Wrinkle-improving composition containing a retinol booster as an active ingredient

Abstract

The present invention relates to a composition for improving wrinkles, which contains retinol and a retinol booster as active ingredients, and a method for providing information for selecting a customized retinol booster for improving wrinkles, which comprises a step of identifying the base of a polymorphic site of a genetic polymorphic marker having a significant association with the degree of skin wrinkles.

Description

[Technical field]

[0001] The present invention relates to a composition for improving wrinkles, which contains retinol and a retinol booster as active ingredients, and a method for providing information for selecting a customized retinol booster for improving wrinkles, which comprises a step of identifying the base of a polymorphic site of a genetic polymorphic marker having a significant association with the degree of skin wrinkles. [Background technology]

[0002] Skin is the outermost tissue of the body, and is constantly in contact with the external environment to protect the human body from harmful factors. It is also an important part that is visible and affects aesthetic aspects. When many signs of aging such as wrinkles and pigmentation appear on the skin, the aesthetic value decreases, so there has always been a need from consumers for solutions to prevent or improve such signs. In particular, wrinkles on the skin are considered to be the most representative indicator of age, so wrinkle-improving cosmetics have become an essential item for women who want youthful skin.

[0003] Retinol is the raw material that is expected to have the best efficacy among existing wrinkle improvement raw materials, but some consumers do not achieve the expected improvement in wrinkles due to problems such as irritation reactions and low reactivity to retinol. In addition, even if a consumer is expected to achieve effective wrinkle improvement due to high reactivity to retinol, if the consumer is unable to recognize this, they may not be able to endure the adaptation period accompanied by irritation, etc., and may discontinue use, resulting in failure to achieve the expected effect. Thus, individual differences in efficacy are a major obstacle that reduces the perceived efficacy and satisfaction of skin improvement products (Patent Document 1).

[0004] Currently, a wide variety of gene polymorphism markers have been disclosed, but most of them are related to skin color, whitening, moisture, ultraviolet rays, etc., and the mechanism of action of few of them has been studied. In particular, it can be said that there are almost no polymorphism markers related to wrinkles for which the mechanism has been studied or customized materials have been developed. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Korean Patent Publication No. 10-2009-0122770 [Non-patent literature]

[0006] [Non-Patent Document 1] S. Kang et al., J. Investigative Dermatology, 105, 549-556, 1995 [Non-Patent Document 2] Wu and Wallace, Genomics 4, 560 (1989) [Non-Patent Document 3] Landegren et al., Science 241, 1077 (1988) [Non-Patent Document 4] Kwoh et al., Proc. Natl. Acad. Sci. USA 86, 1173 (1989) [Non-Patent Document 5] Guatelli et al., Proc. Natl. Acad. Sci. USA 87, 1874 (1990) Summary of the Invention [Problem to be solved by the invention]

[0007] The inventors aimed to establish a classification system for Korean skin characteristics by constructing big data of genetic information and skin type information, and made earnest efforts to identify the genetic characteristics that determine an individual's skin characteristics, establish scientific skin classification standards, develop personalized active ingredients based on the standards, and contribute to the development of customized cosmetics for each skin characteristic through various product subdivisions. As a result, they selected specific single nucleotide polymorphism (SNP) markers that have a significant correlation with skin wrinkles, and confirmed that it is possible to provide a personalized retinol booster that supports wrinkle improvement based on genetic information, thereby completing the present invention. [Means for solving the problem]

[0008] An object of the present invention is to provide a composition for improving wrinkles that contains retinol and a retinol booster as active ingredients.

[0009] Another object of the present invention is to provide a cosmetic composition or a quasi-drug composition containing the above composition as an active ingredient.

