WO2011068147A1 - Amplification de la synthèse de la mélanine et activation de la tyrosinase - Google Patents

Amplification de la synthèse de la mélanine et activation de la tyrosinase Download PDF

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WO2011068147A1
WO2011068147A1 PCT/JP2010/071548 JP2010071548W WO2011068147A1 WO 2011068147 A1 WO2011068147 A1 WO 2011068147A1 JP 2010071548 W JP2010071548 W JP 2010071548W WO 2011068147 A1 WO2011068147 A1 WO 2011068147A1
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soluble fraction
ethanol
tyrosinase
fat
stevia
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PCT/JP2010/071548
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English (en)
Japanese (ja)
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堅次 杉林
嘉寛 徳留
辰彦 金
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株式会社シャローム
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • 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
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9728—Fungi, e.g. yeasts
    • 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
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • 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
    • A61P17/14—Drugs for dermatological disorders for baldness or alopecia
    • 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
    • A61Q5/00—Preparations for care of the hair

Definitions

  • the present invention relates to a technique capable of promoting melanin synthesis, for example, a method for producing a melanin synthesis promoting substance for promoting melanin synthesis, a melanin synthesis promoting substance, an external preparation for skin, and a method for producing a tyrosinase active substance for activating tyrosinase. , Tyrosinase active substance, and pharmaceuticals.
  • Hair color is the color of melanin synthesized by melanocytes (pigment cells) in the hair matrix that produces hair.
  • melanocytes pigment cells
  • white hair the color of melanin synthesized by melanocytes (pigment cells) in the hair matrix that produces hair.
  • oxidation hair dyes color hair by opening the epidermis (cuticle) of the hair and injecting a pigment into the hair. With an oxidative hair dye, the cuticle is opened and a pigment is injected, so that hair dyed with an oxidative hair dye is easily damaged, loses moisture and becomes uncomfortable.
  • hair dyes that improve the flexibility, moist feeling or smoothness of hair by containing a cationic polymer, an amphoteric polymer and an anionic polymer in an oxidative hair dye are disclosed (for example, Patent Document 1).
  • Patent Document 1 Even if the technique described in Patent Document 1 is used, it does not change that hair cannot be dyed unless a cuticle on the surface of the hair is opened and a pigment is injected, so it is difficult to completely eliminate damage to the hair. is there. Also, since the gray hair itself is not lost, when the hair grows, the color difference between the dyed part on the tip and the unstained part on the scalp side (white hair part) becomes conspicuous, creating a sense of incongruity. End up.
  • the object of the present invention is to provide a new technique that can promote the synthesis of melanin.
  • a method for producing a melanin synthesis promoting substance that promotes melanin synthesis the first fractionation step of fractionating an extract of fermented stevia to obtain an ethanol-soluble fraction; And a second fractionation step of further fractionating the ethanol-soluble fraction to obtain a fat-soluble fraction.
  • the inventor of the present application as a result of earnest research, has the effect of promoting the synthesis of melanin in the fat-soluble fraction obtained by further fractionating the ethanol-soluble fraction obtained by fractionating the extract of fermented stevia. I found it. Therefore, for example, by applying the above fat-soluble fraction to the melanocytes of hair matrix cells that produce hair, melanin can be included in the hair, not from the surface of the hair, Whitening can be suppressed.
  • a fermentation step is performed in which yeast and water are added to the dried stevia plant tissue and fermentation is performed, and an extract is obtained from the fermentation stevia obtained by the fermentation step using an aqueous ethanol solution.
  • An extraction step is performed.
  • the fermented stevia extract obtained through the above fermentation step and extraction step is suitable as a fermented stevia extract for obtaining a melanin synthesis promoting substance by the above production method.
  • the ethanol-soluble fraction obtained in the first fractionation step may be a 30-100% ethanol-soluble fraction.
  • the fat-soluble fraction fractionated from the 30-100% ethanol-soluble fraction is particularly excellent as a melanin synthesis promoting substance.
  • a melanin synthesis promoting substance that promotes melanin synthesis was obtained by further fractionating an ethanol-soluble fraction obtained by fractionating an extract of fermented stevia. Contains fat-soluble fraction.
  • the external preparation for skin includes a melanin synthesis promoting substance produced by the method for producing a melanin synthesis promoting substance.
  • the skin external preparation may be a cosmetic, a quasi-drug, a pharmaceutical, or a gray hair preventing pharmaceutical.
