WO2017011984A1 - 贻贝粘蛋白产品治疗和预防黑色素相关疾病中的应用 - Google Patents

贻贝粘蛋白产品治疗和预防黑色素相关疾病中的应用 Download PDF

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Publication number
WO2017011984A1
WO2017011984A1 PCT/CN2015/084494 CN2015084494W WO2017011984A1 WO 2017011984 A1 WO2017011984 A1 WO 2017011984A1 CN 2015084494 W CN2015084494 W CN 2015084494W WO 2017011984 A1 WO2017011984 A1 WO 2017011984A1
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WIPO (PCT)
Prior art keywords
mussel mucin
mussel
mucin
melanin
enzyme
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PCT/CN2015/084494
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English (en)
French (fr)
Inventor
高敏
Original Assignee
江阴市本特塞缪森生命科学研究院有限公司
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Priority to CN202210732149.3A priority Critical patent/CN115634282A/zh
Priority to EP15898540.8A priority patent/EP3326640B1/en
Application filed by 江阴市本特塞缪森生命科学研究院有限公司 filed Critical 江阴市本特塞缪森生命科学研究院有限公司
Priority to JP2017568248A priority patent/JP6778220B2/ja
Priority to DK15898540.8T priority patent/DK3326640T3/da
Priority to KR1020187001331A priority patent/KR102468524B1/ko
Priority to CN201580081661.4A priority patent/CN108135972B/zh
Priority to CA2992315A priority patent/CA2992315C/en
Priority to PCT/CN2015/084494 priority patent/WO2017011984A1/zh
Priority to SG11201800201QA priority patent/SG11201800201QA/en
Priority to MX2018000335A priority patent/MX2018000335A/es
Priority to US15/742,969 priority patent/US10675327B2/en
Priority to BR112018000955-5A priority patent/BR112018000955A2/zh
Priority to AU2015402833A priority patent/AU2015402833B2/en
Publication of WO2017011984A1 publication Critical patent/WO2017011984A1/zh
Priority to HK18114980.3A priority patent/HK1255896A1/zh
Priority to US16/859,458 priority patent/US11090360B2/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1767—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/56—Materials from animals other than mammals
    • A61K35/618—Molluscs, e.g. fresh-water molluscs, oysters, clams, squids, octopus, cuttlefish, snails or slugs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • A61K8/987—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of species other than mammals or birds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0014—Skin, i.e. galenical aspects of topical compositions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/12—Aerosols; Foams
    • 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
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/46—Hydrolases (3)
    • A61K38/48—Hydrolases (3) acting on peptide bonds (3.4)
    • A61K38/482—Serine endopeptidases (3.4.21)
    • A61K38/4826—Trypsin (3.4.21.4) Chymotrypsin (3.4.21.1)

Definitions

  • the present invention relates generally to the fields of pharmaceuticals, cosmetics, medical products, disinfecting products, health care products, foods, and daily chemical products, and more particularly to mussel mucin products and their use in the treatment and prevention of melanin-related diseases.
  • Melanin is a dark brown pigment found in animal skin or hair, produced by a special cell, melanocytes, and stored therein. Melanin is widely found in human skin, mucous membranes, retina, pia mater, gallbladder and ovary. It is because of the presence of melanin that the skin has color. Melanin is a bio-pigment formed by a series of chemical reactions of tyrosine or 3,4-dihydroxyphenylalanine, and is usually present in a polymeric manner.
  • Melanin is essentially a protein. They are found in the middle of cells in the basal layer of the skin (also called “pigment cells”) and are a substance called “melanin biomass". The pigment mother cells secrete melanin pigment. When ultraviolet rays (UVA and UVB) are applied to the skin, UVB acts on the basal layer of the skin, and the skin is in a “self-protective” state. The ultraviolet light stimulates the melanin pigment and activates tyrosine. The activity of the enzyme protects the skin cells. Dopa is the precursor of melanin, which is oxidized by tyrosine and releases melanin. Melanin moves through layers of cellular metabolism and reaches the epidermis of the skin to form freckles, sunburn, and dark spots.
  • melanin is mostly related to endocrine disorders and sun exposure.
  • the number of melanocytes depends mainly on heredity, and is also related to endocrine hormones and nutritional status.
  • the diseases caused by melanin such as freckles, chloasma, melanosis and other pigmentation diseases It is a subject of extensive research in the beauty industry.
  • the inhibition of the pigment during its formation is different from the treatment of the pigment that has been produced.
  • Inhibition of pigmentation lies in the process of stopping the passage of tyrosine to dopa to dihydroxyanthracene until melanin is formed. This process can inhibit pigmentation by inhibiting inflammation, preventing the formation of inflammatory factors, or blocking pigmentation by substances that react with substances in the pigmentation process, or preventing pigmentation by antioxidants. .
  • the formed melanin is decomposed by a reducing substance, so that the pigment eventually disappears.
  • Mussel adhesive protein also known as Mytilus edulis foot protein (Mefp)
  • Mefp Mytilus edulis foot protein
  • Mytilus coruscus A special protein secreted by Perna viridis. Mussels are usually attached to the reefs on the coast in groups. On the bottom of the ship or the ship, there is the ability to withstand wave impact in the offshore. In fact, mussels can be attached extremely strongly to the substrate of any material, such as metal, wood, glass, and the like. The main reason for the above characteristics of mussels is that they can form and store this special mucin in the girth of the foot. The mussels release the mucin through the foot silk to a solid surface such as rock to form a water-resistant combination. Fix yourself.
  • Mussel mucin has two structural features: (1) containing lysine, which has a high loading of positive charge; (2) containing 3,4 dihydroxyphenylalanine (DOPA, dopa). The cells and tissues of the human body are negatively charged.
  • Mussel mucin plays a protective and therapeutic role by tightly binding cells and tissues through the electrostatic interaction between its own positive charge and the negative charge of cells and tissues.
  • dopa oxidation produces ortho-dioxins, which can be cross-linked with unoxidized dopa to form a membrane or a network scaffold, which promotes the protein to adhere more closely and firmly to the surface of the human body, thereby protecting.
  • Mussel mucin is a macromolecular protein that is completely degraded in the human body for about 3-10 days. Its ability to attach to cell tissues is excellent, so that mussel mucin can be stabilized locally and continue to function.
  • mussel mucin has the above characteristics, its current application field is very limited.
  • Commercial mussel mucin products are Cell-Tak from BD Biosciences, MAP Trix from Kollodis, Korea, and Hydrogel from Biopolymer, Sweden. These products are either used directly in the mussel mucin solution state, or are stored as lyophilized powder formulations and dissolved prior to use. Their primary application is limited to microscopic cell adhesion and tissue adhesives. Mussel mucin has also been reported for use in the repair of fetal membranes, as a coating against seawater corrosion, and as a drug-loaded stent for the heart.
  • mussel mucin as a polyphenol protein has a dopa group contained in the molecule which is an important substance involved in melanin synthesis, and is caused by melanin-induced diseases such as chloasma, freckles, and melanosis.
  • the dopa group in the mucin mucin molecule can enhance the activity of tyrosinase, thereby affecting the secretion of melanin, and achieving the purpose of treating and preventing melanin-related diseases.
  • the molecular weight of mussel mucin is as high as 100 kD, and the absorption rate when used directly by transdermal is low, which affects the effect.
  • Minimally invasive skin formation or enzymatic hydrolysis of mussel mucin can improve the efficiency of transdermal action, more effectively treat chloasma, freckles, melanosis, other pigmented skin diseases, and effectively inhibit melanoma.
  • Skin cancer prevents pigmentation that may occur from skin diseases such as sun exposure or acne.
  • Mussel mucin used herein refers to Mytilus edulis Linnaeus, Mytilus coruscus or Perna viridis from the Mytilidae bivalve mollusc. 11 subclasses of mussel mucin, currently known as purified from marine mussels: mefp1, mefp-2, mefp-3, mefp-4, mefp-5, mefp-6, collagen pre-COL- A mixture of one or more of P, pre-COL-D, pre-COL-NG, foot silk matrix proteins PTMP and DTMP.
  • the mussel mucin used herein may have a pH of 1.0 to 7.0 in an aqueous solution, and particularly may be in the range of pH 3.0 to 6.5 to make the therapeutic effect better.
  • the mussel mucin used herein can be obtained by the following preparation method, for example, a method for separating and purifying mussel mucin using mixed adsorption chromatography in Chinese Patent No. ZL200710179491.0, a kind of carboxy using Chinese Patent No. ZL200710179492.5 A method for purifying mussel mucin by methyl ion exchange chromatography, a method for separating and purifying mussel mucin using salting out and dialysis, Chinese Patent No. ZL200910087567.6.
  • the mussel mucin used herein may be in the form of a solution or a lyophilized powder, in particular, the concentration of mussel mucin in the product may be 0.1-15.0 mg/ml, and when the concentration is too low, the effect of mussel mucin Not large, when the concentration is too high, it can cause cytotoxicity, skin irritation, etc., which is not conducive to the treatment of skin melanin.
  • the mussel mucin used herein can also be prepared as a liquid agent by combining it with an excipient.
  • An exemplary mussel mucin liquid preparation is prepared by dissolving or diluting mussel mucin solution mother liquor or lyophilized powder to a certain concentration or pH, and the solution for dissolving or diluting may be water, physiological saline, phosphate solution, vinegar. Acid solution, borate solution, and the like.