[0010] Furthermore, the present invention aims to provide an information providing method for selecting a customized retinol booster for improving wrinkles, comprising a step of identifying a base at a polymorphic site of a single nucleotide polymorphism marker having a significant association with the degree of skin wrinkles. Effect of the Invention

[0011] The genetic polymorphism markers of the present invention having a significance associated with the degree of skin wrinkles can provide information regarding an individual's degree of skin wrinkles, and further, can allow the development of customized ingredients or products that can alleviate the degree of skin wrinkles depending on the information of the genetic polymorphism markers observed in an individual. [Brief description of the drawings]

[0012] [Figure 1] FIG. 1 shows reactivity to retinol depending on whether or not one possesses an allele of a specific gene SNP. [Diagram 2] FIG. 13 shows the wrinkle improvement effect of EDAR (rs3827760) by treatment with a retinol booster (lupeol and sucralfate). [Diagram 3] FIG. 13 shows the wrinkle-improving effect of retinol booster (glutathione) treatment of BNC2 (rs1693073). [Figure 4] FIG. 1 is a schematic diagram showing how, for a customized prescription, the base of a polymorphic site of a gene polymorphic marker having a significant association with the degree of skin wrinkles is identified, and a customized retinol booster for improving wrinkles is selected. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] These will be described in detail below. Note that each description and embodiment disclosed in the present invention is also applicable to other descriptions and embodiments. In other words, all combinations of various elements disclosed in the present invention are included in the present invention. In addition, the present invention is not limited to the following specific description.

[0014] Additionally, those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein which equivalents are intended to be encompassed by the present invention.

[0015] One aspect of the present invention to achieve the object provides a composition for improving wrinkles, comprising retinol and a retinol booster as active ingredients.

[0016] In the present invention, "retinol" is a type of vitamin A, also called pure vitamin. It is a substance that plays an important role in maintaining the original functions of epidermal cells of the skin. It is present in the intestinal mucosa cells of animals and is found in large amounts in green-yellow plants. It can be converted into the active form, retinoic acid, and is known to play an important role in maintaining the original functions of epidermal cells. It is also known to have the effect of promoting cell differentiation by making the DNA in the cell nucleus in skin cells express RNA, promoting the biosynthesis of collagen, a hard protein that exists as a fibrous solid between animal cells, and elastin, which is composed of elastic fibers, thereby reducing wrinkles and increasing skin elasticity.

[0017] The retinol is contained in an amount of about 2000 to 6000 IU, but is not limited thereto, and may be, for example, about 2000 to 6000 IU, about 3500 to 5500 IU, or about 2500 to 4500 IU.

[0018] The present invention has found that the efficacy of retinol on the skin increases in proportion to its concentration up to about 6000 IU, and there is no significant difference above 6000 IU (Non-Patent Document 1), and aims to provide a composition that maximizes the effect on the skin by setting the content at 5000 IU or less as the content that can improve the efficacy of retinol while minimizing skin irritation. In addition, the present invention aims to provide a composition that maximizes the effect of retinol by using various retinol boosters to improve reactivity to retinol.

[0019] In the present invention, the term "retinol booster" refers to a substance that acts together with retinol to enhance the effect of improving wrinkles. Retinol has the effect of reducing skin wrinkles and increasing skin elasticity, but reactivity to retinol varies from person to person. Therefore, the retinol booster may be any substance that acts together with retinol to improve reactivity to retinol and help maximize the effect of improving wrinkles on the skin.

[0020] For example, the retinol booster may be at least one selected from the group consisting of lupeol, sucralfate, and glutathione, but is not limited thereto.

[0021] The above-mentioned "lupeol" refers to a fat-soluble component derived from plants, specifically from legumes, which can directly dissolve sebum, enter pores to kill acne bacteria, and has the function of suppressing various causes of acne, such as sebum production, inflammation, and keratinization, which are the causes of acne.

[0022] The aforementioned "sucralfate" is a drug used to treat stomach ulcers, gastroesophageal reflux disease, stomach inflammation, etc., and to prevent stress ulcers. The salt portion of sucralfate binds to fibroin in the stomach wall to form a membrane, which coats the stomach wall and protects the stomach from gastric acid, bile, etc., thereby relieving stomach pain, and sucralfate functions as an antacid.