  • a method for producing a tyrosinase active substance that activates tyrosinase includes a first fractionation step for fractionating an extract of fermented stevia to obtain an ethanol-soluble fraction, and a second fraction for further fractionating the ethanol-soluble fraction to obtain a fat-soluble fraction. And a process.
  • the present inventor has found that the fat-soluble fraction obtained by further fractionating the ethanol-soluble fraction obtained by fractionating the extract of fermented stevia has a function of activating tyrosinase. It was. Since tyrosinase is an essential enzyme in the melanin synthesis process, it is possible to improve the ability of melanocytes to synthesize melanocytes by applying the above fat-soluble fraction that activates tyrosinase to melanocytes.
  • L-DOPA melanin from tyrosine to L-DOPA (3,4-dihydroxy-L-phenylalanine) is performed by tyrosinase.
  • L-DOPA is a precursor of dopamine, which is a neurotransmitter, and can pass through the blood-brain barrier, and thus is used as a useful drug for Parkinson's disease and the like. Therefore, the above fat-soluble fraction that activates tyrosinase can promote the synthesis of tyrosine to L-DOPA, and may be applicable as a drug suitable for Parkinson's disease in the future.
  • a fermentation step is performed in which yeast and water are added to the dried stevia plant tissue and fermentation is performed, and an extract is obtained from the fermentation stevia obtained by the fermentation step using an aqueous ethanol solution.
  • An extraction step is performed.
  • the ethanol-soluble fraction obtained in the first fractionation step may be a 30-100% ethanol-soluble fraction.
  • the tyrosinase active substance for activating tyrosinase according to an embodiment of the present invention is a fat-soluble fraction obtained by further fractionating an ethanol-soluble fraction obtained by fractionating an extract of fermented stevia. including.
  • a pharmaceutical product according to an embodiment of the present invention includes a tyrosinase active substance produced by the above-described method for producing a tyrosinase active substance.
  • the constituent elements based on the technical idea of the method for producing the melanin synthesis promoting substance and the explanation thereof can be applied to the melanin synthesis promoting substance, the external preparation for skin, the method for producing the tyrosinase active substance, the tyrosinase active substance, and the pharmaceutical. is there.
  • the synthesis of melanin can be promoted by activating tyrosinase, and melanin can be contained in the hair that grows in the future to suppress whitening of the hair.
  • Embodiments include, for example, a method for producing a melanin synthesis promoting substance, a melanin synthesis promoting substance, an external preparation for skin, a method for producing a tyrosinase active substance, a tyrosinase active substance, and a pharmaceutical product.
  • FIG. 1 is an explanatory diagram for explaining a method of manufacturing a melanin synthesis promoting substance that promotes melanin synthesis according to one embodiment.
  • the method for producing a melanin synthesis promoting substance according to this example includes a step of drying and crushing stevia plant tissue (dry crushing step: S100), and a dried stevia obtained by the dry crushing step S100.
  • a step of fermenting yeast and water by adding yeast and water to the plant tissue (fermentation step: S102) and a step of extracting from the fermentation stevia obtained by the fermentation step S102 using an aqueous ethanol solution to obtain an extract (extraction step: S104) ),
  • a step of fractionating the extract obtained in the extraction step (S104) to obtain an ethanol-soluble fraction (first fractionation step: S106), and a first fractionation step (S108).
  • a step of further fractionating the ethanol-soluble fraction to obtain a fat-soluble fraction (second fractionation step: S108), and the reduced pressure of the fat-soluble fraction obtained by the second fractionation step (S108) , Step concentrated to dryness (concentrated to dryness step: S110) and a.
  • the drying and pulverizing step S100 is a step of drying and pulverizing the plant tissue of the stevia plant as a raw material.
  • Stevia plant which is one of the raw materials of this example is a perennial plant belonging to the family Asteraceae originating from Paraguay in South America.
  • the scientific name for Stevia plant is Stevia Rebaudiana Bertoni (scientific name).
  • the melanin synthesis promoting substance is a dry stevia plant tissue powder (hereinafter simply referred to as dry stevia powder) prepared by thoroughly drying the plant tissue of this stevia plant and finely grinding it. Used as Even if it says powder, it is sufficient if it grind
  • Fermentation process S102 is a process of fermenting the dry stevia powder obtained by dry crushing process S100. Fermentation of the dry stevia powder can be performed by adding water and yeast to the dry stevia powder, stirring and leaving it to stand. The amount of water to be added in the fermentation step S102 only needs to be sufficient for fermentation, and an amount sufficient to wet the whole is sufficient. As yeast, it is preferable to use Saccharomyces.
