WO2012144080A1 - Agent d'amélioration de la fonction de barrière de la peau - Google Patents

Agent d'amélioration de la fonction de barrière de la peau Download PDF

Info

Publication number
WO2012144080A1
WO2012144080A1 PCT/JP2011/059985 JP2011059985W WO2012144080A1 WO 2012144080 A1 WO2012144080 A1 WO 2012144080A1 JP 2011059985 W JP2011059985 W JP 2011059985W WO 2012144080 A1 WO2012144080 A1 WO 2012144080A1
Authority
WO
WIPO (PCT)
Prior art keywords
trpv4
skin
receptor activator
trpv
barrier function
Prior art date
Application number
PCT/JP2011/059985
Other languages
English (en)
Japanese (ja)
Inventor
真琴 富永
隆彰 曽我部
尚子 木田
大場　愛
金丸　晶子
福田　寿之
Original Assignee
大学共同利用機関法人自然科学研究機構
ポーラ化成工業株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 大学共同利用機関法人自然科学研究機構, ポーラ化成工業株式会社 filed Critical 大学共同利用機関法人自然科学研究機構
Priority to JP2013510839A priority Critical patent/JP5737663B2/ja
Priority to PCT/JP2011/059985 priority patent/WO2012144080A1/fr
Publication of WO2012144080A1 publication Critical patent/WO2012144080A1/fr

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
    • C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
    • C07H17/08—Hetero rings containing eight or more ring members, e.g. erythromycins
    • 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
    • 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
    • A61K8/602—Glycosides, e.g. rutin
    • 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
    • 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
    • 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

