CN110022849B - Skin whitening composition containing SH3BP 4-inhibiting substance and method for screening SH3BP 4-inhibiting substance - Google Patents
Skin whitening composition containing SH3BP 4-inhibiting substance and method for screening SH3BP 4-inhibiting substance Download PDFInfo
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Abstract
Disclosed are a composition for skin whitening comprising a substance for inhibiting SH3BP4 expression or activity as an active ingredient, and a method for screening a substance for inhibiting SH3BP4 expression or activity. The composition comprising a substance for inhibiting SH3BP4 expression or activity may provide excellent whitening effects by reducing the melanin content in the skin. In addition, according to the above screening method, the relative expression inhibition degree of the SH3BP4 gene can be confirmed, thereby easily screening skin whitening substances.
Description
Technical Field
The present invention relates to a composition for skin whitening and a screening method for skin whitening substances.
Background
Melanin produced by the action of various enzymes such as tyrosinase in melanocytes of the human body is the most important factor for determining the color of human skin, and when the melanin is produced excessively more than necessary, pigmentation disorders such as chloasma, freckles and nevi are induced, resulting in poor cosmetic results. In addition, melanin produced by stress stimulation inside and outside the skin does not disappear with the disappearance of stress before being discharged through keratinization of the skin.
Therefore, studies have been made to inhibit the production of such melanin, and ascorbic acid, kojic acid, arbutin, hydroquinone, glutathione or derivatives thereof, or substances inhibiting tyrosinase activity have been used in cosmetics and pharmaceuticals. However, their use is limited due to insufficient whitening effect of these substances and safety problems to the skin.
[ references in the related art ]
[ patent document ]
(patent document 1) korean patent publication No. 10-2013-0025768 (03.12.2013).
Disclosure of Invention
Technical problem
In one aspect, it is an object of the present invention to provide an identification marker for a novel skin whitening substance for regulating melanin synthesis in the skin.
In another aspect, the present invention is directed to a composition for skin whitening comprising a substance for inhibiting the expression or activity of the skin whitening substance identification marker as an active ingredient.
In another aspect, it is an object of the present invention to provide a method for screening skin whitening substances by using the skin whitening substance discrimination marker.
In another aspect, the present invention is directed to a composition or kit for diagnosing a pigment-related skin condition by using the skin whitening substance discrimination marker.
In another aspect, the present invention is directed to a basic information providing method for diagnosing a pigment-related skin condition by using the skin whitening substance discrimination marker.
Technical scheme
In one aspect, the present invention provides a composition for skin whitening, comprising a substance for inhibiting the expression or activity of SH3BP4 (SH 3 domain binding protein 4) as an active ingredient.
In one aspect, the present invention provides a method for screening a skin whitening substance, comprising the steps of:
(a) Treating melanocytes (melanocytes) with a test substance; and
(b) It was confirmed whether the test substance can inhibit the expression of SH3BP4 (SH 3 domain-binding protein 4) in melanocytes.
In one aspect, the present invention provides a composition for diagnosing a pigment-related skin condition, the composition comprising a reagent for detecting SH3BP4mRNA or SH3BP4 protein.
In one aspect, the present invention provides a kit for diagnosing a pigment-associated skin condition, the kit comprising a reagent for detecting SH3BP4mRNA or SH3BP4 protein.
In one aspect, the present invention provides a method for providing basic information for diagnosing a pigment-associated skin condition, the method comprising confirming an expression amount of mRNA or protein of SH3BP4 from skin cells isolated from a subject.
Advantageous effects
The novel skin whitening substance discrimination marker SH3BP4 according to an aspect of the present invention can reduce the synthesis of melanin in the skin. Therefore, a composition comprising a substance for inhibiting the expression or activity of SH3BP4 as an active ingredient may be effectively used for skin whitening. Further, according to an aspect of the present invention, a screening method for skin whitening substances is to easily and efficiently identify the skin whitening substances by treating a test substance to the skin and confirming the relative expression inhibition degree of SH3BP4 gene. According to the composition or kit for diagnosing a pigment-related skin state and the basic information providing method for diagnosing a pigment-related skin state of the present invention, it is possible to quickly and simply diagnose a pigment-related skin state by detecting the SH3BP4 gene contained in the skin, or to provide basic information of a pigment-related skin state.
Drawings
FIG. 1 is a graph comparing the relative contents of melanin in melanocytes according to the degree of inhibition of SH3BP4 gene expression.