[0023] Glutathione (γ-glutamyl-cysteinyl-glycine, GSH) is a peptide consisting of three amino acids and plays an important role in the synthesis of deoxyribonucleotide precursors of DNA. Glutathione acts on the metabolic processing of estrogens, prostaglandins, and leukotrienes, and acts directly or indirectly on the sulfhydryl groups of proteins to regulate the structure of proteins and to protect cells against oxidative and other stresses.

[0024] The retinol booster of the present invention is contained in an amount of about 0.0001 to 10% by weight based on the total weight of the composition, but is not limited thereto. For example, the retinol booster of the present invention may be contained in an amount of about 0.0001 to 10% by weight, about 0.0005 to 5% by weight, about 0.001 to 5% by weight, about 0.01 to 5% by weight, about 0.0001 to 3% by weight, about 0.0005 to 3% by weight, about 0.001 to 3% by weight, about 0.01 to 3% by weight, about 0.0001 to 1% by weight, about 0.0005 to 1% by weight, about 0.001 to 1% by weight, or about 0.01 to 1% by weight based on the total weight of the composition.

[0025] For purposes of the present invention, the retinol booster provides the benefit of promoting the improvement of skin wrinkles.

[0026] For the purposes of the present invention, the term "skin wrinkles" refers to a state in which the elasticity of a part of an individual's skin is reduced and the skin is overlapped. As the skin overlaps, the wrinkled area has a stronger shadow than the surrounding skin, and therefore is relatively darker than the surrounding skin. In addition, since wrinkles are mostly linear in shape, wrinkles are defined as areas of strong shadow that are connected to each other and reach a certain length or more, but are not limited thereto.

[0027] In the present invention, "about" is used before a specific numerical value. In the present invention, "about" includes not only the exact numerical value following the term "about", but also a range that is approximately that numerical value or close to that numerical value. Considering the context in which the number is used, it can be determined whether the numerical value is close to or approximately the specific numerical value mentioned. As an example, "about" indicates a range of -10% to +10% of the specified numerical value. As another example, "about" indicates a range of -5% to +5% of the specified numerical value. However, the present invention is not limited thereto.

[0028] The composition is characterized by being of a personalized type, specifically, a personalized retinol booster is provided for an individual having a single nucleotide polymorphism (SNP) marker that is significantly associated with a specific degree of skin wrinkles.

[0029] For example, a single nucleotide polymorphism (SNP) marker having a significant association with the degree of skin wrinkles is at least one selected from Table 1, but is not limited thereto.

[0030] In the present invention, "polymorphism" means the presence of two or more alleles at one locus, and a single nucleotide polymorphism (SNP) is a difference in a polymorphic site between individuals. A preferred polymorphic marker has two or more alleles that show an occurrence frequency of 1% or more, more specifically 10% or 20% or more, in a selected population. A "genetic polymorphism marker" generally means a marker in which two or more alleles are observed at the same locus (base), and generally may be major allele / major allele, major allele / minor allele, or minor allele / minor allele depending on the individual. In the present invention, it is used interchangeably with "polymorphic marker" and means the base and base site of a minor allele, or is defined together with the number and base position of a chromosome, but is not limited thereto.

[0031] In the present invention, the term "allele" refers to an allele that exists at the same locus of a homologous chromosome and may be any genotype. An allele is used to indicate polymorphism, for example, an SNP has two alleles. It also refers to a combination of two or more bases that have the same chromosomal number and base position, and the bases include a major allele that occurs frequently in individuals of a specific population, and a minor allele that occurs less frequently than the major allele.

[0032] Specifically, the genetic polymorphism markers of the present invention are significant in association with skin wrinkles, and have two alleles, which can be divided into those having major allele / minor allele, minor allele / minor allele, and major allele / major allele.

[0033] When an individual has an allele of a polymorphic marker having significance associated with skin wrinkles, the effect of improving skin wrinkles varies depending on the reactivity to retinol depending on the allele that the individual has. For example, when an individual has major allele / major allele, major allele / minor allele, or minor allele / minor allele, it is possible to determine whether the individual has skin characteristics that are highly or poorly responsive to retinol, i.e., have a high or low degree of wrinkle improvement, and this can provide information when selecting a customized retinol booster to improve wrinkle improvement for an individual with low reactivity to retinol.