  • the pH of mussel mucin in the final product may be pH 1.0-7.0, and in particular, the therapeutic effect may be better in the range of pH 3.0-6.5.
  • the mussel mucin used herein can also be prepared as a gelling agent in combination with an excipient.
  • An exemplary mussel mucin gel is prepared by mixing a mussel mucin solution or a lyophilized powder with a gel matrix material, which may be selected from the group consisting of cellulose derivatives, carbomers, and seaweeds. Acid salt, tragacanth, gelatin, pectin, carrageenan, gellan gum, starch, xanthan gum, cationic guar gum, agar, non-cellulosic polysaccharide, ethylene polymer, acrylic resin, polyvinyl alcohol or poly One of carboxyvinyl or any combination thereof.
  • the mussel mucin used herein can also be combined with excipients to prepare a gel.
  • An exemplary mussel mucin gel is prepared by mixing a mussel mucin solution or a lyophilized powder with a gel matrix, which may include cellulose derivatives, glycerin, non-cellulosic polysaccharides, propylene glycol. One or any combination thereof.
  • the mussel mucin used herein can also be combined with an excipient to prepare a paste.
  • An exemplary mussel mucin paste is prepared by mixing mussel mucin with a paste matrix material, which may include glycerin, petrolatum, paraffin Wait.
  • the mussel mucin used herein can also be combined with a matrix material to form a dressing and application for the skin surface.
  • the liquid in the exemplary mussel mucin application may be a mussel mucin solution or a cellulose derivative, carbomer and alginate, tragacanth, gelatin, pectin, carrageenan, Gellan gum, starch, xanthan gum, cationic guar gum, agar, non-cellulosic polysaccharide, ethylene polymer, acrylic resin, polyvinyl alcohol or carbomer, gelatin, fish gelatin, pectin, alginate, glycerin One or more combinations of petrolatum, paraffin, polyethylene glycol, vitamins, glutathione.
  • the above mussel mucin solution or combination is immersed in a matrix material such as gauze, non-woven fabric, silk paper, and the application may be, for example, a wound patch, a mask, an eye mask, a hand mask, a foot film, or the like.
  • glycerin, polyethylene glycol, vitamins, glutathione, and the like which are known in the art to increase their moisturizing and anti-oxidation properties can be further added to various preparations including mussel mucin, and the moisturizing and anti-oxidation can be further improved.
  • the mussel mucin used herein can be used as a main raw material to prepare a medicine using a pharmaceutically acceptable carrier.
  • the drug may be a liquid agent, a gel, a gel, a paste, an applicator, or a foaming agent.
  • the medicament can be used for in vitro use, in particular for external use on the skin.
  • the mussel mucin used herein can be used as a main raw material to prepare a medical device.
  • the term medical device as used herein refers to materials that are used directly or indirectly to the human body and other similar or related items.
  • the medical device may be a liquid agent, a gel, a gel, a paste, an applicator, or a foaming agent.
  • the medical device can be used for in vitro use, in particular for external use on the skin.
  • the mussel mucin used herein can be used as a main raw material to prepare a cosmetic using an auxiliary material which is acceptable in the field of cosmetics.
  • the cosmetic may be a liquid, a gel, a gel, a paste, an applicator, or a foaming agent.
  • the cosmetic can be used for in vitro use, in particular for topical application to the skin.
  • the mussel mucin used herein can be used as a main raw material to prepare a disinfecting product using an excipient which is acceptable in the field of disinfecting products.
  • the term disinfecting product as used herein refers to a disinfectant, a disinfecting device, a sanitary article, and a disposable medical article that chemically, physically, or biologically kill or eliminate pathogenic microorganisms in the environment.
  • the disinfecting product may be a liquid agent, a gel, a gel, a paste, an applicator, or a foaming agent.
  • the disinfecting product can be used for in vitro use, in particular for external use on the skin.
  • the mussel mucin used herein can be used as a main raw material, and a health care product or food can be prepared by using an excipient which is acceptable in the field of health care or food.
  • the health care product or food may be a liquid agent, a gel, a gel, a paste, an applicator, or a foaming agent.
  • the health supplement or food can be used for in vitro use or consumption, in particular for external use on the skin.
  • the mussel mucin used herein can be used as a main raw material to prepare a daily chemical product using an auxiliary material acceptable in the field of daily chemical products.
  • the term daily chemical product as used herein refers to technical chemicals that are used daily, including shampoos, shower gels, and the like.
  • the daily chemical product may be a liquid agent, a gel, a gel, a paste, an applicator, or a foaming agent.
  • the daily chemical product can be used for in vitro use, in particular for external use on the skin.
  • Another object of the invention is to provide a mussel mucin product for use in the treatment and prevention of melanin deposition.
  • it is used to treat pigmentation diseases such as chloasma, freckles, and melanosis, inhibit skin cancer represented by melanoma, and prevent pigmentation which may occur due to skin diseases such as sun exposure or acne.
  • treatment refers to the process of interfering with or altering a particular state of health, the activity performed to relieve the pain.
  • inhibition as used herein refers to the process by which tissue or body is stimulated to attenuate the original damage.
  • prevention refers to pre-emptive measures that may deviate from subjectively expected or objective general rules in the course of disease treatment to avoid possible damage.
  • Chloasma refers to the yellow-brown pigmentation of the face. Its multiple symmetrical butterfly shape is distributed on the cheek.
  • Freckles are yellow-brown punctate pigmentation spots that occur on facial skin and are autosomal dominant. It occurs on the face, especially the nose and cheeks, and can affect the exposed parts such as the neck, shoulders, and back of the hand. There is no rash on the non-exposed parts. The damage is light brown or dark brown needle size to mung bean big spotted, round, oval or irregular. Scattered or clustered, isolated and not integrated.
  • Melanosis is a chronic inflammation of the skin caused by long-term exposure of workers to asphalt, coal tar, petroleum products or long-term inhalation of volatiles of such substances, and eventually skin pigmentation occurs.
  • Melanoma is a tumor produced by melanocytes of the skin and other organs. Skin melanoma manifests as a marked change in pigmented skin lesions over months or years.
  • Pigmentation caused by skin diseases is caused by inflammatory skin diseases or by various drugs.
  • solar dermatitis or acne treatment leads to hyperpigmentation.
  • Lichen planus and lichen planus-like drug rash cause pigmentation.
  • a fixed drug eruption can leave a characteristic ring pigmentation spot.
  • Drugs that cause hyperpigmentation include amiodarone, tetracycline, minocycline, bleomycin, cyclophosphamide, antimalarial chloroquine and quinine, chlorpromazine and others that cause gray-blue pigment changes in exposed areas Phenothiazines.
  • mussel mucin can be used independently for the treatment and prevention of melanin deposition.
  • mussel mucin when used independently for the treatment and prevention of melanin deposition, it can be used directly or it can form tiny wounds first. After use.
  • Methods for forming microscopic wounds include: microneedles (which can be made of silicon, silicon dioxide, cellulose, etc.), needle rollers (which can be made of stainless steel, titanium, polytetrafluoroethylene, etc.), lasers (eg, dot lasers) ).
  • mussel mucin can be bound to an enzyme and used after enzymatic degradation.
  • the mussel mucin inhibits the oxidation process of melanin formation and binds to dopa (3,4-dihydroxyphenylalanine) in the formation of melanin to stop the further formation of the pigment.
  • dopa 3,4-dihydroxyphenylalanine
  • Enzymatic hydrolysis of mussel mucin improves the efficiency of transdermal administration and enhances the therapeutic effect.
  • the enzymes used herein may be: (1) trypsin from various sources, such as trypsin extracted from salmon pancreas, bovine pancreas, porcine pancreas, krill; (2) collagen of various origins Enzymes; (3) casein from various sources; (4) chymotrypsin from various sources; (5) carboxypeptidases from various sources; (6) keratinases from various sources; (7) various sources Enterokinase; (8) various sources of chymosin and the like.
  • the enzyme which can be used together with mussel mucin for melanin treatment is not limited to the enzyme described above.
  • mussel mucin can be used in combination with an enzyme for treating melanin.
  • the use forms may include: (1) first spraying the enzyme to degrade the stratum corneum of the skin surface, then spraying the enzyme again, and spraying the mussel mucin immediately, for a period of time. Time; (2) first spraying mussel mucin, then spraying the enzyme to work together for a period of time; (3) mixing mussel mucin with the enzyme for a period of time after spraying; (4) spraying the enzyme first, then spraying the mussel sticky Protein, work together for a while.
  • the form of the mussel mucin and the enzyme-forming product may include: (1) the mussel mucin and the enzyme are in different packages: respectively, or mixed by the user before use; (2) Mussel mucin hydrolyzed peptide mixture: mussel mucin is prepared by using the above various enzymes to prepare a mixture of hydrolyzed peptides, and the product is directly formed by stopping the activity of the enzyme by heating or the like; (3) mussel mucin single hydrolyzed peptide: Mussel mucin is prepared by using the above various enzymes to prepare a mixture of hydrolyzed peptides, and is separated by chromatography or the like to remove the enzyme, remove the peptide not containing the dopa group and remove the peptide of unintended molecular weight, and obtain only the dopa group. Single peptide product, used directly.
  • the molar ratio of mussel mucin to enzyme is in the range of from 0.1:1 to 100:1, preferably in the range of from 1:1 to 50:1.
