 macrophages were stained by the same method as in Experiment 1 except that total macrophages were stained with only rabbit-derived anti-human CD11b antibody (abcam). , Total macrophages were stained and their numbers were counted. The ratio (%) of each macrophage (M1, M2) to the total number of all macrophages was calculated and graphed.
  • Experiment 9 Screening for anti-photoaging and / or pigmentation agents that prevent and / or improve photoaging and / or pigmentation by adjusting or improving the M1 / M2 balance of agents that adjust or improve the M1 / M2 balance. Screening was performed. A total of 19 components including Hypericum erectum extract, Thyme extract, and tranexamic acid methylamide (N-methyl-trans-4- (aminomethyl) cyclohexanecarboxamide) were examined as the agents to be screened.
  • Hypericum extract is an extract of the above-ground part of Hypericum purchased from Ichimaru Falcos.
  • Thyme extract is an extract of whole wild thyme purchased from Koei Kogyo.
  • Tranexamic acid methylamide was synthesized by the method described in Patent Document 14. Hypericum erectum extract was dissolved in 50% ethanol, thyme extract was dissolved in butylene glycol, and tranexamic acid methylamide was dissolved in PBS.
  • THP-1 cells The same THP-1 cells as in Experiment 2 were used in the undifferentiated state of M0 and cultured at 37 ° C overnight.
  • Each of the above-mentioned drugs to be screened was added to the medium so that Hypericum erectum extract was 0.1%, thyme extract was 0.1%, tranexamic acid methylamide was 0.06% and 0.03%, respectively, and the same amount of these solvents was added to the control.
  • the cells were cultured at 37 ° C for two nights. The cells were collected, RNA was extracted, the expression levels of CD86 and GAPDH were quantified by real-time PCR, and CD86 / GAPDH was measured.
  • a drug that lowers the CD86 / GAPDH level more reproducibly than the control (cont) was searched for as an M1 differentiation inhibitor.
  • Fig. 13 From Fig. 13, it is shown that the addition of Hypericum erectum extract, thyme extract, and tranexamic acid methylamide significantly reduces the CD86 / GAPDH level and has an M1 differentiation inhibitory effect, and can be used as an M1 / M2 balance adjusting / improving agent. Is suggested.
  • Experiment 10 Screening of anti-photoaging agents that prevent and / or improve photoaging and / or pigmentation by adjusting or improving the M1 / M2 balance Eucalyptus extract to screen for a larger number of agents .
  • the extract of Oubaku was a dried bark of Phellodendron amurensis, and the extract of eucalyptus was an extract of eucalyptus leaves.
  • the Phellodendron amur extract was dissolved in butylene glycol to a concentration of 0.01%, and the eucalyptus extract was dissolved in 50% ethanol to a concentration of 0.1%. These solvents were used as controls.
  • Experiment 11 Effect of M1 and M2 macrophage supernatants on collagen
  • the supernatants of each macrophage were examined in the same manner as in Experiment 3.
  • Real-time PCR was performed using MMP-1, MMP-2, and IL-1 ⁇ probes (Taqman probe manufactured by Applied Bioosystems) as indicators of collagen degradation, and the expression levels of each mRNA were quantified (Fig. 16a).
  • Experiment 12 Degradation and production of collagen in an ex vivo model using human skin of young and elderly young people (20s to 30s) and elderly people (60s to 80s) of the age shown in Fig. 17a Anti-procollagen antibody (millipore), anti-fragmented collagen (AdipoGen), and Experiment 1 that stain the collagen-cleaved fragment on the 3/4 side after slicing the skin of the derived rice cake into frozen sections as in Experiment 1. Using the same CD68, CD11b, CD206, and CD86 antibodies, positive cells were counted in the same manner as in Experiment 1.
  • Experiment 13 In vitro experiment showing the ability to take up melanin in the dermis M1 macrophages and M2 macrophages obtained by differentiating THP-1 cells were differentiated by the same method as in Experiment 3.
  • a melanin solution (Sigma, Melanin-BioReagent, Synthetic, suitable for cell culture) was dissolved in PBS in each differentiated M1 macrophage, M2 macrophage, and fibroblast (manufactured by Kurabou) to adjust to 0.02% W / V. Added. After 24 hours, the cells were washed with PBS and then photographed under a microscope to collect the cells. The number of recovered cells was measured with Alamer Blue (Life Technologies) and the amount of melanin was measured and quantified as follows.
  • the results are shown in Figures 18a-c.
  • the unit "melanin / almar blue” in the figure is the intensity ((1)) of which the cultured cells were stained with alamar blue and the fluorescence of the supernatant was measured at 544 nm / 590 nm, and then the cells were lysed to lyse melanin. It is a relative value ((2) / (1)) of the intensity ((2)) measured at an absorbance of 475 nm, and indicates the melanin content per cell. From these figures, it was found that M2 phagocytoses a large amount of melanin. This tendency was already seen after 24 hours, but the difference became clearer when the cells were continuously cultured and observed until 5 days later (Fig. 18c).
  • Experiment 14 Ex vivo experiment showing the ability to take up melanin in the dermis
  • Experiment 13 revealed that M2 macrophages phagocytose more melanin than fibroblasts and M1 macrophages, so the dermis layer of human skin counted in Experiment 13 was observed ex vivo using LSM880 (manufactured by Karl Zeiss). It was found that melanin was stained black together with M2 macrophages when observed in the bright field, regardless of whether they were old or young, whereas melanin was not present near M1 macrophages. As a result, it was found ex vivo that M2 macrophages took up more melanin than fibroblasts and M1 macrophages in the dermis, and the numbers were counted and graphed (Fig. 19).
  • the proportion of M1 increases due to photoaging (Experiment 7), and the higher the proportion of M2, the greater the amount of skin collagen (Experiments 11 and 12) and the higher the pigment phagocytosis (Experiments 13 and 14). Therefore, the compounds of the present invention such as tranexamic acid methylamide screened as an M1 differentiation inhibitor, thyme extract, aubaku, and eucalyptus extracts, and the physical stimulus of the present invention having an M1 / M2 balance improving effect are anti-photoaging. It is suggested that the effect of suppressing aging and / or pigmentation is high.
  • photoaging causes an imbalance in the balance of M1 / M2, which causes photoaging in the exposed skin and pigmentation in the dermis, but adjusts the balance of M1 / M2. It is suggested that photoaging and / or dermal pigmentation can be prevented and / or improved by improving or improving.
  • the adjustment or improvement of the balance of M1 / M2 can be performed by giving a stretching stimulus or a pressing stimulus, or using a compound of the present invention such as tranexamic acid methylamide, M1 such as Hypericum erectum extract, Thyme extract, Oubaku, and / or Eucalyptus extract. It is achieved by applying a component having an inhibitory effect on differentiation as an M1 / M2 balance adjusting / improving agent, and is expected to prevent and / or improve photoaging and / or dermal pigmentation.
  • prevention and / or amelioration of photoaging and / or dermis pigmentation evaluation of photoaging and / or dermis pigmentation degree, anti-photoaging and / or dermis pigmentation inhibitor and anti-photoaging and / or dermis. It is possible to search for cosmetic treatments that suppress pigmentation.

