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WO2013058578A2 - Système pour sélectionner une matière de blanchiment de la peau - Google Patents

Système pour sélectionner une matière de blanchiment de la peau Download PDF

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Publication number
WO2013058578A2
WO2013058578A2 PCT/KR2012/008555 KR2012008555W WO2013058578A2 WO 2013058578 A2 WO2013058578 A2 WO 2013058578A2 KR 2012008555 W KR2012008555 W KR 2012008555W WO 2013058578 A2 WO2013058578 A2 WO 2013058578A2
Authority
WO
WIPO (PCT)
Prior art keywords
skin
skin whitening
expression
melanocytes
fibroblasts
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/KR2012/008555
Other languages
English (en)
Korean (ko)
Other versions
WO2013058578A3 (fr
Inventor
이애영
이태룡
최현정
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Amorepacific Corp
Industry Academic Cooperation Foundation of Dongguk University
Original Assignee
Amorepacific Corp
Industry Academic Cooperation Foundation of Dongguk University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from KR1020120114965A external-priority patent/KR102031161B1/ko
Application filed by Amorepacific Corp, Industry Academic Cooperation Foundation of Dongguk University filed Critical Amorepacific Corp
Priority to JP2014536989A priority Critical patent/JP6069334B2/ja
Priority to CN201280051510.0A priority patent/CN103857802B/zh
Publication of WO2013058578A2 publication Critical patent/WO2013058578A2/fr
Publication of WO2013058578A3 publication Critical patent/WO2013058578A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • A61K8/981—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of mammals or bird
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0697—Artificial constructs associating cells of different lineages, e.g. tissue equivalents
    • C12N5/0698—Skin equivalents
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2502/00—Coculture with; Conditioned medium produced by
    • C12N2502/09—Coculture with; Conditioned medium produced by epidermal cells, skin cells, oral mucosa cells
    • C12N2502/091—Coculture with; Conditioned medium produced by epidermal cells, skin cells, oral mucosa cells melanocytes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2502/00—Coculture with; Conditioned medium produced by
    • C12N2502/09—Coculture with; Conditioned medium produced by epidermal cells, skin cells, oral mucosa cells
    • C12N2502/094—Coculture with; Conditioned medium produced by epidermal cells, skin cells, oral mucosa cells keratinocytes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2502/00—Coculture with; Conditioned medium produced by
    • C12N2502/13—Coculture with; Conditioned medium produced by connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
    • C12N2502/1323—Adult fibroblasts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2503/00—Use of cells in diagnostics
    • C12N2503/04—Screening or testing on artificial tissues
    • C12N2503/06—Screening or testing on artificial skin

