WO2018181599A1 - 皮膚バリア増強剤、医薬組成物、薬用化粧品及び美容方法 - Google Patents
皮膚バリア増強剤、医薬組成物、薬用化粧品及び美容方法 Download PDFInfo
- Publication number
- WO2018181599A1 WO2018181599A1 PCT/JP2018/013013 JP2018013013W WO2018181599A1 WO 2018181599 A1 WO2018181599 A1 WO 2018181599A1 JP 2018013013 W JP2018013013 W JP 2018013013W WO 2018181599 A1 WO2018181599 A1 WO 2018181599A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- skin barrier
- skin
- protein
- present
- claudin
- Prior art date
Links
Images
Classifications
-
- 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
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
- A61K31/047—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates having two or more hydroxy groups, e.g. sorbitol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
- A61K31/345—Nitrofurans
-
- 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
- A61K31/7004—Monosaccharides having only carbon, hydrogen and oxygen atoms
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
- A61K8/0208—Tissues; Wipes; Patches
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
- A61K8/04—Dispersions; Emulsions
- A61K8/042—Gels
-
- 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
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
- A61K8/345—Alcohols containing more than one hydroxy group
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/60—Sugars; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- 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
-
- 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
Definitions
- the present invention relates to a skin barrier enhancer, a pharmaceutical composition, a medicated cosmetic and a cosmetic method.
- the epidermis has a moisturizing function that prevents evaporation of moisture in the body and maintains the amount of moisture in the skin, and a skin barrier function that prevents infiltration of mites, dust, pollen, microorganisms, and the like.
- a moisturizing function that prevents evaporation of moisture in the body and maintains the amount of moisture in the skin
- a skin barrier function that prevents infiltration of mites, dust, pollen, microorganisms, and the like.
- the loss of moisture is associated with a decrease in moisturizing function and a decrease in skin barrier function.
- Patent Document 1 discloses that glucose promotes the production of HMGB1, induces the release of HMGB1 to the outside of the cell, enhances the proliferation and migration of normal cells, and promotes tissue repair. ing.
- an object of the present invention is to provide a skin barrier enhancer that can further enhance the skin barrier function.
- Filagrin is a protein that is present in cells that form the granule layer of the epidermis, and is a protein that contributes to the function of maintaining moisture and retaining moisture in the skin.
- Claudin-1 is a membrane protein that constitutes a height junction that connects cells of the epidermis.
- Claudin-1 is a protein that prevents the entry of allergens, microorganisms and the like into cells, and contributes to the function of preventing moisture evaporation on the body surface.
- a liquid containing at least one of a monosaccharide having a pH in the acidic region and a sugar alcohol is Filagrin, which is a skin barrier-related protein. And found to have an effect of promoting production of Claudin-1.
- a pharmaceutical composition comprising the skin barrier enhancer according to any one of [1] to [4] and having a pH of 4.0 or more and 7.0 or less.
- the medicated cosmetic according to [6] which is liquid, gel, or sheet.
- a cosmetic method comprising a step of applying the medicated cosmetic product according to [6] or [7] to the skin or mucous membrane.
- a skin barrier enhancer capable of further enhancing the skin barrier function can be provided.
- 6 is a graph showing the evaluation results of the expression level of Filaggrin protein in Examples 1 to 6 and Comparative Examples 1 to 4. It is a graph which shows the evaluation result of the expression level of Filagrin mRNA of Examples 2, 4, 6 and Comparative Examples 2, 4. 4 is a graph showing the evaluation results of Claudin-1 mRNA expression levels of Examples 2, 4, and 6 and Comparative Examples 2 and 4. FIG. It is a graph which shows the evaluation result of the expression level of Filagrin protein of Example 7 and Comparative Example 7. 6 is a graph showing the evaluation results of cytotoxicity rates of Examples 1 to 6 and Comparative Examples 1 to 4. It is a photograph which shows the external appearance of the pinna part of the mouse
- 6 is a graph showing the evaluation results of the expression level of Filaggrin and Claudin-1 mRNA in Reference Test 1. It is a graph which shows the evaluation result of the expression level of Filaggrin protein of the reference test 2. 6 is a graph showing the evaluation results of the expression level of Claudin-1 protein in Reference Test 3.
- the skin barrier enhancer of the present invention contains at least one of a monosaccharide and a sugar alcohol, and has a pH of 4.0 or more and 7.0 or less.
- the form of the skin barrier enhancing agent is not particularly limited as long as the pH can be measured. Specific examples include liquids, gels, sheets, sols, creams, and emulsions, but are not limited thereto.
- the skin barrier enhancer of the present invention contains at least one of a monosaccharide and a sugar alcohol. By containing at least one of a monosaccharide and a sugar alcohol, it is possible to promote the production of Filaggrin and Claudin-1 by cells and enhance the skin barrier function.
- the skin barrier enhancer enhances or restores the skin barrier function inherent to the skin, such as the function of keeping moisture in the skin and retaining moisture, and the function of preventing the entry of microorganisms and the like into cells.
- the content of at least one of a monosaccharide and a sugar alcohol in the skin barrier enhancer is preferably 1 to 5000 mM, more preferably 6 to 3000 mM, still more preferably 6 to 1000 mM, and more preferably 6 to 1000 mM. Particularly preferred is 165 mM. If the content of at least one of monosaccharide and sugar alcohol is not less than the lower limit, the production of Filaggrin and Claudin-1 is promoted by the skin barrier enhancer, and the skin barrier function is easily enhanced or restored.
- At least one of the monosaccharide and sugar alcohol is not more than the above upper limit, at least one of the monosaccharide and sugar alcohol is uniformly dispersed in the skin barrier enhancer, and Filaggrin and Claudin-1 are sufficiently produced.
- the skin barrier function is sufficiently enhanced or restored.
- monosaccharides include, but are not limited to, monosaccharides such as glucose, fructose, mannose, arabinose, growth, xylose, lyxose, erythrose, threose, galactose, and sorbose.
- the skin barrier enhancer preferably contains glucose as a monosaccharide.
- sugar alcohols include, but are not limited to, mannitol, sorbitol, xylitol, erythritol, maltitol, lactitol and the like.
- the skin barrier enhancer preferably contains mannitol as a sugar alcohol.
- the skin barrier enhancer of the present invention has a pH of 4.0 or more and 7.0 or less, and preferably 5.0 or more and 7.0 or less. If the pH is equal to or higher than the lower limit, skin tissue and cell damage by the skin barrier enhancer is reduced, and transcription and expression of Filagrin mRNA and Claudin-1 mRNA are promoted, and the skin barrier function is enhanced or Easy to recover. When the pH is not more than the above upper limit value, the production of Filaggrin and Claudin-1 is sufficiently promoted, and the skin barrier function is sufficiently enhanced or restored.
- pH is a value measured at 25 ° C. using a pH meter (“F-53” manufactured by Horiba, Ltd.).
- the pH of the skin barrier enhancing agent can be adjusted with a buffer.
- the buffer solution is not particularly limited as long as the pH of the skin barrier enhancer is maintained at 4.0 or more and 7.0 or less.
- Buffers include phosphate buffer, citrate buffer, citrate-phosphate buffer, maleate buffer, malate buffer, succinate buffer, metaphosphate buffer, sorbate buffer, carbonate buffer Solution, trishydroxymethylaminomethane-HCl buffer (Tris-HCl buffer), MES buffer (2-morpholinoethanesulfonic acid buffer), TES buffer (N-tris (hydroxymethyl) methyl-2-aminoethanesulfone) Acid buffer), acetate buffer, MOPS buffer (3-morpholinopropanesulfonic acid buffer), HEPES buffer (4- (2-hydroxyethyl) -1-piperazine ethanesulfonic acid buffer), and other buffer solutions Glycine-hydrochloric acid buffer, glycine-NaOH buffer, glycylgly
- Amino acid-based buffer Tris - borate buffer, borate -NaOH buffer, borate buffer solution such as borate buffer; imidazole buffer and the like are not limited thereto.
