WO2022050318A1 - Utilisation d'extrait de placenta - Google Patents

Utilisation d'extrait de placenta Download PDF

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WO2022050318A1
WO2022050318A1 PCT/JP2021/032175 JP2021032175W WO2022050318A1 WO 2022050318 A1 WO2022050318 A1 WO 2022050318A1 JP 2021032175 W JP2021032175 W JP 2021032175W WO 2022050318 A1 WO2022050318 A1 WO 2022050318A1
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general formula
production
represented
following general
placenta
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Japanese (ja)
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邦義 清水
朋恵 西尾
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国立大学法人九州大学
ドクタープロラボジャパン株式会社
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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
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • 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
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
    • 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
    • A61K31/66—Phosphorus compounds
    • A61K31/661—Phosphorus acids or esters thereof not having P—C bonds, e.g. fosfosal, dichlorvos, malathion or mevinphos
    • 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
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48—Reproductive organs
    • A61K35/50—Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
    • 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
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/36—Carboxylic acids; Salts or anhydrides 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
    • A61K8/30—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
    • A61K8/55—Phosphorus 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
    • A61K8/63—Steroids; Derivatives 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/16—Emollients or protectives, e.g. against radiation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/18—Antioxidants, e.g. antiradicals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • 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
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/04—Preparations for care of the skin for chemically tanning 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
    • A61Q19/08—Anti-ageing preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q5/00—Preparations for care of the hair
    • A61Q5/02—Preparations for cleaning the hair

Definitions

  • the present invention relates to the use of placenta extract.
  • placenta mammalian placenta (placenta) and its extracts are attracting attention from a wide range of age groups, regardless of gender, as new materials that promote both health and beauty (see, for example, Patent Document 1).
  • placenta is used mainly in the form of extracts in the field of health foods for the purpose of supporting health and in the field of cosmetics for the purpose of supporting beauty, and daily health and skin condition. It has been well received by users in terms of.
  • the present invention has been made in view of such circumstances, and provides a new method of using the placenta extract for a clearer purpose.
  • a component that suppresses or promotes melanin production in cosmetics or functional foods a hyaluronic acid production promoting component, a collagen production promoting component, an elastase activity inhibiting component, and an antiallergic substance. It was decided to use placenta extract as an additive material for any of the components selected from the activity promoting component and the tyrosinase activity inhibiting component.
  • the present invention also has the following features.
  • the placenta extract comprises a group of components exhibiting solubility in a non-aqueous specific solvent consisting of a mixture of any one or two or more selected from dimethyl sulfoxide, ethanol, methanol and hexane. matter.
  • the placenta extract is composed of a group of components exhibiting solubility in an aqueous solvent containing water as a main component.
  • the present invention is selected from a component that suppresses or promotes melanin production in cosmetics or functional foods, a hyaluronic acid production promoting component, a collagen production promoting component, an elastase activity inhibiting component, an antiallergic activity promoting component, and a tyrosinase activity inhibiting component. Since the placenta extract is used as an additive material for any of the above components, it is possible to provide a new method of using the placenta extract for a clearer purpose.
  • the placenta extract comprises a group of components exhibiting solubility in a non-aqueous specific solvent consisting of a mixture of any one or two or more selected from dimethyl sulfoxide, ethanol, methanol and hexane. If so, it can exhibit an action of promoting melanin production, and also exhibits a more remarkable effect of inhibiting elastase activity than the elastase activity-inhibiting component contained in the component group exhibiting solubility in an aqueous solvent. Can be done.
  • the placenta extract is composed of a group of components exhibiting solubility in an aqueous solvent containing water as a main component, it has an action of suppressing melanin production and a solubility in a non-aqueous specific solvent. It is possible to exhibit more remarkable actions as compared with the hyaluronic acid production promoting component and the collagen production promoting component contained in the indicated component group.
  • the present invention relates to a method of using the placenta extract, and more specifically, in health foods and cosmetics which have been used in some cases without necessarily clarifying the significance of the addition of the placenta extract. It provides a new usage of placenta extract for various purposes.
  • a component that suppresses or promotes melanin production in cosmetics or functional foods a hyaluronic acid production promoting component, a collagen production promoting component, an elastase activity inhibiting component, an antiallergic activity promoting component, and a tyrosinase activity. It is characteristic in that placenta extract is used as an additive material for any of the components selected from the inhibitory components.