Definitions

  • the present invention relates to a skin lightening material screening system and a method of screening skin lightening material.
  • melanocytes melanocyte-producing cells
  • melanocytes melanocyte-producing cells
  • the hyperpigmentation of the skin differs in the causes and mechanisms of the increase in melanin production by UV rays. In the absence of this, melanin production and dispersion continue to increase. Therefore, in the case of the whitening material developed based on the existing UV irradiation method, in many cases, the clinical trial does not show the effect of inhibiting human skin hyperpigmentation.
  • biopsy of the skin of a subject's hyperpigmentation site to study skin hyperpigmentation is not only costly and laborious, and difficult to obtain human skin tissue. Animal models of microdepressants are also difficult to obtain.
  • An aspect of the present invention is to provide a skin whitening material screening system capable of screening an effective and simple clinically useful skin whitening material, and a method for screening a skin whitening material using the same.
  • Another aspect of the present invention is to provide a skin whitening composition that exhibits an excellent skin whitening effect, including the material screened according to the skin whitening material screening method as an active ingredient.
  • another aspect of the present invention is to provide a composition for skin whitening or external composition for skin comprising the melanogenesis gene involved in the skin itself as an active ingredient.
  • One aspect of the present invention provides a skin whitening material screening system comprising one or more layers comprising one or more cells of fibroblast, keratinocyte and melanocyte hyperplasia.
  • Another aspect of the present invention provides a method for screening a skin whitening substance treated with a candidate substance, wherein at least one cell of fibroblasts, keratinocytes and melanocytes is used.
  • It provides a skin whitening material screening method comprising the step of determining the degree of expression or phosphorylation of one or more of WIF 1, WNT 1, WNT 5a and NFATC 2.
  • Another aspect of the present invention provides a skin whitening composition
  • a skin whitening composition comprising the material screened according to the skin whitening material screening method as an active ingredient or a skin whitening composition and a skin external composition comprising WIF 1 itself as an active ingredient.
  • Skin whitening material screening system is designed to be close to the actual skin tissue can be easily screened clinically whitening material in a small time and cost in vitro.
  • the skin whitening substance screening system according to the present invention has a gene expression level of actual skin tissue showing hyperpigmentation and Since the conditions are similar to the phosphorylation degree of the signal transduction factor, it can be easily screened for skin whitening substances, especially those effective for improving hyperpigmentation. This may be a useful system or kit for the development of new skin whitening substances, specifically hyperpigmentation improving substances, in that it is difficult to obtain human and animal models for hyperpigmentation studies.
  • FIG. 1 is a graph comparing differences in the expression level of WIF 1 gene between normal skin (N) and hyperpigmentation site (L) of a subject.
  • FIG. 2 shows WNT 1 and TOT of normal skin (N) and hyperpigmentation site (L) of a subject.
  • MC melanocytes
  • KC keratinocytes
  • FB fibroblasts
  • 1 is a graph comparing the difference in gene expression.
  • Figure 4 is a result showing the increase in the expression of tyrosinase (tyrosinase) in the whitening screening system including fibroblasts that inhibited WIF 1 expression.
  • NFATC 2 is dephosphorylated in a whitening screening system including fibroblasts that inhibited WIF 1 expression.
  • FIG. 7 is an experimental result showing an increase in the expression of tyrosinase in a whitening screening system containing keratinocytes that inhibited WIF 1 expression.
  • FIG. 8 shows the increased melanosomal migration in the whitening screening system containing keratinocytes that inhibited WIF 1 expression.
  • FIG. 9 shows the results of dephosphorylation of NFATC 2 in a whitening screening system containing keratinocytes that inhibited WIF 1 expression.
  • FIG. 10 is a result showing that WIF 1 expression of melanocytes is increased when human WIF 1 prepared by genetic recombination technology is treated with melanocytes.
  • FIG. 11 is a result showing that the expression of tyrosinase in the melanocytes induced the expression of WIF 1 as shown in FIG.
  • Figure 12 is a result showing that the movement of the melanosomes is reduced when the human WIF 1 made by genetic recombination technology in a whitening screening system including melanocytes.
  • FIG. 13 is a result showing that NFATC 2 is phosphorylated in melanocytes induced with expression of WIF 1 as shown in FIG. 10.
  • the term "skin” refers to a tissue covering the body surface of an animal, and is a broad concept including not only tissues covering the body surface such as the face or body, but also the scalp and hair.
