CN110022849A - The screening method of skin lightening compositions comprising SH3BP4 inhibiting substances and SH3BP4 inhibiting substances - Google Patents
The screening method of skin lightening compositions comprising SH3BP4 inhibiting substances and SH3BP4 inhibiting substances Download PDFInfo
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- CN110022849A CN110022849A CN201780072676.3A CN201780072676A CN110022849A CN 110022849 A CN110022849 A CN 110022849A CN 201780072676 A CN201780072676 A CN 201780072676A CN 110022849 A CN110022849 A CN 110022849A
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Abstract
Include the composition for whitening skin for inhibiting SH3BP4 expression or active substance as effective component and the screening method for inhibiting SH3BP4 expression or active substance the invention discloses a kind of.Described includes that SH3BP4 is expressed or the composition of active substance can provide excellent whitening effect by melanin content in reduction skin for inhibiting.In addition, relative expression's inhibition level of SH3BP4 gene can be confirmed according to above-mentioned screening method, thus easily screening skin-whitening substance.
Description
Technical field
The present invention relates to composition for whitening skin and for the screening method of skin-whitening substance.
Background technique
The melanin that the effect of the various enzymes such as tyrosinase generates in melanocyte in human body is to determine human skin color
Most important factor will induce such as chloasma, freckle and mole pigmentation when it is excessively more than to need that melanin, which generates,
Disease is cosmetically bringing undesirable consequence.Also, because the melanin that the stress stimulation in the inside and outside portion of skin generates is passing through skin
Skin keratinization discharge before, will not with stress disappearance and disappear.
Therefore, carrying out inhibiting the research of this melanin production, and in cosmetics and drug with the use of anti-
Bad hematic acid, kojic acid, arbutin, quinhydrones, glutathione or derivatives thereof or the substance inhibited tyrosinase activity.But due to
Insufficient whitening effect of these substances and safety problem to skin, cause their use to be restricted.
[related fields bibliography]
[patent document]
(patent document 1) Korean Patent Publication No. 10-2013-0025768 (on 03 12nd, 2013).
Summary of the invention
Technical problem
In one aspect, the object of the present invention is to provide a kind of for adjusting the novel skin of the B16 cell in skin
The identification marker of whitening material.
On the other hand, the object of the present invention is to provide a kind of composition for whitening skin, the composition includes conduct
The expression for being used to that the skin-whitening substance to be inhibited to identify marker of active constituent or active substance.
On the other hand, the object of the present invention is to provide one kind identifies marker by using the skin-whitening substance to sieve
The method for looking into skin-whitening substance.
On the other hand, the object of the present invention is to provide one kind identifies marker by using the skin-whitening substance to examine
The compositions or agents box of off-color element related dermal state.
On the other hand, the object of the present invention is to provide one kind identifies marker by using the skin-whitening substance to examine
The essential information providing method of off-color element related dermal state.
Technical solution
On the one hand, the present invention provides a kind of composition for whitening skin, and the composition includes as active constituent
For inhibit SH3BP4 (SH3 Domain Binding protein 4) expression or active substance.
On the one hand, the present invention provides a kind of screening method for skin-whitening substance comprising following steps:
(a) test substances are handled to melanocyte (melanocyte)；With
(b) confirm whether the test substances can inhibit the table of SH3BP4 in melanocyte (SH3 Domain Binding protein 4)
It reaches.
On the one hand, the present invention is provided to diagnose the composition of pigment related dermal state, the composition includes for examining
Survey the reagent of the mRNA or SH3BP4 protein of SH3BP4.
On the one hand, the present invention is provided to diagnose the kit of pigment related dermal state, which includes for examining
Survey the reagent of the mRNA or SH3BP4 protein of SH3BP4.
On the one hand, the present invention is provided to diagnose the method for the essential information of pigment related dermal state offer, this method
Including confirming the mRNA of SH3BP4 or the expression quantity of protein from the Skin Cell that subject separates.
Beneficial effect
Novel skin whitening material identification marker SH3BP4 according to an aspect of the present invention can reduce black in skin
The synthesis of pigment.Therefore, comprising being used to inhibit the expression of the SH3BP4 or the combination of active substance as active constituent
Object is effectively used for skin-whitening.In addition, according to an aspect of the present invention, the screening method for skin-whitening substance is logical
It crosses and test substances is handled in skin to and confirmed relative expression's inhibition level of SH3BP4 gene, Lai Rongyi, effectively identify skin
Skin whitening material.Compositions or agents box for diagnosing pigment related dermal state according to an aspect of the present invention and
For diagnosing the essential information providing method of pigment related dermal state, can by the SH3BP4 gene for including in skin into
Row detection quickly and easily to diagnose pigment related dermal state, or can provide the basis letter of pigment related dermal state
Breath.
Detailed description of the invention
Fig. 1 is to compare figure according to the relative amount of melanin in the melanocyte of SH3BP4 gene expression inhibition degree.
Fig. 2 is the drop that confirmation is expressed according to tyrosinase (TYR) in the melanocyte of SH3BP4 gene expression inhibition degree
The figure of low degree.
Fig. 3 is relatively to scheme according to tyrosinase (TYR) in the melanocyte of SH3BP4 gene expression inhibition degree is active.
Specific embodiment
Hereinafter, the present invention is described in detail.
In the present specification, term " skin " refers to the organ outside covering organism, by epidermis, corium and subcutaneous rouge
Fat layer composition, being not only includes the tissue covered outside entire face or body, further includes the broadest general of scalp and hair
It reads.
One embodiment of the present of invention can provide for skin-whitening method comprising to needing the object of skin-whitening to apply
With for inhibiting the expression of SH3BP4 or the step of active substance.
Another embodiment of the invention can provide composition for whitening skin, and it includes the use as active constituent
In the expression or active substance that inhibit SH3BP4.
Another embodiment of the invention can provide expression for inhibiting SH3BP4 or active substance and use in preparation
Purposes in the composition of skin-whitening.
In addition, another embodiment of the invention can provide the expression or active inhibition of the SH3BP4 for skin-whitening
Substance.
In one embodiment of the invention, it is described for inhibit SH3BP4 expression or active substance can for for
The substance for inhibiting SH3BP4mRNA translation, more specifically, may include the widow combined at least part of SH3BP4mRNA
Nucleotide.