  • the placenta extract is an extract of the placenta (placenta) of mammals.
  • placenta placenta
  • the placenta extract contains the extract or the dried product of the extract.
  • placenta from which the placenta extract is extracted is not particularly limited, and placenta of various mammals including pigs, horses, and humans can be adopted.
  • cosmetics include basic cosmetics, hair tonics, perfumes, toothpastes, shampoos, conditioners, soaps and cleaning agents used for body cleaning, and toiletry products such as bath salts. It also includes medicated cosmetics that claim a preventive effect.
  • makeup cosmetics include foundations, eyebrows, mascara, eye shadows, eyelines, lipsticks, glosses, blushers, white powders, and manicures.
  • Basic cosmetics include, for example, lotions and milky lotions. Examples include facial cleansers, cleansers, serums and creams.
  • functional food is a concept that includes all foods that do not contain pharmaceutical ingredients and are thought to contribute to the maintenance and promotion of health. It also includes foods, foods for specified health use, foods with nutritional function, and foods with health claims such as foods with functional claims.
  • the placenta extract is selected from a component that suppresses or promotes melanin production, a hyaluronic acid production promoting component, a collagen production promoting component, an elastase activity inhibiting component, an antiallergic activity promoting component, and a tyrosinase activity inhibiting component. It is used as an additive material for the ingredients.
  • the use as an additive material means that the placenta extract is added to cosmetics and functional foods so as to contain each of the above components, and the placenta extract is used as an extraction material for each of the above components. That is also included. In the latter case, each extracted component will be added to cosmetics and functional foods.
  • placenta extract is made from any of the components selected from a component that suppresses or promotes melanin production, a hyaluronic acid production promoting component, a collagen production promoting component, an elastase activity inhibiting component, an antiallergic activity promoting component, and a tyrosinase activity inhibiting component.
  • the purpose is to be an additive material.
  • placenta extract itself exerts its function as these components.
  • the action may be substantially exerted by a single substance contained in the placenta extract.
  • the action may be exerted by a plurality of substances.
  • ingredients that suppress or promote melanin production are used externally in the form of cosmetics and / or orally in the form of functional foods (hereinafter referred to as external / oral use modes) in the skin. It suffices if the production of melanin can be suppressed or promoted. Such components can be confirmed by a melanin production test described later.
  • the hyaluronic acid production promoting component and the collagen production promoting component need only be able to promote the production of hyaluronic acid and collagen when used in an external or oral use mode, and can be confirmed by, for example, a collagen or hyaluronic acid production test described later.
  • the elastase activity-inhibiting component only needs to be able to inhibit the activity of elastase and decompose elastin when used in an external or oral use mode, and can be confirmed by, for example, an elastase activity-inhibiting test described later.
  • the anti-allergic activity promoting component may be used as long as it can suppress allergic symptoms when used externally or orally. For example, it is confirmed by measuring ⁇ -hexosaminidase activity as in the anti-allergic activity test described later. be able to.
  • the tyrosinase activity-inhibiting component may inhibit the activity of tyrosinase and prevent the colonization of melanin when used in an external or oral use mode, and can be confirmed by, for example, a tyrosinase inhibitory activity test described later.
  • the placenta extract may also consist of a group of components exhibiting solubility in a non-aqueous specific solvent consisting of a mixture of any one or two or more selected from dimethyl sulfoxide, ethanol, methanol and hexane. good.
  • the placenta extract can be an extract extracted by a non-aqueous specific solvent.
  • the placenta extract consisting of a group of components exhibiting solubility in a non-aqueous specific solvent as each of the above-mentioned components, it is possible to exhibit an action of promoting melanin production, and also in an aqueous solvent. It is possible to exhibit a more remarkable elastase activity inhibitory action as compared with the elastase activity inhibitory component contained in the component group showing solubility.
  • the placenta extract may consist of a group of components exhibiting solubility in an aqueous solvent containing water as a main component.
  • the placenta extract can be an extract extracted with an aqueous solvent.
  • the aqueous solvent is a solvent containing water as a main component, and in addition to water itself, for example, a solvent containing water at a ratio of 50 w / w% or more is included.
  • the components other than water constituting the aqueous solvent may be, for example, a solid component (powder or the like) that is soluble in water, or a liquid that is miscible with water such as alcohol.