- phosphate buffer, citrate buffer, malate buffer, succinate buffer and metaphosphate buffer are preferable.
- the pH of the buffer solution can be adjusted as appropriate using acidic substances such as hydrochloric acid and acetic acid, or basic substances such as sodium hydroxide and ammonia.
- acidic substances such as phosphoric acid, acetic acid and amino acids
- the pH of the skin barrier enhancer is 4.0 or more and 7 You may adjust to below 0.0.
- the concentration of the buffer solution in the skin barrier enhancer is preferably 6 to 165 mM. If the concentration of the buffer solution is equal to or higher than the lower limit, the pH of the skin barrier enhancer can be easily adjusted. When the concentration of the buffer solution is not more than the above upper limit value, the production of Filagrin and Claudin-1 is sufficiently promoted by the skin barrier enhancer, and the skin barrier function is easily enhanced or restored.
- the skin barrier enhancing agent can contain an optional component other than at least one of a monosaccharide and a sugar alcohol, as necessary, as long as the effects of the present invention are not impaired.
- optional components include excipients, thickeners, carriers, fragrances, and pigments. These arbitrary components may be used individually by 1 type, and may be used in combination of 2 or more type. The optional component will be specifically described in the section “(Pharmaceutical composition)” described later.
- the present invention is the use of at least one of a monosaccharide and a sugar alcohol for a skin barrier enhancer, comprising administering at least one of a monosaccharide and a sugar alcohol at a pH of 4.0 or more and 7.0 or less.
- a monosaccharide and a sugar alcohol for a skin barrier enhancer, comprising administering at least one of a monosaccharide and a sugar alcohol at a pH of 4.0 or more and 7.0 or less.
- Said use is the use of a combination of at least one of a monosaccharide and a sugar alcohol and a pH adjuster as an optional component for producing a skin barrier enhancer, and the pH is 4.0 or more and 7.0.
- said use comprising administering at least one of a monosaccharide and a sugar alcohol.
- the pharmaceutical composition of the present invention contains the skin barrier enhancer of the present invention, and has a pH of 4.0 or more and 7.0 or less.
- the content of at least one of the monosaccharide and sugar alcohol in the pharmaceutical composition can be determined in consideration of the use, usage, etc. of the pharmaceutical composition (skin barrier enhancer). For example, in the case of a transdermal absorbent using a pharmaceutical composition, 100 to 5000 mM is preferable, and 1000 to 3000 mM is more preferable. In the case of an injection using a pharmaceutical composition, 1 to 1000 mM is preferable, and 6 to 165 mM is more preferable.
- the pharmaceutical composition promotes the production of Filaggrin and Claudin-1, and the skin barrier function is easily enhanced or restored. If the content of at least one of the monosaccharide and sugar alcohol is not more than the above upper limit, at least one of the monosaccharide and sugar alcohol is uniformly dispersed in the pharmaceutical composition, and the production of Filaggrin and Claudin-1 is sufficiently achieved. It is facilitated and the skin barrier function is likely to be sufficiently enhanced or restored.
- the pH of the pharmaceutical composition is 4.0 or more and 7.0 or less, and preferably 5.0 or more and 7.0 or less.
- the pH is at least the lower limit, the expression of mRNA encoding Filagrin and mRNA encoding Claudin-1 is promoted, and the skin barrier function is likely to be enhanced or restored.
- the pH is not more than the above upper limit value, skin tissue damage due to administration of the pharmaceutical composition is unlikely to occur, the production of Filagrin and Claudin-1 is sufficiently promoted, and the skin barrier function is likely to be sufficiently enhanced or restored.
- the pharmaceutical composition is used as a therapeutic agent for autoimmune diseases such as atopic dermatitis, bronchial asthma, rheumatoid arthritis and collagen disease; nephrotic syndrome, ulcerative colitis, hepatitis, pancreatitis, thyroiditis, cold, stomatitis, gingiva
- Anti-inflammatory drugs for diseases such as organ inflammation such as inflammation and periodontitis
- wound treatment drugs for wounds such as burns, wounds, pressure ulcers, acne, skin rashes and moyake
- genetic diseases such as xeroderma pigmentosum and ichthyosis And the like.
- the effects of the present invention are particularly remarkable when the pharmaceutical composition of the present invention is applied to anti-inflammatory drugs and wound healing drugs.
- the dosage form of the pharmaceutical composition is not particularly limited as long as the pH can be measured.
- Specific examples of dosage forms include oral preparations such as liquids, emulsions, suspensions and syrups; transdermal absorption agents such as ointments, lotions, creams, patches and aerosols; injections, eye drops and suppositories These parenteral agents are exemplified. Among these, the effects of the present invention are remarkably exhibited in a transdermal absorbent such as an ointment.
- the injection may have its water removed by lyophilization after the solution is filled in a vial or the like. When moisture has been removed, the freeze-dried product may be dispersed in physiological saline or the like immediately before use to prepare a solution.
- a pharmaceutical composition it determines suitably according to the kind etc. of disease, for example.
- the disease is a skin wound such as atopic dermatitis, a cut or an abrasion
- a method of directly applying a pharmaceutical composition as a transdermal absorbent to the affected area is exemplified.
- a method of orally administering the pharmaceutical composition as an oral preparation a method of injecting the pharmaceutical composition into an affected tissue as an injection, and the like are exemplified.
- Mucosal regeneration may be promoted by washing the affected area with a pharmaceutical composition under the upper endoscope and applying the pharmaceutical composition to the affected area.
- a healthy mucous membrane can be formed, natural healing can be gradually increased, and mucosal invasion can be prevented. Furthermore, by applying the pharmaceutical composition to the gastric mucosa, regeneration of the gastric mucosa can be promoted, and the barrier function of the epithelial tissue can be enhanced or restored.
- the disease is a disease of the nasal / ocular mucosa such as hay fever
- a method of applying the pharmaceutical composition to the affected part is exemplified.
- the pharmaceutical composition is applied to the nose / ocular mucosa, the nose / ocular mucosa is repaired and forms a strong barrier in the mucosal layer.
- the allergic reaction can be suppressed, and nasal clogging or nasal discharge or tear secretion can be eliminated.
- the disease is inflammation (cold) of the upper respiratory tract mucosa caused by infection with a virus such as a cold or bacteria
- a method of instilling or inhaling the pharmaceutical composition is exemplified.
- inflammation of the upper airway mucosa can be suppressed, the upper airway mucosa can be repaired and regenerated to suppress the progression of infection, and activation of the innate immune response can be promoted.
- the dosage of the pharmaceutical composition is appropriately determined according to the age or weight of the patient or subject, the type / degree of disease, the administration method, and the like.
- the pharmaceutical composition can contain optional components other than the skin barrier enhancer of the present invention (hereinafter also referred to as “pharmaceutical optional components”) as necessary, as long as the effects of the present invention are not inhibited.
- pharmaceutical optional ingredients include pharmacologically acceptable salts, excipients, thickeners, carriers, fragrances, dyes and the like. These pharmaceutical optional ingredients may be used alone or in combination of two or more.
- Examples of pharmacologically acceptable salts include conventional non-toxic salts, that is, acid addition salts and salts with various bases. Specific examples include inorganic acid salts such as hydrochloride, nitrate and sulfate; organic acid salts such as acetate, citrate, fumarate and tartrate; methanesulfonate, p-toluenesulfonate, and the like.
- Amino acid salts such as sulfonate, alanine, leucine and glutamate; inorganic bases such as alkali metal salts (for example, sodium salt, potassium salt) and alkaline earth metal salts (for example, magnesium salt, calcium salt)
- Salts Organic amine salts such as triethylamine salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N, N′-dibenzylethylenediamine salt and the like are exemplified. These salts may be used individually by 1 type, and may be used in combination of 2 or more type. This is because when such a salt is contained, crystallization is easy.
- the excipient can be appropriately selected according to the dosage form of the pharmaceutical composition.