  • Melanocytes in the skin are cells that make melanin, which determines the color of the skin, and are located on the basement membrane, which serves as the boundary between the dermis and the epidermis of the skin. Developmentally, melanocytes depart from neural-crest eel is and finally move through the melanocyte stem cell and the progenitor melanoblast (1 ⁇ 131101) 1 &51; It is known to differentiate into melanocytes, which are pigment cells capable of producing melanin.
  • the differentiated melanocytes are not present in the skin, but are produced by melanin binding to keratinocytes, keratinocytes, and fibroblasts in the dermis. Melanin from melanocytes travels to the surrounding keratinocytes through dendrite and is widely dispersed on the skin surface, protecting the skin from harmful factors such as ultraviolet rays and displaying skin color. Normal skin produces more melanin and increases melanin dispersal than usual when irritation such as ultraviolet rays occurs, then returns melanin production and dispersal back to low levels when the irritation disappears.
  • whitening has been developed by conventional methods that melanocytes do not reflect the actual skin conditions that produce melanin under complex influences from physical and chemical factors of other cells in the skin or the surrounding microenvironment.
  • a skin whitening material screening system comprising one or more layers comprising one or more cells of fibroblasts, keratinocytes and melanocytes.
  • Another aspect of the invention is a fibroblast layer comprising a fibroblast;
  • a skin lightening material screening system comprising a keratinocyte layer comprising keratinocytes and a melanocyte layer comprising melanocytes.
  • the fibroblast layer is stacked on the bottom, the melanocyte layer is stacked on top of it, and the keratinocyte layer may be stacked on top of it, so as to resemble the actual skin condition.
  • the stem is a fibroblast layer comprising fibroblasts; And keratinocytes and melanocytes in the same layer, including layers comprising one or more of keratinocytes and melanocytes.
  • the fibroblast layer may be stacked below, and a layer containing melanocytes and keratinocytes may be stacked thereon.
  • the skin whitening material screening system is a three-dimensional three-dimensional structure of the skin cell layer is stacked, including not only melanocytes but also surrounding cells, it can reflect the actual skin condition well.
  • the skin whitening material screening system according to another aspect of the present invention may include a layer including melanocytes and keratinocytes, which are known to play a more important role in melanogenesis, except for the fibroblast layer.
  • the fibroblast layer of the skin whitening material screening system reproduces the dermal layer and includes a layer containing fibroblasts in collagen.
  • the fibroblasts may be a layer prepared by putting fibroblasts in collagen solution and hardening.
  • the collagen is 0.001 to 30% by weight, specifically 0.01 to 20% by weight, more specifically 0.1 to 3 ⁇ 4 to 10% by weight based on the total weight of the skin lightening material screening system May be included. When included in the above range it is appropriate to show the intended effect of the present invention, it may be appropriate to use the above range in terms of cost-effectiveness.
  • one or more cells of fibroblasts, keratinocytes and melanocytes are selected from one or more selected from birds and pigs such as humans, mice, guinea pigs, and chickens. Contains derived cells.
  • Skin whitening material screening system according to an aspect of the present invention is not only a material that has a skin whitening effect, in particular, a material that improves skin damage caused by ultraviolet rays, in particular, blemishes, spots, gums or gums Effective screening for substances that improve skin hyperpigmentation, such as mushrooms.
  • WIF Kffnt inhibitory factor 1 ACCESSION NP_009122, VERSION NP_009122.2 GI: 111125011
  • WIF 1 is an extracellular signal molecule associated with embryonic growth regulation.
  • the expression of WIF 1 gene was reduced in keratinocytes and fibroblasts, not melanocytes, which are melanocytes, but also affected melanogenesis in melanocytes.
  • melanin diffusion into keratinocytes was increased with the expression of tyrosinase involved in melanogenesis. This increase was found to have a close relationship between WIF 1 and skin hyperpigmentation.
  • the expression of the WIF 1 gene is involved in the expression of GSK-3P (Glycogen synthase kinase-3
  • an aspect of the present invention provides a skin whitening screening system in which WIF 1 expression of fibroblasts, keratinocytes and melanocytes is suppressed, specifically, WIF 1 expression of keratinocytes or fibroblasts.
  • a suppressed skin lightening material screening system in another aspect of the present invention, includes cells in which WIF 1 expression is suppressed using siRNA.
  • the screening system reflects hyperpigmentation skin condition with reduced WIF 1 expression of keratinocytes or fibroblasts, and can almost reproduce the skin condition showing hyperpigmentation. Therefore, in the case of using the screening system, it is possible to effectively select a material that is effective in improving skin whitening, particularly a skin pigmentation effect.