FIG. 2 is a graph for confirming the degree of reduction of Tyrosinase (TYR) expression in melanocytes according to the degree of inhibition of SH3BP4 gene expression.
FIG. 3 is a graph comparing Tyrosinase (TYR) activity in melanocytes according to the degree of inhibition of SH3BP4 gene expression.
Detailed Description
The present invention is described in detail below.
In the present specification, the term "skin" refers to an organ covering the outside of a living body, which is composed of epidermis, dermis and subcutaneous fat layer, and is the broadest concept including not only tissues covering the entire face or the outside of the body but also scalp and hair.
One embodiment of the present invention may provide a method for skin whitening, which includes the step of administering a substance for inhibiting the expression or activity of SH3BP4 to a subject in need of skin whitening.
Another embodiment of the present invention may provide a composition for skin whitening, which includes a substance for inhibiting the expression or activity of SH3BP4 as an active ingredient.
Another embodiment of the present invention may provide use of a substance for inhibiting the expression or activity of SH3BP4 in preparing a composition for skin whitening.
In addition, another embodiment of the present invention may provide an inhibitory substance of the expression or activity of SH3BP4 for skin whitening.
In one embodiment of the present invention, the substance for inhibiting the expression or activity of SH3BP4 may be a substance for inhibiting the translation of SH3BP4mRNA, and more specifically, may comprise an oligonucleotide bound to at least a portion of SH3BP4 mRNA.
Whether the SH3BP4 has a relation with melanin synthesis is not yet disclosed, but the inventors of the present invention found through research that the SH3BP4 can regulate melanin synthesis in skin melanocytes. More specifically, it has been confirmed that when the expression or activity of SH3BP4 is inhibited or decreased in melanocytes of the skin, there is a decrease in the amount of melanin production, or a decrease in the expression and activity of tyrosinase (tyrosinase).
In one embodiment of the present invention, the oligonucleotide may be any one or more of siRNA, shRNA and miRNA. The substance for inhibiting the expression or activity of SH3BP4 may be any one or more of siRNA, shRNA and miRNA that can induce an RNA interference (RNAi) phenomenon. One embodiment of the present invention can inhibit the mRNA expression of the SH3BP4 gene by using the RNAi phenomenon for inducing SH3BP4mRNA interference.
miRNA is a small RNA (endogenous small RNA) present in cells, which is derived from DNA that does not synthesize proteins, and is produced from hairpin-shaped transcripts (hairpin-shaped transcript). miRNA binds to a complementary sequence of 3' -UTR of target mRNA to induce translational inhibition or destabilization of mRNA, and finally acts as an inhibitor (suppressor) to inhibit protein synthesis of target mRNA. It is known that one miRNA can target several mrnas, and that mrnas can also be regulated by several mirnas. Other RNAs for inducing RNAi phenomenon include short interfering RNA (siRNA) which is short RNA of about 19-27mer, and shRNA having short hairpin structure.
In one embodiment of the present invention, the oligonucleotide is double-stranded, specifically, may be any one of siRNA, shRNA or miRNA, and comprises SEQ ID NO:1 and nucleotide sequence shown in SEQ ID NO:1 a hybridizable nucleotide sequence.
5'-GCGGUAUGAUUGAUAAUCU-3'(SEQ ID NO：1)
In one embodiment, the oligonucleotide is double-stranded, specifically, may be any one of siRNA, shRNA and miRNA and comprises SEQ ID NO:1 and SEQ ID NO:2, and the nucleotide sequence shown in SEQ ID NO:1 and SEQ ID NO:2 can hybridize to each other.
Sense 5 'GCGGUAUGAUUGAUAAUCU-3' (SEQ ID NO: 1)
Antisense 5 'AGAUUAUCAAUCAUACCGC-3' (SEQ ID NO: 2)
In one embodiment, the oligonucleotide is double-stranded, specifically, may be any one of siRNA, shRNA or miRNA, and comprises SEQ ID NO:3 and a nucleotide sequence corresponding to the nucleotide sequence shown in SEQ ID NO:3 a hybridizable nucleotide sequence.
5'-GCGUAUGACUUCUUACUCA-3'(SEQ ID NO：3)
In one embodiment, the oligonucleotide is double-stranded, specifically, may be any one of siRNA, shRNA and miRNA and comprises SEQ ID NO:3 and SEQ ID NO:4, and the nucleotide sequence shown in SEQ ID NO:3 and SEQ ID NO:4 can hybridize to each other.