- Specific examples include water such as distilled water, ion-exchanged water and pure water, liquid excipients such as glycerol and lower alcohols having 1 to 6 carbon atoms such as methanol and ethanol; lactose, starch, dextrin, sucrose, etc.
- Examples include solid excipients. These excipients may be used alone or in combination of two or more.
- the thickener can be appropriately selected depending on the dosage form of the pharmaceutical composition. Specific examples include gelatin, xanthan gum, carrageenan and the like. These thickeners may be used individually by 1 type, and may be used in combination of 2 or more type.
- the present invention is the use of at least one of a monosaccharide and a sugar alcohol for a pharmaceutical composition in the treatment of a skin disease, wherein the pH is 4.0 or more and 7.0 or less, and at least one of the monosaccharide and the sugar alcohol is administered.
- Said use comprising: That is, the present invention is the use of a combination of at least one of a monosaccharide and a sugar alcohol and an optional pH adjuster for producing a pharmaceutical composition in the treatment of skin diseases, and having a pH of 4.0.
- the pharmaceutical composition of the present invention can be used in a method for enhancing the skin barrier function.
- the method for enhancing the skin barrier function includes administering at least one of a monosaccharide and a sugar alcohol having an active ingredient amount or more at a pH of 4.0 or more and 7.0 or less.
- the medicated cosmetic of the present invention contains the skin barrier enhancer of the present invention and has a pH of 4.0 or more and 7.0 or less.
- the medicinal cosmetics of the present invention are, for example, lotions, creams, emulsions, foundations and the like, and are intended to prevent skin diseases and moisturize the skin.
- Medicinal cosmetics are classified as quasi-drugs stipulated in the Japanese Pharmaceutical Affairs Law.
- the content of the skin barrier enhancer in medicinal cosmetics can be determined in consideration of the use and mode of administration of medicinal cosmetics.
- the concentration of at least one of monosaccharide and sugar alcohol in medicinal cosmetics is preferably 1 to 1000 mM, more preferably 6 to 165 mM. If the concentration of at least one of the monosaccharide and sugar alcohol is within the above range, the production of Filaggrin and Claudin-1 is promoted, the skin barrier function is sufficiently enhanced or restored, and the repair of the skin tissue is further promoted.
- the pH of the medicated cosmetic product is 4.0 or more and 7.0 or less, and preferably 5.0 or more and 7.0 or less. If the pH is equal to or higher than the lower limit, the expression of Filaggrin mRNA and Claudin-1 mRNA is promoted, and the skin barrier function is easily enhanced or restored. If the pH is not more than the above upper limit value, skin tissue damage due to administration of the pharmaceutical composition is unlikely to occur, the production of Filagrin and Claudin-1 is sufficiently promoted, and the skin barrier function is likely to be sufficiently enhanced or restored.
- the form of the medicated cosmetic is not particularly limited as long as the pH can be measured.
- Specific examples thereof include medicinal cosmetics such as liquid, gel and sheet.
- the method for using medicinal cosmetics is appropriately determined according to, for example, the purpose of beauty.
- medicinal cosmetics intended for skin beauty and moisturizing there are exemplified methods of directly applying liquid, emulsion, gel and sheet medicinal cosmetics to the skin.
- examples include a method of orally applying liquid and gel medicinal cosmetics, a method of injecting liquid medicinal cosmetics as an injection, and the like.
- the medicated cosmetic can contain optional components other than the barrier enhancer of the present invention (hereinafter, also referred to as “cosmetic optional components”) as needed, as long as the effects of the present invention are not impaired.
- Cosmetic optional ingredients include liquid paraffin, ceresin, jiro, lanolin, petrolatum, cetanol, squalene, jojoba oil, stearic acid, palmitic acid, lauryl alcohol, stearyl alcohol, cetyl alcohol, beeswax, methylpolysiloxane, dimethylcyclopolysiloxane, etc.
- Moisturizers such as propanol, glycol, propylene glycol, hyaluronic acid, collagen, polyethylene glycol, cholesteryl hydroxystearate, glycerin, sorbitol; various surfactants; emulsifiers; excipients: thickeners; pH adjusters; Examples include antioxidants; pigments; fragrances; ultraviolet absorbers and the like.
- These cosmetic optional ingredients may be used alone or in combination of two or more.
- the present invention is the use of at least one of a monosaccharide and a sugar alcohol for medicinal cosmetics in skin moisturizing or cosmetics, and the pH is 4.0 or more and 7.0 or less, and at least one of the monosaccharide and the sugar alcohol is administered.
- Said use comprising: That is, the present invention is the use of a combination of at least one of a monosaccharide and a sugar alcohol and a pH adjuster, which is an optional component, for producing medicinal cosmetics for moisturizing skin or beauty, and having a pH of 4.0.
- the use as described above, comprising administering at least one of a monosaccharide and a sugar alcohol at 7.0 or less.
- the cosmetic method of the present invention includes a first step of applying the above-described medicated cosmetic of the present invention to the skin or mucous membrane.
- the application amount of the medicated cosmetic is appropriately determined according to the age, weight, skin condition, application method, etc. of the patient or subject.
- the cosmetic method of the present invention preferably includes, after the first step, a second step of returning the pH of the skin or mucous membrane to a basic region greater than 7.0.
- the time from the first step to the second step is preferably about 15 to 30 minutes. If the interval between the first step and the second step is about 15 to 30 minutes, it is easy to return to the basic region that is the original pH of the skin or mucous membrane.
- Methods for returning the pH to the basic region include a method of rinsing with running water, a method of removing medicinal cosmetics by wiping, a method of applying a cream containing a neutralizing agent to the skin or mucous membrane from the medicinal cosmetics, etc. Illustrated. These methods may be used singly or in combination of two or more. In the cosmetic method of the present invention, it is preferable to repeat the first step and the second step described above.
- the cosmetic method of the present invention includes administering at least one of a monosaccharide and a sugar alcohol having an active ingredient amount or more at a pH of 4.0 or more and 7.0 or less.
- at least one of monosaccharide and sugar alcohol may be applied to the skin or mucous membrane of a patient or a subject.
- the skin barrier enhancer of the present invention contains at least one of a monosaccharide and a sugar alcohol, and has a pH of 4.0 or more and 7.0 or less, so that the production of Filagrin and Claudin-1 in cells is suppressed. It can promote the transcription and expression of Filaggrin mRNA and Claudin-1 mRNA. Therefore, the skin barrier function is sufficiently enhanced or restored by the skin barrier enhancer.
- Examples 1 to 7, Comparative Examples 1 to 7 In a HuMedia-KG2 medium (Kurabo), D-glucose (Product No .: G5767, manufactured by SIGMA) or D-mannitol (Product No .: M9677, manufactured by SIGMA) and the medium so as to have the concentrations shown in Table 1.
- hydrochloric acid (HCl) product number: 080-0106, manufactured by Wako Pure Chemical Industries, Ltd.
- Test Medium a medium containing a skin barrier enhancer adjusted to the pH shown in Table 1 (hereinafter, “ Also referred to as “Test Medium”).
- the pH is a value measured at 25 ° C.
- ⁇ Analysis of the expression level of Filagrin protein 1> Normal human skin keratinocytes (primary cultured normal skin keratinocytes derived from a single donor, manufactured by Takara Bio Inc.) using HuMedia-KG2 medium (pH 7.6, manufactured by Kurabo Industries) in 6 wells (6 wells) It seed
- the culture supernatant was removed by aspiration, and the medium was replaced with a HuMedia-KG2 medium having a pH of 7.6, followed by culturing at 37 ° C. for 3 hours (post-culture).
- the medium was removed and the protein was extracted from the cultured cells.
- the extracted protein was developed by SDS-PAGE. Thereafter, the extracted protein was transferred to a nitrocellulose membrane, and the expression level of Filagrin protein in the cultured cells was analyzed by the Western blot method using a mouse-derived Filagrin antibody. Further, as an endogenous control, the expression level of ⁇ -actin protein in the same sample was analyzed.
