WO2024022318A1 - Use of ergothioneine in promoting collagen generation - Google Patents

Use of ergothioneine in promoting collagen generation Download PDF

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Publication number
WO2024022318A1
WO2024022318A1 PCT/CN2023/109052 CN2023109052W WO2024022318A1 WO 2024022318 A1 WO2024022318 A1 WO 2024022318A1 CN 2023109052 W CN2023109052 W CN 2023109052W WO 2024022318 A1 WO2024022318 A1 WO 2024022318A1
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Prior art keywords
ergothioneine
type iii
collagen
iii collagen
production
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PCT/CN2023/109052
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French (fr)
Chinese (zh)
Inventor
刘少英
王瑞妍
陆震
张一鸣
张晓鸥
毛华
陶文文
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华熙生物科技股份有限公司
北京华熙海御科技有限公司
北京华熙荣熙生物技术研究有限公司
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Publication of WO2024022318A1 publication Critical patent/WO2024022318A1/en

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/175—Amino acids
    • 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
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A61K31/4172—Imidazole-alkanecarboxylic acids, e.g. histidine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A61K8/494—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with more than one nitrogen as the only hetero atom
    • A61K8/4946—Imidazoles or their condensed derivatives, e.g. benzimidazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/18—Antioxidants, e.g. antiradicals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations

Definitions

  • the present application belongs to the technical fields of cosmetics and food, and specifically relates to the use of ergothioneine in promoting collagen production.
  • Collagen also known as collagen, is a biological macromolecule protein synthesized by animal cells. It is widely found in the body of animals, is rich in content, and has a wide variety. It has a significant impact on the normal function and damage repair of cells, tissues and even organs. .
  • the collagen in adult skin tissue is mainly type I and type III collagen.
  • the proportion of type III collagen in fetal skin is significantly higher than that of adults.
  • a decrease in type III collagen is associated with skin aging, resulting in a lack of elasticity and regenerative capacity.
  • Supplementing type III collagen can reduce scar hyperplasia and promote skin regeneration and wound healing.
  • Human type III collagen can effectively promote the transformation of macrophages from M1 to M2, the migration ability of L929 cells, and the angiogenesis of human umbilical vein endothelial cells (HUVECs).
  • type I and type III collagen are not positively correlated. Promoting the production of type I collagen does not mean that it can also promote the production of type III collagen, and vice versa. Most of the relevant research focuses on the production of type I collagen, but there are currently few reports on how to promote the production of type III collagen.
  • Ergothioneine is a natural antioxidant that can protect cells in the human body. It is an important active substance in the body and can be used in food, biomedicine, cosmetics and other fields. Although ergothioneine has been widely used, its efficacy has not been fully elucidated. The development of new efficacy of ergothioneine is still an urgent issue that needs to be researched and solved.
  • this application involves the following aspects:
  • the present application provides a collagen production promoter, which includes ergothioneine as an active ingredient.
  • the collagen includes type III collagen.
  • the administration mode of the collagen production promoter includes in vivo and/or in vitro.
  • the present application provides a use of ergothioneine in promoting the production of type III collagen.
  • the administration mode of ergothioneine includes in vivo and/or in vitro.
  • the present application provides the use of ergothioneine in the preparation of products that promote the production of type III collagen in the skin.
  • the present application also provides the use of ergothioneine in the preparation of a product for preventing or alleviating skin problems associated with type III collagen reduction.
  • the products include medicines, medical devices, cosmetics or food.
  • the present application also provides a method of preventing or alleviating skin problems associated with type III collagen reduction, which includes administering ergothioneine to a subject in need thereof.
  • the present application also provides the use of ergothioneine in preventing or alleviating skin problems associated with type III collagen reduction.
  • ergothioneine has the effect of promoting the production of type III collagen. Therefore, ergothioneine can be widely used as a collagen production accelerator to prepare products that promote the production of type III collagen and anti-skin aging.
  • ergothioneine has the effect of promoting the production of type III collagen, and therefore provides a collagen production accelerator, which includes ergothioneine as a as active ingredient.
  • the collagen production accelerator can use ergothioneine as the only active ingredient to promote the production of type III collagen, or it can also contain other active ingredients to work with ergothioneine to promote the production of type III collagen. role.
  • the collagen production enhancing agent is administered by means of in vivo and/or in vitro administration.
  • the concentration of ergothioneine at the time of application can be 0.0001wt%-20wt%, for example, it can be 0.0001wt%, 0.0002wt%, 0.0003wt%, 0.0004wt%, 0.0005wt% , 0.0006wt%, 0.0007wt%, 0.0008wt%, 0.0009wt%, 0.001wt%, 0.002wt%, 0.003wt%, 0.004wt%, 0.005wt%, 0.006wt%, 0.007wt%, 0.008wt%, 0.009 wt%, 0.01wt%, 0.02wt%, 0.03wt%, 0.04wt%, 0.05wt%, 0.06wt%, 0.07wt%, 0.08wt%, 0.09w
  • the above-mentioned collagen production promoter can be applied to the skin directly or indirectly, for example, it can be applied to the skin by smearing, topical application, injection or oral administration.
  • the above-mentioned collagen production promoter can be made into cosmetics, medical devices, medicines, foods, etc.
  • the collagen production boosters mentioned above can be used alone or in combination with other active ingredients. It can be formulated into cosmetics, medical devices, medicines, foods, etc. to promote the production of type III collagen.
  • the present application also provides the use of ergothioneine in promoting the production of type III collagen.
  • the use of ergothioneine in promoting type III collagen production can be therapeutic or non-therapeutic, such as cosmetic uses.
  • the ergothioneine is administered in vivo and/or in vitro.
  • the concentration of ergothioneine at the time of application can be 0.0001wt%-20wt%, for example, it can be 0.0001wt%, 0.0002wt%, 0.0003wt%, 0.0004wt%, 0.0005wt% , 0.0006wt%, 0.0007wt%, 0.0008wt%, 0.0009wt%, 0.001wt%, 0.002wt%, 0.003wt%, 0.004wt%, 0.005wt%, 0.006wt%, 0.007wt%, 0.008wt%, 0.009 wt%, 0.01wt%, 0.02wt%, 0.03wt%, 0.04wt%, 0.05wt%, 0.06wt%, 0.07wt%, 0.08wt%, 0.09wt%,
  • ergothioneine in the application of promoting the production of type III collagen, ergothioneine can be applied to the skin directly or indirectly, for example, through application, topical application, injection or oral administration.
  • ergothioneine can be made into cosmetics, medical devices, medicines, foods, etc.
  • ergothioneine can be formulated alone or with other active ingredients It can be used in cosmetics, medical devices, medicines, foods, etc. to promote the production of type III collagen.
  • ergothioneine resists skin aging by promoting the production of type III collagen.
  • the ergothioneine is administered in vivo and/or in vitro.
  • the concentration of ergothioneine at the time of application can be 0.0001wt%-20wt%, for example, it can be 0.0001wt%, 0.0002wt%, 0.0003wt%, 0.0004wt%, 0.0005wt% , 0.0006wt%, 0.0007wt%, 0.0008wt%, 0.0009wt%, 0.001wt%, 0.002wt%, 0.003wt%, 0.004wt%, 0.005wt%, 0.006wt%, 0.007wt%, 0.008wt%, 0.009 wt%, 0.01wt%, 0.02wt%, 0.03wt%, 0.04wt%, 0.05wt%, 0.06wt%, 0.07wt%, 0.08wt%, 0.09wt%,
  • the present application also provides the use of ergothioneine in preventing or alleviating skin problems associated with reduced type III collagen.
  • the present application also provides a method of preventing or alleviating skin problems associated with type III collagen reduction, which includes administering ergothioneine to a subject in need thereof.
  • the present application also provides the use of ergothioneine in the preparation of a product for preventing or alleviating skin problems associated with type III collagen reduction.
  • skin problems related to reduced type III collagen can be of various types known in the art Skin problems caused by reduced type III collagen, such as reduced skin elasticity, loss of skin elasticity, increased skin wrinkles, skin sagging, slow skin barrier repair, rough skin, etc.
  • ergothioneine is administered in vivo and/or in vitro.
  • the concentration of ergothioneine at the time of application can be 0.0001wt%-20wt%, for example, it can be 0.0001wt%, 0.0002wt%, 0.0003wt%, 0.0004wt%, 0.0005wt% , 0.0006wt%, 0.0007wt%, 0.0008wt%, 0.0009wt%, 0.001wt%, 0.002wt%, 0.003wt%, 0.004wt%, 0.005wt%, 0.006wt%, 0.007wt%, 0.008wt%, 0.009 wt%, 0.01wt%, 0.02wt%, 0.03wt%, 0.04wt%, 0.05wt%, 0.06wt%, 0.07wt%, 0.08wt%, 0.09wt%,
  • ergothioneine can be applied to the skin directly or indirectly, for example, it can be applied to the skin by smearing, topical application, injection or oral administration.
  • the products can be cosmetics, medical devices, medicines, foods, etc.
  • ergothioneine can be formulated into cosmetics, medical devices, pharmaceuticals, foods, etc., alone or together with other active ingredients, for Prevent or relieve skin problems associated with reduced type III collagen.
  • Ergothioneine in this application is used for therapeutic or non-therapeutic purposes to achieve the above effects.
  • the role of ergothioneine in this application is achieved solely for cosmetic purposes.
  • Human fibroblast HFF cells Take the human fibroblast HFF cells in the logarithmic growth phase, discard the culture medium, wash them once with PBS solution, add 0.25wt% trypsin, and place them in the cell culture incubator for 1 minute to stop digestion of the culture solution. The culture medium was centrifuged at 1000 rpm for 3 min, and then the cells were resuspended in complete DMEM culture medium and counted. Human fibroblast HFF was seeded into a 12-well plate at a density of 3 ⁇ 10 5 cells/ml, 1 ml per well. After incubating for 24 hours in an incubator at 37°C and 5% CO2 , remove the culture medium and wash with PBS.
  • the cells washed in step 1 were divided into 2 groups, namely the blank control group and the ergothioneine group. Repeat each group of 3 holes.
  • the blank control group 1 ml of serum-free culture medium.
  • Ergothioneine group 1 ml of serum-free culture medium containing 0.2wt% ergothioneine.
  • phase contrast microscopy to observe cell growth, morphology, and integrity of cell monolayers, and record cells Changes in morphology and growth. Determine the type III collagen content with reference to the Human Collagen III ELISA kit.
  • the calculation method of type III collagen promotion rate is:
  • the ergothioneine solid powder was prepared into a solution using basic culture medium, and the concentration was adjusted to 0.001wt%.
  • the Human Collagen Type III ELISA kit was used to test the effect of ergothioneine on the synthesis of type III collagen in dermal fibroblasts. The statistical results are shown in Table 3.
  • the positive control substance 100ng/mL IGF-1 significantly promoted the synthesis of type III collagen in fibroblasts (P ⁇ 0.05).
  • Ergothioneine in the sample group significantly promoted the production of type III collagen at a concentration of 0.001% (P ⁇ 0.05).