  • mussel mucin application according to embodiment 1, wherein the mussel mucin may be from a subclass: mefp1, mefp-2, mefp-3, mefp-4, mefp-5, mefp-6, collagen pre-COL-P, pre-COL-D, pre-COL-NG, A mixture of one or more of the foot silk matrix proteins PTMP and DTMP.
  • the mussel mucin application according to embodiment 1, wherein the mussel mucin concentration may be from 0.1 to 15.0 mg/ml.
  • Mussel mucin application according to embodiment 1, wherein the mussel mucin in the final product may be in the range of pH 1.0-7.0, in particular in the range of pH 3.0-6.5.
  • microneedles which may be made of silicon, silica, cellulose, etc.
  • needle rollers which may be made of stainless steel, titanium, poly Made of tetrafluoroethylene, etc.
  • laser such as: dot matrix laser
  • the enzyme may be: (1) trypsin from various sources, such as pancreas extracted from salmon pancreas, bovine pancreas, porcine pancreas, krill. Protease; (2) collagenase from various sources; (3) casein from various sources; (4) chymotrypsin from various sources; (5) carboxypeptidases from various sources; (6) various sources Keratinase; (7) enterokinase from various sources; (8) chymosin from various sources.
  • various sources such as pancreas extracted from salmon pancreas, bovine pancreas, porcine pancreas, krill. Protease
  • collagenase from various sources
  • casein from various sources
  • (4) chymotrypsin from various sources (5) carboxypeptidases from various sources; (6) various sources Keratinase; (7) enterokinase from various sources; (8) chymosin from various sources.
  • the mussel mucin application according to embodiment 7, wherein the combination of the mussel mucin and the enzyme comprises: (1) first spraying the enzyme to degrade the stratum corneum of the skin surface, then spraying the enzyme again, and spraying immediately Mussel mucin, for a period of time; (2) spraying mussel mucin first, then spraying the enzyme for a period of time; (3) mixing the mussel mucin with the enzyme for a period of time; (4) first Spray the enzyme and then spray the mussel mucin for a period of time.
  • mussel mucin application according to any one of embodiments 16-23, wherein mussel mucin is used in combination with an enzyme.
  • a medicament for the treatment of melanin comprising mussel mucin and a pharmaceutically acceptable carrier, wherein the mussel mucin is present in a concentration of from 0.1 to 15.0 mg/ml.
  • a medical device for treating melanin deposition comprising a mussel mucin and a carrier acceptable for use in the field of medical devices, wherein the mussel mucin is present in a concentration of from 0.1 to 15.0 mg/ml.
  • a cosmetic for the treatment of melanin comprising mussel mucin and a carrier acceptable for the cosmetic field, wherein the mussel mucin is present in a concentration of from 0.1 to 15.0 mg/ml.
  • a disinfecting product for the treatment of melanin comprising a carrier acceptable in the field of mussel mucin and a disinfecting product, wherein the mussel mucin is present in a concentration of from 0.1 to 15.0 mg/ml.
  • a health care product/food for the treatment of melanin comprising a mussel mucin and a health care/food acceptable carrier, wherein the mussel mucin is present in a concentration of from 0.1 to 15.0 mg/ml.
  • a daily chemical product for treating melanin deposition comprising a carrier acceptable for the field of mussel mucin and a daily chemical product, wherein the concentration of mussel mucin is from 0.1 to 15.0 mg/ml.
  • the pharmaceutical according to any one of embodiments 25-30, further comprising an enzyme.
  • Example 1 Application of mussel mucin gel cosmetic in the treatment of chloasma.
  • the mussel mucin solution was mixed with polyvinyl alcohol, hydroxypropyl cellulose and glycerin in a mass ratio of 2:2:1:2 to obtain a mussel mucin gel cosmetic having a mussel mucin concentration of 3 mg/g.
  • Example 2 Application of mussel mucin condensation daily chemical products in the treatment of chloasma.
  • the mussel mucin solution was mixed with propylene glycol and glycerin at a mass ratio of 2:2:1 to obtain a mussel mucin condensation daily product having a mussel mucin concentration of 1.5 mg/g.
  • Example 3 Application of mussel mucin liquid medical device in the treatment of chloasma.
  • a mussel mucin solution having a concentration of 20.0 mg/ml was taken, and 9 ml of a 0.1% citric acid solution was added to prepare an aqueous solution having a mussel mucin concentration of 2.0 mg/ml.
  • 1.0 mg of squid enzyme was taken, and 1.0 ml of deionized water was added to prepare a solution having a concentration of 1.0 mg/ml.
  • the selected patients were randomly divided into three groups.
  • the first group was treated with commercially available ecchymosis solution and recorded as group A.
  • the second group uses the above mussel mucin liquid medical device to spray the affected area and records it as group B.
  • the third group uses the mussel mucin liquid medical device and the enzyme composition to spray the affected area, and firstly sprays mussel mucin and then sprays the enzyme. Formulation, recorded as group C.
  • the three groups of patients were used 3 times a day, and the dosage should be evenly applied to the affected area. Determine the melanin value of the affected area and the melanin value at the center of the affected area before use. The melanin value of the affected area was measured every 10-30 days, and the rate of melanin change in the affected area was calculated (see Table 3 for the results).
  • the melanin change rate is calculated using the following formula:
  • Example 4 Application of mussel mucinized hydrogel cosmetics in the treatment of chloasma.
  • the selected patients were sprayed with the above mussel mucin hydrogel cosmetic, and then sprayed with mussel mucin hydrogel cosmetic and then sprayed with enzyme preparation 3 times a day.
  • the melanin value of the affected area was measured every 10-30 days, and the rate of melanin change in the affected area was calculated (see Table 4 for the results).
  • the melanin change rate is calculated using the following formula:
  • the mussel mucin product of the invention can alleviate pigmentation at the chloasma and can be used for the treatment of chloasma.
  • Example 5 Application of mussel mucin liquid medicine in the treatment of freckles.
  • the selected patients were sprayed with the above mussel mucin liquid medicine, and the mussel mucin liquid medicine was sprayed first and then the enzyme preparation was sprayed 3 times a day.
  • the melanin value of the affected area was measured every 10-30 days, and the rate of melanin change in the affected area was calculated (see Table 5 for the results).
  • the melanin change rate is calculated using the following formula:
  • the mussel mucin product of the invention can reduce the pigmentation of freckles patients and can be used for the treatment of freckles.
  • Example 6 Application of mussel mucin condensation daily chemical products in the treatment of freckles.
  • the propylene glycol and glycerol were mixed in a ratio of 1:1 to form a condensation matrix, and another 2.5 ml of a mussel mucin solution having a concentration of 10.0 mg/ml was added to the gel matrix while stirring, and uniformly mixed to form a mussel.
  • the mucin condensation daily product has a mussel mucin concentration of 3.0 mg/ml.
  • the enrolled patients were randomly divided into three groups.
  • the first group used commercially available ecchymosis, which was recorded as group A.
  • the second group directly applied the above-mentioned mussel mucin condensation daily product to the affected area, which was recorded as group B; the third group first used micro-needle to form a minimally invasive surface in the affected area, and then used
  • the mussel mucin condensation daily product was applied to the affected area and recorded as group C.
  • the three groups were used three times a day in an amount to evenly spread the affected area. Determine the melanin value of the affected area and the melanin value at the center of the affected area before use. The melanin value of the affected area was measured every 10-30 days after the mussel protein was used, and the rate of melanin change in the affected area was calculated (see Table 6 for the results).
  • the melanin change rate is calculated using the following formula:
  • the melanin change rate was increased by 3 times after 90 days of using the mussel mucin condensation daily product compared with the commercially available product. After 90 days of combination of mussel mucin condensation daily products and enzymes, the change rate of pigmentation in the affected area reached 42.6%, which significantly improved the freckle pigmentation in the affected area. In addition, compared with mussel mucin products alone, the combination of mussel mucin products and enzymes, the rate of change in pigmentation of the affected area is increased by 2.5 times, which is more conducive to the treatment of freckles.
  • Example 7 Application of mussel mucin condensation cosmetic in the treatment of melanosis.
  • the mussel mucin freeze-dried powder is mixed with gelatin, fish gelatin and glycerin in a ratio of 3:1:1:0.5 to form a mussel mucin condensation cosmetic, wherein the mussel mucin concentration is 5.5 mg/ml. .
  • the pigmentation of the affected area was measured by a colorimeter. After 20 days of continuous use, the average pigment value of the affected area began to decrease, dropping to about 96% of the initial value. By 90 days, the affected area pigment value decreased to 82.6% of the initial value (see Table 7), demonstrating that the use of the mussel mucin product of the present invention can reduce the pigmentation of melanosis.
  • Example 8 Application of mussel mucin gel health supplement in the treatment of melanosis.
  • the mussel mucin solution was mixed with gelatin and glycerin in a ratio of 1:1:1 to form a mussel mucin gel health supplement, wherein the mussel mucin concentration was 10.0 mg/ml.
  • the pigmentation of the affected area was measured by a colorimeter. After 20 days of continuous use, the average pigment value of the affected area began to decrease, dropping to 96% of the initial value. By 90 days, the affected area pigment value decreased to 80% of the initial value (see Table 8), demonstrating that the use of the mussel mucin product of the present invention can reduce the pigmentation of melanosis.
  • Example 9 Application of mussel mucin liquid health care product in the treatment of melanosis.
  • the patient used laser minimally invasive technique to form a minimally invasive surface on the test surface, and then used the above mussel mucin liquid health care product twice a day to completely cover the test area for 90 days.