  • the placenta extract consisting of the component group exhibiting solubility in an aqueous solvent as each of the above-mentioned components, the action of suppressing melanin production and the solubility in a non-aqueous specific solvent are exhibited. It can exhibit more remarkable actions as compared with the hyaluronic acid production promoting component and the collagen production promoting component contained in the component group.
  • the solvent may be a mixed solvent of an aqueous solvent and a non-aqueous specific solvent. That is, the above-mentioned placenta extraction is performed by using a mixed solvent containing water at a ratio of less than 50 w / w% and using a non-aqueous specific solvent or a solid (powder, etc.) that is soluble in the mixed solvent to be prepared. It is possible to prepare a placenta extract having a composition ratio different from that of the product.
  • the present application also provides a method for using the component found from the placenta extract by the present invention as an active ingredient in cosmetics or a health functional ingredient in functional foods for the purpose of promoting the production of melanin. be.
  • melanin can be obtained from the above-mentioned components. It is possible to provide new cosmetics and functional foods that promote or suppress the production of melanin.
  • placenta extract a placenta extract extracted from horse-derived placenta (placenta)
  • placenta horse-derived placenta
  • placenta extract In recent years, various natural ingredients have been used as functional materials to be blended in cosmetics and functional foods. Among them, placenta, which is the placenta of mammals, is attracting attention as a material that contributes to beauty and health maintenance.
  • placenta extract was first prepared in order to confirm the effectiveness by conducting several functional evaluation tests with horse placenta as a representative example.
  • Horse placenta powder was used as an extraction raw material for horse placenta extract.
  • Horse placenta powder is a lyophilized powder of horse placenta.
  • DMSO dimethyl sulfoxide
  • the tube was allowed to stand at room temperature, and the obtained supernatant was used as a horse placenta extract.
  • This horse placenta extract was prepared by adding 1 ml of an extraction solvent to 100 mg of placenta powder, and was referred to as a 100 mg / ml sample.
  • the diluted sample prepared by appropriately diluting the 100 mg / ml sample is, for example, 1 mg / ml sample for a 100-fold diluted sample and 10 ⁇ g / ml for a 10000-fold diluted sample. Express like a sample.
  • Melanin production / suppression ability confirmation test (1st test) Melanin is a general term for substances in which phenolic substances are polymerized into pigments. Melanin present in human skin is in the form of polymers formed by various indole compounds synthesized from tyrosine.
  • melanin The most important role of melanin is UV protection, which suppresses sunlight damage and the development of malignant tumors caused by UV rays.
  • Other known functions of melanin include absorption of reactive oxygen species that are liable to the living body and uptake of metals and drugs.
  • Cells containing melanin gradually dissociate into the upper stratum corneum, and the adhesion between cells is decomposed by the action of lipid-metabolizing enzymes (lipase, etc.) and proteolytic enzymes (elastase, etc.), and finally the stratum corneum is exfoliated and shed.
  • lipid-metabolizing enzymes lipase, etc.
  • proteolytic enzymes elastase, etc.
  • human melanoma cells (G361 cell line) were used as model cells, and cells and samples were co-cultured, and melanin produced in the culture medium and cell viability were measured.
  • human melanoma cells (G361 cell line) were precultured using McCoy's 5A medium (including 1% penicillin-streptomycin and 10% fetal bovine serum (FBS)).
  • McCoy's 5A medium including 1% penicillin-streptomycin and 10% fetal bovine serum (FBS)
  • 0.5 ml was seeded on a 24-well plate at a concentration of 1.0 ⁇ 10 5 cells / ml and cultured overnight in a CO 2 incubator (37 ° C, 5% CO 2 ).
  • each sample of 2.5 ⁇ l of placenta extract diluted to a predetermined concentration (hereinafter also referred to as each diluted sample) are added, and the mixture is returned to the CO 2 incubator.
  • the cells were cultured for 72 hours.
  • the ability to produce and suppress melanin was confirmed by confirming the amount of melanin produced.
  • the medium was removed from the plate that had been cultured for 72 hours, each well was washed with PBS (Phosphate Buffered Saline; phosphate buffered saline), and then 1 ml of 1 M NaOH solution was added to each well.
  • the reaction was carried out at room temperature for 4 hours, and the absorbance at 405 nm was measured with a plate reader to determine the amount of melanin produced.