Abstract

The present application provides a collagen generation promoter comprising ergothioneine as an effective component, use of ergothioneine in promoting type III collagen generation, and use of ergothioneine in preparing a product for combating skin aging or preventing or alleviating skin problems related to type III collagen reduction. Ergothioneine has the effect of promoting type III collagen generation. Thus, as a collagen generation promoter, ergothioneine can be widely used for preparing products for promoting type III collagen generation and products for combating skin aging.

Description

麦角硫因在促进胶原蛋白生成中的用途The use of ergothioneine in promoting collagen production 技术领域Technical field
本申请属于化妆品和食品技术领域，具体地，涉及麦角硫因在促进胶原蛋白生成中的用途。The present application belongs to the technical fields of cosmetics and food, and specifically relates to the use of ergothioneine in promoting collagen production.
背景技术Background technique
胶原蛋白(collagen)又称胶原，是由动物细胞合成的一种生物大分子蛋白质，广泛存在于动物体内，含量丰富，种类繁多，对细胞、组织乃至器官行驶正常功能以及损伤修复都有重大影响。Collagen, also known as collagen, is a biological macromolecule protein synthesized by animal cells. It is widely found in the body of animals, is rich in content, and has a wide variety. It has a significant impact on the normal function and damage repair of cells, tissues and even organs. .
成人皮肤组织的胶原蛋白主要是I型和III型胶原蛋白。相比之下，胎儿皮肤中III型胶原蛋白的比例明显高于成人。III型胶原蛋白的减少与皮肤老化有关，导致缺乏弹性和再生能力。补充III型胶原蛋白可减少疤痕增生，促进皮肤再生和伤口愈合。人III型胶原蛋白可有效促进巨噬细胞从M1型向M2型转变、L929细胞的迁移能力和人脐静脉内皮细胞(HUVECs)的血管生成。The collagen in adult skin tissue is mainly type I and type III collagen. In contrast, the proportion of type III collagen in fetal skin is significantly higher than that of adults. A decrease in type III collagen is associated with skin aging, resulting in a lack of elasticity and regenerative capacity. Supplementing type III collagen can reduce scar hyperplasia and promote skin regeneration and wound healing. Human type III collagen can effectively promote the transformation of macrophages from M1 to M2, the migration ability of L929 cells, and the angiogenesis of human umbilical vein endothelial cells (HUVECs).
然而，I型和III型胶原蛋白的生成并不是正相关的，能够促进I型胶原蛋白生成，并不意味着也能促进III型胶原蛋白生成，反之亦然。相关研究大多集中于I型胶原蛋白的生成，而对于如何能促进III型胶原蛋白的生成，目前的报道很少。However, the production of type I and type III collagen is not positively correlated. Promoting the production of type I collagen does not mean that it can also promote the production of type III collagen, and vice versa. Most of the relevant research focuses on the production of type I collagen, but there are currently few reports on how to promote the production of type III collagen.
麦角硫因(Ergothioneine，EGT)是一种天然抗氧化剂，在人体内可以对细胞起到保护作用，是机体内的重要活性物质，其可以应用于食品、生物医学、化妆品等领域。尽管麦角硫因已得到了广泛地应用，但是其功效尚未完全阐释清楚，开发麦角硫因的新功效仍是目前亟待研究和解决的课题。Ergothioneine (EGT) is a natural antioxidant that can protect cells in the human body. It is an important active substance in the body and can be used in food, biomedicine, cosmetics and other fields. Although ergothioneine has been widely used, its efficacy has not been fully elucidated. The development of new efficacy of ergothioneine is still an urgent issue that needs to be researched and solved.
发明内容Contents of the invention
针对现有技术存在的问题，本申请意外地发现，麦角硫因具有促进Ⅲ型胶原蛋白生成的作用，从而完成了本发明。In view of the problems existing in the prior art, the applicant unexpectedly discovered that ergothioneine has the effect of promoting the production of type III collagen, thus completing the present invention.
具体来说，本申请涉及如下方面： Specifically, this application involves the following aspects:
本申请提供一种胶原蛋白生成促进剂，其包括麦角硫因作为有效成分。The present application provides a collagen production promoter, which includes ergothioneine as an active ingredient.
进一步地，所述胶原蛋白包括III型胶原蛋白。Further, the collagen includes type III collagen.
进一步地，所述的胶原蛋白生成促进剂的施用方式包括体内和/或体外。Further, the administration mode of the collagen production promoter includes in vivo and/or in vitro.
本申请提供一种麦角硫因在促进III型胶原蛋白生成中的用途。The present application provides a use of ergothioneine in promoting the production of type III collagen.
进一步地，所述麦角硫因的施用方式包括体内和/或体外。Further, the administration mode of ergothioneine includes in vivo and/or in vitro.
本申请提供一种麦角硫因在制备促进皮肤中III型胶原蛋白生成的产品中的用途。The present application provides the use of ergothioneine in the preparation of products that promote the production of type III collagen in the skin.
本申请还提供麦角硫因在制备预防或缓解III型胶原蛋白降低相关的皮肤问题的产品中的用途。The present application also provides the use of ergothioneine in the preparation of a product for preventing or alleviating skin problems associated with type III collagen reduction.
进一步地，所述产品包括药品、医疗器械、化妆品或食品。Further, the products include medicines, medical devices, cosmetics or food.
本申请还提供一种预防或缓解III型胶原蛋白降低相关皮肤问题的方法，其包括向有此需要的受试者施用麦角硫因。The present application also provides a method of preventing or alleviating skin problems associated with type III collagen reduction, which includes administering ergothioneine to a subject in need thereof.
本申请还提供麦角硫因在预防或缓解III型胶原蛋白降低相关皮肤问题中的用途。The present application also provides the use of ergothioneine in preventing or alleviating skin problems associated with type III collagen reduction.
本申请发现麦角硫因具有促进III型胶原蛋白生成的作用，因此，麦角硫因可以作为胶原蛋白生成促进剂广泛用于制备促进III型胶原蛋白生成的产品和抗皮肤衰老。This application found that ergothioneine has the effect of promoting the production of type III collagen. Therefore, ergothioneine can be widely used as a collagen production accelerator to prepare products that promote the production of type III collagen and anti-skin aging.
具体实施方式Detailed ways