  • the melanin change rate is calculated using the following formula:
  • Example 10 Application of mussel mucinized hydrogel medical device in the treatment of melanosis.
  • Mussel mucin, carbomer and propylene were mixed at a mass ratio of 1:1:2 to form a mussel mucin hydrogel medical device, wherein the mussel mucin concentration was 2.5 mg/ml.
  • the melanin value of the affected area was measured every 10-30 days after the mussel protein was used, and the rate of melanin change in the affected area was calculated (see Table 10 for the results).
  • the melanin change rate is calculated using the following formula:
  • the mussel mucin product of the invention can reduce pigmentation in patients with melanosis and can be used for the treatment of melanosis.
  • Example 11 Application of mussel mucin liquid cosmetic in the treatment of melanin precipitation after inhibiting burns.
  • the mussel mucin solution was taken, and 0.01% citric acid was added to adjust the solution pH to 4.0 to form a mussel mucin liquid cosmetic, wherein the mussel mucin concentration was 10.0 mg/ml.
  • the burn area of the patients was greater than 2% of the human body surface area. After the subjects voluntarily signed the informed consent form, they participated in the test.
  • the patient used the above-mentioned mussel mucin liquid cosmetic 3 times a day, each time in order to uniformly cover the affected area.
  • the adjacent areas of the same body were selected for comparison, that is, two areas were selected in the same body, one part was used for the mussel mucin liquid cosmetic (test group), and the other part was treated according to the normal procedure (control group). Pigments of the test group, the control group, and healthy skin were measured on Days 0, 7, and 14 (see Table 11). After 14 days of treatment, the skin after using the mussel mucin liquid cosmetic recovered did not show significant melanin deposition, while the unused skin showed significant pigmentation.

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Abstract

本发明公开了贻贝粘蛋白或其制剂在治疗和预防黑色素相关疾病中的应用。具体地公开了贻贝粘蛋白或其制剂在黄褐斑、雀斑、黑变病等色素沉着中的应用；在以黑色素瘤为代表的皮肤癌中的应用；在因皮肤疾病或药物可能产生的色素沉着治疗中的应用。

Description

贻贝粘蛋白产品治疗和预防黑色素相关疾病中的应用 技术领域
本发明大体涉及药品、化妆品、医疗产品、消毒产品、保健品、食品以及日化产品技术领域，更具体地，涉及贻贝粘蛋白产品及其在治疗和预防黑色素相关疾病中的应用。
背景技术
黑色素是动物皮肤或者毛发中存在的一种黑褐色的色素，由一种特殊的细胞即黑色素细胞生成并且储存在其中。黑色素广泛存在于人的皮肤、粘膜、视网膜、软脑膜、胆囊与卵巢等处。正是由于黑色素的存在，皮肤才有了颜色。黑色素是一种生物色素，由酪氨酸或3，4-二羟苯丙氨经过一连串化学反应形成，并且通常是以聚合的方式存在。
黑色素本质上是一种蛋白质。它们存在于皮肤基底层的细胞(也叫做“色素母细胞”)中间，是一种被称作“黑色素原生物质”的物质。色素母细胞分泌麦拉宁色素，当紫外线(UVA和UVB)照射到皮肤上，UVB作用于皮肤基底层，肌肤就会处于“自我防护”状态，通过紫外线刺激麦拉宁色素，激活酪氨酸酶的活性，从而保护皮肤细胞。多巴是黑色素的前身，经酪氨酸氧化而成，释放出黑色素。黑色素又经由细胞代谢的层层移动，到达肌肤表皮层形成雀斑、晒斑、黑斑等形状。
黑色素的产生多与内分泌失调，日光照射有关，黑色素细胞的多少主要取决于遗传，也与内分泌激素及营养状况有关，由黑色素导致的疾病例如雀斑、黄褐斑、黑变病等色素沉着性疾病，是美容界广泛研究的课题。
色素在其生成过程中的抑制有别于已经生成的色素的治疗。抑制色素的沉着在于中止从酪氨酸到多巴再到二羟基吲哚，直至生成黑色素的过程。这一过程可以通过抑制炎症，阻止炎症因子的生成，从而实现色素抑制作用；或是通过可以与色素形成过程中的物质进行反应的物质来阻断色素的生成；或是通过抗氧化阻止色素形成。而色素治疗是通过还原性物质使已经形成的黑色素分解，从而使色素最终消失。
贻贝粘蛋白(Mussel adhesive protein，MAP)，也称作贻贝足丝蛋白(Mytilus edulis foot protein，Mefp)，是海洋贝类紫贻贝(Mytilus edulis Linnaeus)、厚壳贻贝(Mytilus coruscus)、翡翠贻贝(Perna viridis)等分泌的一种特殊的蛋白质。贻贝通常成群地附着在海岸边的礁石 上或者轮船的底部，有在近海耐受波浪冲击的能力。实际上贻贝几乎可以极其牢固地附着在任何材料的基底上，如金属、木材、玻璃等。贻贝具有上述特性的主要原因是其足丝腺内可生成并储存这种特殊的粘蛋白，贻贝通过足丝释放粘蛋白到岩石一类的固体表面上，形成抗水的结合，从而将自己固定。
目前从贻贝中鉴定得到11种粘蛋白亚类，包括mefp1、mefp-2、mefp-3、mefp-4、mefp-5、mefp-6、胶原蛋白pre-COL-P、pre-COL-D、pre-COL-NG、足丝基质蛋白PTMP和DTMP(朱曜曜等，海洋科学进展，2014，32(4)：560-568)。贻贝粘蛋白具有2个结构特点：(1)含有赖氨酸，使蛋白带有高载量正电荷；(2)含3，4二羟基苯丙氨酸(DOPA，多巴)。人体的细胞和组织带有负电荷。贻贝粘蛋白通过自身正电荷与人体的细胞和组织负电荷之间的静电相互作用与细胞和组织紧密结合，发挥防护和治疗的作用。此外，多巴氧化生成邻位二醌，可以和未被氧化的多巴相互交联形成膜或是网状支架，促使蛋白质更加紧密、稳固地附着在人体表面，起到保护作用。贻贝粘蛋白是大分子蛋白质，在人体内完全降解的时间约为3-10天，其附着于细胞组织的能力优异，使贻贝粘蛋白可以稳固于局部，持续发挥作用。