- the concentration of each detected protein band was measured with a densitometer, and the expression levels of Filagrin protein and ⁇ -actin protein were analyzed.
- the ⁇ -actin protein expression level of the same sample was set to 1, and the Filaggrin protein expression level was evaluated based on the relative value of the Filaggrin protein expression level (relative expression level). The results are shown in FIG.
- ⁇ Analysis of expression level of Filagrin mRNA and Claudin-1 mRNA> Except for changing the test medium to those of Examples 2, 4 and 6 and Comparative Examples 2 and 4, pre-culture, main culture and post-culture in the same manner as “ ⁇ Analysis of expression level of Filagrin protein 1>” Culture was performed. After the post-culture, the medium was removed, and total RNA was extracted from the cultured cells. Using the RT-PCR method, Filagrin and Claudin-1 mRNA was amplified from the extracted RNA, and the expression level of each mRNA was analyzed.
- the expression level of each mRNA in Comparative Example 2 is 1, and the relative expression values (relative expression levels) of each mRNA in Examples 2, 4, 6 and Comparative Example 4 are the values of Filaggrin mRNA and Claudin-1 mRNA. The expression level was evaluated. The results are shown in FIGS.
- Example 7 As shown in Table 1 and FIG. 4, in Example 7 to which the present invention was applied, the expression level of Filaglin protein was increased 4.3 times compared to Comparative Example 7.
- Example 7 From the results of Example 7 and Comparative Example 7, it was found that the production of Filaglin protein by cultured cells can be promoted by adjusting the mannitol concentration of the test medium to 110 mM and adjusting the pH to 5.7. From the above results, it was found that application of the present invention promotes the expression of Filaglin and Claudin-1 and can enhance the skin barrier function.
- ⁇ Cytotoxicity test> Normal human skin keratinocytes were cultured in the same manner as in “ ⁇ Analysis of expression level of Filagrin protein 1>”. The degree of damage of cultured cells was analyzed by the LDH method. The damage rate of the cells cultured in each case was evaluated when the degree of damage of the cells treated with Triton X 100 for the cells cultured in the test medium of each case was defined as 100%.
- FIG. 5 is a graph showing the results of evaluating the failure rate of cells cultured in the test media of Examples 1 to 6 and Comparative Examples 1 to 4. As shown in Table 1 and FIG. 5, the failure rate of the cells of Examples 1 to 6 to which the present invention was applied was lower than the failure rate of the cells of Comparative Examples 1 and 2 having a pH of 7.6. On the other hand, the failure rate of the cells of Comparative Examples 3 and 4 having a pH of 3.0 was higher than that of the cells of Comparative Examples 1 and 2 having a pH of 7.6.
- ⁇ Evaluation with atopic dermatitis mouse model An aqueous solution of phosphate buffered saline (PBS) adjusted to a glucose concentration of 20% and a pH of 6.0 was prepared, and used as a skin barrier enhancer to which the present invention was applied. Mite antigen was intradermally administered from Day 0 to Day 14 twice a week in the right auricle of 8-week-old female NC / Nga mice after habituation breeding. In parallel with this, the skin barrier enhancer of the present invention was applied and administered once daily from Day 0 to Day 28. PBS adjusted to pH 7.6 as the Mock group was applied and administered once a day from Day 0 to Day 28.
- PBS phosphate buffered saline
- Photographs of the auricles of the Mock group and the skin barrier enhancer applied to Day 28 were taken, and left and right auricles were collected from mice exsanguinated and euthanized under isoflurane anesthesia, and neutral formalin fixation was performed. It was. Tissue sections were prepared after embedding in paraffin. The sections were stained with a hematoxylin-eosin (HE) solution and immunostained with mouse-derived Filaggrin and Claudin-1 antibodies. As negative control, an auricle section of a mouse in which 8-week-old female NC / Nga mice were raised to Day 28 without treatment was used.
- HE hematoxylin-eosin
- FIG. 6 is a photograph showing the appearance of the auricle portion of a mouse evaluated using an atopic dermatitis mouse model.
- the mouse pinna As a result of observing the mouse pinna with the naked eye, mild ulceration and bleeding were observed at Day 15 in the group (Mock group) in which PBS with a pH of 7.6 was applied to the mouse pinna sensitized with mite antigen. On Day 28, severe bleeding and ulceration were observed.
- the skin barrier enhancing agent of the present invention was applied, although slight skin swelling was observed at Day 28, neither ulcer formation nor bleeding at the auricle was observed at both Day 15 and Day 28.
- FIG. 7 is a photograph showing the result of staining the tissue of the auricle section of a mouse evaluated with an atopic dermatitis mouse model.
- the HE staining result of FIG. 7 infiltration of inflammatory cells in the dermis was strongly observed in the Mock group.
- the skin barrier enhancer of the present invention was applied, the infiltration of inflammatory cells was reduced, and the stratum corneum and epidermis were kept normal.
- the results of immunostaining in FIG. 7 in the Mock group the continuity of expression of Claudin-1 protein was lost and the expression level was decreased.
- mRNA of Filagrin and Claudin-1 was amplified from RNA extracted from each culture solution, and the expression level of each mRNA was analyzed for each culture solution having different main culture time.
- the main culture time is 0 hour
- the expression level of each mRNA is 1, and the expression of each of Filaggrin and Claudin-1 is expressed as a relative value (relative expression level) of each mRNA expression level in each cell during each culture time. The amount was evaluated.
- FIG. 8 is a graph showing the evaluation results of the expression levels of Filaggrin and Claudin-1 mRNA in Comparative Example 2 in each culture time of Reference Test 1.
- Comparative Example 2 the peak of Claudin-1 mRNA expression induction was observed 0.5 hours after the initiation of expression induction.
- the peak of Filagrin mRNA expression induction was observed 2.0 hours after the initiation of expression induction.
- FIG. 9 is a graph showing the evaluation results of the expression level of Filaggrin protein in Reference Test 2. An increase in the expression level of Filaggrin protein was observed 2 hours after the initiation of expression induction. An increase in the expression level of Filagrin protein was observed for up to 24 hours from the start of expression induction.
- FIG. 10 is a graph showing the evaluation results of the expression level of Claudin-1 protein in Reference Test 3. An increase in the expression level of Claudin-1 protein was observed 2 hours after the initiation of expression induction. An increase in the expression level of Claudin-1 protein was observed for up to 24 hours after the start of expression induction.
- the skin barrier enhancer of the present invention can further enhance the expression of the skin barrier-related protein when the pH is 4.0 or more and 7.0 or less (FIGS. 1 and 4). Therefore, the skin barrier enhancer of the present invention can enhance the skin barrier function more effectively than the conventional skin barrier enhancer. Therefore, when the skin barrier enhancer of the present invention is applied to pharmaceuticals and the like, it is possible to expect remarkable effects such as the ability to repair skin tissue more effectively.
- the skin barrier enhancer, pharmaceutical composition and medicinal cosmetics obtained by the present invention can further enhance the skin barrier function, and can be used for moisturizing the skin or beauty and enhancing the skin barrier function.