下面结合实施例进一步说明本申请，应当理解，实施例仅用于进一步说明和阐释本申请，并非用于限制本申请。The present application will be further described below with reference to the examples. It should be understood that the examples are only used to further illustrate and illustrate the present application and are not intended to limit the present application.
除非另外定义，本说明书中有关技术的和科学的术语与本领域内的技术人员所通常理解的意思相同。虽然在实验或实际应用中可以应用与此间所述相似或相同的方法和材料，本文还是在下文中对材料和方法做了描述。在相冲突的情况下，以本说明书包括其中定义为准，另外，材料、方法和例子仅供说明，而不具限制性。以下结合具体实施例对本申请作进一步的说明，但不用来限制本申请的范围。Unless otherwise defined, technical and scientific terms used in this specification have the same meaning as commonly understood by those skilled in the art. Although methods and materials similar or equivalent to those described herein can be used in experimental or practical applications, the materials and methods are described below. In case of conflict, the present specification, including definitions, will control. Additionally, materials, methods, and examples are illustrative only and not limiting. The present application will be further described below with reference to specific embodiments, but shall not be used to limit the scope of the present application.
针对现有技术存在的问题，本申请的发明人发现麦角硫因具有促进III型胶原蛋白生成的作用，因此提供一种胶原蛋白生成促进剂，其包括麦角硫因作 为有效成分。In view of the problems existing in the prior art, the inventor of the present application found that ergothioneine has the effect of promoting the production of type III collagen, and therefore provides a collagen production accelerator, which includes ergothioneine as a as active ingredient.
本领域技术人员可以理解，胶原蛋白生成促进剂可以以麦角硫因作为唯一的有效成分发挥促进III型胶原蛋白生成的作用，也可以含有其他有效成分与麦角硫因共同发挥促进III型胶原蛋白生成的作用。Those skilled in the art can understand that the collagen production accelerator can use ergothioneine as the only active ingredient to promote the production of type III collagen, or it can also contain other active ingredients to work with ergothioneine to promote the production of type III collagen. role.
在一些实施方式中，所述胶原蛋白生成促进剂的施用方式包括体内和/或体外施用。当体外施用时，例如涂抹或外敷时，麦角硫因在施用时的浓度可以为0.0001wt％-20wt％，例如可以为0.0001wt％、0.0002wt％、0.0003wt％、0.0004wt％、0.0005wt％、0.0006wt％、0.0007wt％、0.0008wt％、0.0009wt％、0.001wt％、0.002wt％、0.003wt％、0.004wt％、0.005wt％、0.006wt％、0.007wt％、0.008wt％、0.009wt％、0.01wt％、0.02wt％、0.03wt％、0.04wt％、0.05wt％、0.06wt％、0.07wt％、0.08wt％、0.09wt％、0.1wt％、0.2wt％、0.3wt％、0.4wt％、0.5wt％、0.6wt％、0.7wt％、0.8wt％、0.9wt％、1wt％、2wt％、3wt％、4wt％、5wt％、6wt％、7wt％、8wt％、9wt％、10wt％、11wt％、12wt％、13wt％、14wt％、15wt％、16wt％、17wt％、18wt％、19wt％、20wt％等，以及这些数值之间的任意范围；当体内施用时，例如口服时，麦角硫因在施用时的浓度可以为0.0001wt％-20wt％，例如可以为0.0001wt％、0.0002wt％、0.0003wt％、0.0004wt％、0.0005wt％、0.0006wt％、0.0007wt％、0.0008wt％、0.0009wt％、0.001wt％、0.002wt％、0.003wt％、0.004wt％、0.005wt％、0.006wt％、0.007wt％、0.008wt％、0.009wt％、0.01wt％、0.02wt％、0.03wt％、0.04wt％、0.05wt％、0.06wt％、0.07wt％、0.08wt％、0.09wt％、0.1wt％、0.2wt％、0.3wt％、0.4wt％、0.5wt％、0.6wt％、0.7wt％、0.8wt％、0.9wt％、1wt％、2wt％、3wt％、4wt％、5wt％、6wt％、7wt％、8wt％、9wt％、10wt％、11wt％、12wt％、13wt％、14wt％、15wt％、16wt％、17wt％、18wt％、19wt％、20wt％等，以及这些数值之间的任意范围；例如注射时，麦角硫因在施用时的浓度可以为10μg/L-50g/L，例如可以为10μg/L、100μg/L、1000μg/L、1g/L、10g/L、20g/L、30g/L、40g/L、50g/L等，以及这些数值之间的任意范围。In some embodiments, the collagen production enhancing agent is administered by means of in vivo and/or in vitro administration. When applied in vitro, such as when applied or applied externally, the concentration of ergothioneine at the time of application can be 0.0001wt%-20wt%, for example, it can be 0.0001wt%, 0.0002wt%, 0.0003wt%, 0.0004wt%, 0.0005wt% , 0.0006wt%, 0.0007wt%, 0.0008wt%, 0.0009wt%, 0.001wt%, 0.002wt%, 0.003wt%, 0.004wt%, 0.005wt%, 0.006wt%, 0.007wt%, 0.008wt%, 0.009 wt%, 0.01wt%, 0.02wt%, 0.03wt%, 0.04wt%, 0.05wt%, 0.06wt%, 0.07wt%, 0.08wt%, 0.09wt%, 0.1wt%, 0.2wt%, 0.3wt% , 0.4wt%, 0.5wt%, 0.6wt%, 0.7wt%, 0.8wt%, 0.9wt%, 1wt%, 2wt%, 3wt%, 4wt%, 5wt%, 6wt%, 7wt%, 8wt%, 9wt %, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt%, 16 wt%, 17 wt%, 18 wt%, 19 wt%, 20 wt%, etc., and any range between these values; when administered in vivo, For example, when administered orally, the concentration of ergothioneine when administered can be 0.0001wt%-20wt%, for example, it can be 0.0001wt%, 0.0002wt%, 0.0003wt%, 0.0004wt%, 0.0005wt%, 0.0006wt%, 0.0007wt %, 0.0008wt%, 0.0009wt%, 0.001wt%, 0.002wt%, 0.003wt%, 0.004wt%, 0.005wt%, 0.006wt%, 0.007wt%, 0.008wt%, 0.009wt%, 0.01wt%, 0.02wt%, 0.03wt%, 0.04wt%, 0.05wt%, 0.06wt%, 0.07wt%, 0.08wt%, 0.09wt%, 0.1wt%, 0.2wt%, 0.3wt%, 0.4wt%, 0.5wt %, 0.6wt%, 0.7wt%, 0.8wt%, 0.9wt%, 1wt%, 2wt%, 3wt%, 4wt%, 5wt%, 6wt%, 7wt%, 8wt%, 9wt%, 10wt%, 11wt% , 12wt%, 13wt%, 14wt%, 15wt%, 16wt%, 17wt%, 18wt%, 19wt%, 20wt%, etc., and any range between these values; for example, when injected, the concentration of ergothioneine at the time of administration It can be 10μg/L-50g/L, for example, it can be 10μg/L, 100μg/L, 1000μg/L, 1g/L, 10g/L, 20g/L, 30g/L, 40g/L, 50g/L, etc., and any range between these values.
在一些实施方式中，上述胶原蛋白生成促进剂可以直接或间接地施用于皮肤，例如可以通过涂抹、外敷、注射或口服等方式施用于皮肤。In some embodiments, the above-mentioned collagen production promoter can be applied to the skin directly or indirectly, for example, it can be applied to the skin by smearing, topical application, injection or oral administration.
在一些实施方式中，上述胶原蛋白生成促进剂可以制成化妆品、医疗器械、药品、食品等。例如，上述胶原蛋白生成促进剂可以单独或与其他活性成分一 起配制成化妆品、医疗器械、药品、食品等，用于促进III型胶原蛋白生成。In some embodiments, the above-mentioned collagen production promoter can be made into cosmetics, medical devices, medicines, foods, etc. For example, the collagen production boosters mentioned above can be used alone or in combination with other active ingredients. It can be formulated into cosmetics, medical devices, medicines, foods, etc. to promote the production of type III collagen.