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Abstract

Le problème décrit par la présente invention est de fournir une méthode, un dispositif et un médicament destinés à prévenir et/ou à atténuer le photovieillissement et/ou la pigmentation dermique, une méthode destinée au criblage d'un agent inhibiteur de photovieillissement et/ou de pigmentation dermique, une méthode destinée à l'évaluation d'un traitement cosmétique inhibiteur de photovieillissement et/ou de pigmentation dermique et une méthode destinée à l'évaluation du photovieillissement et/ou de la pigmentation dermique. À cet effet, selon l'invention, le photovieillissement et/ou la pigmentation dermique peuvent être prévenus et/ou atténués par réglage de l'équilibre M1/M2. Le photovieillissement et/ou la pigmentation dermique peuvent être évalués objectivement en utilisation l'équilibre M1/M2 en tant qu'indice, et des médicaments et des traitements cosmétiques destinés à prévenir et/ou à atténuer le photovieillissement et/ou la pigmentation dermique peuvent être recherchés sur la base d'une telle méthode.
PCT/JP2020/017082 2019-04-19 2020-04-20 Méthode et dispositif de prévention et/ou d'atténuation du photovieillissement et/ou de la pigmentation dermique, agent inhibiteur de photovieillissement et/ou de pigmentation dermique, méthode de criblage d'un agent inhibiteur de photovieillissement et/ou de pigmentation dermique, méthode d'évaluation d'un traitement cosmétique inhibiteur de photovieillissement et/ou de pigmentation dermique, et méthode d'évaluation du photovieillissement et/ou de la pigmentation dermique WO2020213743A1 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023063118A1 (fr) * 2021-10-14 2023-04-20 株式会社 資生堂 Inducteur ou activateur de macrophages m2 ou de type m2, procédé pour induire ou activer des macrophages m2 ou de type m2, composition pour prévenir et/ou améliorer la pigmentation dermique et procédé pour prévenir et/ou améliorer la pigmentation dermique