  • step S102 moisture is first added to the dried stevia powder, and yeast (for example, Saccharomyces) is further added, and fermentation is continued for 1 to 4 weeks at room temperature while supplying water. Then, when stevia powder is included in the mouth and the sweetness is no longer felt, it is considered to be “completely” fermented.
  • yeast for example, Saccharomyces
  • Extraction step S104 extraction is performed using an aqueous ethanol solution from the fermented stevia powder (hereinafter simply referred to as “fermentation stevia”) obtained in the fermentation step S102.
  • the extract extracted from the fermented stevia obtained in the fermentation step S102 has a function of promoting melanin synthesis or a function of activating tyrosinase.
  • Extraction is performed by preparing an aqueous solution of about 30% ethanol, adding fermented stevia, and infiltrating at room temperature for several hours to several days with gentle stirring.
  • a filtrate obtained by filtering fermented stevia into about 30% ethanol aqueous solution through a filter paper of about 140 mesh is used as an extract.
  • the extract is further concentrated under reduced pressure at about 60 ° C. to concentrate the extract.
  • the first fractionation step S106 is a step of fractionating the concentrated extract obtained in the extraction step S104 (hereinafter simply referred to as a concentrated extract) to obtain an ethanol-soluble fraction.
  • an ion exchange resin synthetic adsorbent
  • the ethanol-soluble fraction is fractionated from the concentrated extract by passing the concentrated extract through a column packed with DIAION (registered trademark) HP20.
  • the ethanol soluble fraction obtained in the first fractionation step S106 is preferably a 30-100% ethanol soluble fraction, more preferably a 70-100% ethanol soluble fraction, and even more preferably 85%. It is an ethanol soluble fraction.
  • the second fractionation step S108 is a step of further fractionating the ethanol-soluble fraction obtained in the first fractionation step S106 to obtain a fat-soluble fraction.
  • a weak anion exchange resin is used to fractionate the fat-soluble fraction.
  • the fat-soluble fraction is fractionated from the ethanol-soluble fraction by passing the ethanol-soluble fraction through a column packed with DIAION (registered trademark) WA30.
  • the ethanol-soluble fraction obtained in the first fractionation step S106 is passed through a column packed with DIAION (registered trademark) WA30, and then DIAION (registered trademark) WA30. Those that were not adsorbed on the surface were taken as the fat-soluble fraction. Such a fat-soluble fraction functions as a melanin synthesis promoting substance.
  • the concentration / drying step S110 is a step of concentrating and drying the fat-soluble fraction obtained in the second fractionation step S108.
  • the fat-soluble fraction fractionated in the second fractionation step S108 is concentrated to dryness under reduced pressure so that it can be easily stored and transported.
  • the melanin synthesis promoting substance can be produced through the dry pulverization step S100, the fermentation step S102, the extraction step S104, the first fractionation step S106, the second fractionation step S108, and the concentration and drying step S110 described above. .
  • the fat-soluble fraction according to the present example promotes melanin synthesis by activating tyrosinase, which is an enzyme essential in the melanin synthesis process. Therefore, the above fat-soluble fraction also functions as a tyrosinase active substance.
  • FIG. 2 is an explanatory diagram for explaining a biosynthesis pathway of melanin.
  • melanin is synthesized using tyrosine as a starting material.
  • tyrosine supplied from the blood is oxidized by tyrosinase, which is a copper-containing enzyme, and synthesized into L-DOPA.
  • L-DOPA is also oxidized by tyrosinase and synthesized into dopaquinone.
  • Dopaquinone is automatically oxidized and synthesized into dopachrome and melanin.
  • FIG. 3 is an explanatory diagram for explaining an experimental procedure of a comparative experiment of the amount of melanin synthesis using B16 melanoma.
  • FIG. 3 first, in a medium in which the fat-soluble fraction produced by the above production method was added to B16 melanoma, culture was performed at 0.3 ⁇ 10 5 cells / well at 37 ° C. under 5% CO 2 for 96 hours. Went.
  • the culture was carried out under the same conditions as the above culture conditions in a medium in which no fat-soluble fraction was added to B16 melanoma.
  • the B16 melanoma to which the fat-soluble fraction was added and the control B16 melanoma were collected from the culture dish, and the number of viable cells was counted using the Trypan Blue staining method.