本申请还提供麦角硫因在促进III型胶原蛋白生成中的用途。麦角硫因在促进III型胶原蛋白生成中的用途可以为治疗用途，也可以为非治疗目的的用途，例如美容用途。The present application also provides the use of ergothioneine in promoting the production of type III collagen. The use of ergothioneine in promoting type III collagen production can be therapeutic or non-therapeutic, such as cosmetic uses.
在一些实施方式中，所述麦角硫因的施用方式包括体内和/或体外。当体外施用时，例如涂抹或外敷时，麦角硫因在施用时的浓度可以为0.0001wt％-20wt％，例如可以为0.0001wt％、0.0002wt％、0.0003wt％、0.0004wt％、0.0005wt％、0.0006wt％、0.0007wt％、0.0008wt％、0.0009wt％、0.001wt％、0.002wt％、0.003wt％、0.004wt％、0.005wt％、0.006wt％、0.007wt％、0.008wt％、0.009wt％、0.01wt％、0.02wt％、0.03wt％、0.04wt％、0.05wt％、0.06wt％、0.07wt％、0.08wt％、0.09wt％、0.1wt％、0.2wt％、0.3wt％、0.4wt％、0.5wt％、0.6wt％、0.7wt％、0.8wt％、0.9wt％、1wt％、2wt％、3wt％、4wt％、5wt％、6wt％、7wt％、8wt％、9wt％、10wt％、11wt％、12wt％、13wt％、14wt％、15wt％、16wt％、17wt％、18wt％、19wt％、20wt％等，以及这些数值之间的任意范围；当体内施用时，例如口服时，麦角硫因在施用时的浓度可以为0.0001wt％-20wt％，例如可以为0.0001wt％、0.0002wt％、0.0003wt％、0.0004wt％、0.0005wt％、0.0006wt％、0.0007wt％、0.0008wt％、0.0009wt％、0.001wt％、0.002wt％、0.003wt％、0.004wt％、0.005wt％、0.006wt％、0.007wt％、0.008wt％、0.009wt％、0.01wt％、0.02wt％、0.03wt％、0.04wt％、0.05wt％、0.06wt％、0.07wt％、0.08wt％、0.09wt％、0.1wt％、0.2wt％、0.3wt％、0.4wt％、0.5wt％、0.6wt％、0.7wt％、0.8wt％、0.9wt％、1wt％、2wt％、3wt％、4wt％、5wt％、6wt％、7wt％、8wt％、9wt％、10wt％、11wt％、12wt％、13wt％、14wt％、15wt％、16wt％、17wt％、18wt％、19wt％、20wt％等，以及这些数值之间的任意范围；例如注射时，麦角硫因在施用时的浓度可以为10μg/L-50g/L，例如可以为10μg/L、100μg/L、1000μg/L、1g/L、10g/L、20g/L、30g/L、40g/L、50g/L等，以及这些数值之间的任意范围。In some embodiments, the ergothioneine is administered in vivo and/or in vitro. When applied in vitro, such as when applied or applied externally, the concentration of ergothioneine at the time of application can be 0.0001wt%-20wt%, for example, it can be 0.0001wt%, 0.0002wt%, 0.0003wt%, 0.0004wt%, 0.0005wt% , 0.0006wt%, 0.0007wt%, 0.0008wt%, 0.0009wt%, 0.001wt%, 0.002wt%, 0.003wt%, 0.004wt%, 0.005wt%, 0.006wt%, 0.007wt%, 0.008wt%, 0.009 wt%, 0.01wt%, 0.02wt%, 0.03wt%, 0.04wt%, 0.05wt%, 0.06wt%, 0.07wt%, 0.08wt%, 0.09wt%, 0.1wt%, 0.2wt%, 0.3wt% , 0.4wt%, 0.5wt%, 0.6wt%, 0.7wt%, 0.8wt%, 0.9wt%, 1wt%, 2wt%, 3wt%, 4wt%, 5wt%, 6wt%, 7wt%, 8wt%, 9wt %, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt%, 16 wt%, 17 wt%, 18 wt%, 19 wt%, 20 wt%, etc., and any range between these values; when administered in vivo, For example, when administered orally, the concentration of ergothioneine when administered can be 0.0001wt%-20wt%, for example, it can be 0.0001wt%, 0.0002wt%, 0.0003wt%, 0.0004wt%, 0.0005wt%, 0.0006wt%, 0.0007wt %, 0.0008wt%, 0.0009wt%, 0.001wt%, 0.002wt%, 0.003wt%, 0.004wt%, 0.005wt%, 0.006wt%, 0.007wt%, 0.008wt%, 0.009wt%, 0.01wt%, 0.02wt%, 0.03wt%, 0.04wt%, 0.05wt%, 0.06wt%, 0.07wt%, 0.08wt%, 0.09wt%, 0.1wt%, 0.2wt%, 0.3wt%, 0.4wt%, 0.5wt %, 0.6wt%, 0.7wt%, 0.8wt%, 0.9wt%, 1wt%, 2wt%, 3wt%, 4wt%, 5wt%, 6wt%, 7wt%, 8wt%, 9wt%, 10wt%, 11wt% , 12wt%, 13wt%, 14wt%, 15wt%, 16wt%, 17wt%, 18wt%, 19wt%, 20wt%, etc., and any range between these values; for example, when injected, the concentration of ergothioneine at the time of administration It can be 10μg/L-50g/L, for example, it can be 10μg/L, 100μg/L, 1000μg/L, 1g/L, 10g/L, 20g/L, 30g/L, 40g/L, 50g/L, etc., and any range between these values.
在本申请的促进III型胶原蛋白生成的用途中，麦角硫因可以直接或间接地施用于皮肤，例如可以通过涂抹、外敷、注射或口服等方式施用于皮肤。In the application of promoting the production of type III collagen, ergothioneine can be applied to the skin directly or indirectly, for example, through application, topical application, injection or oral administration.
在本申请的促进III型胶原蛋白生成的用途中，麦角硫因可以制成化妆品、医疗器械、药品、食品等。例如，麦角硫因可以单独或与其他活性成分一起配制 成化妆品、医疗器械、药品、食品等，用于促进III型胶原蛋白生成。In the application of promoting the production of type III collagen, ergothioneine can be made into cosmetics, medical devices, medicines, foods, etc. For example, ergothioneine can be formulated alone or with other active ingredients It can be used in cosmetics, medical devices, medicines, foods, etc. to promote the production of type III collagen.
进一步地，麦角硫因通过促进III型胶原蛋白生成而抗皮肤衰老。Furthermore, ergothioneine resists skin aging by promoting the production of type III collagen.
在一些实施方式中，所述麦角硫因的施用方式包括体内和/或体外。当体外施用时，例如涂抹或外敷时，麦角硫因在施用时的浓度可以为0.0001wt％-20wt％，例如可以为0.0001wt％、0.0002wt％、0.0003wt％、0.0004wt％、0.0005wt％、0.0006wt％、0.0007wt％、0.0008wt％、0.0009wt％、0.001wt％、0.002wt％、0.003wt％、0.004wt％、0.005wt％、0.006wt％、0.007wt％、0.008wt％、0.009wt％、0.01wt％、0.02wt％、0.03wt％、0.04wt％、0.05wt％、0.06wt％、0.07wt％、0.08wt％、0.09wt％、0.1wt％、0.2wt％、0.3wt％、0.4wt％、0.5wt％、0.6wt％、0.7wt％、0.8wt％、0.9wt％、1wt％、2wt％、3wt％、4wt％、5wt％、6wt％、7wt％、8wt％、9wt％、10wt％、11wt％、12wt％、13wt％、14wt％、15wt％、16wt％、17wt％、18wt％、19wt％、20wt％等，以及这些数值之间的任意范围；当体内施用时，例如口服时，麦角硫因在施用时的浓度可以为0.0001wt％-20wt％，例如可以为0.0001wt％、0.0002wt％、0.0003wt％、0.0004wt％、0.0005wt％、0.0006wt％、0.0007wt％、0.0008wt％、0.0009wt％、0.001wt％、0.002wt％、0.003wt％、0.004wt％、0.005wt％、0.006wt％、0.007wt％、0.008wt％、0.009wt％、0.01wt％、0.02wt％、0.03wt％、0.04wt％、0.05wt％、0.06wt％、0.07wt％、0.08wt％、0.09wt％、0.1wt％、0.2wt％、0.3wt％、0.4wt％、0.5wt％、0.6wt％、0.7wt％、0.8wt％、0.9wt％、1wt％、2wt％、3wt％、4wt％、5wt％、6wt％、7wt％、8wt％、9wt％、10wt％、11wt％、12wt％、13wt％、14wt％、15wt％、16wt％、17wt％、18wt％、19wt％、20wt％等，以及这些数值之间的任意范围；例如注射时，麦角硫因在施用时的浓度可以为10μg/L-50g/L，例如可以为10μg/L、100μg/L、1000μg/L、1g/L、10g/L、20g/L、30g/L、40g/L、50g/L等，以及这些数值之间的任意范围。