Sense: 5 'GCGUAUGACUUCUUCA-3' (SEQ ID NO: 3)
Antisense 5 'UGAGUAAGAAGUCAUACGC-3' (SEQ ID NO: 4)
The oligonucleotide according to one embodiment of the present invention may be included in a form of being entrapped in a liposome, or being inserted in a vector.
In one embodiment, the concentration of the oligonucleotide may be 1-200nM. More specifically, the concentration of the oligonucleotide may be 5-50nM.
The concentration of the oligonucleotide may be 1nM or more, 2nM or more, 3nM or more, 5nM or more, 7nM or more, 9nM or more, 11nM or more, 15nM or more, 16nM or more, 17nM or more, 18nM or more, 19nM or more, 20nM or more, 21nM or more, 22nM or more, 23nM or more, 24nM or more, 25nM or more, or 50nM or more.
Furthermore, the concentration of the oligonucleotide may be 200nM or less, 190nM or less, 180nM or less, 170nM or less, 150nM or less, 130nM or less, 100nM or less, 80nM or less, 50nM or less, 40nM or less, 30nM or less, 29nM or less, 28nM or less, 27nM or less, 26nM or less, 25nM or less, 24nM or less, 23nM or less, 22nM or less, 21nM or less, 20nM or less, 19nM or less, 18nM or less, 17nM or less, 16nM or less, 15nM or less, 14nM or less, 13nM or less, 12nM or less, 11nM or less, or 10nM or less.
When the concentration of the oligonucleotide is within the above range, the SH3BP4 expression-inhibiting effect is excellent, and it is suitable for skin whitening.
In one example, the mRNA sequence of SH3BP4 (SH 3 domain binding protein 4) is NCBI reference sequence NM _014521.2, more specifically, can be represented by SEQ ID NO:5 in the sequence listing.
The composition according to one embodiment of the present invention may comprise the substance for inhibiting the expression or activity of SH3BP4 in an amount of 0.00001 to 30% by weight based on the total weight of the composition.
Further, the composition according to one embodiment of the present invention may be a skin external composition, and more particularly, may include a cosmetic composition or a pharmaceutical composition.
In one embodiment, the cosmetic composition may be provided in all dosage forms suitable for topical application. For example, the dosage form may be provided as a solution, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an aqueous phase in an oil phase, a suspension, a solid, a gel, a powder, a paste, a foam, or an aerosol composition. Compositions of such dosage forms may be prepared according to conventional methods in the art.
In one embodiment, the cosmetic composition may preferably further comprise other components capable of producing a synergistic effect with the primary effect, within a range not affecting the primary effect, in addition to the above-mentioned substances. The cosmetic composition according to the present invention may include a substance selected from the group consisting of vitamins, high molecular peptides, high molecular polysaccharides, and sphingolipids. Further, in one embodiment, the cosmetic composition may comprise moisturizers, lubricants, surfactants, ultraviolet absorbers, preservatives, bactericides, antioxidants, pH adjusters, organic and inorganic pigments, fragrances, coolants, antiperspirants. The content of the above-mentioned components can be easily selected by those skilled in the art within a range not affecting the object and effect of the present invention, and may be 0.01 to 5% by weight, specifically, 0.01 to 3% by weight based on the total weight of the composition.
Furthermore, in one embodiment, the pharmaceutical composition may be provided in all dosage forms suitable for topical application. For example, administration can be by oral, transdermal, intravenous, intramuscular, subcutaneous injection. In one embodiment, the pharmaceutical composition may be an injection, a skin external preparation, a suspension, an emulsion, a gel, a patch, or a spray, but is not limited thereto. The dosage form can be easily prepared according to a conventional method in the art, and a surfactant, an excipient, a wettable powder, an emulsification promoter, a suspending agent, a salt or buffer for controlling osmotic pressure, a colorant, a fragrance, a stabilizer, a preservative or other commonly used adjuvants can be suitably used.
The effective ingredients of the pharmaceutical composition according to one embodiment of the present invention vary according to the age, sex, body weight, pathological conditions and severity of the subject, the administration route or the judgment of the prescriber. It is within the level of ordinary skill in the art to determine the dosage based on such factors, for example, a daily dosage of from 0.1 mg/kg/day to 5000 mg/kg/day, more specifically from 50 mg/kg/day to 500 mg/kg/day, but not limited thereto.