Definitions

  • the present invention relates to a skin barrier function improving agent and a skin external preparation for improving skin barrier function, and more particularly, to a TRPV (Transient Receptor-potential vanilloid) receptor activator and a skin external preparation containing the same.
  • TRPV Transient Receptor-potential vanilloid
  • the skin has a barrier function to prevent the evaporation of moisture and protect the body from external and physical stimuli in addition to transmitting external stimuli such as heat and pain to the living body.
  • the skin barrier function plays an important role in maintaining the functions of the living body, but it is known that the function decreases due to temperature, humidity, excessive skin friction, lifestyle rhythm modulation, stress, etc. Due to various causes, the skin becomes dry, becomes hypersensitive to irritation, and is susceptible to irritation. Furthermore, if this state progresses, it causes direct damage and allergic reaction due to invasion of bacteria and harmful substances from outside the body, leading to the onset of skin disorders such as atopic dermatitis and sebum deficiency eczema.
  • Patent Document 1 a skin barrier function improving agent
  • Patent Document 2 Various attempts have been made such as a skin roughening agent (see, for example, Patent Document 2), a skin external preparation containing a ceramide synthesis accelerator (see, for example, Patent Document 3), and the like.
  • the non-selective cation channel TRP (Transient receptor potential) channel identified as the causative gene of Drosophila photoreceptor mutants in 1989, is classified into at least nine subfamilies based on amino acid sequence homology. ing.
  • the temperature-sensitive receptors belonging to the TRP channel have been first elucidated for the molecular entity of TRPV1.
  • nine types of temperature-sensitive receptors (TRPV1, TRPV2) opened by respective temperature stimuli from low temperature to high temperature. , TRPV3, TRPV4, TRPM2, TRPM4, TRPM5, TRPM8, TRPA1) have been reported.
  • TRPV4 was originally discovered as an osmotic sensitive receptor activated with low osmotic pressure, and is expressed in various tissues such as sensory nerve, hypothalamus, skin, kidney, lung, inner ear, osmotic stimulation, mechanical It plays an important role in the transmission of pain caused by stimulation and inflammatory pain. Moreover, TRPV4 functioning as a transmembrane calcium influx channel is not only a physical stimulus, but also an endogenous cannabinoid, arachidonic acid metabolite, 4 ⁇ -phorbol ester (4 ⁇ -PDD) (4alpha-phorbol 12,13- It is also activated by compounds such as didecanoate).
  • 4 ⁇ -PDD 4 ⁇ -phorbol ester
  • TRPV4 is considered to exhibit many biological activities from the diversity of expressed tissues and stimulation responses, and is deeply involved in neurological disease symptoms including stimulation and pain recognition and autism.
  • activities related to energy metabolism such as significant weight-inhibitory effects on body weight gain, total fat mass and visceral fat mass (see, for example, Patent Document 4) have been reported in TRPV4-deficient mice.
  • TRPV4 which is present in epidermal cells, has a function as a moisture transpiration sensor, and activation by 4 ⁇ -phorbol ester suppresses transpiration and enhances the skin barrier function. has been reported (for example, see Non-Patent Document 1 and Non-Patent Document 2).
  • TJ and adherence-junction are belt-shaped around cells, and close the gap by adjoining adjacent cells, and are continuous. It is an intercellular adhesion structure that binds cells to each other.
  • TJ is present in the epidermal cells of the granule layer and plays an important role in preventing water and substances from permeating through the cell gap and maintaining the skin barrier function (for example, non-patented Reference 3).
  • AJ adherence-junction
  • TRPV4 agonists such as biphenyl derivatives (see, for example, Patent Document 2), 1,4-diamine derivatives (see, for example, Patent Document 3), piperidine, and pyrrolidine.
  • TRPV4 agonists see, for example, Patent Document 5, Patent Document 6, and Patent Document 7 of derivatives and thiophene derivatives (see, for example, Patent Document 4) relieve symptoms such as pain, rheumatoid arthritis, neuropathy, and hyperalgesia It is known to be effective for improvement.
  • TRPV4 agonist has an action to improve skin barrier function, that activation by 4 ⁇ -phorbol ester suppresses water transpiration and enhances skin barrier function (for example, non-patent literature). 1, see Non-Patent Document 2), and its mechanism of action has not been elucidated.
  • a component that acts on both TJ and AJ and TRPV of the intercellular adhesion structure, particularly, a component having an action of promoting the formation of TJ and / or AJ by activating TRPV4 present in the epidermis is a skin barrier. It has not been known to improve the function and be effective in improving rough skin.
  • 3-O-methylellagic acid 4′-O- ⁇ -L-rhamnopyranoside (hereinafter sometimes referred to as “compound 1”), 3,3′-di-O-methylellagic acid 4′-O- ⁇ -D -Glucopyranoside (hereinafter sometimes referred to as “compound 2”), 3,4,3′-tri-O-methylellagic acid 4′-O- ⁇ -D-glucopyranoside (hereinafter referred to as “compound 3”)
  • 3′-O-methyl-3,4-methylenedioxyellagic acid 4′-O- ⁇ -D-glucopyranoside (hereinafter sometimes referred to as “compound 4”) is a known compound.
  • Non-Patent Document 4 Non-Patent Document 5 for Compound 1
  • Non-Patent Document 6 for Compound 2
  • Non-Patent Document 7 for Compound 3
  • Non-patent for Compound 4 Reference 8 describes.
  • whitening action, stain and dullness improvement action (Non-patent Documents 9 and 10) for compound 1
  • nitric oxide release promoting action or TNF- ⁇ release for compound 2 Promoting action (Non-patent document 11)
  • Nitric oxide release promoting action or TNF- ⁇ release promoting action Ni-patent document 11 for compound 3
  • Antioxidant action Non-patent document 8, Non-patent document 8, 12
  • these compounds have never been studied for TRPV receptor activation, AJ and / or TJ formation promotion, skin barrier function improvement, and the like.
  • the present invention has been made under such circumstances, and is to provide a novel TRPV receptor activator and a skin external preparation containing the activator, which are suitable for improving the skin barrier function. .
  • the present inventors have sought for a skin external preparation suitable for improving the skin barrier function, and as a result of intensive efforts, have made a novel TRPV receptor activator and skin containing the same. It has been found that an external preparation has such characteristics. Furthermore, as a result of further detailed investigation, it was found that a plant extract obtained from a plant belonging to the genus Sarsbergi genus or the genus Rubiaceae belonging to the genus Rubiaceae belongs to the TRPV receptor activation activity. A compound having an activating action was isolated to complete the invention.
  • the present invention is as follows.
  • a TRPV receptor activator comprising one or more selected from the compounds represented by the following formula, or a plant extract containing an effective amount of the same compound as a TRPV receptor activator.
  • the hydroxyl group may be substituted with a methoxy group or an ethoxy group, and the methoxy group may be substituted with a hydroxyl group or an ethoxy group.
  • the compound is 3-O-methylellagic acid 4′-O- ⁇ -L-rhamnopyranoside, 3,3′-di-O-methylellagic acid 4′-O- ⁇ -D-glucopyranoside, 3,4, 3′-tri-O-methylellagic acid 4′-O- ⁇ -D-glucopyranoside or 3′-O-methyl-3,4-methylenedioxyellagic acid 4′-O- ⁇ -D-glucopyranoside, 1>
  • the TRPV receptor activator according to 1>.
  • TRPV receptor activator according to ⁇ 1> or ⁇ 2>, wherein the extract is an extract obtained from a plant belonging to the genus Sarsberga or a plant belonging to the genus Rubiaceae.
  • ⁇ 5> The TRPV receptor activator according to any one of ⁇ 1> to ⁇ 4>, wherein the TRPV receptor is TRPV4.
  • ⁇ 6> A skin external preparation containing an effective amount of the TRPV receptor activator according to any one of ⁇ 1> to ⁇ 5>.
  • ⁇ 7> The skin external preparation according to ⁇ 6>, wherein the effective amount is 0.00001 to 10% by mass as the amount of the compound with respect to the total amount of the skin external preparation.
  • ⁇ 8> The external preparation for skin according to ⁇ 6> or ⁇ 7>, which is for improving the skin barrier function.
  • ⁇ 9> The external preparation for skin according to any one of ⁇ 6> to ⁇ 8>, which is used for promoting the formation of tight junctions and / or adherence junctions.
  • ⁇ 10> The external preparation for skin according to any one of ⁇ 6> to ⁇ 9>, which is a cosmetic or a quasi drug.
  • ⁇ 11> One or more selected from the compounds represented by the following formula, or a TRPV receptor activator comprising a plant extract containing an effective amount of the same compound as a TRPV receptor activator, A method for improving the skin barrier function, comprising a step of applying to a site where the skin is required.
  • hydroxyl group may be substituted with a methoxy group or an ethoxy group, and the methoxy group may be substituted with a hydroxyl group or an ethoxy group.
  • the hydroxyl group may be substituted with a methoxy group or an ethoxy group, and the methoxy group may be substituted with a hydroxyl group or an ethoxy group.