In some embodiments, the ergothioneine is administered in vivo and/or in vitro. When applied in vitro, such as when applied or applied externally, the concentration of ergothioneine at the time of application can be 0.0001wt%-20wt%, for example, it can be 0.0001wt%, 0.0002wt%, 0.0003wt%, 0.0004wt%, 0.0005wt% , 0.0006wt%, 0.0007wt%, 0.0008wt%, 0.0009wt%, 0.001wt%, 0.002wt%, 0.003wt%, 0.004wt%, 0.005wt%, 0.006wt%, 0.007wt%, 0.008wt%, 0.009 wt%, 0.01wt%, 0.02wt%, 0.03wt%, 0.04wt%, 0.05wt%, 0.06wt%, 0.07wt%, 0.08wt%, 0.09wt%, 0.1wt%, 0.2wt%, 0.3wt% , 0.4wt%, 0.5wt%, 0.6wt%, 0.7wt%, 0.8wt%, 0.9wt%, 1wt%, 2wt%, 3wt%, 4wt%, 5wt%, 6wt%, 7wt%, 8wt%, 9wt %, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt%, 16 wt%, 17 wt%, 18 wt%, 19 wt%, 20 wt%, etc., and any range between these values; when administered in vivo, For example, when administered orally, the concentration of ergothioneine when administered can be 0.0001wt%-20wt%, for example, it can be 0.0001wt%, 0.0002wt%, 0.0003wt%, 0.0004wt%, 0.0005wt%, 0.0006wt%, 0.0007wt %, 0.0008wt%, 0.0009wt%, 0.001wt%, 0.002wt%, 0.003wt%, 0.004wt%, 0.005wt%, 0.006wt%, 0.007wt%, 0.008wt%, 0.009wt%, 0.01wt%, 0.02wt%, 0.03wt%, 0.04wt%, 0.05wt%, 0.06wt%, 0.07wt%, 0.08wt%, 0.09wt%, 0.1wt%, 0.2wt%, 0.3wt%, 0.4wt%, 0.5wt %, 0.6wt%, 0.7wt%, 0.8wt%, 0.9wt%, 1wt%, 2wt%, 3wt%, 4wt%, 5wt%, 6wt%, 7wt%, 8wt%, 9wt%, 10wt%, 11wt% , 12wt%, 13wt%, 14wt%, 15wt%, 16wt%, 17wt%, 18wt%, 19wt%, 20wt%, etc., and any range between these values; for example, when injected, the concentration of ergothioneine at the time of administration It can be 10μg/L-50g/L, for example, it can be 10μg/L, 100μg/L, 1000μg/L, 1g/L, 10g/L, 20g/L, 30g/L, 40g/L, 50g/L, etc., and any range between these values.
本申请还提供麦角硫因在预防或缓解III型胶原蛋白降低相关的皮肤问题中的用途。The present application also provides the use of ergothioneine in preventing or alleviating skin problems associated with reduced type III collagen.
本申请还提供一种预防或缓解III型胶原蛋白降低相关皮肤问题的方法，其包括向有此需要的受试者施用麦角硫因。The present application also provides a method of preventing or alleviating skin problems associated with type III collagen reduction, which includes administering ergothioneine to a subject in need thereof.
本申请还提供麦角硫因在制备预防或缓解III型胶原蛋白降低相关的皮肤问题的产品中的用途。The present application also provides the use of ergothioneine in the preparation of a product for preventing or alleviating skin problems associated with type III collagen reduction.
其中，与III型胶原蛋白降低相关的皮肤问题可以是本领域已知的各种类型 的由III型胶原蛋白降低引起的皮肤问题，例如皮肤弹性降低、皮肤弹性丧失、皮肤皱纹增加、皮肤松弛、皮肤屏障修复缓慢、皮肤粗糙等。Among them, skin problems related to reduced type III collagen can be of various types known in the art Skin problems caused by reduced type III collagen, such as reduced skin elasticity, loss of skin elasticity, increased skin wrinkles, skin sagging, slow skin barrier repair, rough skin, etc.
在一些实施方式中，麦角硫因的施用方式包括体内和/或体外。当体外施用时，例如涂抹或外敷时，麦角硫因在施用时的浓度可以为0.0001wt％-20wt％，例如可以为0.0001wt％、0.0002wt％、0.0003wt％、0.0004wt％、0.0005wt％、0.0006wt％、0.0007wt％、0.0008wt％、0.0009wt％、0.001wt％、0.002wt％、0.003wt％、0.004wt％、0.005wt％、0.006wt％、0.007wt％、0.008wt％、0.009wt％、0.01wt％、0.02wt％、0.03wt％、0.04wt％、0.05wt％、0.06wt％、0.07wt％、0.08wt％、0.09wt％、0.1wt％、0.2wt％、0.3wt％、0.4wt％、0.5wt％、0.6wt％、0.7wt％、0.8wt％、0.9wt％、1wt％、2wt％、3wt％、4wt％、5wt％、6wt％、7wt％、8wt％、9wt％、10wt％、11wt％、12wt％、13wt％、14wt％、15wt％、16wt％、17wt％、18wt％、19wt％、20wt％等，以及这些数值之间的任意范围；当体内施用时，例如口服时，麦角硫因在施用时的浓度可以为0.0001wt％-20wt％，例如可以为0.0001wt％、0.0002wt％、0.0003wt％、0.0004wt％、0.0005wt％、0.0006wt％、0.0007wt％、0.0008wt％、0.0009wt％、0.001wt％、0.002wt％、0.003wt％、0.004wt％、0.005wt％、0.006wt％、0.007wt％、0.008wt％、0.009wt％、0.01wt％、0.02wt％、0.03wt％、0.04wt％、0.05wt％、0.06wt％、0.07wt％、0.08wt％、0.09wt％、0.1wt％、0.2wt％、0.3wt％、0.4wt％、0.5wt％、0.6wt％、0.7wt％、0.8wt％、0.9wt％、1wt％、2wt％、3wt％、4wt％、5wt％、6wt％、7wt％、8wt％、9wt％、10wt％、11wt％、12wt％、13wt％、14wt％、15wt％、16wt％、17wt％、18wt％、19wt％、20wt％等，以及这些数值之间的任意范围；例如注射时，麦角硫因在施用时的浓度可以为10μg/L-50g/L，例如可以为10μg/L、100μg/L、1000μg/L、1g/L、10g/L、20g/L、30g/L、40g/L、50g/L等，以及这些数值之间的任意范围。In some embodiments, ergothioneine is administered in vivo and/or in vitro. When applied in vitro, such as when applied or applied externally, the concentration of ergothioneine at the time of application can be 0.0001wt%-20wt%, for example, it can be 0.0001wt%, 0.0002wt%, 0.0003wt%, 0.0004wt%, 0.0005wt% , 0.0006wt%, 0.0007wt%, 0.0008wt%, 0.0009wt%, 0.001wt%, 0.002wt%, 0.003wt%, 0.004wt%, 0.005wt%, 0.006wt%, 0.007wt%, 0.008wt%, 0.009 wt%, 0.01wt%, 0.02wt%, 0.03wt%, 0.04wt%, 0.05wt%, 0.06wt%, 0.07wt%, 0.08wt%, 0.09wt%, 0.1wt%, 0.2wt%, 0.3wt% , 0.4wt%, 0.5wt%, 0.6wt%, 0.7wt%, 0.8wt%, 0.9wt%, 1wt%, 2wt%, 3wt%, 4wt%, 5wt%, 6wt%, 7wt%, 8wt%, 9wt %, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt%, 16 wt%, 17 wt%, 18 wt%, 19 wt%, 20 wt%, etc., and any range between these values; when administered in vivo, For example, when administered orally, the concentration of ergothioneine when administered can be 0.0001wt%-20wt%, for example, it can be 0.0001wt%, 0.0002wt%, 0.0003wt%, 0.0004wt%, 0.0005wt%, 0.0006wt%, 0.0007wt %, 0.0008wt%, 0.0009wt%, 0.001wt%, 0.002wt%, 0.003wt%, 0.004wt%, 0.005wt%, 0.006wt%, 0.007wt%, 0.008wt%, 0.009wt%, 0.01wt%, 0.02wt%, 0.03wt%, 0.04wt%, 0.05wt%, 0.06wt%, 0.07wt%, 0.08wt%, 0.09wt%, 0.1wt%, 0.2wt%, 0.3wt%, 0.4wt%, 0.5wt %, 0.6wt%, 0.7wt%, 0.8wt%, 0.9wt%, 1wt%, 2wt%, 3wt%, 4wt%, 5wt%, 6wt%, 7wt%, 8wt%, 9wt%, 10wt%, 11wt% , 12wt%, 13wt%, 14wt%, 15wt%, 16wt%, 17wt%, 18wt%, 19wt%, 20wt%, etc., and any range between these values; for example, when injected, the concentration of ergothioneine at the time of administration It can be 10μg/L-50g/L, for example, it can be 10μg/L, 100μg/L, 1000μg/L, 1g/L, 10g/L, 20g/L, 30g/L, 40g/L, 50g/L, etc., and any range between these values.
在本申请的预防或缓解III型胶原蛋白降低相关皮肤问题中的用途、制备预防或缓解与III型胶原蛋白降低相关的皮肤问题的产品中的用途或预防或缓解III型胶原蛋白降低相关皮肤问题的方法中，麦角硫因可以直接或间接地施用于皮肤，例如可以通过涂抹、外敷、注射或口服等方式施用于皮肤。Use in the present application for preventing or alleviating skin problems related to the reduction of type III collagen, use in preparing products for preventing or alleviating skin problems related to the reduction of type III collagen, or preventing or alleviating skin problems related to the reduction of type III collagen In the method, ergothioneine can be applied to the skin directly or indirectly, for example, it can be applied to the skin by smearing, topical application, injection or oral administration.