Abstract
Description
本願は、2017年3月31日に、日本に出願された特願2017-071379号に基づき優先権を主張し、その内容をここに援用する。
種々の皮膚疾患、アトピー性皮膚炎、乾癬及び接触性皮膚炎等にみられる炎症症状においては、皮膚からの水分の消失が顕著に認められている。水分の消失には保湿機能の低下及び皮膚バリア機能の低下が関与している。
以上の状況のもと特許文献1は、グルコースがHMGB1の産生を促進し、HMGB1の細胞外への放出を誘導し、正常細胞の増殖、遊走を高め、組織の修復を促進することを開示している。
そこで、本発明は、皮膚バリア機能をより一層増強することができる皮膚バリア増強剤を提供することを目的とする。
本発明者らは、皮膚組織の修復を促進する皮膚バリア増強剤について鋭意検討した結果、酸性領域にpHを有する単糖及び糖アルコールの少なくとも一方を含有する液体が、皮膚バリア関連タンパク質であるFilaggrin及びClaudin-1の産生を促進する作用を有することを見出した。本発明者らは、これらの皮膚バリア関連タンパク質の産生が促進されれば、皮膚バリア機能を増強することができると考え、本発明を完成させた。
即ち、本発明は、以下の[1]~[6]の態様を有する。
[1] 単糖及び糖アルコールの少なくとも一方を含有し、pHが4.0以上7.0以下である皮膚バリア増強剤。
[2] 前記単糖としてグルコースを含有する[1]に記載の皮膚バリア増強剤。
[3] 前記糖アルコールとしてマンニトールを含有する[1]に記載の皮膚バリア増強剤。
[4] 前記pHが5.0以上7.0以下である[1]~[3]のいずれか1項に記載の皮膚バリア増強剤。
[5] [1]~[4]のいずれか1項に記載の皮膚バリア増強剤を含有し、pHが4.0以上7.0以下である医薬組成物。
[6] [1]~[4]のいずれか1項に記載の皮膚バリア増強剤を含有し、pHが4.0以上7.0以下である薬用化粧品。
[7] 液体状、ゲル状又はシート状である[6]に記載の薬用化粧品。
[8] [6]又は[7]に記載の薬用化粧品を、皮膚又は粘膜に塗布するステップを含む美容方法。
本発明の皮膚バリア増強剤は、単糖及び糖アルコールの少なくとも一方を含有し、pHが4.0以上7.0以下である。皮膚バリア増強剤の形態としては、pHを測定可能な形態であれば特に制限されない。具体例として液体状、ゲル状、シート状、ゾル、クリーム及び乳液等が例示されるが、これらに限定されない。
単糖としては、グルコース、フルクトース、マンノース、アラビノース、グロース、キシロース、リキソース、エリトロース、トレオース、ガラクトース及びソルボース等の単糖が例示されるが、これらに限定されない。これらの中でも皮膚バリア増強剤は、単糖としてグルコースを含有することが好ましい。
糖アルコールとしては、マンニトール、ソルビトール、キシリトール、エリトリトール、マルチトール、ラクチトール等が例示されるが、これらに限定されない。これらの中でも皮膚バリア増強剤は、糖アルコールとしてマンニトールを含有することが好ましい。
本発明の皮膚バリア増強剤は、pHが4.0以上7.0以下であり、5.0以上7.0以下であることが好ましい。pHが前記下限値以上であれば、皮膚バリア増強剤による皮膚組織及び細胞の障害が低減され、かつ、FilaggrinのmRNA及びClaudin-1のmRNAの転写及び発現が促進され、皮膚バリア機能が増強又は回復されやすい。pHが前記上限値以下であれば、Filaggrin及びClaudin-1の産生が十分に促進され、皮膚バリア機能が十分に増強又は回復される。
なお、本明細書において、pHとは、pH計測器(堀場製作所製「F-53」)を用いて、25℃で測定される値である。
緩衝液は、皮膚バリア増強剤のpHを4.0以上7.0以下に維持するものであれば、特に限定されない。緩衝液としては、リン酸緩衝液、クエン酸緩衝液、クエン酸-リン酸緩衝液、マレイン酸緩衝液、リンゴ酸緩衝液、コハク酸緩衝液、メタリン酸緩衝液、ソルビン酸緩衝液、炭酸緩衝液、トリスヒドロキシメチルアミノメタン-HCl緩衝液(トリス塩酸緩衝液)、MES緩衝液(2-モルホリノエタンスルホン酸緩衝液)、TES緩衝液(N-トリス(ヒドロキシメチル)メチル-2-アミノエタンスルホン酸緩衝液)、酢酸緩衝液、MOPS緩衝液(3-モルホリノプロパンスルホン酸緩衝液)、HEPES緩衝液(4-(2-ヒドロキシエチル)-1-ピペラジンエタンスルホン酸緩衝液)、等の緩衝液;グリシン-塩酸緩衝液、グリシン-NaOH緩衝液、グリシルグリシン-NaOH緩衝液及びグリシルグリシン-KOH緩衝液等のアミノ酸系緩衝液;トリス-ホウ酸緩衝液、ホウ酸-NaOH緩衝液、ホウ酸緩衝液等のホウ酸系緩衝液;イミダゾール緩衝液等が例示されるがこれらに限定されない。これらの中でも、リン酸緩衝液、クエン酸緩衝液、リンゴ酸緩衝液、コハク酸緩衝液及びメタリン酸緩衝液好ましい。緩衝液のpH調整は、塩酸及び酢酸等の酸性物質又は水酸化ナトリウム、アンモニア等の塩基性物質を用いて適宜調整することができる。
なお、上述の緩衝液を皮膚バリア増強剤に添加せずに、リン酸、酢酸、アミノ酸等の酸性物質を皮膚バリア増強剤に添加することによって、皮膚バリア増強剤のpHを4.0以上7.0以下に調整してもよい。
皮膚バリア増強剤は、本発明の効果を阻害しない範囲で、必要に応じて、単糖及び糖アルコールの少なくとも一方以外の任意成分を含有できる。
任意成分としては、賦形剤、増粘剤、担体、香料、色素等が例示される。これらの任意成分は、1種単独で用いられてもよいし、2種以上が組み合わされて用いられてもよい。任意成分については、後述の「(医薬品組成物)」の項で具体的に説明する。
すなわち、本発明は、皮膚バリア増強剤を製造するための、単糖及び糖アルコールの少なくとも一方と、任意成分であるpH調整剤との組み合わせの使用であり、pHが4.0以上7.0以下で、単糖及び糖アルコールの少なくとも一方を投与することを含む、前記使用である。
本発明の医薬組成物は、本発明の皮膚バリア増強剤を含有し、pHが4.0以上7.0以下である。
医薬組成物中の単糖及び糖アルコールの少なくとも一方の含有量は、医薬組成物(皮膚バリア増強剤)の用途、用法等を勘案して決定できる。例えば、医薬組成物を用いた経皮吸収剤であれば、100~5000mMが好ましく、1000~3000mMがより好ましい。医薬組成物を用いた注射剤であれば、1~1000mMが好ましく、6~165mMがより好ましい。単糖及び糖アルコールの少なくとも一方の含有量が前記下限値以上であれば、医薬組成物によって、Filaggrin及びClaudin-1の産生が促進され、皮膚バリア機能が増強又は回復されやすい。単糖及び糖アルコールの少なくとも一方の含有量が前記上限値以下であれば、単糖及び糖アルコールの少なくとも一方が医薬組成物中に均一に分散して、Filaggrin及びClaudin-1の産生が十分に促進され、皮膚バリア機能が十分に増強又は回復されやすい。
疾患が口腔内又は胃腸である場合、医薬組成物を経口剤として経口投与する方法、医薬組成物を注射剤として患部組織に注入する方法等が例示される。上部内視鏡下で患部を医薬組成物で洗浄し、患部に医薬組成物を塗布することにより、粘膜再生を促してもよい。これにより、健全な粘膜を形成でき、自然治癒を漸増させ、粘膜侵襲を防ぐことができる。さらに、医薬組成物を胃粘膜に塗布することで、胃粘膜の再生を促し、上皮組織のバリア機能を増強又は回復できる。
疾患が花粉症等の鼻・眼粘膜部の疾患である場合、医薬組成物を患部に塗布する方法が例示される。医薬組成物を鼻・眼粘膜部に塗布すると、鼻・眼粘膜部は、修復され、かつ粘膜層に強固なバリアを形成する。このため、鼻・眼粘膜部にアレルゲンが多数付着してもアレルギー反応を押さえ、鼻詰まりが生じたり、鼻汁又は涙が多量に分泌されたりするのを解消できる。
疾患が風邪等のウィルス、細菌の感染による上気道粘膜の炎症(感冒)である場合、医薬組成物を点鼻したり吸入させたりする方法が例示される。これにより、上気道粘膜の炎症を抑え、上気道粘膜を修復して再生して感染進行を抑制でき、かつ自然免疫応答の賦活化を促進できる。
なお、医薬組成物の投与量は、患者又は被験者の年齢、体重、疾患の種類・程度、投与方法等に応じて適宜決定される。
医薬品任意成分としては、薬理学的に許容される塩、賦形剤、増粘剤、担体、香料、色素等が例示される。これらの医薬品任意成分は、1種単独で用いられてもよいし、2種以上が組み合わされて用いられてもよい。
すなわち、本発明は、皮膚疾患の治療における医薬組成物を製造するための、単糖及び糖アルコールの少なくとも一方と、任意成分であるpH調整剤との組み合わせの使用であり、pHが4.0以上7.0以下で、単糖及び糖アルコールの少なくとも一方を投与することを含む、前記使用である。
本発明の医薬組成物は、皮膚バリア機能の増強方法に使用できる。皮膚バリア機能の増強方法は、有効成分量以上の単糖及び糖アルコールの少なくとも一方をpHが4.0以上7.0以下で投与することを含む。
本発明の薬用化粧品は、本発明の皮膚バリア増強剤を含有し、pHが4.0以上7.0以下である。本発明の薬用化粧品は、例えば、ローション、クリーム、乳液、ファンデーション等であって、皮膚疾患の予防及び皮膚の保湿を目的とする。なお、薬用化粧品は、日本国の薬事法に定められた医薬部外品に分類されるものである。