Another embodiment of the present invention may provide a screening method for skin whitening substances by using the SH3BP 4.
In one embodiment, the method comprises the steps of: (a) treating the melanocytes (melanocytes) with a test substance; and (b) confirming whether the test substance inhibits the expression of SH3BP4 in melanocytes. Wherein, whether the expression of SH3BP4 can be inhibited in the step (b) or not can be whether the translation of SH3BP4mRNA can be inhibited or not.
In one embodiment, the step (b) may include a step of confirming a relative expression inhibition degree of SH3BP4 before and after the test substance is treated by the melanocyte. Further, according to one embodiment, the step (b) may include a step of confirming a relative expression inhibition degree of the melanocytes of the treated test substance and the melanocytes of the untreated test substance.
Furthermore, according to an embodiment, the screening method may further include: a step of determining the test substance as a skin whitening substance when the test substance can inhibit the expression of SH3BP4 after confirming the result of the step (b).
In the present specification, the term "relative expression inhibition degree" may be a degree to which the expression of SH3BP4 in melanocytes before the treatment of a test substance is inhibited as compared with the expression level of SH3BP4 in melanocytes after the treatment of the test substance. Alternatively, the "relative expression inhibition degree" may be a degree of inhibition of expression when the expression level of SH3BP4 of melanocytes of treated test substance is compared with the expression level of SH3BP4 of melanocytes of untreated test substance. The expression level includes expression amount and expression quality (quality). In addition, the expression level, for example, may include the expression level of the protein. More specifically, the expression level may include the degree of melanin synthesis in melanocytes, or the expression level or degree of synthesis of tyrosinase.
In one embodiment of the present invention, the step of determining the skin whitening substance may include: when the "degree of inhibition of relative expression" is two times or more, it is judged as a skin whitening substance. That is, when the expression level of SH3BP4 in the melanocytes before the treatment of the test substance is inhibited by about 1.1 to 2 times or more as compared with the expression level of SH3BP4 in the melanocytes after the treatment of the test substance, it can be judged as a skin whitening substance. Or, when the expression level of SH3BP4 in melanocytes of a treated test substance is inhibited by 1.1-2 times or more as compared with the expression level of SH3BP4 in melanocytes of an untreated test substance, it can be judged as a skin whitening substance.
For example, the relative expression inhibition degree of SH3BP4 after the treatment of the test substance may be 1.1-fold or more, 1.2-fold or more, 1.3-fold or more, 1.4-fold or more, 1.5-fold or more, 1.6-fold or more, 1.7-fold or more, 1.8-fold or more, 1.9-fold or more, or 2-fold or more, compared to before the treatment of the test substance, but is not limited thereto. The expression level is the result measured under conditions ensuring statistical significance. The term "statistically significant" refers to the case where a significant difference is observed by a biometric analysis, and includes the case where the p-value at the time of quantification is less than 0.05.
In step (a) according to one embodiment, the test substance may be transfected into a liposome or vector of the cell, and the cell may be a melanocyte, i.e., a melanocyte. The transfection may be performed by using a microinjection method, a calcium phosphate coprecipitation method, an electroporation method, a liposome application method, or the like, but is not limited thereto.
In addition, in one embodiment, the expression level of SH3BP4 may be determined using known techniques, such as, but not limited to, polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), western blotting, or immunoblotting (immuneblot).
In addition, one embodiment of the present invention may provide a biomarker for diagnosing the degree of pigment-related skin condition. Further, an embodiment of the present invention may provide a composition for diagnosing a pigment-related skin condition, or a kit for diagnosing a pigment-related skin condition, by using the biomarker.
Another embodiment of the present invention may provide a basic information providing method for diagnosing a pigment-related skin state, or a method for diagnosing a pigment-related skin state.
Specifically, one embodiment of the present invention can provide a composition for diagnosing a pigment-associated skin condition, which comprises the reagent for detecting SH3BP4mRNA or SH3BP4 protein.
In addition, another embodiment of the present invention may provide use of a reagent for detecting mRNA of SH3BP4 or SH3BP4 protein in preparing a composition for diagnosing a pigment-related skin condition.
In addition, another embodiment of the present invention can provide a detection reagent for SH3BP4mRNA or SH3BP4 protein for diagnosing pigment-related skin conditions.