虽然贻贝粘蛋白具有以上特点，但目前其产品应用领域非常有限。商品化的贻贝粘蛋白产品有美国BD Biosciences公司的Cell-Tak，韩国Kollodis的MAP Trix和瑞典Biopolymer的Hydrogel。这些产品或者是以贻贝粘蛋白溶液状态直接使用，或者是以冻干粉制剂保存而在使用前溶解，它们的主要应用限于微观的细胞粘附和组织粘合剂。也有报道贻贝粘蛋白用于胎膜修复、作为抗海水腐蚀涂层、心脏载药支架等应用。
发明内容
本发明人发现贻贝粘蛋白作为一种多酚蛋白质，其分子中含有的多巴基团是参与黑色素合成的重要物质，在黄褐斑、雀斑、黑变病等黑色素引起的疾病中，贻贝粘蛋白分子中的多巴基团可增强酪氨酸酶的活性，进而影响黑色素的分泌，达到治疗和预防黑色素相关疾病的目的。
本发明人进一步发现贻贝粘蛋白分子量高达100kD，经皮直接使用时的吸收率低，影响作用效果。采用皮肤表面形成微创或加入酶水解贻贝粘蛋白可提高经皮作用的效率，更有效治疗黄褐斑、雀斑、黑变病、其他色素沉着性皮肤病，有效抑制以黑色素瘤为代表的皮肤癌，预防因日晒或痤疮等皮肤疾病可能产生的色素沉着。
本发明的一个目的是提供一种贻贝粘蛋白产品。
在本文中使用的贻贝粘蛋白是指从贻贝科(Mytilidae)双壳类软体动物中的紫贻贝(Mytilus edulis Linnaeus)、厚壳贻贝(Mytilus coruscus)或翡翠贻贝(Perna viridis)等海洋贻贝中纯化获得的、目前已知的贻贝粘蛋白11个亚类：mefp1、mefp-2、mefp-3、mefp-4、mefp-5、mefp-6、胶原蛋白pre-COL-P、pre-COL-D、pre-COL-NG、足丝基质蛋白PTMP和DTMP中的一种或几种的混合物。在本文中使用的贻贝粘蛋白在水溶液中的酸碱度可以是pH 1.0-7.0，特别是可以在pH 3.0-6.5的范围内以使其治疗效果更佳。
在本文中使用的贻贝粘蛋白可以采用以下制备方法获得，例如中国专利号ZL200710179491.0的一种使用混合吸附色谱分离纯化贻贝粘蛋白的方法，中国专利号ZL200710179492.5的一种使用羧甲基离子交换色谱纯化贻贝粘蛋白的方法，中国专利号ZL200910087567.6的一种使用盐析和透析分离纯化贻贝粘蛋白的方法等。
在本文中使用的贻贝粘蛋白可以是溶液或冻干粉形式，特别是贻贝粘蛋白在产品中的浓度可以是0.1-15.0mg/ml，当浓度过低时，贻贝粘蛋白的功效不大，当浓度过高时，可引起细胞毒性、皮肤刺激等作用，从而不利于皮肤黑色素的治疗。
在本文中使用的贻贝粘蛋白也可以与辅料结合而制备成液体剂。示例性的贻贝粘蛋白液体剂是将贻贝粘蛋白溶液母液或冻干粉溶解或稀释至一定浓度或pH值制得，溶解或稀释用溶液可以是水、生理盐水、磷酸盐溶液、醋酸盐溶液、硼酸盐溶液等等。最终产品中贻贝粘蛋白的酸碱度可以是pH 1.0-7.0，特别是可以在pH 3.0-6.5的范围内治疗效果更佳。
在本文中使用的贻贝粘蛋白也可以与辅料结合而制备成凝胶剂。示例性的贻贝粘蛋白凝胶剂是将贻贝粘蛋白溶液或冻干粉与凝胶基质材料混合制得，所述凝胶基质材料可以是选自纤维素衍生物、卡波姆和海藻酸盐、西黄蓍胶、明胶、果胶、卡拉胶、结冷胶、淀粉、黄原胶、阳离子瓜尔胶、琼脂、非纤维素多糖、乙烯聚合物、丙烯酸树脂、聚乙烯醇或聚羧乙烯中之一或其任意组合。
在本文中使用的贻贝粘蛋白也可以与辅料结合而制备成凝露。示例性的贻贝粘蛋白凝露是由贻贝粘蛋白溶液或冻干粉与凝露基质混合制得，所述凝露基质可以包括纤维素衍生物、甘油、非纤维素多糖、丙二醇中之一或其任意组合。
在本文中使用的贻贝粘蛋白也可以与辅料结合而制备成糊剂。示例性的贻贝粘蛋白糊剂是由贻贝粘蛋白与糊剂基质材料混合制得，所述糊剂基质材料可以包括甘油、凡士林、石蜡 等。
在本文中使用的贻贝粘蛋白还可以与基质材料结合形成用于皮肤表面的敷料和敷贴。示例性的贻贝粘蛋白敷贴中的液体可以是贻贝粘蛋白溶液，或是其与纤维素衍生物、卡波姆和海藻酸盐、西黄蓍胶、明胶、果胶、卡拉胶、结冷胶、淀粉、黄原胶、阳离子瓜尔胶、琼脂、非纤维素多糖、乙烯聚合物、丙烯酸树脂、聚乙烯醇或聚羧乙烯、明胶、鱼胶、果胶、海藻酸盐、甘油、凡士林、石蜡、聚乙二醇、维生素、谷胱甘肽中的一种或多种组合。上述贻贝粘蛋白溶液或组合浸泡在基质材料如纱布、无纺布、蚕丝纸上，所述敷贴可以是例如创口贴、面膜、眼膜、手膜、脚膜等。
可以向上述包括贻贝粘蛋白的各种制剂中进一步添加甘油、聚乙二醇、维生素、谷胱甘肽等本领域公知的可增加其保湿、抗氧化等能力的成分进一步提高其保湿、抗氧化的能力。
可以向上述包括贻贝粘蛋白的各种制剂中进一步添加芦荟、葡萄籽、茶叶、人参、蜗牛等提取物，氨基酸、胶原蛋白、透明质酸等成分，增加其营养皮肤和皮肤附属器官的能力，增加滑润程度。
以上所有制剂都可以采用本领域周知的方法制备，详细操作步骤可参照例如《制剂学》。
在本文中使用的贻贝粘蛋白可以作为主要原料，采用药学上可接受的载体制备药品。所述药品可以是液体剂、凝胶剂、凝露、糊剂、敷贴、泡沫剂。所述药品可用于体外使用，特别是可用于皮肤外用。
在本文中使用的贻贝粘蛋白可以作为主要原料，制备医疗器械。本文中使用的术语医疗器械是指直接或间接用于人体的材料及其他类似或相关的物品。所述医疗器械可以是液体剂、凝胶剂、凝露、糊剂、敷贴、泡沫剂。所述医疗器械可用于体外使用，特别是可用于皮肤外用。
在本文中使用的贻贝粘蛋白可以作为主要原料，采用化妆品领域可接受的辅料制备化妆品。所述化妆品可以是液体剂、凝胶剂、凝露、糊剂、敷贴、泡沫剂。所述化妆品可用于体外使用，特别是可用于皮肤外用。
在本文中使用的贻贝粘蛋白可以作为主要原料，采用消毒产品领域可接受的辅料制备消毒产品。本文中使用的术语消毒产品是指用化学、物理、生物的方法杀灭或者消除环境中病原微生物的消毒剂、消毒器械、卫生用品和一次性使用的医疗用品。所述消毒产品可以是液体剂、凝胶剂、凝露、糊剂、敷贴、泡沫剂。所述消毒产品可用于体外使用，特别是可用于皮肤外用。
在本文中使用的贻贝粘蛋白可以作为主要原料，采用保健品或食品领域可接受的辅料制备保健品或食品。所述保健品或食品可以是液体剂、凝胶剂、凝露、糊剂、敷贴、泡沫剂。所述保健品或食品可用于体外使用或食用，特别是可用于皮肤外用。
在本文中使用的贻贝粘蛋白可以作为主要原料，采用日化产品领域可接受的辅料制备日化产品。本文中使用的术语日化产品是指平日常用的科技化学制品，包括洗发水、沐浴露等。所述日化产品可以是液体剂、凝胶剂、凝露、糊剂、敷贴、泡沫剂。所述日化产品可用于体外使用，特别是可用于皮肤外用。
本发明的另一个目的是提供贻贝粘蛋白产品在治疗和预防黑色素沉着中的应用。特别是用于治疗黄褐斑、雀斑、黑变病等形成色素沉着的疾病，抑制以黑色素瘤为代表的皮肤癌，预防因日晒或痤疮等皮肤疾病可能产生的色素沉着。
在本文中使用的术语治疗是指干预或改变特定健康状态的过程，为解除病痛所进行的活动。在本文中使用的术语抑制是指组织或机体受刺激后，使原有损害减弱的过程。
在本文中使用的术语预防是指预先做好疾病治疗过程中可能出现偏离主观预期轨道或客观普遍规律的应对措施，避免产生可能存在的损害。
黄褐斑是指面部的黄褐色色素沉着。其多对称蝶形分布于颊部。
雀斑是发生在面部皮肤上的黄褐色点状色素沉着斑，属于常染色体显性遗传。其好发于面部，特别是鼻部和两颊，可累及颈、肩、手背等暴露部位，非暴露部位无皮疹。损害为浅褐或暗褐色针头大小到绿豆大斑疹，圆形、卵圆形或不规则。散在或群集分布，孤立不融合。
黑变病是由于工人长期接触沥青、煤焦油、石油类产品或长期吸入这类物质的挥发物而致皮肤慢性炎症，最终发生皮肤色素沉着。
黑色素瘤是由皮肤和其他器官的黑素细胞产生的肿瘤。皮肤黑素瘤表现为色素性皮损在数月或数年中发生明显改变。
皮肤病引起的色素沉着是由炎症性皮肤病所致或由各种药物引起。例如日光性皮炎或痤疮治疗后导致色素沉着。扁平苔藓和扁平苔藓样药疹导致色素沉着。固定型药疹可留下特征性环形色素沉着斑。可引起色素沉着的药物包括乙胺碘呋酮、四环素、美满霉素、争光霉素、环磷酰胺、抗疟药氯喹和喹宁、氯丙嗪和其他可引起暴露部位皮肤灰蓝色色素改变的吩噻嗪类药物。
根据本发明的一个方面，贻贝粘蛋白可以独立使用用于黑色素沉着的治疗和预防。当贻贝粘蛋白独立使用用于黑色素沉着的治疗和预防时，可以直接使用，也可以先形成微小创面 后使用。形成微小创面的方法包括：微针(可以由硅、二氧化硅、纤维素等制成)、滚针(可以由不锈钢、钛、聚四氟乙烯等制成)、激光(如：点阵激光)。