薬用化粧品の使用方法としては、例えば、美容の目的等に応じて適宜決定される。皮膚の美容及び保湿を目的とする薬用化粧品である場合、液体状、乳液状、ゲル状及びシート状の薬用化粧品を直接皮膚に塗布する方法等が例示される。
口腔内の美容を目的とする場合、液体状及びゲル状等の薬用化粧品を経口的に塗布する方法、液体状の薬用化粧品を注射剤として注入する方法等が例示される。
化粧品任意成分としては、流動パラフィン、セレシン、じろう、ラノリン、ワセリン、セタノール、スクワレン、ホホバ油、ステアリン酸、パルミチン酸、ラウリルアルコール、ステアリルアルコール、セチルアルコール、ミツロウ、メチルポリシロキサン、ジメチルシクロポリシロキサン等の油剤;プロパノール、グリコール、プロピレングリコール、ヒアルロン酸、コラーゲン、ポリエチレングリコール、ヒドロキシステアリン酸コレステリル、グリセリン、ソルビトール等の保湿剤;各種界面活性剤;乳化剤;賦形剤:増粘剤;pH調整剤;酸化防止剤;色素;香料;紫外線吸収剤等が例示される。これらの化粧品任意成分は、1種単独で用いられてもよいし、2種以上が組み合わされて用いられてもよい。
すなわち、本発明は、皮膚の保湿又は美容における薬用化粧品を製造するための、単糖及び糖アルコールの少なくとも一方と、任意成分であるpH調整剤との組み合わせの使用であり、pHが4.0以上7.0以下で、単糖及び糖アルコールの少なくとも一方を投与することを含む、前記使用である。
本発明の美容方法は、上述した本発明の薬用化粧品を皮膚又は粘膜に塗布する第1ステップを含む。薬用化粧品の塗布量は、患者又は被験者の年齢、体重、皮膚の状態、塗布の方法等に応じて適宜決定される。
本発明の美容方法は、上述した第1ステップと第2ステップを繰り返し行うことが好ましい。
以上説明したように本発明の皮膚バリア増強剤は、単糖及び糖アルコールの少なくとも一方を含有し、pHが4.0以上7.0以下であるから、細胞のFilaggrin及びClaudin-1の産生を促進することができ、FilaggrinのmRNA及びClaudin-1のmRNAの転写及び発現を促進することができる。したがって、皮膚バリア増強剤によって皮膚バリア機能が十分に増強又は回復される。
HuMedia-KG2培地(クラボウ社製)に、表1の濃度となるようにD-グルコース(製品番号:G5767、SIGMA社製)又はD-マンニトール(製品番号:M9647、SIGMA社製)と、培地のpHを調製するために塩酸(HCl)(製品番号:080-0106、和光純薬社製)とを添加し、表1に示すpHに調製された皮膚バリア増強剤を含有する培地(以下、「試験用培地」とも記す。)を調製した。なお、pHは、pH計測器(堀場製作所製「F-53」)を用いて、25℃で測定される値である。
実施例1~7及び比較例1~4,7の試験用培地を用いて、以下に記すFilaggrinタンパク質の発現量の解析と、Filaggrinタンパク質のmRNA及びClaudin-1タンパク質のmRNAの各発現量の解析を行い、その結果を図1~4に示す。
正常ヒト皮膚角化細胞(単一ドナー由来の初代培養の正常皮膚角化細胞、タカラバイオ株式会社製)をHuMedia-KG2培地(pH7.6、クラボウ社製)を用いて、6穴(6well)マイクロプレートに1,000,000cells/wellの細胞密度となるように播種し、37℃で24時間培養した(前培養)。前培養後、培養液を吸引除去し、培地を実施例1~6及び比較例1~4の試験培地と置換し、37℃で30分間培養した(本培養)。本培養後、培養上清を吸引除去し、培地をpHが7.6であるHuMedia-KG2培地と置換し、37℃で3時間培養した(後培養)。後培養後、培地を除去し、培養した細胞からタンパク質を抽出した。抽出したタンパク質をSDS-PAGEにより展開した。その後、抽出したタンパク質をニトロセルロース膜に転写し、マウス由来のFilaggrin抗体を用いたwestern blot法によって、培養細胞内のFilaggrinタンパク質の発現量を解析した。また、内在性コントロールとして、同一サンプルのβ-actinタンパク質の発現量を解析した。検出されたそれぞれのタンパク質のバンドの濃度をデンシトメーターで測定し、Filaggrinタンパク質及びβ-actinタンパク質の発現量を解析した。前記同一サンプルのβ-actinタンパク質の発現量を1とし、Filaggrinタンパク質の発現量の相対値(相対発現量)で、Filaggrinタンパク質の発現量を評価した。結果を図1に示す。
試験培地を実施例2,4,6及び比較例2,4の試験培地に変更した以外は、「<Filaggrinタンパク質の発現量の解析1>」と同様の手法にて前培養、本培養及び後培養を行った。後培養後、培地を除去し、培養した細胞からTotal RNAを抽出した。RT-PCR法を用いて、抽出したRNAからFilaggrin及びClaudin-1のmRNAを増幅し、各mRNAの発現量を解析した。比較例2の各mRNAの発現量を1とし、実施例2,4,6及び比較例4の各mRNAの発現量の相対値(相対発現量)で、FilaggrinのmRNA及びClaudin-1のmRNAの発現量を評価した。結果を図2,3に示す。
実施例7及び比較例7の試験用培地を用いた以外は、「<Filaggrinタンパク質の発現量の解析1>」と同様の手法にて実施例7及び比較例7のFilaggrinタンパク質の発現量の解析を行った。なお、比較例7のFilaggrinタンパク質の発現量を1とし、実施例7のFilaggrinタンパク質の発現量の比較例7のFilaggrinタンパク質の発現量に対する相対値(相対発現量)で実施例7のFilaggrinタンパク質の発現量を評価した。結果を図4に示す。
実施例1~6及び比較例2,4の結果から、試験培地のグルコース濃度を110mMとし、pHを3.0以上7.0以下に調整することにより、培養細胞内のFilagglin及びClaudin-1の各mRNAの発現を促進できることが判った。
比較例3,4の結果から、グルコース濃度を110mMとし、pHを3.0に調整することにより、Filagglinタンパク質のmRNAの発現は促進されるが、Filagglinタンパク質は検出されないことが判った。
以上の結果から、本発明を適用することで、Filagglin及びClaudin-1の発現が促進され、皮膚バリア機能を増強できることが判った。
「<Filaggrinタンパク質の発現量の解析1>」の項と同様にして、正常ヒト皮膚角化細胞を培養した。培養した細胞の障害の程度をLDH法によって解析した。各例の試験培地で培養した細胞をTriton X 100で処理した細胞の障害の程度を100%としたときの、各例で培養した細胞の障害率を評価した。
表1及び図5に示すように、本発明を適用した実施例1~6の細胞の障害率は、pHが7.6である比較例1,2の細胞の障害率より低かった。一方、pHが3.0である比較例3,4の細胞の障害率は、pHが7.6である比較例1,2の細胞の障害率より高かった。
すなわち、pHが5.0以上である試験培地で本発明を適用することによって、皮膚バリア機能が回復されて、細胞障害が低減されることが判った。
グルコース濃度を20%に、pHを6.0にそれぞれ調製したリン酸緩衝生理食塩水(PBS)の水溶液を作製し、本発明を適用した皮膚バリア増強剤とした。
馴化飼育後の8週令メスNC/Ngaマウスの右耳介部に、ダニ抗原を週2回の頻度でDay0からDay14まで皮内投与した。これと並行して、本発明の皮膚バリア増強剤をDay0からDay28まで1日1回、毎日塗布投与した。Mock群としてpHを7.6に調製したPBSを、Day0からDay28まで1日1回、毎日塗布投与した。
Day28にMock群及び皮膚バリア増強剤を塗布した群の耳介部の写真を撮影し,イソフルラン麻酔下で放血、安楽死させたマウスから左右の耳介部を採取し、中性ホルマリン固定を行なった。パラフィン包埋後、組織切片を作製した。同切片をヘマトキシリン・エオジン(HE)溶液で染色するとともに、マウス由来のFilaggrin及びClaudin-1抗体を用いて免疫染色を行なった。Negative controlには、8週令メスNC/Ngaマウスを無処置のままDay28まで飼育したマウスの耳介切片を用いた。
正常ヒト皮膚角化細胞をHuMedia-KG2培地(pH7.6、クラボウ社製)を用いて、6穴(6well)マイクロプレートに1,000,000cells/wellの細胞密度となるように播種し、37℃で24時間培養した(前培養)。前培養後、培養液を吸引除去し、培地を比較例2の試験培地と置換し、37℃で本培養した。本培養の時間を0,0.5,1.0,2.0,3.0,4.0,5.0時間としたときの各培養液から、培地を除去し、培養した細胞からTotal RNAを抽出した。RT-PCR法を用いて、各培養液から抽出したRNAからFilaggrin及びClaudin-1のmRNAを増幅し、各mRNAの発現量を本培養の時間が異なる各培養液について解析した。本培養の時間が0時間であるときの各mRNAの発現量を1とし、各培養時間の細胞の各mRNAの発現量の相対値(相対発現量)でFilaggrin及びClaudin-1の各mRNAの発現量を評価した。