Whether the SH3BP4 has relationship not yet to disclose with B16 cell, but the present inventor passes through research hair
It is existing, the synthesis of melanin in the adjustable melanocyte in skin of SH3BP4.More specifically, it has already been proven that, it is black when skin
When the expression of SH3BP4 described in plain cell or activity inhibited or reduction, Melanin productions are reduced or tyrosinase
(tyrosinase) expression and activity has reduction.
In one embodiment of the invention, the oligonucleotides can be any one in siRNA, shRNA and miRNA
Kind is a variety of.The expression for inhibiting SH3BP4 or active substance can interfere (RNAi) phenomenon to can induce RNA
Any one or more in siRNA, shRNA and miRNA.One embodiment of the present of invention can be by utilizing for inducing
The RNAi phenomenon of SH3BP4mRNA interference, to inhibit the mRNA of SH3BP4 gene to express.
MiRNA is a kind of tiny RNA (endogenous tiny RNA) being present in cell, derives from and does not synthesize protein
DNA is generated by hairpin transcript (hairpin-shaped transcript).MiRNA is by the 3'-UTR with said target mrna
Complementary series be combined, to induce the Translational repression or stabilization removal of mRNA, eventually as inhibiting factor (repressor)
To inhibit the protein of said target mrna to synthesize.It is known that a miRNA can target several mRNA, and mRNA can also be by several miRNA institutes
It adjusts.Short interfering rna (siRNA) and tool for inducing other RNA of RNAi phenomenon to have, as 19-27mer or so short rna
There is the shRNA of short hair clip (short hairpin) structure.
In one embodiment of the invention, the oligonucleotides is double-strand, specifically, can for siRNA, shRNA or
Any one of miRNA, and include nucleotide sequence shown in SEQ ID NO:1 and with the interfertile nucleosides of SEQ ID NO:1
Acid sequence.
5'-GCGGUAUGAUUGAUAAUCU-3'(SEQ ID NO:1)
In one embodiment, the oligonucleotides is double-strand, specifically, can be in siRNA, shRNA and miRNA
It is any and include nucleotide sequence shown in SEQ ID NO:1 and SEQ ID NO:2, SEQ ID NO:1 and the SEQ ID
Nucleotide sequence shown in NO:2 can mutually hybridize.
Justice: 5'-GCGGUAUGAUUGAUAAUCU-3'(SEQ ID NO:1)
Antisense: 5'-AGAUUAUCAAUCAUACCGC-3'(SEQ ID NO:2)
In one embodiment, the oligonucleotides is double-strand, specifically, can be in siRNA, shRNA or miRNA
It is any, and include nucleotide sequence shown in SEQ ID NO:3 and with the interfertile nucleotides sequence of the SEQ ID NO:3
Column.
5'-GCGUAUGACUUCUUACUCA-3'(SEQ ID NO:3)
In one embodiment, the oligonucleotides is double-strand, specifically, can be in siRNA, shRNA and miRNA
It is any and include nucleotide sequence shown in SEQ ID NO:3 and SEQ ID NO:4, SEQ ID NO:3 and the SEQ ID
Nucleotide sequence shown in NO:4 can mutually hybridize.
Justice: 5'-GCGUAUGACUUCUUACUCA-3'(SEQ ID NO:3)
Antisense: 5'-UGAGUAAGAAGUCAUACGC-3'(SEQ ID NO:4)
The oligonucleotides according to an embodiment of the invention can be trapped in liposome or be inserted in carrier
Form be included.
In one embodiment, the concentration of the oligonucleotides can be 1-200nM.More specifically, the oligonucleotides
Concentration can be 5-50nM.
The concentration of the oligonucleotides can for 1nM or more than, 2nM or more than, 3nM or more than, 5nM or more than, 7nM or with
Upper, 9nM or more than, 11nM or more than, 15nM or more than, 16nM or more than, 17nM or more than, 18nM or more than, 19nM or with
Upper, 20nM or more than, 21nM or more than, 22nM or more than, 23nM or more than, 24nM or more than, 25nM or more than or 50nM or
More than.
In addition, the concentration of the oligonucleotides can be 200nM or following, 190nM or following, 180nM or following, 170nM
Or following, 150nM or following, 130nM or following, 100nM or following, 80nM or following, 50nM or following, 40nM or following,
30nM or following, 29nM or following, 28nM or following, 27nM or following, 26nM or following, 25nM or following, 24nM or following,
23nM or following, 22nM or following, 21nM or following, 20nM or following, 19nM or following, 18nM or following, 17nM or following,
16nM or following, 15nM or following, 14nM or following, 13nM or following, 12nM or following, 11nM or following or 10nM or with
Under.
The concentration of the oligonucleotides within the above range when, the expression inhibiting excellent effect of SH3BP4, and be suitble to use
In skin-whitening.
In one embodiment, the mRNA sequence of the SH3BP4 (SH3 Domain Binding protein 4) is NCBI reference sequences
NM_014521.2, more specifically, can be indicated by the nucleotide sequence of following SEQ ID NO:5.
The composition according to an embodiment of the invention may include the 0.00001-30 weight for accounting for composition total weight
The expression for inhibiting SH3BP4 of amount % or active substance.
In addition, the composition according to an embodiment of the invention, can be Dermatologic preparation composition, more specifically
Ground, it may include cosmetic composition or pharmaceutical composition.
In one embodiment, the cosmetic composition can provide as all dosage forms suitable for part.For example, can mention
For being mutually dispersed in the lotion obtained in water phase for solution, by oil, the lotion for obtaining Aqueous dispersions in oily phase, suspension, consolidating
The dosage form of body, gel, powder, paste, foam or aerosol composition.According to the conventional method of this field, can prepare such
The composition of dosage form.
In one embodiment, the cosmetic composition, in addition to the foregoing, in the model for not influencing main effect
In enclosing, it is preferable that can also include the other compositions that can generate synergistic effect with main effect.Cosmetics according to the present invention
Composition may include the substance in vitamin, macromolecule peptide, macromolecule polysaccharide and sphingolipid.In addition, in one embodiment
In, the cosmetic composition may include moisturizer, lubricant, interfacial agent, ultraviolet absorbing agent, preservative, fungicide,
Antioxidant, pH adjusting agent, organic and inorganic pigment, fragrance, coolant, antiperspirant.Those skilled in the art can be in not shadow
The content of mentioned component is easily chosen in the range of sound the object of the invention and effect, which can account for composition total weight
0.01-5 weight % specifically can account for 0.01-3 weight %.