[0034] In the present invention, "rs_id" refers to rs-ID, which is an independent index assigned to all SNPs initially registered by NCBI, which began accumulating SNP information in 1998. These rs_ids listed in the tables refer to SNP markers that are polymorphic markers of the present invention.

[0035] The single nucleotide polymorphism marker may be one or more single nucleotide polymorphism markers selected from the single nucleotide polymorphism markers shown in Table 1. The single nucleotide polymorphism markers shown in Table 1 may be used to determine the degree of association with the degree of skin wrinkles.

[0036] The degree of skin wrinkles of the single nucleotide polymorphism markers of the present invention was determined by measuring the frequency of each marker. Such significance is characterized by a p-value, such as, but not limited to, a p-value of less than 0.05, less than 0.01, less than 0.001, less than 0.0001, less than 0.00001, less than 0.000001, less than 0.0000001, less than 0.00000001, or less than 0.000000001. Specifically, the p-value is less than 0.01, more specifically, the p-value is less than 0.001, and even more specifically, the p-value is less than 0.0001, but not limited to these.

[0037] The single nucleotide polymorphism (SNP) marker of the present invention is at least one selected from the markers shown in Table 1, but is not limited thereto. The single nucleotide polymorphism (SNP) marker is at least one, and is used in a combination of at least two or the like, which is a number that can determine wrinkles, but is not limited thereto.

[0038] The marker is a SNP itself, a polynucleotide consisting of a continuous DNA sequence of 5 to 100 bases including the position of the SNP, or a polynucleotide consisting of a complementary sequence thereof, but is not limited to these.

[0039] For example, the single nucleotide polymorphism marker having a significant association with the degree of wrinkles is at least one single nucleotide polymorphism marker present in a gene encoding an EDAR or BNC2 protein, specifically, rs3827760 or rs16935073, but is not limited thereto.

[0040] For example, when the SNP ID is rs3827760, the Chr.Position (GRCh ver.37) is described as "2:109513601" and the Allele is described as G>A, this indicates that the 109513601st base of human chromosome 2 is G or A, and the base to the left of the ">" in the allele indicates the major allele, and the base to the right indicates the minor allele.

[0041] As a specific example, the marker selected from Table 1 is at least one polynucleotide selected from the group consisting of a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 109513601st base of human chromosome 2, which is G or A (rs3827760), a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 16795790th base of human chromosome 9, which is A or C (rs16935073), and complementary polynucleotides thereof. The markers are only partially exemplified in Table 1, but may be selected in the same manner in other chromosomes.

[0042] For the purposes of the present invention, the "degree of skin wrinkles" refers to, but is not limited to, the result of calculating the ratio of all measurement sites to the total of all target areas when a linear area of ​​a certain length or more due to skin wrinkles becomes darker relative to the surrounding skin. Specifically, it means determining whether wrinkles are likely to occur using at least one single nucleotide polymorphism (SNP) marker selected from Table 1. More specifically, a skin type that is likely to cause wrinkles refers to a skin type that ages quickly and has a high rate of wrinkles occurring in a given skin area.

[0043] The alleles of the present invention have the same number of chromosomes in each individual, and include major and minor alleles of SNPs. As the minor alleles of the bases at the polymorphic site of the polymorphic marker increase by one, the major alleles decrease by one, and as the major alleles increase by one, the minor alleles decrease by one. However, the range in which the minor alleles and major alleles increase or decrease is within the range of three types: i) major allele / major allele, ii) major allele / minor allele, and iii) minor allele / minor allele, and the alleles increase or decrease within the range of the three types, but are not limited thereto.