  • FIG. 4 is an explanatory diagram for explaining the number of viable cells of the B16 melanoma to which the fat-soluble fraction was added and the control B16 melanoma.
  • error bars added to the bar graph indicate standard deviations (mean ⁇ SD).
  • the number of viable cells of the control B16 melanoma (1.37 ⁇ 10 6 cells / well (standard deviation 0.12)) and the number of viable cells of the B16 melanoma to which the fat-soluble fraction was added (1.17). Since there is almost no difference in ⁇ 10 6 / well (standard deviation 0.15)), it can be seen that the addition of the fat-soluble fraction does not promote the death of B16 melanoma.
  • 1N NaOH was added to the cultured B16 melanoma collected from the culture dish, and incubated at 75 ° C. for 90 minutes to dissolve the cultured B16 melanoma to obtain a lysate. Then, the absorbance of the solution was measured at a wavelength of 405 nm to calculate the concentration of melanin. In addition, 405 nm is an absorption wavelength of melanin. In addition, a calibration curve was created here using certified synthetic melanin.
  • FIG. 5 is an explanatory diagram for explaining the amount of melanin between a B16 melanoma to which a fat-soluble fraction is added and a control B16 melanoma.
  • FIG. 5 (a) shows the amount of melanin synthesis per cell.
  • (B) shows the amount of melanin synthesis per well.
  • error bars added to the bar graph indicate standard deviation (mean ⁇ SD).
  • the control B16 melanoma contains 30.34pg (standard deviation 4.83) melanin per cell
  • the B16 melanoma to which the fat-soluble fraction was added was 1 Contains 55.00 pg (standard deviation 6.93) of melanin per cell.
  • the B16 melanoma to which the fat-soluble fraction was added as compared with the control B16 melanoma synthesized about 1.6 times the amount of melanin per cell.
  • the control B16 melanoma contains 41.27 ⁇ g (standard deviation 3.47) of melanin per well, and the B16 melanoma added with the fat-soluble fraction is 1 well. It contains 63.48 ⁇ g (standard deviation 5.28) of melanin. Therefore, it can be seen that the B16 melanoma to which the fat-soluble fraction was added as compared with the control B16 melanoma synthesized about 1.5 times as much melanin per well.
  • the fat-soluble fraction produced by the above production method promotes the synthesis of melanin. Therefore, it becomes possible to improve the melanin synthetic ability of a melanocyte by applying the fat-soluble fraction (melanin synthesis acceleration
  • FIG. 6 is an explanatory diagram for explaining an experimental procedure for measuring tyrosinase activity in a cell-free system.
  • the fat-soluble fraction was added to 0.1 M PBS (Phosphate Buffered Saline) (pH 6.) while irradiating ultrasonic waves so as to be 100 pg / mL, 100 ng / mL, and 100 ⁇ g / mL. Dissolved in 8).
  • PBS Phosphate Buffered Saline
  • FIG. 7 is a diagram showing an equation for evaluating the tyrosinase activation rate
  • FIG. 8 is an explanatory diagram for explaining the tyrosinase activity rate of the fat-soluble fraction.
  • the absorbance of a mixed solution of mushroom tyrosinase and L-DOPA to which a fat-soluble fraction at 0 minutes was added (hereinafter simply referred to as a mixture of fat-soluble fractions), fat at 20 minutes
  • Absorbance of a mixture of soluble fractions absorbance of a mixture of mushroom tyrosinase and L-DOPA (hereinafter simply referred to as a control mixture) added with control at 0 minutes
  • control mixture By substituting the absorbance, the tyrosinase activation rate (%) of the fat-soluble fraction can be calculated.
  • the fat-soluble fraction had a higher tyrosinase activity rate than the control (100% (standard deviation 9.90)).
  • the tyrosinase activity rate when the fat-soluble fraction was added at 100 pg / mL was 105.99% (standard deviation 10.46), and the tyrosinase activity rate at 100 ng / mL was 114.30% (standard deviation 14.4).
  • the tyrosinase activity rate when added at 100 ⁇ g / mL was 118.06% (standard deviation 8.02). Therefore, it was found that the fat-soluble fraction activates tyrosinase in a concentration-dependent manner.
  • the fat-soluble fraction produced by the above production method activates tyrosinase in a cell-free system.
  • FIG. 9 is an explanatory diagram for explaining an experimental procedure for measuring tyrosinase activity in cells.