培地を比較例5の試験培地を用い、本培養の時間を0,2.0,4.0,6.0,24.0時間としたこと以外は、「<Filaggrinタンパク質の発現量の解析1>」の項と同様にして、各培養時間におけるFilaggrinタンパク質の発現量を評価した。
図9は、参考試験2のFilaggrinタンパク質の発現量の評価結果を示すグラフである。発現誘導を開始してから2時間でFilaggrinタンパク質の発現量の増加が認められた。Filaggrinタンパク質の発現量の増加は、発現誘導を開始してから24時間までの間認められた。
培地を比較例6の試験培地を用い、マウス由来のClaudin-1抗体を用いた以外は、「<参考例2>」の項と同様の手法によって、各培養時間におけるClaudin-1タンパク質の発現量を評価した。
図10は、参考試験3のClaudin-1タンパク質の発現量の評価結果を示すグラフである。発現誘導を開始してから2時間でClaudin-1タンパク質の発現量の増加が認められた。Claudin-1タンパク質の発現量の増加は、発現誘導を開始してから24時間までの間認められた。
参考試験2,3の結果でも示したようにpHが7.6である比較例2,5,6でも、グルコースを55~165mM含有することにより、皮膚バリア関連タンパク質の発現量及び産生量の増加が認められた(図8~図10)。しかし本発明の皮膚バリア増強剤は、pHが4.0以上7.0以下であることにより、皮膚バリア関連タンパク質の発現をより一層増強することができる(図1,図4)。よって、本発明の皮膚バリア増強剤は、従来の皮膚バリア増強剤より効果的に皮膚バリア機能を増強することができる。したがって、本発明の皮膚バリア増強剤を、医薬品等に適用すれば、皮膚組織を一層効果的に修復することができる等の顕著な効果を期待できる。
Claims (8)
- 単糖及び糖アルコールの少なくとも一方を含有し、pHが4.0以上7.0以下である皮膚バリア増強剤。
- 前記単糖としてグルコースを含有する請求項1に記載の皮膚バリア増強剤。
- 前記糖アルコールとしてマンニトールを含有する請求項1に記載の皮膚バリア増強剤。
- 前記pHが5.0以上7.0以下である請求項1~3のいずれか一項に記載の皮膚バリア増強剤。
- 請求項1~4のいずれか一項に記載の皮膚バリア増強剤を含有し、pHが4.0以上7.0以下である医薬組成物。
- 請求項1~4のいずれか一項に記載の皮膚バリア増強剤を含有し、pHが4.0以上7.0以下である薬用化粧品。
- 液体状、ゲル状又はシート状である請求項6に記載の薬用化粧品。
- 請求項6又は7に記載の薬用化粧品を、皮膚又は粘膜に塗布するステップを含む美容方法。
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP18776267.9A EP3610863A4 (en) | 2017-03-31 | 2018-03-28 | MEANS FOR IMPROVING SKIN BARRIER, MEDICAL COMPOSITION, MEDICAL COSMETIC AND BEAUTY CARE PROCEDURE |
CN201880022486.5A CN110461317A (zh) | 2017-03-31 | 2018-03-28 | 皮肤屏障增强剂、药物组合物、药用化妆品及美容方法 |
US16/493,482 US20210128437A1 (en) | 2017-03-31 | 2018-03-28 | Agent for enhancing skin barrier, medicinal composition, medicated cosmetic, and beauty care method |
KR1020197027511A KR20190116475A (ko) | 2017-03-31 | 2018-03-28 | 피부 장벽 증강제, 의약 조성물, 약용 화장품 및 미용 방법 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2017071379A JP2018172328A (ja) | 2017-03-31 | 2017-03-31 | 皮膚バリア増強剤、医薬組成物、薬用化粧品、及び美容方法 |
JP2017-071379 | 2017-03-31 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2018181599A1 true WO2018181599A1 (ja) | 2018-10-04 |
Family
ID=63676287
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2018/013013 WO2018181599A1 (ja) | 2017-03-31 | 2018-03-28 | 皮膚バリア増強剤、医薬組成物、薬用化粧品及び美容方法 |
Country Status (7)
Country | Link |
---|---|
US (1) | US20210128437A1 (ja) |
EP (1) | EP3610863A4 (ja) |
JP (1) | JP2018172328A (ja) |
KR (1) | KR20190116475A (ja) |
CN (1) | CN110461317A (ja) |
TW (1) | TW201838620A (ja) |
WO (1) | WO2018181599A1 (ja) |
Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH11322575A (ja) * | 1998-03-17 | 1999-11-24 | Shiseido Co Ltd | 肌荒れ改善・防止用皮膚外用剤 |
JP2000103728A (ja) * | 1998-07-28 | 2000-04-11 | Shiseido Co Ltd | 皮膚バリア―機能回復促進剤 |
JP2006045186A (ja) * | 2004-07-05 | 2006-02-16 | Fancl Corp | ラメラ構造再生剤及び皮膚外用剤 |
JP2006315962A (ja) * | 2005-05-10 | 2006-11-24 | Geo Co Ltd | システイン含有水性組成物 |
WO2011061932A1 (ja) * | 2009-11-18 | 2011-05-26 | 株式会社ロッテ | セラミド及びコラーゲンの合成促進剤並びにコラーゲンの糖化抑制剤 |
WO2013111729A1 (ja) * | 2012-01-23 | 2013-08-01 | 株式会社Cac | 医薬組成物及びこれを用いた薬用化粧品 |
JP2017071379A (ja) | 2015-10-09 | 2017-04-13 | 国立大学法人 名古屋工業大学 | 陸上走行可能な飛行体 |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH10287550A (ja) * | 1997-04-10 | 1998-10-27 | Nippon Rideia Oririi Kyokai | 皮膚外用剤 |
EP0943324A3 (en) * | 1998-03-17 | 2002-01-02 | Shiseido Company, Ltd. | External skin treatment composition |
US20040044077A1 (en) * | 2000-12-28 | 2004-03-04 | Chika Katagiri | Agents for inhibiting or restoring skin damage caused by drying and method for evaluating the same |
FR2963555A3 (fr) * | 2010-08-03 | 2012-02-10 | Sephar Soc D Etudes Pharma | Nouvelle composition cosmetique et ses applications dans les corrections des epidermes deshydrates, inflammatoires et necessitant un apport nutritionnel |
-
2017
- 2017-03-31 JP JP2017071379A patent/JP2018172328A/ja active Pending
-
2018
- 2018-03-28 EP EP18776267.9A patent/EP3610863A4/en not_active Withdrawn
- 2018-03-28 KR KR1020197027511A patent/KR20190116475A/ko not_active Application Discontinuation
- 2018-03-28 US US16/493,482 patent/US20210128437A1/en not_active Abandoned