  • the cell viability was measured by the MTT method. Remove medium from plates that have been cultured for 72 hours and add 1 ml of fresh medium to each well. Then, 200 ⁇ l of MTT stain (5 mg / ml in PBS) was added to each well, and the cells were statically cultured in a CO 2 incubator for 4 hours.
  • FIG. 1 is a graph showing the melanin production ratio with respect to the control
  • FIG. 1 (a) shows the melanin production ratio by the placenta extract (hereinafter, also referred to as placenta water-based extract) extracted with an aqueous solvent
  • FIG. (B) shows the melanin production ratio by the placenta extract (hereinafter referred to as placenta non-aqueous extract) extracted with a non-aqueous specific solvent.
  • the placenta aqueous extract contains a component that suppresses melanin production, in other words, a component that contributes to skin whitening.
  • the placenta non-aqueous extract contains a component that promotes melanin production and has an effect of maintaining the black color of hair.
  • the placenta water-based extract and the placenta non-water-based extract have the opposite properties of "suppression” and “promotion” in the production of melanin, respectively. It can be said that it is a really interesting point that became clear.
  • Collagen is a structural protein that forms connective tissues such as animal skin. Collagen contributes to the maintenance of the structure of the skin and is important for keeping the skin youthful and preventing wrinkles.
  • Collagen is produced by fibroblasts present in the dermis of the skin, and if the fibroblasts can promote collagen production, youthful skin can be maintained. Therefore, the collagen production promoting component of fibroblasts is considered to be useful as an anti-aging cosmetic material.
  • Hyaluronic acid is mainly produced by fibroblasts, which are the main cells of the dermis of the skin, and cells of other connective tissues. It is the main component of the extracellular matrix and is involved in various cell-cell interactions. In addition, it is present in synovial fluid at a high concentration and is responsible for normal water retention and lubrication of joints.
  • FIG. 2 is a graph showing the hyaluronic acid production ratio with respect to the control
  • FIG. 2 (a) shows the hyaluronic acid production ratio by the placenta aqueous extract
  • FIG. 2 (b) shows the hyaluronic acid production ratio by the placenta non-aqueous extract. Is shown.
  • FIG. 3 is a graph showing the collagen production ratio to the control
  • FIG. 3 (a) shows the collagen production ratio by the placenta aqueous extract
  • FIG. 3 (b) shows the collagen production ratio by the placenta non-aqueous extract.
  • both the aqueous and non-aqueous placenta extracts contained a component that promotes collagen production.
  • the placenta aqueous extract contains a component that promotes hyaluronic acid and collagen production
  • the placenta non-aqueous extract contains a component that promotes collagen production. rice field.
  • Elastase activity inhibition test The cross-linked structure of collagen and elastin present in the dermis plays an important role in maintaining the elasticity of the skin. Elastase, which is an elastin-degrading enzyme, is induced by stimulation with ultraviolet rays or the like, and when elastin is decomposed, the crosslinked structure becomes brittle and the elasticity of the skin is lost. In this test, the skin elasticity retention function of placenta extract is evaluated by measuring the elastase inhibitory activity.
  • cytolytic solution was used in the experiment.
  • human skin fibroblasts (NHDF-Ad) precultured until they became confluent were washed with PBS and resuspended in 1.0 ⁇ 10 6 cells / ml with the cytolytic solution. Then, sonication was performed, and the mixture was centrifuged at 4 ° C. and 13,000 rpm for 15 minutes, and the supernatant was collected as a cytolytic solution.
  • the absorbance at 405 nm was measured with a microplate reader, and the elastase activity was calculated.
  • phosphoramidone final concentration 0.04 mM
  • elastase inhibitor was used as a positive control.
  • FIG. 4 is a graph showing the elastase activity ratio with respect to the control
  • FIG. 4 (a) shows the elastase activity ratio with the placenta aqueous extract
  • FIG. 4 (b) shows the elastase activity ratio with the placenta non-aqueous extract.
  • aqueous and non-aqueous placenta extracts contained a component that inhibits elastase activity.
  • the placenta aqueous extract is probably used at a concentration of 100 ⁇ g / ml or more, and based on the result shown in FIG. 4 (a), it is used at a concentration of 400 ⁇ g / ml or more, and the placenta non-aqueous extract is used. Therefore, it is expected that the use of the placenta extract at a concentration of 40 ⁇ g / ml or more, more effectively at a concentration of 400 ⁇ g / ml or more, can contribute to the maintenance of skin elasticity using the placenta extract as an active ingredient for inhibiting elastase activity. ..