  • a novel TRPV receptor activator and a skin external preparation containing the activator can be provided.
  • TRPV4 activator (4 (alpha) -PDD) by TER value measurement It is a figure which shows TJ and / or AJ formation promotion effect of TRPV4 activator (4 ⁇ -PDD) by FITC-Dextran permeation amount measurement. It is a figure which shows TRPV4 knockdown efficiency by si-RNA transfection. It is a figure which shows the expression level of the cell adhesion related gene of a normal human newborn foreskin epidermal keratinocyte (NHEK) by TRPV4 knockdown. It is a figure which shows the time-dependent change of the TER value in a TRPV4 knockdown cell.
  • NHEK normal human newborn foreskin epidermal keratinocyte
  • FIG. It is a figure which shows the increase effect (control) of the intracellular calcium ion concentration of the compound 3. It is a figure which shows the result of the TER value measurement (TJ and / or AJ shape promotion effect) of compound 1. It is a figure which shows the result of TER value measurement (TJ and / or AJ shape promotion effect) of compound 3. It is a figure which shows the result of the TER value measurement (TJ and / or AJ shape promotion effect) of compound 4. It is a figure which shows the increase effect
  • TRPV receptor activator of the present invention Nine types of temperature-sensitive receptors (TRPV1, TRPV2, TRPV3, TRPV4, TRPM2, TRPM4, TRPM5, TRPM8, and TRPA1) that are opened by a temperature stimulus from a low temperature to a high temperature are known as TRP channels. Of these, TRPV1, TRPV3, and TRPV4 have been reported to be present in epidermal cells. TRPV is a molecular entity of a transmembrane calcium ion channel, and is considered to be deeply involved in the skin barrier function by adjusting the intracellular calcium ion concentration. In particular, it is suggested that TRPV1 and TRPV4 are deeply involved in moisture transpiration and skin barrier function.
  • the TRPV receptor activator of the present invention is a receptor activator that acts directly or indirectly on TRPV.
  • the TRPV receptor activator of the present invention is a receptor activator that acts on TRPV4.
  • the TRPV receptor activator of the present invention has an action of activating the receptor.
  • the TRPV receptor activator of the present invention adjusts the concentration of calcium ions in intercellular lipids and epidermal cells, and improves the skin barrier function.
  • the TRPV receptor activator of the present invention exerts a TJ and / or AJ formation promoting action by directly or indirectly acting on TRPV4 and improves the skin barrier function.
  • the TRPV receptor activator of the present invention is a substance that exerts a TJ and / or AJ formation promoting action through the TRPV4 activation action, and may exhibit an additive or synergistic skin barrier function improving action. Be expected.
  • the TRPV receptor activator of the present invention may have an effect other than the TRPV receptor activation effect.
  • a TRPV receptor activator can be used even if it has an effect other than TRPV receptor activation as long as the TRPV receptor activation effect is not hindered.
  • the TRPV receptor activator of the present invention comprises one or more selected from the compounds represented by the following formula, or a plant extract containing an effective amount of the same compound as a TRPV receptor activator. is there.
  • the hydroxyl group may be substituted with a methoxy group or an ethoxy group, and the methoxy group may be substituted with a hydroxyl group or an ethoxy group.
  • Preferred examples of the compound include 3-O-methylellagic acid 4′-O- ⁇ -L-rhamnopyranoside (compound 1), 3,3′-di-O-methylellagic acid 4′-O- ⁇ -D-glucopyranoside ( Compound 2), 3,4,3′-tri-O-methylellagic acid 4′-O- ⁇ -D-glucopyranoside (Compound 3), or 3′-O-methyl-3,4-methylenedioxyellagic acid 4 '-O- ⁇ -D-glucopyranoside (Compound 4).
  • the compound include a chemical substance purified and isolated from a plant containing the compound, and an extract containing an effective amount of the compound derived from animals and plants as a TRPV receptor activator. Only seeds can be contained, or two or more kinds can be contained in combination.
  • the animal or plant-derived extract of the present invention is an animal or plant-derived extract, specifically, the extract itself, a fraction of the extract, a purified fraction, an extract or a fraction, purification. This is a general term for the product from which the solvent is removed.
  • preferred examples include extracts obtained from plants belonging to the genus Cranaceae and Rubiaceae, and more preferable are extracts of Cranalis genus C., Rubiaceae ganbi- A plant extract obtained from the above can be suitably exemplified.
  • the giant crape myrtle is a evergreen small Takagi native to tropical Asia. In Japan, it is cultivated in Okinawa and Kyushu.
  • the leaf of the giant leafhopper is used to produce banaba tea by drying the leaves and used as a herbal medicine, a supplement, and the like.
  • the Rubiaceae genus Gambir is a plant native to the coast of the Malacca Strait.
  • the dry water extract obtained from Gambir leaves is called asenyaku (Asenyaku), which has anti-fibrinolytic activity and antithrombotic activity. The action, the antitumor action, etc. are known.
  • the extract obtained from the above-mentioned plant in Japan can be produced using a plant grown in Japan or sold in Japan as a herbal medicine raw material, etc.
  • Commercial extracts sold by companies that handle plant extracts such as companies can be purchased and used.
  • the plant part used for producing the extract obtained from the plant is not particularly limited, and whole grass can be used.
  • the plant body, the above-ground part, the rhizome part, the tree trunk part, the leaf part, and the stem are used. It is also possible to use only parts such as parts, flower spikes and flower buds.
  • a part used for extraction of a plant body or the like is processed in advance so as to improve extraction efficiency by crushing or chopping.
  • an extract 1 to 30 parts by mass of a solvent is added to 1 part by mass of a plant used for extraction of a plant or the like, and several days at room temperature or several at a temperature near the boiling point. Immerse for hours. After the immersion, the solution can be cooled to room temperature, insoluble matter can be removed if desired, and then the solvent can be removed by concentration under reduced pressure. Thereafter, the desired extract can be obtained by fractional purification by column chromatography packed with silica gel or ion exchange resin. The above compound is obtained by subjecting the extract obtained as described above to a water extraction method or an organic solvent extraction method, if desired, and preparative purification using column chromatography packed with silica gel or an ion exchange resin. Can be obtained.
  • the extraction solvent is preferably a polar solvent, and alcohols such as water, ethanol, isopropyl alcohol, and butanol, and polyvalent alcohols such as 1,3-butanediol and polypropylene glycol.
  • alcohols such as water, ethanol, isopropyl alcohol, and butanol
  • polyvalent alcohols such as 1,3-butanediol and polypropylene glycol.
  • Preferred examples include one or two or more selected from ketones such as amino acids, acetone and methyl ethyl ketone, and ethers such as diethyl ether and tetrahydrofuran.
  • mineral salts such as hydrochloride, sulfate, nitrate, phosphate, carbonate, maleate, fumarate, oxalate, citrate, lactate, tartrate, methanesulfonate, para Organic acid salts such as toluene sulfonate and benzene sulfonate, alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as calcium salt and magnesium salt, triethylamine salt, triethanolamine salt, ammonium salt, Suitable examples include organic amine salts such as monoethanolamine salts and piperidine salts, and basic amino acid salts such as lysine salts and alginates.
  • any method capable of evaluating the TRPV activation action can be applied without particular limitation.
  • a method for evaluating the activation effect of TRPV using the calcium ion concentration as an index is preferred.
  • a method for evaluating the activation effect of TRPV4 is preferred. This is because TRPV4 has been confirmed to exist in epidermal cells and is expected to be involved in the skin barrier function.
  • the activity of TRPV4 is measured using the intracellular calcium ion concentration in cells in which TRPV4 expression is confirmed, more preferably in cells overexpressing TRPV4 as an index.
  • a method for discriminating the chemical action is preferred.
  • any method can be used without particular limitation as long as it is a method for measuring the intracellular calcium ion concentration.
  • Drosophila painless is a Ca 2+ -requiring channel activated by noxious heat .
  • J Neurosci., 2008 Oct 1; 28 (40): 9929-38 preferably the calcium imaging method or the patch clamp method.
  • the intracellular calcium ion concentration in cells in which TRPV4 expression has been confirmed or cells overexpressing TRPV4 is confirmed by the calcium imaging method or patch clamp method, and the like.
  • the degree of calcium ion concentration is increased in the presence of the test substance compared to the absence of the test substance (control)
  • This efficacy is preferably 100% in the presence of the test substance in the presence of the test substance, compared to 100% in the presence of the test substance, compared to the increase in intracellular calcium ion concentration in the absence of the test substance.