[0044] The composition of the present invention is used as a cosmetic composition and can be formulated into various forms. The cosmetic composition of the present invention can be prepared into a dosage form selected from the group consisting of a solution, an ointment for external use, a cream, a foam, a nutritious lotion, a softening lotion, a pack, a softening lotion, a milky lotion, a makeup base, an essence, a soap, a liquid cleanser, a bath additive, a sunscreen cream, a sun oil, a suspension, an emulsion, a paste, a gel, a lotion, a powder, a surfactant-containing cleanser, an oil, a powder foundation, an emulsion foundation, a wax foundation, a patch, and a spray, but is not limited thereto.

[0045] In addition, the cosmetic composition of the present invention may further contain one or more cosmetically acceptable carriers that are blended in general skin cosmetics, and may appropriately contain, as usual ingredients, for example, oils, water, surfactants, moisturizers, lower alcohols, thickeners, chelating agents, colorants, preservatives, fragrances, etc., but are not limited to these.

[0046] The cosmetically acceptable carrier contained in the cosmetic composition of the present invention varies depending on the formulation.

[0047] When the dosage form of the present invention is an ointment, paste, cream or gel, the carrier component may be an animal oil, a vegetable oil, a wax, a paraffin, a starch, tragacanth, a cellulose derivative, a polyethylene glycol, a silicone, a bentonite, a silica, a talc, a zinc oxide or a mixture thereof.

[0048] When the dosage form of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or mixtures thereof may be used as a carrier component, and particularly in the case of a spray, a propellant such as hydrochlorofluorocarbon, propane / butane or dimethyl ether may be further included.

[0049] When the dosage form of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, for example, water, ethanol, isopropanol, diethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil may be used, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, sesame oil, glycerin aliphatic esters, polyethylene glycol or fatty acid esters of sorbitan may be used.

[0050] When the dosage form of the present invention is a suspension, the carrier component may be a liquid diluent such as water, ethanol, or propylene glycol; a suspending agent such as ethoxylated stearyl alcohol, polyoxyethylene sorbitol ester, or polyoxyethylene sorbitan ester; or microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tragacanth.

[0051] When the dosage form of the present invention is a soap, the carrier component may be an alkali metal salt of a fatty acid, a fatty acid hemiester salt, a fatty acid protein hydrolysate, an isethionate salt, a lanolin derivative, a fatty alcohol, a vegetable oil, glycerin, a sugar, or the like.

[0052] In another embodiment of the present invention, the composition provides a quasi-drug composition for reducing or improving skin wrinkles.

[0053] In the present invention, the term "quasi-drug" refers to an article used for the purpose of diagnosing, curing, improving, mitigating, treating, or preventing a disease in humans or animals, which has a milder effect than a drug. For example, according to the Korean Pharmaceutical Affairs Act, quasi-drugs exclude articles used for medicinal purposes, and include products used for the treatment or prevention of diseases in humans or animals, and products that have a mild effect on the human body or that do not have a direct effect.

[0054] In one embodiment, the quasi-drug composition of the present invention may be prepared in a form selected from the group consisting of body cleansers, shampoos, conditioners, foams, soaps, masks, ointments, creams, lotions, essences, and sprays, but is not limited thereto.

[0055] In yet another aspect, the present invention provides an information providing method for selecting a customized retinol booster for wrinkle improvement, comprising the steps of: (a) obtaining a biological sample from an individual; (b) amplifying or hybridizing with a probe a polymorphic site of a single nucleotide polymorphism marker, which is rs3827760 or rs16935073, from the biological sample; and (c) confirming the base of the amplified or hybridized polymorphic site in the (b) step.

[0056] In the present invention, the term "individual" refers to a subject to be diagnosed for the degree of skin wrinkles. DNA can be obtained from samples such as hair, urine, blood, various body fluids, isolated tissues, isolated cells, and saliva of the specimen, but is not limited thereto.

[0057] The method for obtaining genomic DNA in the step (a) may be any method known to those skilled in the art.

[0058] The step of amplifying the polymorphic site of the single nucleotide polymorphism marker from the DNA obtained in step (a) or hybridizing with a probe may be performed by any method known to those skilled in the art. For example, the target nucleic acid may be amplified by PCR and purified. Other methods that may be used include ligase chain reaction (LCR) (Non-Patent Documents 2 and 3), transcription amplification (Non-Patent Document 4), self-sustained sequence replication (Non-Patent Document 5), and nucleic acid sequence-based amplification (NASBA).