根据本发明的另一个方面，贻贝粘蛋白可以与酶结合，经酶降解后使用。其中贻贝粘蛋白抑制黑色素形成的氧化过程，与黑色素形成过程中的多巴(3，4-二羟基苯丙氨酸)结合中止色素的进一步形成过程。此外，由于贻贝粘蛋白分子量大，经皮给药困难，能被利用的程度低，采用酶水解贻贝粘蛋白提高经皮给药的效率，增强治疗效果。
在本文中使用所的酶可以是：(1)各种来源的胰蛋白酶，例如从鳕鱼胰脏、牛胰脏、猪胰脏、磷虾中提取的胰蛋白酶；(2)各种来源的胶原酶；(3)各种来源的酪蛋白酶；(4)各种来源的靡蛋白酶；(5)各种来源的羧肽酶；(6)各种来源的角蛋白酶；(7)各种来源的肠激酶；(8)各种来源的凝乳酶等。可以与贻贝粘蛋白共同用于黑色素治疗的酶不局限于以上所述酶。
根据本发明，贻贝粘蛋白可以与酶组合用于治疗黑色素，使用形式可以包括：(1)先喷涂酶降解皮肤表面角质层，然后再次喷涂酶，并立刻喷涂贻贝粘蛋白，共同作用一段时间；(2)先喷涂贻贝粘蛋白，然后喷涂酶，共同作用一段时间；(3)将贻贝粘蛋白与酶混合一段时间后喷涂使用；(4)先喷涂酶，然后喷涂贻贝粘蛋白，共同作用一段时间。
根据本发明，贻贝粘蛋白与酶形成产品的形式可以包括：(1)贻贝粘蛋白与酶在不同的包装物内：分别使用，或是在使用前用使用者自行混合；(2)贻贝粘蛋白水解肽混合物：贻贝粘蛋白采用上述各种酶制备成水解肽的混合物，采用加热等手段中止酶的活性后形成产品，直接使用；(3)贻贝粘蛋白单一水解肽：贻贝粘蛋白采用上述各种酶制备成水解肽的混合物，采用色谱等分离方法，去除酶，去除不含有多巴基团的肽并去除非预期分子量的肽，获得仅含有多巴基团的单一肽产品，直接使用。
根据本发明，当用于治疗黑色素时，贻贝粘蛋白与酶的摩尔比在0.1∶1-100∶1的范围内，优选在1∶1-50∶1的范围内。
具体实施方式
本发明的实施方式包括：
1、贻贝粘蛋白在治疗和预防黑色素沉着中的应用。
2、根据实施方式1的贻贝粘蛋白应用，其中所述贻贝粘蛋白可以是来自亚类：mefp1、mefp-2、mefp-3、mefp-4、mefp-5、mefp-6、胶原蛋白pre-COL-P、pre-COL-D、pre-COL-NG、 足丝基质蛋白PTMP和DTMP中的一种或几种的混合物。
3、根据实施方式1的贻贝粘蛋白应用，其中所述贻贝粘蛋白浓度可以是0.1-15.0mg/ml。
4、根据实施方式1的贻贝粘蛋白应用，其中所述贻贝粘蛋白可以是以液体剂、凝胶剂、凝露、糊剂、敷贴或泡沫剂使用。
5、根据实施方式1的贻贝粘蛋白应用，其中最终产品中的贻贝粘蛋白可以是在pH 1.0-7.0的范围内，特别是可以在pH 3.0-6.5的范围内。
6、根据实施方式1-5中任一项的贻贝粘蛋白应用，其中所述黑色素沉着可以是黄褐斑、雀斑、黑变病等色素沉着，黑色素瘤为代表的皮肤癌，因日晒或痤疮等皮肤疾病产生的色素沉着。
7、根据实施方式1的贻贝粘蛋白应用，其中所述贻贝粘蛋白可以单独使用或与酶组合使用。
8、根据实施方式7的贻贝粘蛋白应用，其中所述贻贝粘蛋白可以直接使用，或可以先形成微小创面后使用。
9、根据实施方式8的贻贝粘蛋白应用，其中形成微小创面的方法可以包括：微针(可以由硅、二氧化硅、纤维素等制成)、滚针(可以由不锈钢、钛、聚四氟乙烯等制成)、激光(如：点阵激光)。
10、根据实施方式7的贻贝粘蛋白应用，其中所述酶可以是：(1)各种来源的胰蛋白酶，例如从鳕鱼胰脏、牛胰脏、猪胰脏、磷虾中提取的胰蛋白酶；(2)各种来源的胶原酶；(3)各种来源的酪蛋白酶；(4)各种来源的靡蛋白酶；(5)各种来源的羧肽酶；(6)各种来源的角蛋白酶；(7)各种来源的肠激酶；(8)各种来源的凝乳酶等。
11、根据实施方式7的贻贝粘蛋白应用，其中所述贻贝粘蛋白与所述酶组合使用形式可以包括：(1)先喷涂酶降解皮肤表面角质层，然后再次喷涂酶，并立刻喷涂贻贝粘蛋白，共同作用一段时间；(2)先喷涂贻贝粘蛋白，然后喷涂酶，共同作用一段时间；(3)将贻贝粘蛋白与酶混合一段时间后喷涂使用；(4)先喷涂酶，然后喷涂贻贝粘蛋白，共同作用一段时间。
12、根据实施方式7的贻贝粘蛋白应用，其中所述贻贝粘蛋白与酶可以在不同的包装物内。
13、根据实施方式7的贻贝粘蛋白应用，其中所述贻贝粘蛋白与酶组合使用可以是以贻贝粘蛋白水解肽混合物的形式使用。
14、根据实施方式7的贻贝粘蛋白应用，其中所述贻贝粘蛋白与酶组合使用可以是以贻贝粘蛋白单一水解肽的形式使用。
15、根据实施方式7的贻贝粘蛋白应用，其中所述贻贝粘蛋白与酶的摩尔比可以在0.1∶1-100∶1的范围内，优选可以在1∶1-50∶1的范围内。
16、贻贝粘蛋白作为活性成分在用于治疗和预防黑色素沉着的组合物中的应用，其中所述组合物是以液体剂、凝胶剂、凝露、糊剂、敷贴或泡沫剂使用。
17、根据实施方式16的贻贝粘蛋白应用，其中所述组合物是皮肤外用组合物。
18、贻贝粘蛋白作为活性成分在在用于治疗黑色素的药品中的应用。
19、贻贝粘蛋白作为活性成分在在用于治疗黑色素的医疗器械中的应用。
20、贻贝粘蛋白作为活性成分在在用于治疗黑色素的化妆品中的应用。
21、贻贝粘蛋白作为活性成分在在用于治疗黑色素的消毒产品中的应用。
22、贻贝粘蛋白作为活性成分在在用于治疗黑色素的保健品或食品中的应用。
23、贻贝粘蛋白作为活性成分在在用于治疗黑色素的日化产品中的应用。
24、根据实施方式16-23中任一项的贻贝粘蛋白应用，其中贻贝粘蛋白与酶组合使用。
25、一种用于治疗黑色素沉着的药品，包括贻贝粘蛋白和药学上可接受的载体，其中贻贝粘蛋白的浓度为0.1-15.0mg/ml。
26、一种用于治疗黑色素沉着的医疗器械，包括贻贝粘蛋白和医疗器械领域可接受的载体，其中贻贝粘蛋白的浓度为0.1-15.0mg/ml。
27、一种用于治疗黑色素沉着的化妆品，包括贻贝粘蛋白和化妆品领域可接受的载体，其中贻贝粘蛋白的浓度为0.1-15.0mg/ml。
28、一种用于治疗黑色素沉着的消毒产品，包括贻贝粘蛋白和消毒产品领域可接受的载体，其中贻贝粘蛋白的浓度为0.1-15.0mg/ml。
29、一种用于治疗黑色素沉着的保健品/食品，包括贻贝粘蛋白和保健品/食品领域可接受的载体，其中贻贝粘蛋白的浓度为0.1-15.0mg/ml。
30、一种用于治疗黑色素沉着的日化产品，包括贻贝粘蛋白和日化产品领域可接受的载体，其中贻贝粘蛋白的浓度为0.1-15.0mg/ml。
31、根据实施方式25-30中任一项的药品，进一步包括酶。
下面将结合具体实施例对本发明作进一步说明。需要指出的是，由本发明中的贻贝粘蛋白或贻贝粘蛋白的各种制剂形成的药品、医疗器械、化妆品、消毒产品、保健品或食品、日化产品在施用于受试者后，都可以应用于上文所述的适应症并展现出上文所述的功能，在本发明范围内的所有剂型均已测试，下文中，仅仅是为说明，只在实施例中描述了其中一少部分，然而不应将其理解为对本发明的限制。
除非特殊说明，否则本发明中所使用的试剂都是市售可购买的。
实施例1：贻贝粘蛋白凝胶剂化妆品在黄褐斑治疗中的应用。
取贻贝粘蛋白溶液与聚乙烯醇、羟丙基纤维素、甘油按照质量比为2∶2∶1∶2混合，获得贻贝粘蛋白浓度3mg/g的贻贝粘蛋白凝胶剂化妆品。
收集黄褐斑病例10例，要求入选患者患处为颊部，斑片颜色为深褐色，经皮肤科专家确诊后入组试验。在患处涂抹上述贻贝粘蛋白凝胶剂化妆品，每日1次，每次使用以能均匀涂抹在患处表面为准。每10天用色素测定仪测定患处色素值，10例患者在涂抹贻贝粘蛋白凝胶剂化妆品50天后，患处色素值下降9％(参见表1)，证明长期使用贻贝粘蛋白产品可以减化黄褐斑处色素沉积。
表1：
第0天患处色素平均值 308±10.7
使用10天后色素平均值 303.4±9.9
使用20天后色素平均值 299.2±9.5
使用30天后色素平均值 293.6±8.2
使用40天后色素平均值 286.6±5.8
使用50天后色素平均值 280.6±6.8
实施例2：贻贝粘蛋白凝露日化产品在黄褐斑治疗中的应用。
取贻贝粘蛋白溶液与丙二醇、甘油按照质量比为2∶2∶1混合，获得贻贝粘蛋白浓度为1.5mg/g的贻贝粘蛋白凝露日化产品。
收集黄褐斑病例10例，要求入选患者患处为颊部，斑片颜色为深褐色，经皮肤科专家确诊后入组试验。在患处涂抹上述贻贝粘蛋白凝露日化产品，每日2次，每次使用以能均匀涂 抹在患处表面为准。每10天用色素测定仪测定患处色素值，10例患者在涂抹贻贝粘蛋白凝露日化产品50天后，患处色素值下降7.3％(参见表2)，证明长期使用贻贝粘蛋白产品可以减化黄褐斑处色素沉积。
表2：
第0天患处色素平均值 316.0±13.6
使用10天后色素平均值 313.8±9.3
使用20天后色素平均值 309.4±7.8
使用30天后色素平均值 302.3±7.2
使用40天后色素平均值 298.6±6.9
使用50天后色素平均值 293.9±8.2