  • the present inventors have found that the proto-oncogene precursor WNT KWingless-type MMTV integration site family, member 1, ACCESSION NP_005421, VERSION NP_005421.1 GI: 4885655) and protein WNT 5a in skin with hyperpigmentation (Wingl ess-type MMTV integration site family, member 5a, ACCESSION NP_001243034, VERSION NP_001243034.1 GI: 371502087) Increased gene expression, NFATC2 (nuclear factor of activated T cells, cytoplasmic, calcineur in-dependent 2) ) Is dephosphorylated, and in one aspect of the present invention, one or more cells of islet infant cells, keratinocytes and melanocytes are cells in which WNT 1 and WNT 5a expression is increased or NFATC 2 is dephosphorylated. Provides a whitening material screening system. Since the screening system can reproduce the skin condition showing the hyperpigmentation layer close to reality,
  • candidates such as natural products, extracts thereof, or chemical synthetic materials were treated only with melanocytes to evaluate the whitening efficacy.
  • the candidate substance grafted in the cell culture should be treated directly to the cell, and the melanocytes cannot be treated with the candidate substance applied to the actual formulation, such as cream or lotion. Therefore, candidates can only be clinically tested using conventional methods. It can be determined whether the formulation containing the whitening substance has a clinically whitening effect.
  • the skin whitening substance screening system according to the present invention can evaluate the presence or absence of the whitening effect by using the candidate substance in the culture medium as in the conventional method, as well as using a formulation such as cream or lotion containing the candidate substance. You can also evaluate the whitening effect.
  • the skin whitening material screening system for evaluating the whitening effect on the formulation is made of artificial skin by laminating a plurality of layers including at least one of fibroblasts, melanocytes and keratinocytes and then exposing to air.
  • the skin whitening screening system according to the present invention has a physiological and chemical characteristic similar to that of actual skin, and the evaluation of the whitening effect can be performed using a formulation that is actually used. It allows for efficient and simple sorting in less time and at less cost.
  • One aspect of the present invention is directed to one or more of WIF 1, WNT 1, WNT 5a and NFATC 2 of at least one of fibroblasts, keratinocytes and melanocytes in the skin lightening material screening system treated with the candidate substance. It provides a skin whitening material screening method comprising the step of checking the degree of expression or phosphorylation.
  • WIF 1, WNT 1, WNT 5a and NFATC 2 are all related to the expression and diffusion of melanin pigment. Specifically, decreased expression of WIF 1, increased expression of TOT 1 or WNT 5a, or increased dephosphorylation of NFATC 2 resulted in an increase in the expression of tyrosinase, an enzyme involved in melanogenesis, and keratinocytes.
  • the method for screening a skin whitening substance may further include determining a substance for increasing the expression level of WIF 1 as a skin whitening substance after confirming the expression level of WIF 1.
  • a method for screening a skin whitening material is provided, after determining the expression level of wr 1 or WNT 5a, determining a material that reduces the expression level of WNT 1 or WNT 5a as a skin whitening material. It may further include.
  • a method for screening a skin whitening substance may further include determining a substance that increases the phosphorylation degree of NFATC 2 as a skin whitening substance after determining the phosphorylation degree of NFATC 2.
  • Scoop Skin whitening substance screening system comprising at least one of fibroblasts, melanocytes and keratinocytes, which have reduced expression of WIF.l, increased expression of WNT 1 or WNT 5a, or dephosphorylation of FATC 2. Since it is reproduced in the actual skin condition where the skin degradation was performed, how the candidate substance changes the degree of expression of the WIF 1, WNT 1 or WNT 5a gene or the phosphorylation of the signal transducing factor NFATC 2 in the above-regulated system. By evaluating whether the candidate substance is a substance having a whitening effect, in particular, whether the substance can improve skin hyperpigmentation.
  • compositions for skin whitening comprising a material screened according to the skin whitening material screening method as an active ingredient.
  • a material screened according to the skin whitening material screening method as an active ingredient.
  • Substances that increase the expression of WIF 1, decrease the expression of WNT 1 or WNT 5a, or increase the degree of dephosphorylation of NFATC 2 may be considered to have a skin whitening effect, specifically, a substance that can improve skin hyperpigmentation.
  • a composition comprising the same as an active ingredient may exhibit a skin whitening effect, specifically, a skin hyperpigmentation improvement effect.