[0059] In the above-mentioned method, the base of the polymorphic site in step (b) may be determined by, but is not limited to, sequencing analysis, hybridization using microarray, allele specific PCR, dynamic allele-specific hybridization (DASH), PCR extension analysis, SSCP, PCR-RFLP analysis or TaqMan method, SNPlex platform (Applied Biosystems), mass spectrometry (e.g., Sequenom's MassARRAY system), mini-sequencing, Bio-Plex system (BioRad), CEQ and SNPstream system (Beckman), Molecular Inversion Probe array technology (e.g., Affymetrix GeneChip), and BeadArray Technologies (e.g., Illumina GoldenGate and Infinium assays). The above method, or other methods available to one skilled in the art, identifies at least one allele at a polymorphic marker, including a microsatellite, SNP, or other type of polymorphic marker. The determination of the base at such a polymorphic site can be specifically performed by a SNP chip.

[0060] The above method further provides information that, when the base at the polymorphic site amplified or hybridized in step (i)(c) contains at least one base of a minor allele determined by the single nucleotide polymorphism marker, the reactivity with retinol is low and treatment with an additional retinol booster is required, or, when the base at the polymorphic site amplified or hybridized in step (ii)(c) contains at least one base of a major allele determined by the single nucleotide polymorphism marker, the reactivity with retinol is low and treatment with an additional retinol booster is required, but is not limited thereto.

[0061] The retinol booster is at least one selected from the group consisting of lupeol, sucralfate and glutathione, but is not limited thereto.

[0062] For the purpose of the present invention, the gene having the significance associated with the degree of wrinkles can provide information for selecting a specific retinol booster for each individual.For example, in an individual having at least one single nucleotide polymorphism marker present in the gene encoding EDAR or BNC2 protein, treatment with at least one selected from the group consisting of lupeol, sucralfate and glutathione as a retinol booster can improve wrinkle improvement by retinol, and provide the optimal combination of retinol boosters according to the genetic characteristics of each individual.

[0063] This suggests that it may be possible to select specific single nucleotide polymorphism (SNP) markers that have a significant correlation with skin wrinkles, providing information that will enable the selection of a personalized retinol booster that can maximize the anti-wrinkle effects of retinol based on genetic information.

[0064] For example, a method for providing information is provided that, when the base of rs3827760 contains at least one minor allele, further selects a step of treating with lupeol and sucralfate, or, when the base of rs16935073 contains at least one major allele, further selects a step of treating with glutathione, but is not limited thereto.

[0065] In addition to the above method, (i) when the bases of the polymorphic site amplified or hybridized in step (c) contain two bases of a major allele determined by the single nucleotide polymorphism marker, the reactivity with retinol is high, providing information for treating only retinol alone without treatment with an additional retinol booster, or (ii) when the bases of the polymorphic site amplified or hybridized in step (c) contain two bases of a minor allele determined by the single nucleotide polymorphism marker, the reactivity with retinol is high, providing information for treating only retinol alone without treatment with an additional retinol booster, but the present invention is not limited thereto.

[0066] For example, a method of providing information is provided that allows a step of treating with retinol alone to be further selected when the rs3827760 base contains two major alleles, or a method of providing information is provided that allows a step of treating with retinol alone to be further selected when the rs16935073 base contains two minor alleles, but is not limited to these.

[0067] In the present invention, the term "SNP chip" refers to a type of DNA microarray that can simultaneously confirm each base of hundreds of thousands of SNPs.

[0068] The TaqMan method includes the steps of: (1) designing and preparing primers and TaqMan probes so as to amplify the desired DNA fragment; (2) labeling the probes of different alleles with FAM dye and VIC dye (Applied Biosystems); (3) using the DNA as a template and carrying out PCR using the primers and probes; (4) analyzing and confirming the TaqMan analysis plate after the PCR reaction is completed using a nucleic acid analyzer; and (5) determining the genotype of the polynucleotide of step (1) from the analysis results.