在本申请的制备预防或缓解与III型胶原蛋白降低相关的皮肤问题的产品中的用途中，所述产品可以为化妆品、医疗器械、药品、食品等。例如，麦角硫因可以单独或与其他活性成分一起配制成化妆品、医疗器械、药品、食品等，用于 预防或缓解与III型胶原蛋白降低相关的皮肤问题。In the use of the present application in preparing products for preventing or alleviating skin problems related to the reduction of type III collagen, the products can be cosmetics, medical devices, medicines, foods, etc. For example, ergothioneine can be formulated into cosmetics, medical devices, pharmaceuticals, foods, etc., alone or together with other active ingredients, for Prevent or relieve skin problems associated with reduced type III collagen.
本申请中的麦角硫因是出于治疗或非治疗目的来实现以上作用。例如仅出于美容的目的实现麦角硫因在本申请中的作用。Ergothioneine in this application is used for therapeutic or non-therapeutic purposes to achieve the above effects. For example, the role of ergothioneine in this application is achieved solely for cosmetic purposes.
实施例Example
实施例中使用的实验材料和实验设备的相关信息如表1所示。Relevant information about the experimental materials and experimental equipment used in the examples is shown in Table 1.
表1
Table 1
实施例1Example 1
实验步骤：Experimental steps:
1铺板1 planking
取对数生长期的人成纤维细胞HFF，弃去培养基，用PBS液清洗一遍，加入0.25wt％胰酶，在细胞培养箱放置1min，培养液终止消化。将培养液在1000rpm离心3min，然后用完全DMEM培养液重悬细胞，进行计数。按3×105细胞/ml的密度将人成纤维细胞HFF接种到12孔板中，每孔1ml。在37℃、5％CO2条件下于培养箱孵育24h后，去除培养液，PBS清洗。Take the human fibroblast HFF cells in the logarithmic growth phase, discard the culture medium, wash them once with PBS solution, add 0.25wt% trypsin, and place them in the cell culture incubator for 1 minute to stop digestion of the culture solution. The culture medium was centrifuged at 1000 rpm for 3 min, and then the cells were resuspended in complete DMEM culture medium and counted. Human fibroblast HFF was seeded into a 12-well plate at a density of 3×10 5 cells/ml, 1 ml per well. After incubating for 24 hours in an incubator at 37°C and 5% CO2 , remove the culture medium and wash with PBS.
2细胞分组2 cell grouping
将步骤1清洗后的细胞分为2组，分别为空白对照组和麦角硫因组。每组3孔重复。其中，空白对照组：1ml的无血清培养液。麦角硫因组：1ml含0.2wt％麦角硫因的无血清培养液。The cells washed in step 1 were divided into 2 groups, namely the blank control group and the ergothioneine group. Repeat each group of 3 holes. Among them, the blank control group: 1 ml of serum-free culture medium. Ergothioneine group: 1 ml of serum-free culture medium containing 0.2wt% ergothioneine.
将上述各组混匀后，放置培养箱中继续培养72h。After mixing the above groups, place them in an incubator and continue culturing for 72 hours.
3检测3 detection
使用相差显微镜观察细胞生长情况、形态和细胞单层的完整性，记录细胞 形态和生长的变化。参照人Human Collagen III ELISA试剂盒测定III型胶原蛋白含量。Use phase contrast microscopy to observe cell growth, morphology, and integrity of cell monolayers, and record cells Changes in morphology and growth. Determine the type III collagen content with reference to the Human Collagen III ELISA kit.
III型胶原蛋白促进率的计算方式为：
The calculation method of type III collagen promotion rate is:
实验结果：Experimental results:
如表2所示，与空白对照组相比，麦角硫因组的III型胶原蛋白促进率显著提高，这说明麦角硫因可显著促进III型胶原蛋白的生成。As shown in Table 2, compared with the blank control group, the type III collagen promotion rate in the ergothioneine group was significantly increased, which shows that ergothioneine can significantly promote the production of type III collagen.
表2
Table 2
注：*代表P<0.05，**代表P<0.01，***代表P<0.001Note: * represents P<0.05, ** represents P<0.01, *** represents P<0.001
实施例2Example 2
1.样品制备1. Sample preparation
将麦角硫因固体粉末用基础培养液配制成溶液，并调节浓度至0.001wt％。The ergothioneine solid powder was prepared into a solution using basic culture medium, and the concentration was adjusted to 0.001wt%.
2.人原代真皮成纤维细胞的III型胶原蛋白生成量测试2. Test of type III collagen production of human primary dermal fibroblasts
当体外培养的成纤维细胞生长至80-90％汇合时，用胰酶将其消化并接种于96孔板。细胞在96孔板中贴附培养48h后，用步骤(1)中配制的麦角硫因样品处理细胞(N＝3)，同时设置含100ng/mL IGF-1的处理组为阳性对照组。48h后，取细胞培养上清液用于III型胶原蛋白的ELISA测试，所测得样品组的III型胶原蛋白含量与空白对照组进行统计比较，评估麦角硫因对III型胶原蛋白生成量的影响。When the in vitro cultured fibroblasts grow to 80-90% confluence, they are digested with trypsin and seeded in a 96-well plate. After the cells were adherently cultured in a 96-well plate for 48 hours, the cells (N=3) were treated with the ergothioneine sample prepared in step (1), and the treatment group containing 100ng/mL IGF-1 was set as a positive control group. After 48 hours, the cell culture supernatant was taken for the ELISA test of type III collagen. The measured type III collagen content of the sample group was statistically compared with the blank control group to evaluate the effect of ergothioneine on the production of type III collagen. Influence.
3.统计方法3. Statistical methods
所有结果以Mean±SD表示，计量资料差异比较采用unpaired t检验。P<0.05视为差异有统计学意义。All results are expressed as Mean ± SD, and unpaired t test was used to compare differences in measurement data. P<0.05 was considered as a statistically significant difference.
4.实验结果4.Experimental results
利用Human Collagen Type III ELISA试剂盒测试麦角硫因对真皮成纤维细胞III型胶原蛋白合成量的影响，数据统计结果如表3所示。阳性对照品100ng/mL IGF-1对成纤维细胞的III型胶原蛋白合成量有显著促进作用(P<0.05)。 样品组麦角硫因在0.001％浓度下对III型胶原蛋白生成量具有显著促进作用(P<0.05)。The Human Collagen Type III ELISA kit was used to test the effect of ergothioneine on the synthesis of type III collagen in dermal fibroblasts. The statistical results are shown in Table 3. The positive control substance 100ng/mL IGF-1 significantly promoted the synthesis of type III collagen in fibroblasts (P<0.05). Ergothioneine in the sample group significantly promoted the production of type III collagen at a concentration of 0.001% (P<0.05).
表3
table 3