  • a medium in which the fat-soluble fraction produced by the above production method was added to B16 melanoma to a concentration of 100 ⁇ g / mL, 0.3 ⁇ 10 5 cells / well, 37 ° C. Culture was performed for 96 hours under 5% CO 2 .
  • the culture was carried out under the same conditions as the above culture conditions in a medium in which no fat-soluble fraction was added to B16 melanoma.
  • each of the B16 melanoma to which the fat-soluble fraction was added and the control B16 melanoma were washed with PBS.
  • Each of the B16 melanoma washed with PBS and added with the fat-soluble fraction and the control B16 melanoma were dissolved using a cell lysis protein extraction reagent (for example, Cell-LyEX1 manufactured by Toyo B-Net Co., Ltd.).
  • the cell lysate obtained using the cell lysis protein extraction reagent was collected and centrifuged at 15000 g at 4 ° C. for 15 minutes to obtain a supernatant.
  • the protein of the obtained supernatant was quantified (Lowly method), and it was confirmed that the serum contained in the medium was completely removed.
  • the supernatant of the cell lysate of B16 melanoma to which the obtained fat-soluble fraction was added (hereinafter simply referred to as the supernatant of the fat-soluble fraction), the supernatant of the cell lysate of B16 melanoma as a control, 2 mM, L-DOPA was added to each, and kept at 37 ° C., and absorbance was measured at a wavelength of 475 nm every 10 minutes until 60 minutes had elapsed.
  • FIG. 10 is a diagram showing an equation for evaluating the tyrosinase activation rate
  • FIG. 11 is an explanatory diagram for explaining the tyrosinase activity rate of the supernatant of the fat-soluble fraction.
  • the formula shown in FIG. 10 shows the absorbance of the mixture of the fat-soluble fraction supernatant and L-DOPA at 0 minutes, and the absorbance of the mixture of the fat-soluble fraction supernatant and L-DOPA at 60 minutes.
  • the tyrosinase activation rate (%) of the supernatant can be calculated.
  • the tyrosinase activity rate of the fat-soluble fraction was 131.55% (standard deviation 19.67).
  • Min supernatant was found to activate 131.55% of tyrosinase compared to control supernatant.
  • the fat-soluble fraction produced by the above production method activates tyrosinase even in cells. Since tyrosinase is an essential enzyme in the melanin synthesis process, it is possible to improve the ability of melanocytes to synthesize melanocytes by applying the above fat-soluble fraction that activates tyrosinase to melanocytes. Therefore, by applying the above fat-soluble fraction to the melanocytes of hair matrix cells that produce hair, melanin can be included in the hair, not from the surface of the hair. It becomes possible to suppress.
  • L-DOPA is a precursor of dopamine, which is a neurotransmitter, and can pass through the blood-brain barrier, and thus is used as a useful drug for Parkinson's disease and the like. Therefore, the above fat-soluble fraction that activates tyrosinase can promote the synthesis of tyrosine to L-DOPA, and may be applied as a drug (medicine) suitable for Parkinson's disease in the future. There is.
  • the details of the method for producing the melanin synthesis promoting substance and the method for producing the tyrosinase active substance according to this example and the experimental results demonstrating the effects of the melanin synthesis promoting substance and the tyrosinase active substance produced by the production method are shown. From the experimental results described above, it was shown that the obtained melanin synthesis promoting substance has a surprising melanin synthesis promoting effect, and that the obtained tyrosinase active substance has a surprising tyrosinase active effect.
  • either one or both of the above melanin synthesis promoting substance and tyrosinase active substance can be used in a wide range of skin external preparations such as cosmetics, quasi-drugs, pharmaceuticals, and gray hair prevention medicines.
  • skin external preparations such as cosmetics, quasi-drugs, pharmaceuticals, and gray hair preventives suitable for those with white hair or similar symptoms or those with neurological diseases such as Parkinson's disease or similar symptoms. I can expect.
  • the term “white hair preventive drug” described in the present specification is a coined word, but means an agent that can promote the expression of the background color of the body hair such as hair (original hair color).
  • the effect of preventing or delaying whitening of the hair or increasing the color can be expected due to the tyrosinase activity action or melanin synthesis promoting action of the agent.
  • the color of body hair fades with age, and often turns white at the end, but depending on the example, the effect of preventing, delaying, or reversing the progress of such aging changes may be achieved. It can be expected.