[0069] The sequencing analysis can be performed by a conventional method for determining base sequences, and can be performed by an automated gene analyzer. Allele-specific PCR refers to a PCR method in which the base at which the SNP is located is set as the 3' end, and a primer set including a designed primer is used to amplify a DNA fragment at which the SNP is located. The principle of this method is that, for example, when a specific base is substituted from A to G, a primer containing the A as the 3' end base and a reverse primer that amplifies a DNA fragment of an appropriate size are designed and a PCR reaction is performed; if the base at the SNP position is A, the amplification reaction is performed normally and a band at the desired position is observed, and if the base is substituted with G, the primer binds to the template DNA complementary to it, but the 3' end side does not bind complementary to it, so the amplification reaction is not performed correctly. DASH can be performed by a conventional method, specifically, by a method such as PRINCE.

[0070] On the other hand, PCR extension analysis can be performed by first amplifying a DNA fragment containing the base where the single nucleotide polymorphism is located with a primer pair, then dephosphorylating all nucleotides added to the reaction to inactivate it, and then adding an SNP-specific extension primer, a dNTP mixture, a dideoxynucleotide, a reaction buffer, and a DNA polymerase to carry out a primer extension reaction. Here, the extension primer considers the base adjacent to the 5' side of the base where the SNP is located as the 3' end, and the dNTP mixture excludes nucleic acids having the same base as the dideoxynucleotide, and the dideoxynucleotide is selected from one of the bases that indicate SNP. For example, when A is substituted with G, if a mixture of dGTP, dCTP, and dTTP and ddATP are added to the reaction, the primer is extended by DNA polymerase at the base where the substitution has occurred, and the primer extension reaction is terminated by ddATP at the position where the A base first appears after several bases. If the substitution does not occur, the extension reaction terminates at that position, and the type of base indicating the SNP can be identified by comparing the lengths of the extended primers as described above.

[0071] As for the detection method, when the extension primer or dideoxynucleotide is fluorescently labeled, the SNP can be detected by detecting fluorescence using a gene analyzer used for general base sequence determination (e.g., Model 3700 manufactured by ABI, etc.), and when the extension primer and dideoxynucleotide are unlabeled, the SNP can be detected by measuring the molecular weight using the matrix assisted laser desorption ionization-time of flight (MALDI-TOF) method. EXAMPLES

[0072] The present invention will be described in more detail below with reference to examples. However, these examples are merely illustrative of the present invention and are not intended to limit the present invention. EXAMPLES

[0073] Derivation of gene polymorphism markers with significant association with skin wrinkles We attempted to find genomic sites (gene mutations) that cause differences in the degree (value) of facial skin wrinkles according to genetic information in Korean women. The present invention aims to find genomic sites (gene mutations) related to wrinkles, and utilizes a microarray genotyping chip (manufactured by Illumina) that allows screening at the whole genome level without preselecting candidate genes.

[0074] In the present invention, an image-based skin diagnostic device (PIE Janus3) was used to evaluate the degree of wrinkles, and the results were adjusted for age, BMI, and principal component values ​​of the population to minimize external effects that affect the degree (value) of skin wrinkles.

[0075] To confirm the association between genomic sites (gene mutations) and the degree of skin wrinkles (number), linear regression analysis was used to quantify the correlation significance and genetic effect.

[0076] 1-1: Method for assessing the degree of skin wrinkles Healthy Korean women in their 20s to 70s were recruited to derive gene polymorphism markers that can explain the degree of general skin wrinkles. In addition, all subjects who were to undergo skin measurement were washed with cleansing or soap, and waited for 30 minutes without applying any products to allow the skin to adapt to the measurement environment, after which the degree of skin wrinkles was evaluated. An image-based skin diagnostic device (PIE Janus3) was used for the evaluation (measurements and analysis were performed according to the device manufacturer's instructions). Specifically, the subjects were made to wear a headband, a black cloth was placed over their clothes, their face was fixed to the forehead and chin rests built into the device, and they were asked to maintain the measurement position with their eyes closed, and high-resolution images of the subjects' faces were taken with the camera built into the Janus 3 device. Using a program provided by Janus 3 itself, the areas on the sides of the outer eyes and the area under the eyes were defined based on the areas of both eyes, and all wrinkles within these areas were extracted and quantified.