  • an increase in concentration of 20% or more, more preferably an increase of concentration of 30% or more is observed, it is determined to be effective.
  • the improvement effect of the skin barrier function is remarkable as the degree is remarkable.
  • the increase rate of intracellular calcium ion concentration by the test substance relative to ionomycin As for the increase rate of intracellular calcium ion concentration by the test substance relative to ionomycin (positive control), 3 or 5 cells whose intracellular calcium ion concentration increased by addition of the test substance were selected at random, and each test substance was selected.
  • the initial value, ie, the calcium ion concentration before addition is subtracted from the calcium ion concentration after addition to calculate the amount of increase in calcium ion concentration due to the test substance, and then the initial value, ie, addition, from the calcium ion concentration after addition of ionomycin (positive control).
  • TRPV overexpressing cells are used.
  • TRPV1 overexpressing cells for example, according to the method described in JP-A-2009-082053, and for the production of TRPV4 overexpressing cells, TRPV4 overexpression is carried out by the method described below based on the above method. Cells can be made.
  • any cell expressing TRPV4 can be applied without particular limitation, and more preferable is a TRPV4 expression vector.
  • Cells in which TRPV4 is overexpressed by introducing ⁇ into a host cell are preferred.
  • host cells overexpressing TRPV4 include bacterial cells, plant cells, animal cells, insect cells, etc. More preferably, HEK293 cells, CHO cells, COS-7 cells, NIH3T3 cells and the like are suitable. It can be illustrated.
  • the host cell preferably incorporates a TRPV4 expression vector, allows TRPV4 to be efficiently expressed, and is easy to culture.
  • a cDNA encoding TRPV4 can be used without any particular limitation. More preferred is a cDNA cell vector pcDNA3 (manufactured by Invitrogen). And a nucleic acid obtained by ligating a TRPV4-encoded nucleic acid.
  • nucleic acid encoded by TRPV4 a nucleic acid encoded by all nucleotide sequences can be used, for example, an mRNA extracted and converted into cDNA by allowing reverse transcriptase to act on it, It is also possible to select a suitable primer for only the portion where the TRPV4 main function is coded, amplify it by PCR, and use this to ligate. Particularly preferred is the oligonucleotide shown in SEQ ID NO: 1, which can be obtained by subjecting the DNA or cDNA extracted using the primers of SEQ ID NO: 2 and SEQ ID NO: 3 to a PCR reaction. ,Obtainable.
  • the cells overexpressing TRPV4 that can be used for evaluation of the component having TRPV activating action of the present invention are characterized by the functional expression of TRPV4 in the cells, expression at the protein level, co-introduction of fluorescent substances, and the like. , You can choose.
  • An appropriate selection medium can be used for selection of a cell that has been introduced into a host cell so that the nucleic acid encoding TRPV4 can be expressed in the cell.
  • a material obtained by adding a growth factor such as FBS to DMEM or RPMI medium can be suitably exemplified.
  • Examples of the medium used for culturing cells that overexpress TRPV4 include components suitable for growth of cells that overexpress TRPV4, such as glucose, amino acids, peptones, vitamins, cell growth promoting factors (for example, , Cell growth factors, hormones, binding proteins, cell adhesion factors, lipids), serum (eg, FBS, FCS, etc.), calcium chloride, magnesium chloride, etc.
  • the medium may be a commercially available medium.
  • the medium used for culturing cells overexpressing TRPV4 is not particularly limited as long as it is a medium suitable for such cells, and examples thereof include MEM medium, DMEM medium, RPMI 1640 medium, and the like. For example, when the host cell used is HEK293 cells, a DMEM medium containing high glucose and 10% by mass FBS is used.
  • the TRPV receptor activator in the present invention activates TRPV, particularly, TRPV4 present in epidermal cells, and thereby promotes the formation of TJ and / or AJ, thereby exhibiting a skin barrier function improving action.
  • the interaction regarding the improvement of the skin barrier function between TRPV (particularly TRPV4) and TJ and / or AJ of the cell-cell adhesion structure is confirmed by the following test results. That is, when the TRPV4 activation by TRPV4 ligand 4 ⁇ -PDD (4 ⁇ -phorbol ester) is confirmed to have the same action as TJ and / or AJ formation promoting action, it has the TRPV4 activation action. It can be said that the component exhibits the skin barrier function improving action by the TJ and / or AJ formation promoting action.
  • a biotin-labeled diffusion train using a three-dimensional epidermis model or living skin described in Non-Patent Document 3 is used.
  • TER transepithelial electric resistance
  • JP-A-2007-174931 a method for measuring transepithelial electric resistance described in JP-A-2007-174931 can be exemplified, but from the viewpoint of simplicity of operation and measurement accuracy, FITC- The dextran permeability test method and the method of measuring the transepithelial cell electrical resistance value (TER value) can be preferably exemplified.
  • the degree of intercellular mass transfer in the epidermal keratinized cell layer membrane constructed on the support is determined based on the intercellular substance permeability or transepithelial cell electrical resistance value.
  • TER value is measured, etc., and the degree of intercellular mass transfer is suppressed in the presence of the test substance compared to the absence of the test substance (control), or TER is in the absence of the test substance
  • control the degree of intercellular mass transfer is suppressed in the presence of the test substance compared to the absence of the test substance (control)
  • TER is in the absence of the test substance
  • control it is judged that the TJ and / or AJ of the epidermal keratinocyte layer membrane has been densely constructed, and as a result applied to the in vivo skin It is determined that the substance permeation suppressing action of the stratum corneum or the epidermal granule layer is improved, thereby improving the skin barrier function.
  • the external preparation for skin of the present invention is characterized by containing a TRPV receptor activator as an essential component.
  • the TRPV receptor activator in the present invention is a receptor activator that acts directly or indirectly on TRPV.
  • a TRPV4 receptor activator is a preferred example.
  • the TRPV receptor activator of the present invention has an effect of activating TRPV, particularly, TRPV4.
  • the TRPV receptor activator of the present invention may be any of purified and isolated chemical substances, extracts derived from animals and plants, mixed purified products such as fractionated purified products thereof.
  • the skin external preparation of the present invention may contain only one TRPV receptor activator or a combination of two or more thereof.
  • the skin external preparation of the present invention activates TRPV by blending a TRPV receptor activator, promotes the formation of TJ and / or AJ, and exhibits an effect of improving (improving) the skin barrier function.
  • the skin (between cells) barrier function improving effect is a barrier function of skin (or between skin cells) [for example, transdermal moisture transpiration suppression function (function to prevent moisture transpiration from skin) and substances This is an effect of improving the permeability suppression function (function of preventing the invasion of a substance into the skin, etc.), and can also be referred to as a rough skin prevention or improvement effect.
  • the TRPV receptor activator of the present invention is 0.00001% by mass to 10% by mass, more preferably 0.0001% by mass to 5% by mass, and still more preferably, as the total amount of the compound with respect to the total amount of the external preparation for skin. It is preferable to contain 0.001 to 3% by mass. This is because when the content of the TRPV receptor activator is too small, the TRPV activation action, and the skin barrier function improving action through the TJ and / or AJ formation promoting action by TRPV activation are not exhibited, This is because even if the amount is too large, the effect reaches a limit and the degree of freedom of the system may be impaired.
  • the external preparation for skin of the present invention can contain optional components usually used in medicines, quasi drugs, foods, and cosmetics, in addition to the essential components.
  • such optional ingredients include hydrocarbons such as squalane, petrolatum and microcrystalline wax, jojoba oil, carnauba wax, esters such as octyldodecyl oleate, olive oil, beef tallow, Triglycerides such as coconut oil, fatty acids such as stearic acid, oleic acid and retinoic acid, lower alcohols such as ethanol and isopropanol, higher alcohols such as oleyl alcohol, stearyl alcohol and octyldodecanol, sulfosucci Anionic surfactants such as acid esters and sodium polyoxyethylene alkyl sulfates, amphoteric surfactants such as alkylbetaine salts, cationic surfactants such as dialkylammonium salts, sorbitan fatty acid esters
  • Non-ionic surfactants such as lend adduct, polyoxyethylene alkyl ether, polyoxyethylene fatty acid ester, polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, polyethylene glycol, dipropylene glycol, glycerin, Contains polyhydric alcohols such as 1,3-butanediol, 1,2-pentanediol, thickening / gelling agents, antioxidants, UV absorbers, colorants, preservatives, powders, etc. be able to.