In addition, in one embodiment, described pharmaceutical composition can provide as all dosage forms suitable for part.For example,
It can be administered by oral, transdermal, vein, muscle, subcutaneous injection.In one embodiment, described pharmaceutical composition can be
Injection, skin preparations for extenal use, suspending agent, emulsion, gelling agent, patch or spray, but not limited to this.The dosage form can basis
The conventional method of this field is easily prepared, and interfacial agent, excipient, wettable powder, emulsification rush can be suitably used
Into agent, suspending agent, the salt for controlling osmotic pressure or buffer, colorant, fragrance, stabilizer, preservative, preservative agent or other
Usual auxiliaries.
The effective component of pharmaceutical composition according to an embodiment of the invention is according to the age of object, gender, weight, disease
The judgement of reason situation and severity, administration route or prescriber and change.According to these because usually determining dosage in this field
In the level of those of ordinary skill, for example, its daily dose can be -5000mg/kg/ days 0.1mg/kg/ days, more specifically,
It can be -500mg/kg/ days 50mg/kg/ days, but not limited to this.
Another embodiment of the invention can provide the screening for skin-whitening substance by using the SH3BP4
Method.
In one embodiment, it the described method comprises the following steps: (a) melanocyte (melanocyte) processing being tested
Substance；And (b) whether exact p-value substance can inhibit the expression of SH3BP4 in melanocyte.Wherein, in the step (b)
Whether can inhibit the expression of SH3BP4, can be that whether can inhibit the translation of SH3BP4mRNA.
In one embodiment, the step (b) may include, before and after handling test substances to melanocyte
The step of relative expression's inhibition level of SH3BP4 confirms.In addition, according to one embodiment, the step (b) may include,
Relative expression's inhibition level of the melanocyte of melanocyte and untreated test substances through handling test substances is carried out true
The step of recognizing.
In addition, the screening method may further include according to one embodiment: the knot of the step (b) to be confirmed
After fruit, when the test substances can inhibit the expression of SH3BP4, the test substances are determined as to the step of skin-whitening substance
Suddenly.
In this specification, term " relative expression's inhibition level " can be, in the melanocyte before handling test substances
SH3BP4 expression compared with the expression for handling the SH3BP4 in the melanocyte after test substances, expression
Suppressed degree.Also or, " relative expression's inhibition level " can be, the table of the SH3BP4 of the melanocyte through handling test substances
The degree of expression by inhibitation when up to level compared with the expression of the SH3BP4 of the melanocyte of untreated test substances.Institute
Stating expression includes expression quantity and expression quality (quality).In addition, expression, such as, it may include the expression of protein
It is horizontal.More specifically, the expression may include the degree of the B16 cell in melanocyte or the expression of tyrosinase
Horizontal or synthesis degree.
In one embodiment of the present of invention, the judgment step of the skin-whitening substance can include: when " relative expression inhibits
When degree " is twice or twice or more, then it is judged as skin-whitening substance.That is, the melanocyte before processing test substances
In SH3BP4 expression compared with the expression for handling the SH3BP4 in the melanocyte after test substances, work as table
When up to 1.1-2 times or more of suppressed restriction, it can determine whether as skin-whitening substance.Also or, when the melanocyte through handling test substances is thin
The expression of SH3BP4 is compared with the expression of SH3BP4 in the melanocyte of untreated test substances in born of the same parents, when expression by
When inhibiting about 1.1-2 times or more, it can determine whether as skin-whitening substance.
For example, the relative expression of the SH3BP4 inhibits after processing test substances compared with before processing test substances
Degree, can for 1.1 times or more, 1.2 times or more, 1.3 times or more, 1.4 times or more, 1.5 times or more, 1.6 times or
Above, 1.7 times or more, 1.8 times or more, 1.9 times or more or 2 times or more, but not limited to this.The expression
It is measured result under conditions of ensuring statistical significance.The concept of statistical significance refers to be analyzed by biometric
The case where meaningful difference that method is shown, including it is quantitative when situation of the p value less than 0.05.
In the step of according to one embodiment (a), the test substances can be transfected into the liposome or carrier of cell
In, and cell can be melanocyte, i.e. melanocyte.The transfection can be total by using microinjection, calcium phosphate
The precipitation method, electroporation or liposome are carried out using method etc., but not limited to this.
In addition, in one embodiment, the expression of the SH3BP4 can be determined using known technology, for example, poly-
Synthase chain reaction method (RT-PCR), enzyme-linked immunosorbent assay method (ELISA), western blot method or immunoblotting
(immune blot) method, but not limited to this.
In addition, one embodiment of the present of invention can provide the biomarker for diagnosing pigment related dermal state degree
Object.In addition, one embodiment of the present of invention can provide by being used to diagnose pigment related dermal using the biomarker
The composition of state, or the kit for diagnosing pigment related dermal state.
Another embodiment of the invention can provide the essential information providing method for diagnosing pigment related dermal state,
Or the diagnostic method of pigment related dermal state.
Specifically, one embodiment of the present of invention can provide the composition for diagnosing pigment related dermal state, the group
Closing object includes described for detecting the reagent of the mRNA or SH3BP4 protein of SH3BP4.
In addition, another embodiment of the invention can provide the examination of the mRNA or SH3BP4 protein for detecting SH3BP4
Agent is preparing the purposes in the composition for diagnosing pigment related dermal state.
In addition, another embodiment of the invention can provide the SH3BP4's for diagnosing the relevant skin condition of pigment
The detection reagent of mRNA or SH3BP4 protein.
In addition, another embodiment of the invention can provide, the essential information for diagnosing pigment related dermal state is mentioned
Method for method or for diagnosing pigment related dermal state, the method includes right from the Skin Cell that subject separates
The step of expression quantity of SH3BP4 is confirmed.At this point, in one embodiment, SH3BP4 expression quantity can be for SH3BP4's
The expression quantity of mRNA or SH3BP4 protein, and the skin of the Skin Cell isolated from subject and Normal group
Cell can be melanocyte.