  • rat basophil leukemia cells (RBL-2H3) were used as basophil model cells, and the granule release inhibitory activity (anti-allergic activity) by the placenta extract was evaluated.
  • the granules contain ⁇ -hexosaminidase, and the amount of enzyme released from the granules is measured by measuring the amount of enzyme in the cell supernatant, and the amount of enzyme is the enzyme in the cell supernatant. It was measured by the intensity of activity.
  • rat basophil leukemia cells (RBL-2H3) were prepared at a concentration of 5 ⁇ 10 5 cells / ml, and 100 ⁇ l was seeded on a 96-well plate. Two such plates, one for measuring ⁇ -hexosaminidase activity and one for MTT test, were prepared and precultured in a CO 2 incubator (37 ° C, 5% CO 2 ) for 40 hours.
  • p-Nitrophenyl-N-acetyl- ⁇ -glucosaminide p-Nitrophenyl-N-acetyl-beta-
  • 50 ⁇ l of the supernatant was separated.
  • 50 ⁇ l of a glucosamine) solution was added as a substrate, and the mixture was reacted at room temperature for 1 to 3 hours after being shielded from light with an aluminum foil.
  • 100 ⁇ l of the reaction terminator was added to all the wells to stop the enzyme reaction, and the absorbance at 405 nm was measured with a plate reader to measure the ⁇ -hexosaminidase activity.
  • MTT stain 5 mg / ml in medium
  • 100 ⁇ l of MTT stain 5 mg / ml in medium
  • the cells were statically cultured at 37 ° C. for 4 hours.
  • the medium was removed, 1 ml of hydrochloric acid-isopropanol solution was added to each well, the light was shielded, and the mixture was left at room temperature for 4 hours.
  • the absorbance at 570 nm was measured with a microplate reader and used as an index of cell viability.
  • FIG. 5 is a graph showing the ⁇ -hexosaminidase activity ratio to the control. As can be seen from FIG. 5, the ⁇ -hexosaminidase activity decreased in both the aqueous and non-aqueous placenta extracts when the 400 ⁇ g / ml sample, which was in the high concentration range, was added.
  • the placenta extract according to this embodiment contains a component showing antiallergic activity.
  • Tyrosinase inhibitory activity test The melanin pigment that causes age spots and freckles on the skin is produced by the action of an enzyme called tyrosinase in melanocytes. Tyrosinase is activated by ultraviolet rays to produce melanin, and it is known that inhibition of the action of this enzyme prevents the fixation of melanin and obtains a whitening effect.
  • the test method includes 33.3 ⁇ l of each diluted sample, 333 ⁇ l of 2.5 mM substrate (L-tyrosine or L-DOPA), 600 ⁇ l of 0.1 M phosphate buffer (pH 6.8), and 33 ⁇ l of tyrosinase (1380 unit / ml). ) was mixed, and immediately after that, the change over time in the absorbance at 475 nm was recorded by a spectrophotometer. The absorbance at 475 nm was attributed to the formation of dopachrome after the enzyme-substrate reaction, and the increase in absorbance per hour was calculated to determine the enzyme activity value (L-tyrosine: 10 minutes measurement, L-DOPA: 3 minutes). measurement).
  • the relative tyrosinase activity at the time of adding each diluted sample was calculated with the tyrosinase activity value at the time of adding the control (solvent) as 100%.
  • Kojic acid final concentration: 17 ⁇ g / ml was used as a positive control.
  • FIG. 6A is a graph showing the effect of the aqueous or non-aqueous placenta extract of the tyrosinase activity rate using L-DOPA as a substrate
  • FIG. 6B is a graph showing the effect of the tyrosinase activity rate using L-tyrosine as a substrate. It is a graph which showed the influence of the water-based or non-water-based placenta extract of.
  • the aqueous or non-aqueous placenta extract according to the present embodiment contains a component showing tyrosinase inhibitory activity, and this effect tends to be more remarkable in the placenta non-aqueous placenta extract. It was suggested that it was in.
  • the components isolated by the present inventors from the placenta non-aqueous extract are as follows.
  • HS-6 Represented by 3 ⁇ , 6 ⁇ -dihydroxycholest-5-ene (3 ⁇ , 6 ⁇ -dihydroxycholest-5-ene).