  • the agent utilizing the present invention may be referred to as an anti-aging agent for preventing aging.
  • Either one or both of the melanin synthesis promoting substance and the tyrosinase active substance according to the present invention is a pharmaceutical product for preventing white hair (an anti-aging hair agent for preventing hair aging) that returns the hair color to the background color in cosmetics.
  • combination of melanin of this specification, and the tyrosinase active substance which activates tyrosinase does not necessarily need to process in time series along the order described as a flowchart. It is also possible to proceed in parallel.
  • the present invention is used for, for example, a method for producing a melanin synthesis promoting substance that promotes melanin synthesis, a melanin synthesis promoting substance, an external preparation for skin, a method for producing a tyrosinase active substance that activates tyrosinase, a tyrosinase active substance, and a pharmaceutical. be able to.

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Abstract

La présente invention a pour objet une nouvelle technique par laquelle la synthèse de la mélanine peut être amplifiée. La présente invention concerne spécifiquement un procédé de production d'un amplificateur de la synthèse de la mélanine. Un mode de réalisation du procédé comprend : une étape de séchage / pulvérisation (S100) dans laquelle les tissus végétaux de stevia servant de matière première sont séchés et pulvérisés ; une étape de fermentation (S102) dans laquelle la poudre de stevia séchée obtenue dans l'étape de séchage / pulvérisation (S100) est fermentée ; une étape d'extraction (S104) dans laquelle l'extraction à partir de la poudre de stevia fermentée (désignée ci-après sous le nom de « stevia fermentée ») est mise en œuvre à l'aide d'une solution aqueuse d'éthanol, ce qui permet d'obtenir un extrait liquide ; une première étape de fractionnement (S106) dans laquelle l'extrait liquide de la stevia fermentée est fractionné, ce qui permet d'obtenir une fraction soluble dans l'éthanol ; et une seconde étape de fractionnement (S108) dans laquelle la fraction soluble dans l'éthanol est encore fractionnée, ce qui permet d'obtenir une fraction liposoluble.
PCT/JP2010/071548 2009-12-03 2010-12-02 Amplification de la synthèse de la mélanine et activation de la tyrosinase WO2011068147A1 (fr)

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JP2010017510A JP5017386B2 (ja) 2009-12-03 2010-01-28 メラニン合成促進物質の製造方法、メラニン合成促進物質、皮膚外用剤、チロシナーゼ活性物質の製造方法、チロシナーゼ活性物質、および医薬品

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013093880A1 (fr) * 2011-12-23 2013-06-27 L'oreal Utilisation de stéviol, d'un dérivé glycoside de stéviol, ou d'un de leurs isomères, pour prévenir, réduire et/ou traiter une altération du teint de la peau
WO2012116990A3 (fr) * 2011-03-01 2014-05-01 Dsm Ip Assets B.V. Nouvelle utilisation
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JP6013180B2 (ja) * 2012-12-28 2016-10-25 株式会社シャローム 美白剤

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WO2012116990A3 (fr) * 2011-03-01 2014-05-01 Dsm Ip Assets B.V. Nouvelle utilisation
WO2013093880A1 (fr) * 2011-12-23 2013-06-27 L'oreal Utilisation de stéviol, d'un dérivé glycoside de stéviol, ou d'un de leurs isomères, pour prévenir, réduire et/ou traiter une altération du teint de la peau
FR2984742A1 (fr) * 2011-12-23 2013-06-28 Oreal Utilisation de steviol, d'un derive glycoside du steviol, ou d'un de leurs isomeres, pour prevenir, reduire et/ou traiter une alteration du teint de la peau.
WO2014080666A1 (fr) * 2012-11-26 2014-05-30 株式会社シャローム Composé diterpénique, agent de blanchiment de la peau et procédé de production d'un composé diterpénique
CN104024421A (zh) * 2012-11-26 2014-09-03 株式会社夏洛美 二萜化合物、美白剂及二萜化合物的制造方法
JP5933748B2 (ja) * 2012-11-26 2016-06-15 株式会社シャローム ジテルペン化合物、美白剤およびジテルペン化合物の製造方法
JPWO2014080666A1 (ja) * 2012-11-26 2017-01-05 株式会社シャローム ジテルペン化合物、美白剤およびジテルペン化合物の製造方法
CN104024421B (zh) * 2012-11-26 2018-01-02 株式会社夏洛美 二萜化合物、美白剂及二萜化合物的制造方法

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