  • Another aspect of the present invention provides a skin whitening composition or an external skin composition comprising WIF 1 itself, which is a gene according to the present invention, thereby inhibiting the expression of tyrosinase of WIF 1. It may have an effect of improving skin whitening and hyperpigmentation by inhibiting melanin diffusion into keratinocytes.
  • a composition for skin whitening includes a cosmetic composition, a pharmaceutical composition, or a food composition.
  • composition shown in Table 1 it can be prepared by the conventional method.
  • composition shown in Table 2 below it can be produced in a nutritious recipe.
  • Nutritional creams may be prepared by conventional methods according to the compositions shown in Table 3 below. ⁇ 66> [Table 3]
  • Packs can be prepared by conventional methods according to the compositions shown in Table 4 below.
  • Ointments can be prepared by conventional methods according to the compositions shown in Table 5 below.
  • the TOT 1 and TOT 5a in the skin tissue of the hyperpigmentation site Expression is increased compared to normal skin tissue. This confirms that WNT 1 and WNT 5a are associated with skin hyperpigmentation.
  • HMGs human melanocyte growth supplement
  • PMA phorbol 12-myri state 13-acetate
  • WIF 1 was expressed in fibroblasts (FB) and keratinocytes (KC), but not in melanocytes (MC). Through this, it can be seen that WIF 1 is expressed in fibroblasts and keratinocytes, but not melanocytes.
  • Example 2 A three-dimensional system prepared by using keratinocytes and melanocytes in which strings were suppressed was used as Example 2, and a system prepared using normal keratinocytes and melanocytes was used as Comparative Example 2.
  • Example 3 After treatment with WIF 1 made from a human recombinant gene at 50 ng / ml in the culture medium was mixed keratinocytes with melanocytes with increased expression of WIF1 was prepared in substantially the same manner as in Example 1 A system prepared by winding on one artificial dermis was used as Example 3.
  • Example 1 Whether or not Example 1, a system according to the present invention, reflects well the actual skin condition was evaluated in comparison with Comparative Example 1 as follows.
  • Example 1 which contains fibroblasts that inhibited the expression of WIF 1, had a higher level of expression of tyrosinase, an enzyme that plays an important role in melanogenesis, as compared to the actual skin hyperpigmentation site. Significant increase was confirmed by Western blotting method through protein electrophoresis (see FIG. 4). In addition, staining keratinocytes with fluorescent antibodies that recognize keratin 14, which is expressed only in keratinocytes, and tyrosinase-related protein Ktyrosinase-related protein 1, TRP, expressed in melanocytes.
  • Example 5 Evaluation of a system containing keratinocytes inhibited the expression of WIF 1 Whether or not Example 2, a system according to the present invention, reflects the actual skin condition was evaluated in comparison with Comparative Example 2 as follows.
  • Example 2 including keratinocytes inhibiting the expression of WIF 1 was carried out in substantially the same manner as in Experiment 4, and thus played an important role in melanin production as in the actual skin hyperpigmentation site. It was confirmed that the expression of tyrosinase, an enzyme, was significantly increased compared to Comparative Example 2 (see FIG. 7).
  • Example 2 melanin diffusion is increased like the skin hyperpigmentation site (see Fig. 8). In this case, it was confirmed that p-NFATC 2, a phosphorylated form of NFATC 2, was reduced.
  • the system according to the present invention manufactured in three dimensions using melanocytes and keratinocytes is similar to the actual skin condition, and it can be confirmed that the skin hyperpigmentation site can be reproduced.
  • WIF 1 was processed by Western blotting using protein electrophoresis for the system of Example 3 comprising melanocytes treated with WIF 1 made using a human recombinant gene to be 50 ng / ml in the cell culture medium. It was confirmed that the expression of increased (see Figure 10). In this case, it was confirmed that the expression of tyrosinase was reduced in the system of Example 3 in substantially the same manner as in Experimental Example 4 (see FIG. 11). In addition, as compared with Comparative Example 1, it was confirmed that the melanin diffusion in Example 3 is reduced (see Fig. 12), it was also confirmed that the phosphorylation of NFATC 2 is increased (see Fig. 13).
  • WIF 1 and NFATC 2 are closely related to melanin pigment production, and thus the candidate material is treated in the system according to the present invention, and then the expression level of WIF 1 and its lower signaling factor NFATC 2 are reduced. By assessing the degree of phosphorylation, it can be seen that the skin whitening material can be screened.
  • the skin whitening material screening system can reproduce the actual skin, thereby effectively screening the material exhibiting clinically excellent skin whitening effect even in vitro. Furthermore, this is the actual skin tone of human Because it reflects the job well, it can be used to study the mechanism or improvement of pigment deficiency such as vitiligo or white hair.