  • test method is the same as in the first test described above, but the sample to be added is not a placenta extract, but is added to the culture solution after dissolving in a dimethyl sulfoxide solution so that each of these components has a final concentration of 100 ⁇ M. did.
  • FIG. 7 (a) is a graph showing the cell viability in each sample
  • FIG. 7 (b) is the extracellular melanin production ratio per cell to the control
  • FIG. 7 (c) is the intracellular melanin production per cell to the control. It is a graph which shows the ratio.
  • Table 1 shows a table of extracellular / intracellular melanin production ratios per cell, including cell viability. In the "type” shown in Table 1, "A + " indicates that the amount of melanin produced increases at a rate of more than 20% with respect to the control, and "A-" indicates that the amount of melanin produced increases by 20% or less with respect to the control.
  • HS-3 (3 ⁇ -hydroxycholesta-4-ene-6-on), HS-7 (cholesta-3,5-diene-7-on), HS-10 (palmitic acid) and HSE-2. It was confirmed that (thymine) significantly increased cell viability compared to control (DMSO) (HS-3: 109.4%, HS-7: 113.3%, HS-10: 110.3%, HSE- 2: 114.3%). Based on these results, it can be confirmed that HS-3, HS-6, HS-7, HS-8, HS-10, HSE-2 and HAS-1 do not show cytotoxicity to B16 melanoma cells. rice field.
  • the intracellular melanin production ratio per cell is controlled by the addition of HS-3, HS-4, HS-8, HS-10, HSE-2 and HAS-1 (DMSO).
  • HS-3, HS-4, HS-8, HS-10, HSE-2 and HAS-1 DMSO
  • HSE-2 HSE-2
  • HAS-1 HAS-1
  • the addition of HS-1, HS-2, HS-6, HS-7 and HAS-2 increased the extracellular melanin production ratio per cell as compared with control (DMSO).
  • HS-1 cholesterol
  • HS-2 cholesterol-7-on
  • HS-3 (3 ⁇ -hydroxycholesta-4-en-6-one) showed a decrease in melanin production at a rate of more than 20% relative to the control both intracellularly and extracellularly. In particular, extracellularly, a decrease in melanin production equivalent to that of arbutin (100 ⁇ M) was observed.
  • HS-4 (5 ⁇ , 6 ⁇ -epoxy-reductase-3 ⁇ -ol) showed a decrease in melanin production both intracellularly and extracellularly, albeit at a rate of 20% or less with respect to the control.
  • HS-6 (3 ⁇ , 6 ⁇ -dihydroxycholesta-5-ene) showed a decrease in melanin production at a rate of more than 20% relative to the control extracellularly, especially to the same extent as arbutin (100 ⁇ M). On the contrary, in the cells, an increase in melanin production was observed, albeit at a rate of 20% or less with respect to the control.
  • HS-7 (Cholesta-3,5-diene-7-on) showed an increase in melanin production of more than 20% relative to the control both intracellularly and extracellularly.
  • HS-8 cholesterol esterate
  • HS-10 palmitic acid showed a decrease in melanin production at a rate of more than 20% relative to the control, both intracellularly and extracellularly.
  • HSE-2 thymine
  • HAS-1 uridine showed a decrease in melanin production both intracellularly and extracellularly, both intracellularly and extracellularly, at a rate of 20% or less with respect to the control.
  • HAS-2 uridine monophosphate
  • cholesterol (HS-1), cholesterol-7-one (HS-2), 3 ⁇ , 6 ⁇ -dihydroxycholesta-5-ene (HS-6), cholesterol-3 , 5-Diene-7-one (HS-7), uridine monophosphate (HAS-2) components have been shown to promote melanin production compared to control (DMSO). More specifically, the components of cholesterol (HS-1), cholesterol-7-on (HS-2), and cholesterol-3,5-diene-7-on (HS-7) are both intracellular and extracellular. In addition, it was shown that each component of 3 ⁇ , 6 ⁇ -dihydroxycholester-5-ene (HS-6) and uridine monophosphate (HAS-2) promotes the production of melanin in the cell. This can be said to indicate that these ingredients are useful as active ingredients and health functional ingredients for the purpose of promoting the production of melanin in cosmetics and functional foods.
  • 3 ⁇ -hydroxycholesta-4-ene-6-one (HS-3), 5 ⁇ , 6 ⁇ -epoxycholestane-3 ⁇ -ol (HS-4), 3 ⁇ , 6 ⁇ -dihydroxycholesta-5- En (HS-6), cholesteryl oleate (HS-8), palmitic acid (HS-10), thymine (HSE-2), uridine (HAS-1), uridine monophosphate (HAS-2) are melanin It was shown to suppress the production of.