- 2018-03-28 CN CN201880022486.5A patent/CN110461317A/zh active Pending
- 2018-03-28 WO PCT/JP2018/013013 patent/WO2018181599A1/ja active Application Filing
- 2018-03-29 TW TW107110947A patent/TW201838620A/zh unknown
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH11322575A (ja) * | 1998-03-17 | 1999-11-24 | Shiseido Co Ltd | 肌荒れ改善・防止用皮膚外用剤 |
JP2000103728A (ja) * | 1998-07-28 | 2000-04-11 | Shiseido Co Ltd | 皮膚バリア―機能回復促進剤 |
JP2006045186A (ja) * | 2004-07-05 | 2006-02-16 | Fancl Corp | ラメラ構造再生剤及び皮膚外用剤 |
JP2006315962A (ja) * | 2005-05-10 | 2006-11-24 | Geo Co Ltd | システイン含有水性組成物 |
WO2011061932A1 (ja) * | 2009-11-18 | 2011-05-26 | 株式会社ロッテ | セラミド及びコラーゲンの合成促進剤並びにコラーゲンの糖化抑制剤 |
WO2013111729A1 (ja) * | 2012-01-23 | 2013-08-01 | 株式会社Cac | 医薬組成物及びこれを用いた薬用化粧品 |
JP2013147480A (ja) | 2012-01-23 | 2013-08-01 | Cac:Kk | 医薬組成物及びこれを用いた薬用化粧品 |
JP2017071379A (ja) | 2015-10-09 | 2017-04-13 | 国立大学法人 名古屋工業大学 | 陸上走行可能な飛行体 |
Non-Patent Citations (4)
Title |
---|
NAKAMURA KIYOKA: "Skin of acidity recovery effect, Antibacterial effect of Capryloyl glycine and its application to cosmetics", FRAGRANCE JOURNAL, no. 2011, February 2011 (2011-02-01), pages 36 - 40, XP009516271, ISSN: 0288-9803 * |
See also references of EP3610863A4 |
TEZUKA TADASHI: "Special EDITION/ Percutaneous absorption and barrier function and permeability of liposome skin", FRAGRANCE JOURNAL, no. 87, 1987, pages 27 - 32, XP009516272, ISSN: 0288-9803 * |
YAMADA,K. ET AL.: "Topical Glucose induces Claudin-1 and Filaggrin Expression in a Mouse Model of Atopic Dermatitis and in Keratinocyte Culture, Exerting Anti-inflammatory Effects by Repairing Skin Barrier Function", ACTA DERMATO-VENEREOLOGICA, vol. 98, no. 1, 12 January 2018 (2018-01-12), pages 19 - 25, XP055612895 * |
Also Published As
Publication number | Publication date |
---|---|
EP3610863A1 (en) | 2020-02-19 |
CN110461317A (zh) | 2019-11-15 |
KR20190116475A (ko) | 2019-10-14 |
US20210128437A1 (en) | 2021-05-06 |
EP3610863A4 (en) | 2020-12-16 |
TW201838620A (zh) | 2018-11-01 |
JP2018172328A (ja) | 2018-11-08 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JPH08208488A (ja) | 皮膚外用剤 | |
US11376231B2 (en) | Compositions of bioactive fulvate fractions and uses thereof | |
TW201912156A (zh) | 組成物在製備治療睡眠障礙的藥劑的用途 | |
JP5891651B2 (ja) | 角層細胞分化正常化用皮膚外用剤 | |
JP5685315B2 (ja) | ニコチン酸アデニンディヌクレオチドリン酸又はその誘導体を含む薬学又は化粧料組成物 | |
JP4300370B2 (ja) | 上皮改善剤 | |
WO2018181599A1 (ja) | 皮膚バリア増強剤、医薬組成物、薬用化粧品及び美容方法 | |
WO2017098396A1 (en) | Synergistic combination of pyrrolidone carboxylic acid and/or salts thereof and hyaluronic acid and/or salts thereof, for use in the treatment and/or prevention of dryness and irritation of the mucosae, and related pharmaceutical formulations | |
TWI468184B (zh) | 藥學組成物及使用其之藥用化妝品 | |
JP5455248B2 (ja) | 保湿作用を有する皮膚外用剤 | |
WO2017114379A1 (zh) | 一种治疗多发性硬化症的鼻用凝胶组合物 | |
JP6484547B2 (ja) | 掻痒改善剤 | |
JP6656890B2 (ja) | フィラグリン産生促進剤 | |
JP7396585B2 (ja) | Tslp遺伝子発現抑制用、il-33遺伝子発現抑制用、又はフィラグリン産生促進用組成物 | |
KR20130056825A (ko) | 면역억제제 및 트랜스글루타미나제 2 억제제를 포함하는 아토피 피부염의 예방, 치료 또는 개선용 조성물 | |
JP3187636B2 (ja) | コラーゲン代謝改善剤 | |
JPH10316575A (ja) | 表皮角質化促進剤 | |
JP2006273756A (ja) | セラミド合成促進剤、コラゲナーゼ阻害剤及びコラーゲン合成促進剤 | |
JP2006117552A (ja) | 光老化防止用皮膚外用剤 | |
JP2022155642A (ja) | 医薬品または化粧品組成物 | |
JP5923410B2 (ja) | コラーゲンゲル収縮促進剤 | |
WO2014131096A1 (pt) | Formulação obtida pela associação do antiviral aciclovir, com vitamina b6 e anti-histamínico para tratamento do herpes e meio de aplicação | |
CN116829165A (zh) | 用于预防、治疗或减轻病毒感染性疾病或呼吸道疾病的包含副干酪乳杆菌来源的囊泡的组合物 | |
JP2016074634A (ja) | 経口用肌のキメ改善剤 | |
JPS5931712A (ja) | メナジオンを原料とする安定なタムシ治療剤の製造法 |
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: 18776267 Country of ref document: EP Kind code of ref document: A1 |
|
ENP | Entry into the national phase |
Ref document number: 20197027511 Country of ref document: KR Kind code of ref document: A |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2018776267 Country of ref document: EP |
|
ENP | Entry into the national phase |
Ref document number: 2018776267 Country of ref document: EP Effective date: 20191031 |