In one embodiment, for diagnosing the essential information providing method of pigment related dermal state or for diagnosing pigment
The method of related dermal state further includes, by SH3BP4 expression quantity and the normal control from the Skin Cell that subject separates
The step of SH3BP4 expression quantity is compared in group Skin Cell.In addition, in one embodiment, the method also includes, when
The SH3BP4 expression quantity from the Skin Cell that subject separates is higher than normally to the SH3BP4 table in group photograph Skin Cell
Up to when measuring, exist with pigment related dermal state abnormal come the step of providing information or diagnose.For example, if described from tested
Expression quantity of the SH3BP4 expression quantity than the SH3BP4 in Normal group Skin Cell in the Skin Cell of person's separation is about higher by
At 1.1-2 times or more, it is abnormal to can determine whether that pigment related dermal state exists.More specifically, for example, described from the subject
The expression quantity of SH3BP4 in isolated Skin Cell is higher by 1.1 times or more, 1.2 compared with Normal group Skin Cell
Times or it is above, 1.3 times or more, 1.4 times or more, 1.5 times or more, 1.6 times or more, 1.7 times or more, 1.8 times or
Above, 1.9 times or more or 2 times or more when, it is abnormal to can determine whether that pigment related dermal state exists.The expression quantity be
Ensure the result measured under conditions of statistical significance.The concept of statistical significance refers to be had by what biology was shown
The case where difference of meaning, including it is quantitative when situation of the p value less than 0.05.
In addition, another embodiment of the invention can provide comprising mRNA the or SH3BP4 protein for detecting SH3BP4
Reagent for diagnosing the kit of pigment related dermal state, which may further include specification, the explanation
Book describes the above-mentioned essential information providing method for diagnosing pigment related dermal state or for pigment related dermal shape
The diagnostic method of state.
In addition, in one embodiment, it is described for diagnosing the composition of pigment related dermal state or for diagnosing color
SH3BP4mRNA detection reagent included in the kit of plain related dermal state may include, with SH3BP4 specific binding
One of primer and probe is a variety of.In one embodiment, the SH3BP4 protein detection reagents may include with
One of antibody and probe of SH3BP4 specific binding are a variety of.
Hereinafter, composition and effect of the invention is described in more details in reference implementation example.But reality below
It applies example to be only used for helping to understand the present invention, is not intended to limit scope of the invention and range.
[experimental example 1]
By following experiment, to be identified through the reduction that the pigment for the expression for inhibiting SH3BP4 is formed.
Prepare the primary melanocyte of people (the moderately pigmented by separating in the pigmented tissue of moderate
Human primary Melanocyte) (be purchased from Thermofisher company, https: //www.thermofisher.com/
Order/catalog/product/C1025C? ICID=search-product).
In order to reduce the expression of SH3BP4 in the melanocyte, introducing is shown into specificity to SH3BP4mRNA sequence
Following two SH3BP4 siRNA.
Embodiment 1:si-SH3BP4#1
Justice: 5'-GCGGUAUGAUUGAUAAUCU-3'(SEQ ID NO:1)
Antisense: 5'-AGAUUAUCAAUCAUACCGC-3'(SEQ ID NO:2).
Embodiment 2:si-SH3BP4#2
Justice: 5'-GCGUAUGACUUCUUACUCA-3'(SEQ ID NO:3)
Antisense: 5'-UGAGUAAGAAGUCAUACGC-3'(SEQ ID NO:4).
Specifically, in the primary melanocyte of the people (Human primary Melanocyte) as melanocyte strain
On, by the SH3BP4 siRNA of the embodiment 1 by being transfected into the cell using liposome (liposome), until SH3BP4
SiRNA concentration reaches 20nM.The liposome has used Lipofectaminer RNAiMAX reagent (Thermo Fisher
Scientific, https: //www.thermofisher.com/order/catalog/product/13778150).It is described
The SH3BP4 siRNA of embodiment 2 is also by method identical with above-described embodiment 1, by being turned using liposome (liposome)
Contaminate it is intracellular, until SH3BP4 siRNA concentration reaches 20nM.
As negative control group, using being functionally proved to be any gene for not reducing the mankind, cell proliferation
And the smallest siRNA of influence to cell activity, i.e. Silencer Select Negative Control No.1siRNA
(Thermo Fisher Scientific, https: //www.thermofisher.com/order/catalog/product/
4390843? ICID=search-product).The siRNA of the negative control group is also according to identical with above-described embodiment 1
Method, by being transfected into the cell using liposome (liposome), until siRNA concentration reaches 20nM.
By the cell of the Examples 1 and 2 of each diameter transfection, negative control group, at 37 DEG C, 5%CO2It is trained in incubator
It supports about 7 days.
After each cell is washed twice with 5ml PBS buffer solution, 1ml PBS buffer solution is added, is then scraped with scraper plate
Under all cells cell granulations (cell is made with the speed of 13000rpm centrifuge separation (centrifugation) 5 minutes
Pellet), and by the color of observation cell granulations compare the degree of pigment formation.In addition, the cell granulations are dissolved
After in the 1N NaOH that volume is 2001 μ l, by the absorbance at measurement 450nm, to measure the amount of melanin.
As a result as shown in Figure 1, it may be determined that in the Examples 1 and 2 for the expression for inhibiting SH3BP4 using SH3BP4 siRNA
With negative control phase group ratio, the melanin amount contained in melanocyte substantially reduces about 2 times or more.It means that when inhibiting
In skin when the expression or activity of SH3BP4, skin whitening effects can be presented.
[experimental example 2]
According to following experiment, by using the primary melanocyte of people identical with [experimental example 1], Examples 1 and 2
The siRNA of SH3BP4 siRNA and negative control, to determine the reduction by the tyrosinase expression for inhibiting the expression of SH3BP4.
Firstly, according to method identical with [experimental example 1], by utilizing liposome by the SH3BP4 of Examples 1 and 2
The siRNA and siRNA of negative control group is transfected into the cell respectively, until SH3BP4 siRNA and siRNA concentration respectively reaches
20nM。
By the respectively Examples 1 and 2 through transfecting, negative control group cell, at 37 DEG C, 5%CO2It is cultivated in incubator
About 3 days.