  • the external preparation for skin of the present invention can be produced without difficulty by treating these components according to a conventional method except that the TRPV receptor activator of the present invention is contained.
  • the skin external preparation of the present invention can be produced by treating these essential components and optional components according to a conventional method and processing them into lotions, emulsions, essences, creams, pack cosmetics, cleansing agents and the like.
  • any dosage form is possible, but since the active ingredient penetrates the skin and exerts its effect, the lotion and emulsion that are well-familiar with the skin , Cream, essence and the like are more preferable.
  • TER value measurement test using TRPV receptor activator> Frozen normal human epidermal keratinocytes (NHEK) (manufactured by Kurashiki Boseki Co., Ltd.) are thawed and cultured at 37 ° C. with 5% CO 2 in 0.15 mM-calcium ion-containing culture solution (Humdia-KG2: Kurashiki Boseki Co., Ltd.). Culturing was performed under a carbon stream.
  • NHEK normal human epidermal keratinocytes
  • TRPV3 and TRPV4 have been confirmed in epidermal cells that are deeply involved in the skin barrier function. From the results of FIG. 1, in the TRPV4 inactive temperature region, NHEK cultured in the presence of TRPV4 ligand 4 ⁇ -PDD has a statistically significant increase in TER value as compared to NHEK cultured in the absence. Was confirmed. On the other hand, in NHEK cultured in the presence of camphor of the TRPV3 ligand, an increase in TER value was not observed as compared with the absence. It was shown that the increase in the TER value, that is, the action of improving the skin barrier function by promoting TJ and / or AJ formation is exhibited by specific activation of TRPV4.
  • FITC-Dextran substance permeation test using TRPV receptor activator> the medium of the sample 9 hours after the medium exchange used for measurement was removed.
  • a standard curve group (100 mg / mL to 0 mg / mL) is prepared on an 96-well plate using an Apical Buffer.
  • the recovered basal buffer was added to each 96 wells in 200 mL, and then measured with a spectrophotometer (excitation 485/535 nm). The results are shown in FIG.
  • si-RNA transfection test> Frozen normal human epidermal keratinocytes (NHEK) (manufactured by Kurashiki Boseki Co., Ltd.) are thawed and cultured until they become sub-confluent, then detached and collected with trypsin EDTA, and 7.5 ⁇ 10 5 cells / cell in a 6-well plate. The cells were seeded in a well and cultured at 37 ° C. in a 5% carbon dioxide stream for 2 hours. In addition, si-RNA transfection (volume is described for 1 well) was performed according to the following procedure.
  • NHEK normal human epidermal keratinocytes
  • Opti-MEM 5 ⁇ L of 10 ⁇ M si-RNA (On TargetTplus SMARTpool L-004195-00-0005, Human TRPV4, NM 147204, Target Sequence: SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 and SEQ ID NO: 7 And then 15 ⁇ L of HiperFect was added and mixed by vortexing, and then allowed to stand at room temperature for 10 minutes. After dripping into the well and diffusing gently, culturing at 37 ° C.
  • TRPV4 knockdown efficiency by si-RNA was determined by performing RT-PCR using Hs TRPV4 1 SG QuantiTect Primer Assay (Qiagen: QT00077217, sequence not disclosed) as a primer to determine the expression level of TRPV4 mRNA. confirmed. The results are shown in FIGS. These si-RNA transfection tests were performed using a commercially available si-RNA ON-TARGET plus SMART pool Human TRPV4 (manufactured by Thermo scientific).
  • the results of the above tests 1 to 3 indicate that activation of TRPV, especially, TRPV4 promotes the formation of TJ and / or AJ and improves the skin barrier function.
  • a component having a TRPV receptor activating action exerts a skin barrier function improving action through a TJ and / or AJ formation promoting action and is useful as a skin barrier function improving agent.
  • TRPV4 Expression of TRPV4 in human normal epidermal keratinocytes>
  • NHEK human normal epidermal keratinocytes
  • polyAb anti- ⁇ -catenin Chemicon
  • mAb anti-E-cadherin Takara Bio
  • polyAb anti-TRPV4 prepared against the N-terminal peptide of TRPV4
  • NHEK membrane fraction was immunoprecipitated (IP) using an anti- ⁇ -catenin antibody, and a sample of the resulting complex was separated and immunoblotted (IB) using an anti-N-terminal TRPV4 antibody.
  • Is. Black triangles indicate the position of TRPV4 co-precipitated with ⁇ -catenin.
  • the right lane is replotted with anti-E-cadherin antibody.
  • Black triangles indicate E-cadherin. That is, TRPV4 was coimmunoprecipitated with ⁇ -catenin and E-cadherin present in the cell membrane.
  • ⁇ -catenin and E-cadherin are the main constituent proteins of AJ, suggesting that TRPV4 is present as part of the AJ complex in human keratinocytes.
  • Rho family of GTPases has been reported to play an important role in the differentiation of epidermal keratinocytes.
  • the Rho family is activated by increasing intracellular calcium ion concentration and mediates keratinocyte differentiation including actin formation, intercellular adhesion formation and superficial actin formation.
  • the TRP ion channel causes calcium ions to flow from outside the cell into the cell and promotes an increase in the intracellular calcium ion concentration.
  • the expression level of active Rho was evaluated in NHEK cultured at 33 ° C. where TRPV3 and TRPV4 are activated and 28 ° C.
  • NHEK was cultured at 33 ° C. or 28 ° C. for 24 hours. Thereafter, NHEK cultured at 28 ° C. was treated with 10 ⁇ M 4 ⁇ -PDD or 3 mM camphor and cultured for 8 hours. For each of these NHEKs, the expression level of active Rho was evaluated using Western blotting. The results are shown in FIG. The expression level of active Rho is shown in the upper panel. 15 ⁇ g of total protein was loaded as input (lower panel). The expression of the active form of NHEK cultured at 28 ° C. was lower than the expression of the active form of NHEK cultured at 33 ° C.
  • the expression level of active Rho at 28 ° C. was remarkably increased by the addition of 4 ⁇ -PDD, which is a TRPV4 activator, but not increased by camphor, which is a TRPV3 activator. This indicates that below the activation temperature threshold of TRPV3 and TRPV4 is insufficient to enhance Rho activity, but is specifically compensated by chemical activation of TRPV4, and TRPV4 mediated intracellular calcium It was suggested that ion influx is involved in Rho activation.
  • occludin a TJ-related protein
  • mAb anti-Occludin manufactured by Invitrogen
  • FIG. 1 One of the TJ-related proteins, occludin (red in the original figure), was localized along the intercellular adhesion in NHEK cultured at 33 ° C. 48 hours after differentiation induction, whereas it was 28 ° C. In, the localization of occludin at the cell-cell junction was intermittent.
  • ⁇ Test Example 6 Effect on barrier function in human keratinocytes and human skin tissue by temperature change and TRPV4 activation>
  • the barrier function in the skin is roughly divided into a stratum corneum barrier function and an epidermal cell (TJ) barrier function.
  • TJ epidermal cell
  • TER transepithelial electrical resistance
  • Millicell- Measurements were made using ERS (Millipore). NHEK subjected to differentiation induction by a calcium ion switch was cultured at 33 ° C. ( ⁇ ) or 28 ° C.
  • NHEK cultured at 28 ° C. was added to 4 ⁇ -PDD ( ⁇ ) or camphor ( ⁇ ) was added to enhance TRPV4 and TRPV3 activation.
  • the results are shown in FIG. The arrow indicates the timing of enhancement.
  • Data are the mean ⁇ SEM of 5 independent experiments. D. It shows with. (**: P ⁇ 0.01, NS: not significant.)
  • a significant difference test was performed by the Bonferroni method. A value of P ⁇ 0.05 was considered significant.
  • the human skin tissue used at this time was the skin tissue (Biopredic and Juntendo Urayasu Hospital) excised by surgical operation after obtaining the consent of the patient through informed consent. These human skin tissues are pre-cultured using Skin Long Term Culture Medium (manufactured by Biopredic), and then stripped with C40SH02 adhesive tape (manufactured by Seikagaku Corporation) to completely remove the stratum corneum. As a result, the barrier was destroyed.
  • FIG. (A) shows the barrier recovery rate of the skin tissue at 1.5, 4 and 24 hours after barrier destruction. Each label indicates the following. ⁇ : 33 ° C. water administration, ⁇ : 28 ° C. water administration, ⁇ : 28 ° C.
  • FIG. (B) shows an immunofluorescent image of a cross section of skin tissue 24 hours after destruction of the stratum corneum barrier.
  • the arrow indicates occludin (green in the original figure), which is one of the major TJ-related proteins localized in the epidermal granule layer.
  • a white triangle indicates a substance permeation marker (red in the original drawing) that has passed through the position of occludin.
  • permeation of the substance permeation marker was blocked at the outermost layer of the epidermal granular layer where occludin was localized.
  • TRPV4 regulates not only the TJ barrier function but also the stratum corneum barrier function. From the above results, that is, at a low temperature below the TRPV4 activation temperature threshold, it was shown that recovery of the skin barrier is delayed, but is specifically compensated by activation of TRPV4.