Claims (10)

  1. 一种胶原蛋白生成促进剂，其包括麦角硫因作为有效成分。A collagen production promoter including ergothioneine as an active ingredient.
  2. 根据权利要求1所述的胶原蛋白生成促进剂，所述胶原蛋白包括III型胶原蛋白。The collagen production promoter according to claim 1, wherein the collagen includes type III collagen.
  3. 根据权利要求1或2所述的胶原蛋白生成促进剂，其施用方式包括体内和/或体外。The collagen production accelerator according to claim 1 or 2, its administration mode includes in vivo and/or in vitro.
  4. 麦角硫因在促进III型胶原蛋白生成中的用途。The use of ergothioneine in promoting the production of type III collagen.
  5. 根据权利要求4所述的用途，所述麦角硫因的施用方式包括体内和/或体外。According to the use according to claim 4, the administration mode of the ergothioneine includes in vivo and/or in vitro.
  6. 麦角硫因在制备促进皮肤中III型胶原蛋白生成的产品中的用途。Use of ergothioneine in the preparation of products that promote the production of type III collagen in the skin.
  7. 麦角硫因在制备预防或缓解III型胶原蛋白降低相关的皮肤问题的产品中的用途。Use of ergothioneine in the preparation of products for preventing or alleviating skin problems associated with reduced type III collagen.
  8. 根据权利要求7所述的用途，所述产品包括药品、医疗器械、化妆品或食品。According to the use of claim 7, the product includes pharmaceuticals, medical devices, cosmetics or food.
  9. 一种预防或缓解III型胶原蛋白降低相关皮肤问题的方法，其包括向有此需要的受试者施用麦角硫因。A method of preventing or alleviating skin problems associated with type III collagen reduction, comprising administering ergothioneine to a subject in need thereof.
  10. 麦角硫因在预防或缓解III型胶原蛋白降低相关皮肤问题中的用途。 The use of ergothioneine in preventing or alleviating skin problems associated with reduced type III collagen.
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Citations (2)