Further, another embodiment of the present invention may provide a basic information providing method for diagnosing a pigment-associated skin condition or a method for diagnosing a pigment-associated skin condition, the method including the step of confirming an expression amount of SH3BP4 from skin cells isolated from a subject. At this time, in one example, the SH3BP4 expression amount may be an expression amount of mRNA of SH3BP4 or SH3BP4 protein, and the skin cells isolated from the subject and the skin cells of the normal control group may be melanocytes.
In one embodiment, the basic information providing method for diagnosing a pigment-associated skin state or the method for diagnosing a pigment-associated skin state further comprises the step of comparing the expression level of SH3BP4 in the skin cells isolated from the subject with the expression level of SH3BP4 in skin cells of a normal control group. Further, in one embodiment, the method further comprises the step of providing information or making a diagnosis that there is an abnormality in the pigment-associated skin state when the SH3BP4 expression level in the skin cells isolated from the subject is higher than the SH3BP4 expression level in normal control skin cells. For example, if the expression level of SH3BP4 in the skin cells isolated from the subject is about 1.1-2 times or more higher than the expression level of SH3BP4 in skin cells of a normal control group, it can be judged that there is an abnormality in the pigment-associated skin state. More specifically, for example, when the expression level of SH3BP4 in the skin cells isolated from the subject is 1.1-fold or more, 1.2-fold or more, 1.3-fold or more, 1.4-fold or more, 1.5-fold or more, 1.6-fold or more, 1.7-fold or more, 1.8-fold or more, 1.9-fold or more, or 2-fold or more higher than that of the skin cells of the normal control group, the skin condition related to the pigment can be judged to be abnormal. The expression level is the result of measurement under conditions ensuring statistical significance. The concept of statistical significance refers to the case of significant differences exhibited by biology, including the case where the p-value at the time of quantification is less than 0.05.
In addition, another embodiment of the present invention may provide a kit for diagnosing a pigment-related skin condition, comprising a reagent for detecting SH3BP4mRNA or SH3BP4 protein, which may further include an instruction manual describing the above-described basic information providing method for diagnosing a pigment-related skin condition or a diagnostic method for a pigment-related skin condition.
In addition, in one embodiment, the composition for diagnosing a pigment-related skin state or the SH3BP4mRNA detecting reagent contained in the kit for diagnosing a pigment-related skin state may include one or more of a primer and a probe that specifically bind to SH3BP 4. In one embodiment, the SH3BP4 protein detection reagent may comprise one or more of an antibody and a probe that specifically binds to SH3BP 4.
Hereinafter, the constitution and effect of the present invention will be described in more detail with reference to examples. However, the following examples are only for the purpose of facilitating understanding of the present invention, and do not limit the scope and the scope of the present invention.
[ Experimental example 1 ]
The decrease in pigmentation by inhibiting the expression of SH3BP4 was confirmed by the following experiment.
Human primary melanocytes (modified segmented Human primary melanocytes) isolated from moderately pigmented tissues (purchased from Thermofish Co., https:// www. Thermofisher. Com/order/catalog/product/C1025 CICICICICICICICICICICICICID = search-product) were prepared.
In order to reduce the expression of SH3BP4 in the melanocytes, the following two SH3BP4 sirnas exhibiting specificity for SH3BP4mRNA sequences were introduced.
Example 1 si-SH3BP4#1
5 'positive-sense GCGGUAUGAUAUAAUCU-3' (SEQ ID NO: 1)
Antisense 5 'AGAUUAUCAAUCAUACCGC-3' (SEQ ID NO: 2).
Example 2 si-SH3BP4#2
Sense: 5 'GCGUAUGACUUCUUCA-3' (SEQ ID NO: 3)
Antisense 5 'UGAGUAAGAAGUCAUACGC-3' (SEQ ID NO: 4).
Specifically, the SH3BP4 siRNA of example 1 was transfected into cells by using liposome (liposome) on the Human primary Melanocyte (Human primary Melanocyte) as a Melanocyte line until the SH3BP4 siRNA concentration reached 20nM. The liposomes used LipofectamineRNAiMAX reagent (Thermo Fisher Scientific, https:// www. Thermolisher. Com/order/catalog/product/13778150). The SH3BP4 siRNA of example 2 was also transfected into cells by using liposome (liposome) by the same method as example 1 above until the SH3BP4 siRNA concentration reached 20nM.