  • the plant extract obtained from the genus Coleoptera can be produced according to the quasi-drug raw material standard 2006, or a plant extract commercially available from Maruzen Pharmaceutical Co., Ltd., Yamakawa Trading Co., Ltd., etc. can be purchased. Can also be used.
  • a plant extract obtained from Yamakawa Trading Co., Ltd. and obtained from the genus Coleoptera was cultivated. After concentrating the plant extract (9950 g, manufactured by Yamakawa Trading Co., Ltd.) obtained from the crape myrtle family, the mixture is divided into liquid and liquid, and the resulting aqueous layer fraction is diaionized.
  • the column adsorbing part was sequentially eluted with 50% methanol and 100% methanol (manufactured by Wako Pure Chemical Industries, Ltd.) through HP-20 (manufactured by Mitsubishi Chemical Corporation).
  • the substance peak was isolated by preparative HPLC with respect to 100% methanol elution (745 mg) out of the elution part.
  • the sorting conditions are as follows.
  • Example 2 ⁇ Evaluation test of intercellular barrier function of compounds 1 to 4> Compounds 1 to 4 purified according to the above procedure were subjected to a TRPV4 activating effect evaluation test by calcium imaging. The results for compounds 1 and 3 are shown in FIGS.
  • FIGS. the results of the intercellular barrier function evaluation test of normal human epidermal keratinocytes by TER measurement performed on the compounds 1, 3, and 4 according to the test method described in Test Example 1 are shown in FIGS. .
  • the vertical axis represents the TER value ( ⁇ ⁇ m 2 ), and the horizontal axis represents the TER measurement time after differentiation induction by the calcium switch. From these results, the compound of the present invention showed a marked increase in TER value as compared with the control DMSO-added group, and the effect of promoting the intercellular barrier function was recognized.
  • the plant extract of the present invention (the extract obtained from the honey beetle genus Coleoptera or Rubiaceae genus Gambir) can be produced according to the quasi-drug raw material standard 2006, or is commercially available from Maruzen Co., Ltd. It is also possible to purchase and use plant extracts. In this example, a plant extract purchased from Maruzen Co., Ltd. was used.
  • Human TRPV4-expressing cells were prepared according to the following experimental procedure. PCR was performed using cDNA encoding human TRPV4 [SEQ ID NO: 1 (GenBank accession number: NM 021625) 90 bp to 2705 bp polynucleotide] and oligonucleotides SEQ ID NO: 2 and SEQ ID NO: 3 as primers. The product was amplified and inserted into the MCS BamHI and XhoI sites of the product name: pcDNA3 (manufactured by Invitrogen), which is a vector for mammalian cells, to obtain a human TRPV4 expression vector.
  • pcDNA3 manufactured by Invitrogen
  • Human TRPV4 expression vector obtained (corresponding to 0.5 ⁇ g) and DsRed (corresponding to 0.1 ⁇ g), positive reagent (trade name, catalog number: 11514-015, manufactured by Invitrogen) 6 ⁇ L, OPTI-MEM (registered) (Trademark) I Reduced-Serum Medium (catalog number: 11058021, manufactured by Invitrogen) 100 ⁇ L was mixed to obtain a mixture 1. Further, 4 ⁇ L of Lipofectamine (registered trademark, catalog number: 18324-012, manufactured by Invitrogen) and 100 ⁇ L of OPTI-MEM were mixed to obtain a mixture 2.
  • HEK293 cells (5 ⁇ 10 5 cells / 35 mm diameter dish) were cultured in DMEM medium containing 10% by mass FBS to 37% confluence at 37 ° C. in a 5% carbon dioxide stream. Thereafter, the mixture of the mixture 1 and the mixture 2 was added to the obtained cells.
  • the human TRPV4 expression vector was introduced into HEK293 cells to obtain TRPV4-expressing cells. Further, in the same manner, a vector without human TRPV4 inserted was prepared and introduced into HEK293 cells to prepare Mock cells.
  • TRPV4-expressing cells or Mock cells prepared according to Example 4 Incubation of TRPV4-expressing cells or Mock cells prepared according to Example 4 in a DMEM medium containing 10% FBS containing 1-20 ⁇ g / ml FURA 2-AM (manufactured by Invitrogen) for 60-90 minutes ( FURA 2-AM was introduced into TRPV4-expressing cells or Mock cells.
  • TRPV4-expressing cells or Mock cells after introduction of FURA 2-AM were placed in a chamber (RC-26G; manufactured by Warner Instruments) of an inverted microscope of calcium imaging apparatus and washed with solution A.
  • the fluorescence intensity at an excitation wavelength of 340 nm and the fluorescence intensity at an excitation wavelength of 380 nm were measured while circulating the test solution in a chamber. Thereafter, a fluorescence intensity ratio (fluorescence intensity at 340 nm / fluorescence intensity at 380 nm) between the fluorescence intensity at the excitation wavelength of 340 nm and the fluorescence intensity at the excitation wavelength of 380 nm when the test solution was used was calculated.
  • the enhancement effect of the test substance on the change in intracellular calcium ion concentration was analyzed by IPLab software (manufactured by Scanalytics). Further, the same evaluation was performed using 4 ⁇ -phorbol ester (4 ⁇ -PDD) which is a TRPV4 ligand as a positive control. The results are shown in FIGS.
  • the positive control 4 ⁇ -PDD showed a marked increase in intracellular calcium ion concentration, confirming the objectivity of this evaluation system.
  • the plant extract of the present invention significantly increased intracellular calcium ion concentration in TRPV4-expressing cells.
  • Mock cells FIGGS. 23 and 24.
  • the Acacia yak extract as a plant extract of the present invention was added to the calcium ion free solution B [composition: 140 mM NaCl, 5 mM KCl, 2 mM MgCl 2 , 5 mM EGTA, 10 mM glucose, 10 mM HEPES (pH 7.4)].
  • Example 6 ⁇ Evaluation of TJ and AJ formation promoting action 1: Measurement of TER value of plant extract in the present invention> According to the test method described in Test Example 1, the TER value of the plant extract of the present invention was measured. The results are shown in FIG. In FIG. 29, the vertical axis represents the TER value ( ⁇ m 2 ), and the horizontal axis represents the measurement time. From the results shown in FIG. 29, it was confirmed that the plant extract of the present invention exhibited a TJ and / or AJ function promoting effect showing a marked increase in TER value.
  • Example 7 FITC-Dextran substance permeation test of plant extract in the present invention> According to the test method described in Test Example 2 above, FITC-Dextran substance permeability of the plant extract of the present invention was evaluated. The results are shown in FIG. From the results shown in FIG. 30, the plant extract of the present invention showed a remarkable suppression of FITC-Dextran substance permeability and an action of promoting the formation of TJ and / or AJ.
  • the plant extract of the present invention showed both TJ and AJ formation in the TJ and / or AJ formation promoting activity evaluation (TER value measurement and FITC-Dextran substance permeation test). It showed a promoting effect. From the above results, the plant extract of the present invention has a TRPV activation action and a TJ and / or AJ formation promoting action, and a skin barrier by the TJ and / or AJ formation promoting action via the TRPV activation action. Has a function improvement effect.
  • Example 8 ⁇ Composition of the present invention> ⁇ Manufacture of a composition (skin external preparation) containing a component having an action of improving the skin barrier function of the present invention>
  • a lotion cosmetic which is a composition (external preparation for skin) containing a component having an action of improving the skin barrier function of the present invention
  • the prescription ingredients were stirred at 80 ° C. to solubilize, and then cooled under stirring to obtain a lotion cosmetic [cosmetic 1 or cosmetic 2 (skin external preparation 1 or 2)].
  • the same operation was performed to prepare Comparative Example 1 in which the “component having an action of improving skin barrier function” of the composition of the present invention (external preparation for skin) was replaced with “water”.
  • Example 9 ⁇ Skin roughness improvement test of the external preparation for skin of the present invention> A paneler was used to evaluate the rough skin improving effect of the rough skin model prepared by tape stripping for the cosmetic 1, cosmetic 2, and comparative example 1, which are the external preparations of the present invention. That is, four 1cm x 1cm parts are defined on the left and right forearms, tape stripping is carried out 15 times for each part, and transepidermal water transpiration (TEWL) is measured with "Tevameter” manufactured by Integral. Measured. Thereafter, 50 ⁇ L of the sample was applied once a day, this operation was continued for 6 days, and TEWL was measured again on the 7th day.
  • TEWL transepidermal water transpiration
  • the TEWL improvement rate (%) was calculated by subtracting the TEWL value on the seventh day from the TEWL value on the first day, dividing by the TEWL value on the first day, and multiplying by 100. The n number was 15. The results are shown in Table 3. This shows that the skin external preparation of this invention is excellent in the rough skin improvement effect.
  • the present invention is a component having a skin barrier function improving action suitable as a cosmetic raw material (including quasi-drugs), and can be applied to a skin external preparation such as cosmetics.