实施例3：贻贝粘蛋白液体医疗器械在黄褐斑治疗中的应用。
取浓度为20.0mg/ml的贻贝粘蛋白溶液1ml，加入9ml 0.1％柠檬酸溶液，配制成贻贝粘蛋白浓度为2.0mg/ml的水溶液。取鳕鱼酶1.0mg，加入1.0ml去离子水配成浓度为1.0mg/ml的溶液。
收集30例经皮肤科专家确诊的黄褐斑患者进行试验。患者患处有成片的黄褐。
入选患者随机分为三组，第一组采用市售祛斑液涂抹患处，记为组A。第二组采用上述贻贝粘蛋白液体医疗器械喷涂患处，记为组B；第三组采用贻贝粘蛋白液体医疗器械与酶的组合物喷涂患处，使用时先喷涂贻贝粘蛋白再喷涂酶制剂，记为组C。三组患者使用时均每日3次，用量需能均匀涂抹患处。使用前测定患处黑色素值，以及患处周边无色斑处黑色素值。每10-30天测定患处黑色素值，并计算患处黑色素变化率(结果见表3)。
黑色素变化率采用以下公式计算：
(使用前黑色素值-使用后黑色素值)/(使用前黑色素值-正常部位黑色素值)×100％
表3：
Figure PCTCN2015084494-appb-000001
Figure PCTCN2015084494-appb-000002
结果表明：与市售祛斑产品相比，使用贻贝粘蛋白液体医疗器械90天后，患处色素值变化率超过一倍，祛斑效果优于市售产品。在组合使用贻贝粘蛋白液体医疗器械与酶90天后，患处色素值变化率达到28.2％，起到明显改善患处黄褐斑的效果。另外，与单独使用贻贝粘蛋白产品相比，组合使用贻贝粘蛋白产品与酶，患处色素值变化率提高一倍，更有利于黄褐斑的治疗。
实施例4：贻贝粘蛋白化水凝胶化妆品在黄褐斑治疗中的应用。
取羧甲基纤维素钠10g，加入20ml去离子水，90℃温浴30min至完全溶解得到凝胶基质，另取浓度为10.0mg/ml的贻贝粘蛋白溶液2.5ml，边搅拌边加入到凝胶基质中，混合均匀后形成贻贝粘蛋白化水凝胶化妆品，贻贝粘蛋白浓度为1.1mg/ml。取鳕鱼酶1.0mg，加入1.0ml去离子水配成浓度为1.0mg/ml的溶液。
收集20例经皮肤科专家确诊的黄褐斑患者进行试验。患者患处有成片的黄褐。
入选患者采用上述贻贝粘蛋白水凝胶化妆品喷涂患处，使用时先喷涂贻贝粘蛋白水凝胶化妆品再喷涂酶制剂，每日3次。使用前测定患处黑色素值，以及患处周边无色斑处黑色素值。每10-30天测定患处黑色素值，并计算患处黑色素变化率(结果参见表4)。
黑色素变化率采用以下公式计算：
(使用前黑色素值-使用后黑色素值)/(使用前黑色素值-正常部位黑色素值)×100％
表4：
使用时间(天) 黑色素变化率(％)
10 9.6±1.6
30 18.2±1.9
60 29.6±2.7
90 38.4±1.8
结果表明：使用贻贝粘蛋白水凝胶化妆品与酶10天后，患处色素值已经开始下降，并且随着使用时间的延长，患处黑色素变化率增加，至使用90天后，患处色素变化超过1/3。本发明的贻贝粘蛋白产品能减轻黄褐斑处色素沉着，可用于黄褐斑的治疗。
实施例5：贻贝粘蛋白液体药品在雀斑治疗中的应用。
取浓度为10.0mg/ml的贻贝粘蛋白溶液1ml，加入9ml 0.1％柠檬酸溶液，配制成贻贝粘蛋白浓度为1.0mg/ml的液体药品。取胰蛋白酶1.0mg，加入1.0ml去离子水配成浓度为1.0mg/ml的溶液。
收集20例经皮肤科专家确诊的雀斑患者进行试验。患者患处有成片的褐色斑点。
入选患者采用上述贻贝粘蛋白液体药品喷涂患处，使用时先喷涂贻贝粘蛋白液体药品再喷涂酶制剂，每日3次。使用前测定患处黑色素值，以及患处周边无色斑处黑色素值。每10-30天测定患处黑色素值，并计算患处黑色素变化率(结果见表5)。
黑色素变化率采用以下公式计算：
(使用前黑色素值-使用后黑色素值)/(使用前黑色素值-正常部位黑色素值)×100％
表5：
使用时间(天) 黑色素变化率(％)
10 6.4±1.4
30 21.6±2.8
60 28.7±2.9
90 35.2±3.6
结果表明：使用贻贝粘蛋白液体药品与酶10天后，患处色素值已经开始下降，并且随着使用时间的延长，患处黑色素变化率增加，至使用90天后，患处色素变化达到35.2％，证明本发明的贻贝粘蛋白产品能降低雀斑患者色素沉着，可用于雀斑的治疗。
实施例6：贻贝粘蛋白凝露日化产品在雀斑治疗中的应用。
取丙二醇和丙三醇按1∶l比例混合形成凝露基质，另取浓度为10.0mg/ml的贻贝粘蛋白溶液2.5ml，边搅拌边加入到凝露基质中，混合均匀后形成贻贝粘蛋白凝露日化产品，贻贝粘蛋白浓度为3.0mg/ml。
收集30例经皮肤科专家确诊的雀斑患者进行试验。患者患处有成片的褐色斑块。
入选患者随机分成三组，第一组使用市售祛斑凝露，记为组A。第二组直接使用上述贻贝粘蛋白凝露日化产品涂抹患处，记为组B；第三组先采用微针在患处形成微创面，再使用 该贻贝粘蛋白凝露日化产品涂抹患处，记为组C。三组使用方法均为每日3次，用量以能均匀涂抹患处。使用前测定患处黑色素值，以及患处周边无色斑处黑色素值。使用贻贝蛋白后每10-30天测定患处黑色素值，并计算患处黑色素变化率(结果见表6)。
黑色素变化率采用以下公式计算：
(使用前黑色素值-使用后黑色素值)/(使用前黑色素值-正常部位黑色素值)×100％
表6：
Figure PCTCN2015084494-appb-000003
结果表明：与市售产品相比，使用贻贝粘蛋白凝露日化产品90天后黑色素变化率提高3倍。在组合使用贻贝粘蛋白凝露日化产品与酶90天后，患处色素值变化率达到42.6％，起到明显改善患处雀斑色素沉着的效果。另外，与单独使用贻贝粘蛋白产品相比，组合使用贻贝粘蛋白产品与酶，患处色素值变化率提高2.5倍，更有利于雀斑的治疗。
实施例7：贻贝粘蛋白凝露化妆品在黑变病治疗中的应用。
取贻贝粘蛋白冻干粉，与明胶、鱼胶和甘油，按照3∶1∶1∶0.5的比例混合制剂，形成贻贝粘蛋白凝露化妆品，其中贻贝粘蛋白浓度为5.5mg/ml。
收集12例经皮肤科专家确诊的黑变病患者，患者每日使用2次上述贻贝粘蛋白凝露化妆品，用量以能完全覆盖测试区域，连续使用90天。
用色素测定仪测定患处色素情况，连续使用20天后，患处平均色素值开始下降，下降至初始值的96％左右。至90天时，患处色素值下降至初始值的82.6％(参见表7)，证明使用本发明的贻贝粘蛋白产品可以降低黑变病的色素沉着。
表7：
第0天患处色素平均值 328.0±23.7
使用20天后色素平均值 317.7±19.5
使用40天后色素平均值 303.4±15.8
使用60天后色素平均值 292.6±16.9
使用80天后色素平均值 285.9±18.1
使用90天后色素平均值 271.2±28.8
实施例8：贻贝粘蛋白凝胶保健品在黑变病治疗中的应用。
取贻贝粘蛋白溶液，与明胶和甘油按照1∶1∶1的比例混合，形成贻贝粘蛋白凝胶保健品，其中贻贝粘蛋白浓度为10.0mg/ml。
收集12例经皮肤科专家确诊的黑变病患者，患者每日使用2次上述贻贝粘蛋白凝胶保健品，用量以能完全覆盖测试区域，连续使用90天。
用色素测定仪测定患处色素情况，连续使用20天后，患处平均色素值开始下降，下降至初始值的96％。至90天时，患处色素值下降至初始值的80％(参见表8)，证明使用本发明的贻贝粘蛋白产品可以降低黑变病的色素沉着。
表8：
第0天患处色素平均值 353.3±43.1
使用20天后色素平均值 341.7±36.4
使用40天后色素平均值 316.6±25.6
使用60天后色素平均值 300.3±24.8
使用80天后色素平均值 290.9±19.6
使用90天后色素平均值 282.2±24.9
实施例9：贻贝粘蛋白液体保健品在黑变病治疗中的应用。
取浓度为10.0mg/ml的贻贝粘蛋白溶液10ml，加入10ml 0.001％乙酸溶液，配制成贻贝粘蛋白浓度为5.0mg/ml的水溶液。
收集12例经皮肤科专家确诊的黑变病患者进行试验。患者以激光微创技术在测试表面形成微创面，再使用上述贻贝粘蛋白液体保健品，每日使用2次，用量以能完全覆盖测试区域，连续使用90天。
使用前测定患处黑色素值，以及患处周边无色斑处黑色素值。每10-30天测定患处黑色素值，并计算患处黑色素变化率(结果见表9)。
黑色素变化率采用以下公式计算：
(使用前黑色素值-使用后黑色素值)/(使用前黑色素值-正常部位黑色素值)×100％
表9：
使用时间(天) 黑色素变化率(％)
10 4.2±1.1
30 25.6±3.8
60 37.1±3.6
90 42.2±3.7
结果表明：使用贻贝粘蛋白液体保健品后，患处色素值已经开始下降，并且随着使用时间的延长，患处黑色素变化率增加，至使用90天后，患处色素变化达到42.2％，证明本发明的贻贝粘蛋白产品能降低黑变病患者色素沉着，可用于黑变病的治疗。