By using RIPA buffer, the cell membrane of each cell is ruptured, and is extracted using centrifugal separator
Supernatant containing protein.Then, pass through the protein of about 10 μ g of BCA (dihomocinchonine acid) standard measure.By with Quantitative Western
Matter carries out Western blot experiment.Wherein, by using tyrosinase specific antibody (catalog number (Cat.No.) Sc-20035；Santa
Cruz Biotechnology) and SH3BP4 specific antibody (catalog number (Cat.No.) 17691-1-AP；Proteintech Group), come
Observe the amount of tyrosinase and SH3BP4 protein.Tyrosinase and SH3BP4 protein expression quantity are, by as house keeper's base
The protein expression quantity of the beta-actin (catalog number (Cat.No.) 3280, Abcam) of cause, to be standardized.
Then, containing by the way that in the supernatant using the separated protein of above-mentioned RIPA, 10 μ g proteins are added
Enter in 100 μ 10mM1- dihydroxyphenylalanines (solvent is 50mM sodium phosphate buffer (pH6.8)), it is small in 37 DEG C of reactions 1
When, it measures the absorbance at 490nm and analyzes the activity of tyrosinase (tyrocynase).
As a result black as shown in Fig. 2, being inhibited in the Examples 1 and 2 of the expression of SH3BP4 by using SH3BP4 siRNA
The expression quantity of the intracellular tyrosinase of element occurs reduction.In addition, as shown in figure 3, junket can be confirmed compared with negative control
The activity (TYR activity) of propylhomoserin enzyme significantly reduces about 2 times or more.It means that when the expression or work that inhibit SH3BP4 in skin
When property, skin whitening effects can be presented.
The Formulation Example of composition according to an embodiment of the invention is described below, can also be applied various in it
His dosage form, its object is to the present invention is described in detail, but are not intended to limit the present invention.
[Formulation Example 1] nutrition toner
According to conventional methods, nutrition toner is prepared according to composition shown in following table 1.
[table 1]
| Raw material name content | Content (weight %) |
| Glycerol | 3.0 |
| Butanediol | 3.0 |
| Propylene glycol | 3.0 |
| Carboxyl vinyl polymer | 0.1 |
| Liposome comprising embodiment 1 or 2 | 0.01 |
| Beeswax | 4.0 |
| Polysorbate60 | 1.5 |
| Caprylic/capric triglyceride | 5.0 |
| Saualane | 5.0 |
| Sorbitan sesquioleate | 1.5 |
| Cetostearyl alcohol | 1.0 |
| Triethanolamine | 0.2 |
| Preservative, fragrance | In right amount |
| Pure water | Surplus |
| It amounts to | 100 |
[Formulation Example 2] nutritional emulsions
According to conventional methods, nutritional emulsions are prepared according to composition shown in following table 2.
[table 2]
[Formulation Example 3] nourishing cream
According to conventional methods, nourishing cream is prepared according to composition shown in following table 3.
[table 3]
| Raw material name content | Content (weight %) |
| Glycerol | 3.5 |
| Butanediol | 3.0 |
| Atoleine | 7.0 |
| Beta glucan | 7.0 |
| Carbomer | 0.1 |
| Liposome comprising embodiment 1 or 2 | 0.01 |
| Caprylic/capric triglyceride | 3.0 |
| Saualane | 5.0 |
| Cetearyl glucoside | 1.5 |
| Sorbitan stearate | 0.4 |
| Polysorbate60 | 1.2 |
| Triethanolamine | 0.1 |
| Preservative, fragrance | In right amount |
| Pure water | Surplus |
| It amounts to | 100 |
[preparation embodiment 4] injection
According to conventional methods, injection is prepared according to composition shown in following table 4.
[table 4]
| Raw material name content | Weight ratio (every 1ml) |
| Sodium chloride | 9.0mg |
| Sodium carboxymethylcellulose | 30.0mg |
| Polysorbas20 | 1.0mg |
| Liposome comprising embodiment 1 or 2 | 1.0mg |
| Distilled water for injection | Surplus |
[preparation embodiment 5] ointment
According to conventional methods, ointment is prepared according to composition shown in following table 5.
| [table 5] raw material name content | Content (weight %) |
| Glycerol | 8.0 |
| Butanediol | 4.0 |
| Atoleine | 15.0 |
| Beta glucan | 7.0 |
| Carbomer | 0.1 |
| Embodiment 1 or 2 | 0.01 |
| Caprylic/capric triglyceride | 3.0 |
| Saualane | 1.0 |
| Cetearyl glucoside | 1.5 |
| Sorbitan stearate | 0.4 |
| Cetostearyl alcohol | 1.0 |
| Preservative | In right amount |
| Fragrance | In right amount |
| Pigment | In right amount |
| Beeswax | 4.0 |
| Pure water | Surplus |
| It amounts to | 100 |
<110>Amorepacific Corporation
<120>skin lightening compositions comprising SH3BP4 inhibiting substances and the screening method of SH3BP4 inhibiting substances
<130> OF17P111PCT
<150> KR 10-2016-0122317
<151> 2016-09-23
<160> 5
<170> KoPatentIn 3.0
<210> 1
<211> 19
<212> RNA
<213> Artificial Sequence
<220>
<223> sense siRNA of SH3BP4 #1
<400> 1
gcgguaugau ugauaaucu 19
<210> 2
<211> 19
<212> RNA