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Publication number Priority date Publication date Assignee Title
US20020042438A1 (en) * 2000-06-26 2002-04-11 L'oreal Use of ergothioneine and/or its derivatives as an anti-glycation agent
JP4865083B1 (en) * 2010-11-19 2012-02-01 幸子 清水 Method for producing ergothioneine

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020042438A1 (en) * 2000-06-26 2002-04-11 L'oreal Use of ergothioneine and/or its derivatives as an anti-glycation agent
JP4865083B1 (en) * 2010-11-19 2012-02-01 幸子 清水 Method for producing ergothioneine

Non-Patent Citations (2)

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Title
HSEU YOU-CHENG, VUDHYA GOWRISANKAR YUGANDHAR, CHEN XUAN-ZAO, YANG YI-CHEN, YANG HSIN-LING: "The Antiaging Activity of Ergothioneine in UVA-Irradiated Human Dermal Fibroblasts via the Inhibition of the AP-1 Pathway and the Activation of Nrf2-Mediated Antioxidant Genes", OXIDATIVE MEDICINE AND CELLULAR LONGEVITY, HINDAWI PUBLISHING CORPORATION, US, vol. 2020, 12 February 2020 (2020-02-12), US , pages 1 - 13, XP093133608, ISSN: 1942-0900, DOI: 10.1155/2020/2576823 *
KO, H.J. ET AL.: "Ergothioneine Alleviates Senescence of Fibroblasts Induced by UVB Damage of Keratinocytes via Activation of the Nrf2/HO-1 Pathway and HSP70 in Keratinocytes.", EXPERIMENTAL CELL RESEARCH, vol. 400, no. 1, 9 February 2021 (2021-02-09), XP086503109, ISSN: 0014-4827, DOI: 10.1016/j.yexcr.2021.112516 *

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