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2009268640A (ja) * 2008-05-02 2009-11-19 Kao Corp 皮膚内部の弾性計測方法
JP2012229924A (ja) * 2011-04-25 2012-11-22 Shiseido Co Ltd 過酸化脂質の定量分析方法
JP2013083504A (ja) * 2011-10-07 2013-05-09 Pola Chem Ind Inc スクリ−ニング方法
JP2014041043A (ja) * 2012-08-22 2014-03-06 Rohto Pharmaceut Co Ltd 皮膚状態の評価方法
JP2014080386A (ja) * 2012-10-16 2014-05-08 Shiseido Co Ltd 美白方法及びメラニン合成抑制剤のスクリーニング方法
JP2014215286A (ja) * 2013-04-30 2014-11-17 株式会社シャネル化粧品技術開発研究所 メラニン生成又は色素沈着を調節するための候補化合物のスクリーニング方法
JP2017080341A (ja) * 2015-10-30 2017-05-18 株式会社 資生堂 美容装置

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008280271A (ja) * 2007-05-09 2008-11-20 Oriza Yuka Kk 皮膚光老化予防剤
CN103505374B (zh) * 2012-06-20 2016-02-10 郑锦芬 一种抗皮肤光老化制剂及其应用
CN103830118A (zh) * 2014-03-26 2014-06-04 清华大学 红景天苷在制备预防皮肤光老化护肤品的用途及使用方法
CN105998021B (zh) * 2016-05-18 2019-04-05 西安交通大学第一附属医院 甲基莲心碱防治皮肤光老化的应用
CN106726695A (zh) * 2017-02-11 2017-05-31 佛山市芊茹化妆品有限公司 一种抗光老化面贴膜
KR101934793B1 (ko) * 2017-05-26 2019-01-03 주식회사 아토큐앤에이 고욤나무 잎 추출물을 유효성분으로 함유하는 피부 광노화 예방 또는 개선용 조성물

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2009268640A (ja) * 2008-05-02 2009-11-19 Kao Corp 皮膚内部の弾性計測方法
JP2012229924A (ja) * 2011-04-25 2012-11-22 Shiseido Co Ltd 過酸化脂質の定量分析方法
JP2013083504A (ja) * 2011-10-07 2013-05-09 Pola Chem Ind Inc スクリ−ニング方法
JP2014041043A (ja) * 2012-08-22 2014-03-06 Rohto Pharmaceut Co Ltd 皮膚状態の評価方法
JP2014080386A (ja) * 2012-10-16 2014-05-08 Shiseido Co Ltd 美白方法及びメラニン合成抑制剤のスクリーニング方法
JP2014215286A (ja) * 2013-04-30 2014-11-17 株式会社シャネル化粧品技術開発研究所 メラニン生成又は色素沈着を調節するための候補化合物のスクリーニング方法
JP2017080341A (ja) * 2015-10-30 2017-05-18 株式会社 資生堂 美容装置

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BECKER, L. ET AL.: "Age-dependent shift on Macrophage Polarisation Causes Inflammation- Mediated Degeneration of Enteric Nervous System", GUT, vol. 67, no. 5, May 2018 (2018-05-01), pages 827 - 836, XP055750666, Retrieved from the Internet <URL:https://pubmed.ncbi.nlm.nih.gov/28228489> [retrieved on 20200630] *
MANABE, ICHIRO: "Inflammaging and age-associated diseases", vol. 54, no. 2, 2017, pages 105 - 113, XP055750665, Retrieved from the Internet <URL:https://doi.org/10.3143/geriatrics.54.105> *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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