<213> Artificial Sequence
<220>
<223> antisense siRNA of SH3BP4 #1
<400> 2
agauuaucaa ucauaccgc 19
<210> 3
<211> 19
<212> RNA
<213> Artificial Sequence
<220>
<223> sense siRNA of SH3BP4 #2
<400> 3
gcguaugacu ucuuacuca 19
<210> 4
<211> 19
<212> RNA
<213> Artificial Sequence
<220>
<223> antisense siRNA of SH3BP4 #2
<400> 4
ugaguaagaa gucauacgc 19
<210> 5
<211> 5193
<212> RNA
<213> Artificial Sequence
<220>
<223> mRNA of SH3BP4
<400> 5
gggaccaccc tccgcccgcc gaggcggggg cccagcgcgc ccggcactct cggcggtccg 60
ggcccctcgc cactaccgcc gccgccgccg ccgtgagtcc cgcggagccg cgcgcgcccc 120
cggctgggcc gagccgctgg ccgacgagcg gagcctcagg agccggcggg gacgccatgc 180
gagccagcgt ctcccttctc tcctggacag aaggccgtgt cctgggactt ctctgatggc 240
gagaggctgc ggctgtacca ggaagaaaca tattgccgag tggatgccgc cgcgcagcgt 300
gtttgcttga ggcagaagct tcagcatctg ctgggataac tggaggaaga aatatgaagc 360
cttagcggct ttacccggga agcgagtttc gagatggcgg ctcagcggat ccgagcggcc 420
aactccaatg gcctccctcg ctgcaagtca gaggggaccc tgattgacct gagcgaaggg 480
ttttcagaga cgagctttaa tgacatcaaa gtgccttctc ccagtgcctt gctcgtagac 540
aaccccacac ctttcggaaa tgcaaaggaa gtgattgcga tcaaggacta ttgccccacc 600
aacttcacca cactgaagtt ctccaagggc gaccatctct acgtcttgga cacatctggc 660
ggtgagtggt ggtacgcaca caacaccacc gaaatgggct acatcccctc ctcctatgtg 720
cagcccttga actaccggaa ctcaacactg agtgacagcg gtatgattga taatcttcca 780
gacagcccag acgaggtagc caaggagctg gagctgctcg ggggatggac agatgacaaa 840
aaagtaccag gcagaatgta cagtaataac cctttctgga atggggtcca gaccaatcca 900
tttctgaatg ggaacgtgcc cgtcatgccc agcctggatg agctgaatcc caaaagtact 960
gtggatttgc tcctttttga cgcaggtaca tcctccttca ccgaatccag ctcagccacc 1020
acgaatagca ctggcaacat cttcgatgag cttccagtca caaacggact ccacgcagag 1080
ccgccggtca ggcgggacaa ccccttcttc agaagcaagc gctcctacag tctctcggaa 1140
ctctccgtcc tccaagccaa gtccgatgct cccacatcgt cgagtttctt caccggcttg 1200
aaatcacctg cccccgagca atttcagagc cgggaggatt ttcgaactgc ctggctaaac 1260
cacaggaagc tggcccggtc ttgccacgac ctggacttgc ttggccaaag ccctggttgg 1320
ggccagaccc aagccgtgga gacaaacatc gtgtgcaagc tggatagctc cgggggtgct 1380
gtccagcttc ctgacaccag catcagcatc cacgtgcccg agggccacgt cgcccctggg 1440
gagacccagc agatctccat gaaagccctg ctggaccccc cgctggagct caacagtgac 1500
aggtcctgca gcatcagccc tgtgctggag gtcaagctga gcaacctgga ggtgaaaacc 1560
tctatcatct tggagatgaa agtgtcagcc gagataaaaa atgacctttt tagcaaaagc 1620
acagtgggcc tccagtgcct gaggagcgac tcgaaggaag ggccatatgt ctccgtcccg 1680
ctcaactgca gctgtgggga cacggtccag gcacagctgc acaacctgga gccctgtatg 1740
tacgtggctg tcgtggccca tggcccaagc atcctctacc cttccaccgt gtgggacttc 1800
atcaataaaa aagtcacagt gggtctctac ggccctaaac acatccaccc atccttcaag 1860
acggtagtga ccatttttgg gcatgactgt gccccaaaga cgctcctggt cagcgaggtc 1920
acacgccagg cacccaaccc tgccccggtg gccctgcagc tgtgggggaa gcaccagttc 1980
gttttgtcca ggccccagga tctcaaggtc tgtatgtttt ccaatatgac gaattacgag 2040
gtcaaagcca gcgagcaggc caaagtggtg cgaggattcc agctgaagct gggcaaggtg 2100
agccgcctga tcttccccat cacctcccag aaccccaacg agctctctga cttcacgctg 2160
cgggttcagg tgaaggacga ccaggaggcc atcctcaccc agttttgtgt ccagactcct 2220
cagccacccc ctaaaagtgc catcaagcct tccgggcaaa ggaggtttct caagaagaac 2280
gaagtcggga aaatcatcct gtccccgttt gccaccacta caaagtaccc gactttccag 2340
gaccgcccgg tgtccagcct caagtttggt aagttgctca agactgtggt gcggcagaac 2400
aagaaccact acctgctgga gtacaagaag ggcgacggga tcgccctgct cagcgaggag 2460
cgggtcaggc tccggggcca gctgtggacc aaggagtggt acatcggcta ctaccagggc 2520
agggtgggcc tcgtgcacac caagaacgtg ctggtggtcg gcagggcccg gcccagcctg 2580
tgctcgggcc ccgagctgag cacctcggtg ctgctggagc agatcctgcg gccctgcaaa 2640
ttcctcacgt acatctatgc ctccgtgagg accctgctca tggagaacat cagcagctgg 2700
cgctccttcg ctgacgccct gggctacgtg aacctgccgc tcaccttttt ctgccgggca 2760
gagctggata gtgagcccga gcgggtggcg tccgtcctag aaaagctgaa ggaggactgt 2820
aacaacactg agaacaaaga acggaagtcc ttccagaagg agcttgtgat ggccctactg 2880
aagatggact gccagggcct ggtggtcaga ctcatccagg actttgtgct cctgaccacg 2940
gctgtagagg tggcccagcg ctggcgggag ctggctgaga agctggccaa ggtctccaag 3000
cagcagatgg acgcctacga gtctccccac cgggacagga acggggttgt ggacagcgag 3060
gccatgtgga agcctgcgta tgacttctta ctcacctgga gccatcagat cggggacagc 3120
taccgggatg tcatccagga gctgcacctg ggcctggaca agatgaaaaa ccccatcacc 3180
aagcgctgga agcacctcac tgggactctg atcttggtga actccctgga cgttctgaga 3240
gcagccgcct tcagccctgc ggaccaggac gacttcgtga tttgaatggg tcccctcccc 3300
tcctgctgct ctggagtgca agccctcttc tgccctgcgt gccctgctgt caccgcggag 3360
ctgaagaggg aggaaggggc ggctgctcag acagatttag ggcccgccag ctaggctaca 3420
cccatcatgc gccgccctcc tccatcgagg gagaggcctg aagggactgc ctactgcagc 3480
tcgttgccaa tcacatagct ttctatttgt taagtataaa tttaaattta aaatcacttt 3540
tttaacgaat ggggggaagg gatctatgag aaaggtggta tctaattttt ttatggacca 3600
taaaggttta aaagaaaata ggggcacagg ctgttgaggt ttttatgttg ttatagacct 3660
ttttaaatta tgttagagat gtatataggt atttaaaggt cactgggagc gtttctgatt 3720