实施例10：贻贝粘蛋白化水凝胶医疗器械在黑变病治疗中的应用。
取贻贝粘蛋白、卡波姆、丙二按质量比为1∶1∶2比例混合形成贻贝粘蛋白水凝露医疗器械，其中贻贝粘蛋白浓度为2.5mg/ml。
收集12例经皮肤科专家确诊的黑变病患者进行试验。患者以滚针微创技术在测试表面形成微创面，再使用贻贝粘蛋白水凝胶医疗器械，每日使用2次，用量以能完全覆盖测试区域，连续使用90天。
使用前测定患处黑色素值，以及患处周边无色斑处黑色素值。使用贻贝蛋白后每10-30天测定患处黑色素值，并计算患处黑色素变化率(结果见表10)。
黑色素变化率采用以下公式计算：
(使用前黑色素值-使用后黑色素值)/(使用前黑色素值-正常部位黑色素值)×100％
表10：
使用时间(天) 黑色素变化率(％)
10 10.3±1.6
30 23.6±3.1
60 31.8±2.8
90 39.2±3.5
结果表明：使用贻贝粘蛋白水凝胶医疗器械10天后，患处色素值已经开始下降，并且随着使用时间的延长，患处黑色素变化率增加，至使用90天后，患处色素变化达到39.2％，证明本发明的贻贝粘蛋白产品能降低黑变病患者色素沉着，可用于黑变病的治疗。
实施例11：贻贝粘蛋白液体化妆品在抑制烧伤后黑色素沉淀治疗中的应用。
取贻贝粘蛋白溶液，加入0.01％柠檬酸调节溶液pH为4.0，形成贻贝粘蛋白液体化妆品，其中贻贝粘蛋白的浓度为10.0mg/ml。
收集烧伤后愈合期患者5例，患者烧伤面积大于2％人体表面积，经受试者自愿签署知情同意书后参加试验。患者每天使用上述贻贝粘蛋白液体化妆品3次，每次用量以能均匀覆盖患处为准。试验时选择同体相邻区域进行对照，即同体选择两块区域，一部分使用上述贻贝粘蛋白液体化妆品(试验组)，另一部分按照正常中程序进行治疗(对照组)。使用第0天、7天和14天分别测定试验组、对照组及健康皮肤的色素(参见表11)。经过14天治疗，使用该贻贝粘蛋白液体化妆品的烧伤康复后皮肤未见明显黑色素沉着，而未使用的皮肤可见明显色素沉着。
表11：
  健康的皮肤色素 试验组的皮肤色素 对照组的皮肤色素
第0天 138.7±5.8 139.6±6.1 139.6±6.1
第7天 138.7±5.8 142.6±4.3 157.4±5.5
第14天 138.7±5.8 145.2±4.9 170.2±6.0

Claims (22)

  1. 贻贝粘蛋白在治疗和预防黑色素沉着中的应用。
  2. 根据权利要求1的贻贝粘蛋白应用，其中所述贻贝粘蛋白是来自亚类：mefp1、mefp-2、mefp-3、mefp-4、mefp-5、mefp-6、胶原蛋白pre-COL-P、pre-COL-D、pre-COL-NG、足丝基质蛋白PTMP和DTMP中的一种或几种的混合物。
  3. 根据权利要求1的贻贝粘蛋白应用，其中所述贻贝粘蛋白浓度是0.1-15.0mg/ml。
  4. 根据权利要求1的贻贝粘蛋白应用，其中所述贻贝粘蛋白是以液体剂、凝胶剂、凝露、糊剂、敷贴或泡沫剂使用。
  5. 根据权利要求1的贻贝粘蛋白应用，其中最终产品中的贻贝粘蛋白是在pH 1.0-7.0的范围内，特别是可以在pH 3.0-6.5的范围内。
  6. 根据权利要求1-5中任一项的贻贝粘蛋白应用，其中所述黑色素沉着是黄褐斑、雀斑、黑变病等色素沉着，黑色素瘤为代表的皮肤癌，因日晒或痤疮等皮肤疾病产生的色素沉着。
  7. 根据权利要求1的贻贝粘蛋白应用，其中所述贻贝粘蛋白单独使用或与酶组合使用。
  8. 根据权利要求7的贻贝粘蛋白应用，其中所述贻贝粘蛋白直接使用，或先形成微小创面后使用。
  9. 根据权利要求8的贻贝粘蛋白应用，其中形成微小创面的方法包括微针、滚针、激光。
  10. 根据权利要求7的贻贝粘蛋白应用，其中所述酶是：各种来源的胰蛋白酶；各种来源的胶原酶；各种来源的酪蛋白酶；各种来源的靡蛋白酶；各种来源的羧肽酶；各种来源的角蛋白酶；各种来源的肠激酶；以及各种来源的凝乳酶。
  11. 根据权利要求7的贻贝粘蛋白应用，其中所述贻贝粘蛋白与所述酶组合使用形式包括：先喷涂酶降解皮肤表面角质层，然后再次喷涂酶，并立刻喷涂贻贝粘蛋白，共同作用一段时间；先喷涂贻贝粘蛋白，然后喷涂酶，共同作用一段时间；将贻贝粘蛋白与酶混合一段时间后喷涂使用；先喷涂酶，然后喷涂贻贝粘蛋白，共同作用一段时间。
  12. 根据权利要求7的贻贝粘蛋白应用，其中所述贻贝粘蛋白与所述酶组合使用包括贻贝粘蛋白与酶在不同的包装物内；贻贝粘蛋白水解肽混合物的形式；贻贝粘蛋白单一水解肽的形式。
  13. 根据权利要求7的贻贝粘蛋白应用，其中所述贻贝粘蛋白与酶的摩尔比在0.1∶1-100∶1的范围内，优选在1∶1-50∶1的范围内。
  14. 贻贝粘蛋白作为活性成分在用于治疗和预防黑色素沉着的组合物中的应用，其中所述组合物是以液体剂、凝胶剂、凝露、糊剂、敷贴或泡沫剂使用。
  15. 根据权利要求15的贻贝粘蛋白应用，其中所述组合物是皮肤外用组合物。
  16. 贻贝粘蛋白作为活性成分在在用于治疗和预防黑色素沉着的药品中的应用。
  17. 贻贝粘蛋白作为活性成分在在用于治疗和预防黑色素沉着的医疗器械中的应用。
  18. 贻贝粘蛋白作为活性成分在在用于治疗和预防黑色素沉着的化妆品中的应用。
  19. 贻贝粘蛋白作为活性成分在在用于治疗和预防黑色素沉着的消毒产品中的应用。
  20. 贻贝粘蛋白作为活性成分在在用于治疗和预防黑色素沉着的保健品或食品中的应用。
  21. 贻贝粘蛋白作为活性成分在在用于治疗和预防黑色素沉着的日化产品中的应用。
  22. 根据权利要求16-23中任一项的贻贝粘蛋白应用，其中贻贝粘蛋白与酶组合使用。
PCT/CN2015/084494 2015-07-20 2015-07-20 贻贝粘蛋白产品治疗和预防黑色素相关疾病中的应用 WO2017011984A1 (zh)

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JP2017568248A JP6778220B2 (ja) 2015-07-20 2015-07-20 メラニンに関連する疾患の治療及び予防におけるイガイ接着タンパク質製品の適用
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KR1020187001331A KR102468524B1 (ko) 2015-07-20 2015-07-20 멜라닌과 관련된 질환의 치료 및 예방에서의 홍합 접착 단백질 제품의 용도
CN201580081661.4A CN108135972B (zh) 2015-07-20 2015-07-20 贻贝粘蛋白产品治疗和预防黑色素相关疾病中的应用
MX2018000335A MX2018000335A (es) 2015-07-20 2015-07-20 Aplicaciones de los productos de la proteina adhesiva del mejillon para el tratamiento y la prevencion de enfermedades relacionadas con la melanina.
CN202210732149.3A CN115634282A (zh) 2015-07-20 2015-07-20 贻贝粘蛋白产品治疗和预防黑色素相关疾病中的应用
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BR112018000955-5A BR112018000955A2 (zh) 2015-07-20 2015-07-20 Application of mussel mucin product in the treatment and prevention of melanin - related diseases
AU2015402833A AU2015402833B2 (en) 2015-07-20 2015-07-20 Applications of mussel adhesive protein product in treatment and prevention of diseases related to melanin
HK18114980.3A HK1255896A1 (zh) 2015-07-20 2018-11-22 貽貝粘蛋白產品治療和預防黑色素相關疾病中的應用
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