As a Negative Control group, siRNA that had been functionally confirmed to have not reduced any gene in humans and had minimal effect on cell proliferation and cell activity, namely Silene Select Negative Control No.1siRNA (Thermo Fisher Scientific, https:// www. Thermo Fisher. Com/order/catalog/product/4390843ICID = search-product) was used. The siRNA of the negative control group was also transfected into cells by using liposome (liposome) according to the same method as in example 1 above until the siRNA concentration reached 20nM.
The cells of examples 1 and 2, negative control group, transfected by each protocol were, at 37 ℃,5% CO 2 The culture was carried out in an incubator for about 7 days.
After the cells were washed twice with 5ml of PBS buffer, 1ml of PBS buffer was added, and then all the cells were scraped off with a scraper, centrifuged (centrifugation) at 13000rpm for 5 minutes to prepare cell pellets (cell pellet), and the degree of pigment formation was compared by observing the color of the cell pellets. Further, the amount of melanin was determined by measuring the absorbance at 450nm after dissolving the cell pellet in 1N NaOH in a volume of 2001. Mu.l.
As a result, as shown in fig. 1, it was confirmed that the amount of melanin contained in melanocytes was significantly reduced by about 2-fold or more, compared to the negative control group in examples 1 and 2 in which SH3BP4 siRNA was used to inhibit the expression of SH3BP 4. This means that when the expression or activity of SH3BP4 in the skin is inhibited, a skin whitening effect can be exhibited.
[ Experimental example 2 ]
The reduction of tyrosinase expression by inhibiting SH3BP4 expression was determined according to the following experiment by using the same human primary melanocytes as [ experimental example 1 ], SH3BP4 sirnas of examples 1 and 2, and sirnas of negative control.
First, the SH3BP4 sirnas of examples 1 and 2 and the sirnas of the negative control group were transfected into cells by using liposomes, respectively, according to the same method as [ experimental example 1 ] until the SH3BP4 siRNA and the siRNA concentrations reached 20nM, respectively.
The cells of each of the transfected examples 1 and 2, negative control group were subjected to 5% CO at 37 ℃ 2 The culture was carried out in an incubator for about 3 days.
The cell membrane of each cell was disrupted by using RIPA buffer, and the supernatant containing the protein was extracted using a centrifuge. Then, about 10. Mu.g of protein was quantified by BCA (bicinchoninic acid) method. Western blot experiments were performed by using the quantified protein. Wherein the amounts of tyrosinase and SH3BP4 protein were observed by using tyrosinase-specific antibody (catalog No. Sc-20035. The expression levels of tyrosinase and SH3BP4 protein were normalized by the protein expression level of β -actin (Cat. No. 3280, abcam), which is a housekeeping gene.
Then, in the supernatant containing the protein isolated by using the above RIPA, 10. Mu.g of the protein was added to 100. Mu.10 mM 1-dihydroxyphenylalanine (solvent is 50mM sodium phosphate buffer (pH 6.8)), reacted at 37 ℃ for 1 hour, the absorbance at 490nm was measured and the activity of tyrosinase (tyrocynase) was analyzed.
As a result, as shown in FIG. 2, in examples 1 and 2 in which the expression of SH3BP4 was inhibited by using SH3BP4 siRNA, there was a decrease in the expression amount of tyrosinase in melanocytes. In addition, as shown in fig. 3, it was confirmed that the activity of tyrosinase (TYR activity) was significantly reduced by about 2 times or more compared to the negative control. This means that when the expression or activity of SH3BP4 in the skin is inhibited, a skin whitening effect can be exhibited.
The following describes dosage form examples of the composition according to one embodiment of the present invention, and various other dosage forms may be applied, which are intended to describe the invention in detail, but not to limit the invention.
[ dosage form example 1 ] nutrient astringent
The nutritional lotions were prepared according to the conventional methods according to the compositions shown in table 1 below.
[ TABLE 1 ]
| Name content of raw materials | Content (wt%) |
| Glycerol | 3.0 |
| Butanediol | 3.0 |
| Propylene glycol | 3.0 |
| Carboxyvinyl polymer | 0.1 |
| Liposomes comprising the liposomes of example 1 or 2 | 0.01 |
| Beeswax (Cera flava) | 4.0 |
| Polysorbate 60 | 1.5 |
| Caprylic/capric triglyceride | 5.0 |
| Squalane | 5.0 |
| Sorbitan sesquioleate | 1.5 |
| Cetostearyl alcohol | 1.0 |
| Triethanolamine | 0.2 |
| Antiseptic and spice | Proper amount of |
| Purified water | Balance of |
| Total of | 100 |
Dosage form example 2 nutritional emulsions
The nutritional emulsions were prepared according to conventional methods, following the compositions shown in table 2 below.