Abstract

La présente invention concerne un nouvel activateur du récepteur TRPV (récepteur transitoire à potentiel sensible aux vanilloïdes) qui convient pour l'amélioration d'une fonction de barrière de la peau. Selon l'invention, un activateur de récepteur TRPV comprend au moins un composé choisi parmi les composés représentés respectivement par les formules (1) à (4) ou un extrait de plante contenant ledit au moins un composé en une quantité efficace pour agir comme activateur du récepteur TRPV. (Dans les formules, un groupe hydroxy peut être substitué par un groupe méthoxy ou un groupe éthoxy; et un groupe méthoxy peut être substitué par un groupe hydroxy ou un groupe éthoxy.)
PCT/JP2011/059985 2011-04-22 2011-04-22 Agent d'amélioration de la fonction de barrière de la peau WO2012144080A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP2013510839A JP5737663B2 (ja) 2011-04-22 2011-04-22 皮膚バリア機能改善剤
PCT/JP2011/059985 WO2012144080A1 (fr) 2011-04-22 2011-04-22 Agent d'amélioration de la fonction de barrière de la peau

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/JP2011/059985 WO2012144080A1 (fr) 2011-04-22 2011-04-22 Agent d'amélioration de la fonction de barrière de la peau

Publications (1)

Publication Number Publication Date
WO2012144080A1 true WO2012144080A1 (fr) 2012-10-26

Family

ID=47041226

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2011/059985 WO2012144080A1 (fr) 2011-04-22 2011-04-22 Agent d'amélioration de la fonction de barrière de la peau

Country Status (2)