cccggccaca ctttgcattt caacactcag cccggaaaga tgctcgttcg gttgttggac 3780
ctctttcact ccctgcgtgt aagaaggtga atcacgtggg aaaaagtggc ttttcagtaa 3840
acgggtacag ctcattcttt ctgagaaggc cccaggtcct gctccctcct cggatttgat 3900
tgtcttccgt gctttgcctc actcgtagta aatgaccatc catagaatat gtgaatcttt 3960
ggtgagcttc agtgggcaga gtgaagtccc gcattagcat ttaggtgccc tgagctgttt 4020
ctgccaatag attagaaagc agccatgagt tgacagtctt tagggcccct gccagtgtgc 4080
aattagtcat tgacaagaac aatgccattt gagagtgagg tggtccctgc tgctacgagg 4140
ccattgtact gttttttcct tgaggtcaaa gcagtgcttc ccatagagtt tgctgcctct 4200
tctgtggaca ggaagaaaac ttcatgaccg aatcagagcc ttggtggcca ctgactctcg 4260
tgcttattgc agatgctgtg gttggcctca caagcaacgc cttatgctga tgtgcagagg 4320
tgccagctgc catttgccaa actctgcatt tcatttcatc taaggcttaa cccctcttcc 4380
ttcctggtgt acctgtgtct cctcggaagg aagtcatagt ttagatgaaa ccattttttg 4440
tacaatgtaa agatcatctg agcaagatga gcattttgta aaaatgaaaa tgtgactcac 4500
ataaaatcag gaacttgaca cagtgttgca ttaataactt tagggtgcag acatgctgtg 4560
tgaatctcac aatgcgtcgt agatgtcgcg tgttggaagg gagcaggagg aaggactgat 4620
actggcaaat cagtagagtg aggtgatcct tagcaacgtg ccaggacact tcctgtgtgc 4680
ctgcagttgt cagggaccat ttgggatccc gaatctcatt ctctaaaact gctttcttga 4740
aacatgttac ttccttagta taatcaatgt atactccctt actggcctga aacgttgtat 4800
agctacttat tcagatactg aagaccaacg gactgaaaaa aagaacaaac attagctatt 4860
ttatgctgca agaaccagga cacacaattc gccaatcatc ccaccatata accttcgatt 4920
gtgcttctca actccacccc ataatttctc ccagagacca tctatcacct tttccccaaa 4980
gaagaaacaa aaccagttgc accttaaacc atggatattt tttcctcagg ggctttaaat 5040
agtttcctat gcaacgtgtc ttgtagcaca aataaaattc tacaaaagtt gcagtaaatt 5100
ttatttggat attttaacct gttaagtgtg tgtgtgtttt ctgtacccaa ccagacttta 5160
aataaaacaa acatgaaacc taaaaaaaaa aaa 5193
Claims (23)
1. a kind of composition for whitening skin, it includes be used to inhibit SH3BP4 (SH3 structural domain knot as effective component
Hop protein 4) expression or active substance.
2. composition for whitening skin according to claim 1, which is characterized in that described for inhibiting SH3BP4's
Expression or active substance include the substance for inhibiting SH3BP4 mRNA to translate.
3. composition for whitening skin according to claim 2, which is characterized in that described for inhibiting SH3BP4's
Expression or active substance are the oligonucleotides combined at least part of SH3BP4 mRNA.
4. composition for whitening skin according to claim 3, which is characterized in that the oligonucleotides be siRNA,
Any one or more in shRNA and miRNA.
5. composition for whitening skin according to claim 3, which is characterized in that the oligonucleotides is double-strand,
And include nucleotide sequence shown in SEQ ID NO:1 and with the interfertile nucleotide sequence of SEQ ID NO:1.
6. composition for whitening skin according to claim 3, which is characterized in that the oligonucleotides be double-strand simultaneously
Comprising nucleotide sequence shown in SEQ ID NO:1 and SEQ ID NO:2, shown in the SEQ ID NO:1 and SEQ ID NO:2
Nucleotide sequence mutually hybridize.
7. composition for whitening skin according to claim 3, which is characterized in that the oligonucleotides is double-strand,
And include nucleotide sequence shown in SEQ ID NO:3 and with the interfertile nucleotide sequence of SEQ ID NO:3.
8. composition for whitening skin according to claim 3, which is characterized in that the oligonucleotides be double-strand simultaneously
Comprising nucleotide sequence shown in SEQ ID NO:3 and SEQ ID NO:4, shown in the SEQ ID NO:3 and SEQ ID NO:4
Nucleotide sequence mutually hybridize.
9. according to composition for whitening skin described in any one in claim 3-8, which is characterized in that the widow
Nucleotide is to be trapped in liposome or be inserted in the form in carrier and be included.
10. composition for whitening skin according to claim 1, which is characterized in that the composition is for skin
The cosmetic composition of skin.
11. composition for whitening skin according to claim 1, which is characterized in that the composition is medicine group
Close object.
12. composition for whitening skin according to claim 1, which is characterized in that the composition includes to account for group
The expression for being used to inhibit SH3BP4 of the 0.00001-30 weight % of conjunction object total weight or active substance.
13. a kind of screening method for skin-whitening substance, this method comprises the following steps:
(a) test substances are handled to melanocyte；With
(b) confirm whether the test substances can inhibit the expression of SH3BP4 in melanocyte (SH3 Domain Binding protein 4).
14. the screening method according to claim 13 for skin-whitening substance, which is characterized in that the step (b)
In whether inhibit SH3BP4 is expressed as the translation of SH3BP4 mRNA whether can be inhibited.