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Abstract

L'invention concerne un système pour sélectionner une matière de blanchiment de la peau comprenant une ou plusieurs couches qui comprennent une ou plusieurs cellules de fibroblastes, kératinocytes et mélanocytes, et un procédé pour sélectionner une matière de blanchiment de la peau à l'aide du système. De plus, l'invention présente une composition pour un blanchiment de la peau qui comprend une matière sélectionnée par le procédé de sélection d'une matière de blanchiment de la peau comme ingrédient actif.
PCT/KR2012/008555 2011-10-18 2012-10-18 Système pour sélectionner une matière de blanchiment de la peau Ceased WO2013058578A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP2014536989A JP6069334B2 (ja) 2011-10-18 2012-10-18 皮膚美白物質スクリーニング構造体
CN201280051510.0A CN103857802B (zh) 2011-10-18 2012-10-18 皮肤美白物质筛选系统

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR20110106614 2011-10-18
KR10-2011-0106614 2011-10-18
KR1020120114965A KR102031161B1 (ko) 2011-10-18 2012-10-16 피부 미백 물질 스크리닝 시스템
KR10-2012-0114965 2012-10-16

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WO2013058578A2 true WO2013058578A2 (fr) 2013-04-25
WO2013058578A3 WO2013058578A3 (fr) 2013-07-04

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015160147A1 (fr) * 2014-04-16 2015-10-22 (주)아모레퍼시픽 Substance destinée à améliorer la peau, régulant l'expression de la cadhérine 11 ou de la n-cadhérine, et procédé pour cribler celle-ci
JP2015204780A (ja) * 2014-04-21 2015-11-19 日本メナード化粧品株式会社 美白成分のスクリーニング方法

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KR20100039077A (ko) * 2008-10-07 2010-04-15 숙명여자대학교산학협력단 Ｎｄｒｇ2 유전자를 이용한 미백 활성 물질의 스크리닝 방법

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KR20150119741A (ko) * 2014-04-16 2015-10-26 (주)아모레퍼시픽 캐드헤린11 또는 n-캐드헤린 발현을 조절하는 피부 개선 물질 및 이를 스크리닝하는 방법
KR102212623B1 (ko) * 2014-04-16 2021-02-09 (주)아모레퍼시픽 캐드헤린11 또는 n-캐드헤린 발현을 조절하는 피부 개선 물질 및 이를 스크리닝하는 방법
JP2015204780A (ja) * 2014-04-21 2015-11-19 日本メナード化粧品株式会社 美白成分のスクリーニング方法

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