Country Link
JP (1) JP5737663B2 (fr)
WO (1) WO2012144080A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015140470A3 (fr) * 2014-03-18 2016-01-21 Greenpharma Applications cosmétiques et pharmaceutiques de l'acide gallique et de dérivés de l'acide gallique
WO2016067688A1 (fr) * 2014-10-29 2016-05-06 昭和電工株式会社 Agent améliorant la fonction barrière de la peau et composition pour améliorer la fonction barrière de la peau
JP2016069341A (ja) * 2014-09-30 2016-05-09 株式会社東洋新薬 皮膚バリア機能改善剤、細胞間接着構造の形成促進剤、タイトジャンクション形成促進剤及びｔｒｐｖ４遺伝子の発現亢進剤
FR3031455A1 (fr) * 2015-01-14 2016-07-15 Greenpharma Sas Compose derive de l’acide gallique et applications
JP2017100987A (ja) * 2015-12-01 2017-06-08 株式会社ナールスコーポレーション フィラグリン遺伝子発現促進剤

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH07126143A (ja) * 1993-11-05 1995-05-16 Mikimoto Pharmaceut Co Ltd 化粧料
JP2001048766A (ja) * 1999-08-02 2001-02-20 Mikimoto Pharmaceut Co Ltd ヒスタミン遊離抑制剤
JP2002275184A (ja) * 2001-03-21 2002-09-25 Kampou Science Foundation エラグ酸誘導体および薬用物質
JP2003292432A (ja) * 2002-04-04 2003-10-15 Noevir Co Ltd 皮膚外用剤
WO2006137129A1 (fr) * 2005-06-21 2006-12-28 Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo Preparation cutanee a usage externe
JP2007277149A (ja) * 2006-04-06 2007-10-25 Kao Corp インボルクリン発現促進剤
JP2010511376A (ja) * 2006-12-08 2010-04-15 ポーラ化成工業株式会社 皮膚バリア機能の鑑別法、これを用いた皮膚バリア機能強化素材のスクリーニング法、該皮膚バリア機能強化素材、及び該皮膚バリア機能強化素材を含有してなる化粧料

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH07126143A (ja) * 1993-11-05 1995-05-16 Mikimoto Pharmaceut Co Ltd 化粧料
JP2001048766A (ja) * 1999-08-02 2001-02-20 Mikimoto Pharmaceut Co Ltd ヒスタミン遊離抑制剤
JP2002275184A (ja) * 2001-03-21 2002-09-25 Kampou Science Foundation エラグ酸誘導体および薬用物質
JP2003292432A (ja) * 2002-04-04 2003-10-15 Noevir Co Ltd 皮膚外用剤
WO2006137129A1 (fr) * 2005-06-21 2006-12-28 Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo Preparation cutanee a usage externe
JP2007277149A (ja) * 2006-04-06 2007-10-25 Kao Corp インボルクリン発現促進剤
JP2010511376A (ja) * 2006-12-08 2010-04-15 ポーラ化成工業株式会社 皮膚バリア機能の鑑別法、これを用いた皮膚バリア機能強化素材のスクリーニング法、該皮膚バリア機能強化素材、及び該皮膚バリア機能強化素材を含有してなる化粧料

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015140470A3 (fr) * 2014-03-18 2016-01-21 Greenpharma Applications cosmétiques et pharmaceutiques de l'acide gallique et de dérivés de l'acide gallique
JP2016069341A (ja) * 2014-09-30 2016-05-09 株式会社東洋新薬 皮膚バリア機能改善剤、細胞間接着構造の形成促進剤、タイトジャンクション形成促進剤及びｔｒｐｖ４遺伝子の発現亢進剤
WO2016067688A1 (fr) * 2014-10-29 2016-05-06 昭和電工株式会社 Agent améliorant la fonction barrière de la peau et composition pour améliorer la fonction barrière de la peau
JPWO2016067688A1 (ja) * 2014-10-29 2017-08-10 昭和電工株式会社 皮膚バリア機能改善剤及び皮膚バリア機能改善用組成物
CN107106586A (zh) * 2014-10-29 2017-08-29 昭和电工株式会社 皮肤屏障功能改善剂和皮肤屏障功能改善用组合物
FR3031455A1 (fr) * 2015-01-14 2016-07-15 Greenpharma Sas Compose derive de l’acide gallique et applications
JP2017100987A (ja) * 2015-12-01 2017-06-08 株式会社ナールスコーポレーション フィラグリン遺伝子発現促進剤

Also Published As

Publication number Publication date
JPWO2012144080A1 (ja) 2014-07-28
JP5737663B2 (ja) 2015-06-17

Similar Documents

Publication Publication Date Title
JP5872158B2 (ja) 皮膚美白方法、並びに、皮膚シミ形成抑制及び／又は除去因子のスクリーニング方法
JP5737663B2 (ja) 皮膚バリア機能改善剤
US20160095815A1 (en) Substance for restoring normal co-expression and interaction between the lox and nrage proteins
US20220160607A1 (en) Anti-aging cosmetic compositions
JP7214268B2 (ja) 皮膚用組成物
JP2012152127A (ja) 皮膚バリア機能回復剤
KR20180032506A (ko) 티오레독신 결합단백질 유래 펩타이드 또는 이를 암호화 하는 폴리뉴클레오타이드를 유효성분으로 함유하는 노화 줄기세포의 역노화용 조성물 및 이의 용도
JP2011115152A (ja) スクリーニング法
CN111356468A (zh) 包含黃漆木提取物作为有效成分的用于预防或治疗纤维化疾病的组合物
JP6696097B2 (ja) タルミ改善剤のスクリーニング法
JP2011093830A (ja) 皮膚バリア機能改善剤
JP2011011981A (ja) 化粧料
JP5932279B2 (ja) スクリーニング方法
JP5806859B2 (ja) スクリーニング方法
JP6113981B2 (ja) スクリ−ニング方法
TW201242610A (en) Medicament for improving skin barrier function
CN107951765B (zh) 紫笋茶提取物及其应用
JP2022119185A (ja) 抗光老化のための薬剤及び美容方法，並びに抗光老化剤のスクリーニング方法
JP2013083504A5 (fr)
JP6405194B2 (ja) 多層色素沈着改善剤
Hong et al. Synthetic adiponectin-derived peptide enhances skin barrier function
CN112969920A (zh) 评估物质或组合物预防、减缓或消除皮肤或嘴唇老化迹象的能力的方法
EP3437663A1 (fr) Composition pour stimuler la différenciation de cellules cutanées et procédé de criblage de matériaux stimulant la différenciation de cellules cutanées
Korver Quantitative visualisation of epidermal cell populations; A study in healthy and psoriatic skin and its appendages.
Epernon Caffeic acid improves skin barrier function as Peroxisome proliferator-activated receptor (PPAR)-α agonist

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 11864071

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2013510839

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 11864071

Country of ref document: EP

Kind code of ref document: A1