15. the screening method according to claim 13 for skin-whitening substance, which is characterized in that the step (b)
Relative expression's inhibition level of SH3BP4 before and after including the steps that handling test substances to melanocyte confirms.
16. the screening method that screening according to claim 13 is used for skin-whitening substance, which is characterized in that the step
(b) include to through handle test substances melanocyte and untreated test substances melanocyte relative expression's inhibition level
The step of being confirmed.
17. the screening method that screening according to claim 15 is used for skin-whitening substance, which is characterized in that the screening
Method further includes,, will be described when the test substances inhibit the expression of SH3BP4 after the result of the step (b) to be confirmed
Test substances are determined as the step of skin-whitening substance.
18. a kind of for diagnosing the composition of pigment related dermal state, the composition includes the mRNA for detecting SH3BP4
Or the reagent of SH3BP4 protein.
19. a kind of for diagnosing the essential information providing method of pigment related dermal state, this method includes separating from subject
Skin Cell in the step of expression quantity of SH3BP4 (SH3 Domain Binding protein 4) is confirmed, the table of the SH3BP4
Up to the expression quantity for the mRNA or SH3BP4 protein that amount is SH3BP4.
20. according to claim 19 for diagnosing the essential information providing method of pigment related dermal state, feature
Be, this method further include by from the Skin Cell that the subject separates SH3BP4 expression quantity and Normal group skin
The step of SH3BP4 expression quantity in cell is compared.
21. according to claim 20 for diagnosing the essential information providing method of pigment related dermal state, feature
It is, this method further includes being higher than when from the SH3BP4 expression quantity in the Skin Cell that the subject separates normally to a group photograph
When SH3BP4 expression quantity in Skin Cell, it will thus provide pigment related dermal state has the step of abnormal information.
22. a kind of for diagnosing the kit of pigment related dermal state, which includes the mRNA for detecting SH3BP4
Or the reagent of SH3BP4 protein.
23. according to claim 22 for diagnosing the kit of pigment related dermal state, which is characterized in that the reagent
Box further includes the specification for recording method described in any one of 9-21 according to claim 1.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020160122317A KR20180032954A (en) | 2016-09-23 | 2016-09-23 | A composition for skin brightening comprising SH3BP4 inhibiting materials and a method for screening SH3BP4 inhibiting materials |
| KR10-2016-0122317 | 2016-09-23 | ||
| PCT/KR2017/008953 WO2018056583A1 (en) | 2016-09-23 | 2017-08-17 | Composition for skin whitening comprising sh3bp4-inhibiting material, and method for screening sh3bp4-inhibiting material |
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|---|---|
| CN110022849A true CN110022849A (en) | 2019-07-16 |
| CN110022849B CN110022849B (en) | 2022-11-15 |
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| CN201780072676.3A Active CN110022849B (en) | 2016-09-23 | 2017-08-17 | Skin whitening composition containing SH3BP 4-inhibiting substance and method for screening SH3BP 4-inhibiting substance |
Country Status (3)
| Country | Link |
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| KR (1) | KR20180032954A (en) |
| CN (1) | CN110022849B (en) |
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| CN110343755B (en) * | 2018-04-03 | 2022-08-09 | 伽蓝（集团）股份有限公司 | Method for screening active substance for regulating skin pigmentation and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6379887B1 (en) * | 1998-09-10 | 2002-04-30 | The General Hospital Corporation | Gene for inhibiting melanin biosynthesis |
| US20060228370A1 (en) * | 2003-07-18 | 2006-10-12 | Charite-Universitaets-Medezin Berlin | 7a5/Prognostin and use thereof for the diagnostic and therapy of tumors |
| US20090175827A1 (en) * | 2006-12-29 | 2009-07-09 | Byrom Mike W | miR-16 REGULATED GENES AND PATHWAYS AS TARGETS FOR THERAPEUTIC INTERVENTION |
| US20090203574A1 (en) * | 2006-05-31 | 2009-08-13 | Elaine Fuchs | Method for Modulating Hair Growth |
| US20140134636A1 (en) * | 2011-06-23 | 2014-05-15 | Amorepacific Corporation | Composition for controlling chromogenesis including microrna |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR101207561B1 (en) * | 2009-12-15 | 2012-12-04 | 주식회사 코리아나화장품 | siRNA oligonucleotide which inhibits Tryrosinase expression and Cosmetic composition comprising the same |
| KR102212623B1 (en) * | 2014-04-16 | 2021-02-09 | (주)아모레퍼시픽 | Skin-improving material controlling expression of cadherin11 or N-cadherin and method for screening the material |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6379887B1 (en) * | 1998-09-10 | 2002-04-30 | The General Hospital Corporation | Gene for inhibiting melanin biosynthesis |
| US20060228370A1 (en) * | 2003-07-18 | 2006-10-12 | Charite-Universitaets-Medezin Berlin | 7a5/Prognostin and use thereof for the diagnostic and therapy of tumors |
| US20090203574A1 (en) * | 2006-05-31 | 2009-08-13 | Elaine Fuchs | Method for Modulating Hair Growth |
| US20090175827A1 (en) * | 2006-12-29 | 2009-07-09 | Byrom Mike W | miR-16 REGULATED GENES AND PATHWAYS AS TARGETS FOR THERAPEUTIC INTERVENTION |
| US20140134636A1 (en) * | 2011-06-23 | 2014-05-15 | Amorepacific Corporation | Composition for controlling chromogenesis including microrna |
Non-Patent Citations (5)
| Title |
|---|
| HAH YS，等: "Induction of melanogenesis by rapamycin in human MNT-1 melanoma cells", 《ANN DERMATOL》 * |
| KIM YM, 等: "SH3BP4 is a negative regulator of amino acid-Rag GTPase-mTORC1 signaling", 《MOL CELL》 * |
| KYU-HAN KIM，等: "Novel inhibitory function of miR-125b in melanogenesis", 《PIGMENT CELL MELANOMA RES》 * |
| 何洁凝等: "RNA 干扰的研究进展", 《生命科学研究》 * |
| 张德华: "《蛋白质与酶工程》", 30 September 2015, 合肥工业大学出版社 * |
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| CN110022849B (en) | 2022-11-15 |
| WO2018056583A1 (en) | 2018-03-29 |
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