[ TABLE 2 ]
[ dosage form example 3 ] nourishing cream
A nourishing cream was prepared according to a conventional method with the composition shown in the following table 3.
[ TABLE 3 ]
| Name content of raw materials | Content (wt%) |
| Glycerol | 3.5 |
| Butanediol | 3.0 |
| Liquid paraffin | 7.0 |
| Beta-glucan | 7.0 |
| Carbomer | 0.1 |
| Liposomes comprising the liposomes of example 1 or 2 | 0.01 |
| Caprylic/capric triglyceride | 3.0 |
| Squalane | 5.0 |
| Cetearyl glucoside | 1.5 |
| Sorbitan stearate | 0.4 |
| Polysorbate 60 | 1.2 |
| Triethanolamine | 0.1 |
| Antiseptic and spice | Proper amount of |
| Purified water | Balance of |
| Total of | 100 |
[ PREPARATION EXAMPLE 4 ] injection
Injections were prepared according to the conventional method with the composition shown in the following table 4.
[ TABLE 4 ]
| Name content of raw materials | Weight ratio (per 1 ml) |
| Sodium chloride | 9.0mg |
| Sodium carboxymethylcellulose | 30.0mg |
| Tween 20 | 1.0mg |
| Liposomes comprising the liposomes of example 1 or 2 | 1.0mg |
| Distilled water for injection | Balance of |
[ PREPARATION EXAMPLE 5 ] ointments
Ointments were prepared according to the conventional method with the composition shown in the following table 5.
| [ TABLE 5 ] raw material name content | Content (wt%) |
| Glycerol | 8.0 |
| Butanediol | 4.0 |
| Liquid paraffin | 15.0 |
| Beta-glucans | 7.0 |
| Carbomer | 0.1 |
| Examples 1 and 2 | 0.01 |
| Caprylic/capric triglyceride | 3.0 |
| Squalane | 1.0 |
| Cetearyl glucoside | 1.5 |
| Sorbitan stearate | 0.4 |
| Cetostearyl alcohol | 1.0 |
| Preservative | Proper amount of |
| Perfume | Proper amount of |
| Pigment | Proper amount of |
| Beeswax (Cera flava) | 4.0 |
| Purified water | Balance of |
| Total of | 100 |
<110> Kabushiki Althaea
<120> skin whitening composition containing SH3BP 4-inhibiting substance and method for screening SH3BP 4-inhibiting substance
<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 (5)
1. Use of a substance for inhibiting the expression or activity of SH3BP4 in the preparation of a composition for skin whitening, wherein the substance for inhibiting the expression or activity of SH3BP4 comprises a peptide consisting of SEQ ID NO:1 and SEQ ID NO:2 by hybridization; or comprises a sequence defined by SEQ ID NO:3 and SEQ ID NO:4 in sequence listing.
2. The use according to claim 1, characterized in that the substance inhibiting the expression or activity of SH3BP4 is an oligonucleotide included in a form entrapped within liposomes or inserted in a carrier.
3. Use according to claim 1, characterized in that the composition is a cosmetic composition for the skin.
4. Use according to claim 1, wherein the composition is a pharmaceutical composition.
5. Use according to claim 1, characterized in that the composition comprises from 0.00001 to 30% by weight, relative to the total weight of the composition, of a substance for inhibiting the expression or activity of SH3BP 4.
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| KR10-2016-0122317 | 2016-09-23 | ||
| 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 |
| 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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| WO2007140390A2 (en) * | 2006-05-31 | 2007-12-06 | The Rockefeller University | 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 |
| KR101207561B1 (en) * | 2009-12-15 | 2012-12-04 | 주식회사 코리아나화장품 | siRNA oligonucleotide which inhibits Tryrosinase expression and Cosmetic composition comprising the same |
| KR101938548B1 (en) * | 2011-06-23 | 2019-01-15 | (주)아모레퍼시픽 | Composition for regulating expression of pigmentation-related genes containing microRNA |
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