  • 3 ⁇ -hydroxycholesta-4-en-6-one (HS-3), 5 ⁇ , 6 ⁇ -epoxycholestane-3 ⁇ -ol (HS-4), cholesteryl oleart (HS-8),
  • the components of palmitic acid (HS-10), timine (HSE-2), and uridine (HAS-1) can suppress the production of melanin both intracellularly and extracellularly, especially HS-3, HS-8, and HS.
  • HSE-2, HAS-1 significantly suppress the production of extracellular melanin
  • 3 ⁇ , 6 ⁇ -dihydroxycholesta-5-ene (HS-6), uridine monophosphate (HAS-2).
  • Each component of is shown to suppress the production of melanin extracellularly. This can be said to indicate that these ingredients are useful as active ingredients and health functional ingredients for the purpose of suppressing the production of melanin and, by extension, whitening in cosmetics and functional foods.
  • HS-6 6 ⁇ -dihydroxycholesta-5-ene
  • HAS-2 uridine monophosphate
  • a component that suppresses or promotes melanin production in cosmetics or functional foods a hyaluronic acid production promoting component, a collagen production promoting component, an elastase activity inhibiting component, and an antiallergic substance. Since the placenta extract is used as an additive material for any of the components selected from the activity promoting component and the tyrosinase activity inhibiting component, it is necessary to provide a new method of using the placenta extract for a clearer purpose. Can be done.

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Abstract

L'invention concerne un nouveau procédé d'utilisation d'extrait de placenta pour un objectif plus clair que dans le passé.　L'utilisation d'extrait de placenta comme matière additive pour tout constituant choisi parmi des constituants qui suppriment ou favorisent la production de mélanine, des constituants qui favorisent la production d'acide hyaluronique, des constituants qui favorisent la production de collagène, des constituants qui inhibent l'activité de l'élastase, des constituants qui favorisent l'activité anti-allergique, et des constituants qui inhibent l'activité de la tyrosinase dans un produit cosmétique ou un aliment fonctionnel. L'extrait de placenta a en outre la caractéristique d'être composé d'un groupe de constituants qui présente une solubilité dans un solvant spécifique non aqueux ou d'être composé d'un groupe de constituants qui présente une solubilité dans un solvant aqueux dont le constituant principal est l'eau.
PCT/JP2021/032175 2020-09-02 2021-09-01 Utilisation d'extrait de placenta WO2022050318A1 (fr)

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Citations (5)

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JPH09255552A (ja) * 1996-03-27 1997-09-30 Shiseido Co Ltd 皮膚外用剤
JP2004529975A (ja) * 2001-04-06 2004-09-30 ニューヨーク・ユニバーシティ メラニン産生を制御する方法および組成物
JP2013034423A (ja) * 2011-08-05 2013-02-21 Guthy-Renker Japan Co Ltd 健康食品
JP2014141475A (ja) * 2012-12-27 2014-08-07 Daiichi Sankyo Co Ltd 新規美白剤
JP2017012104A (ja) * 2015-07-02 2017-01-19 ドリーム＆リンク株式会社 経口組成物

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH09255552A (ja) * 1996-03-27 1997-09-30 Shiseido Co Ltd 皮膚外用剤
JP2004529975A (ja) * 2001-04-06 2004-09-30 ニューヨーク・ユニバーシティ メラニン産生を制御する方法および組成物
JP2013034423A (ja) * 2011-08-05 2013-02-21 Guthy-Renker Japan Co Ltd 健康食品
JP2014141475A (ja) * 2012-12-27 2014-08-07 Daiichi Sankyo Co Ltd 新規美白剤
JP2017012104A (ja) * 2015-07-02 2017-01-19 ドリーム＆リンク株式会社 経口組成物

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ANONYMOUS: "Conference report of the 35th Annual Meeting of the Japanese Society of Aesthetic Dermatology", FUREGURANSU JANARU - FRAGRANCE JOURNAL., FUREGURANSU JANARUSHA, TOKYO, JP, vol. 47, no. 9, 31 August 2017 (2017-08-31), JP , pages 100 - 101, XP009534705, ISSN: 0288-9803 *
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