[0077] Gene collection was performed by collecting saliva, and in order to effectively collect genes, all subjects refrained from ingesting any food or drink, including water, for 30 minutes prior to collection.

[0078] Subjects were excluded from the study if they: (1) were pregnant, breastfeeding, or planning to become pregnant within the next 6 months; (2) had used topical skin preparations containing steroids for the treatment of skin diseases for more than 1 month; (3) had participated in the same study for less than 6 months; (4) had sensitive or hypersensitive skin; (5) had abnormal skin findings such as moles, acne, erythema, or telangiectasia in the study area; (6) had used the same or similar cosmetics or medicines in the study area within 3 months from the start of the study; (7) had undergone treatment in the study area (dermabrasion, Botox, or other skin treatment) or were planning to undergo treatment in the next 6 months; (8) had a chronic wasting disease (e.g., asthma, diabetes, high blood pressure), (9) had atopic dermatitis, or (10) were otherwise deemed by the principal investigator to be difficult to participate in the study.

[0079] 1-2: Gene collection and analysis methods For gene analysis, human genomic DNA was extracted from saliva using the QIAamp mini prep kit (QIAGEN). The quality was confirmed by an absorbance (OD 260 / 280) of 1.7, a concentration of 50 ng / ul, and a band test using 1xTAE 1% agarose gel. Gene analysis was performed only on samples that passed the quality check.

[0080] Genetic analysis was performed using Illumina's microarray genotyping chip. Specifically, the subjects' genes were analyzed using the company's global screening array product.

[0081] Genetic analysis experiments using Illumina's microarray genotyping chips were performed according to the provided instructions, and the genomic DNA amplification, DNA fragmentation, precipitation, hybridization, staining, washing, coating, and scanning processes were performed using the provided reagents.

[0082] After the experiment, the microarray genotyping chips were scanned using iScan Control Software (Illumina). After the scan, an idat file was automatically generated, and data quality control (sample call rate 98%, marker call rate 98%) and gene information confirmation were performed using the GenomeStudio (Illumina) program.

[0083] In this experiment, only data that had undergone data quality control after genetic analysis was used.

[0084] 1-3: Derivation of gene polymorphism markers related to the degree of skin wrinkles To derive gene polymorphism markers with significant association with the degree of skin wrinkles, linear regression analysis was performed between the degree of skin wrinkles and gene polymorphism markers in 17,019 Korean women using the PLINK v 1.90 program. For the analysis, PLINK v 1.90 and SNP & Variation suite (Golden Helix, Inc., Bozeman, Montana, USA) programs were used.

[0085] In order to obtain further information on gene polymorphism markers using the gene polymorphism markers confirmed by the experiment, imputation analysis was performed using the Beagle v 5.1 program. The reference data for this analysis was information registered in the 1000 Genomes project, an international open genome data database.

[0086] Imputation is a statistical technique for inferring unanalyzed gene polymorphism marker information based on gene polymorphism marker information obtained through experiments.

[0087] To control the quality of the analyzed polymorphic markers, only those polymorphic markers with minor allele frequencies above 0.01 and Hardy-Weinberg equilibrium levels above 0.000001 were used.

[0088] The significance of gene polymorphism markers showing an association with the degree of skin wrinkles was evaluated by linear regression analysis F-statistics. The criterion was set at P-value<0.00000005. Furthermore, gene polymorphism markers showing a weak level of association with the degree of skin wrinkles were further selected if they met the somewhat relaxed criterion of 0.00000005≦P-value<0.0001.

[0089] In order to minimize external effects that affect the degree of skin wrinkles and to obtain the effect of genetic information, the degree of skin wrinkles was adjusted for age, whether or not the patient had undergone menopause, BMI information, and information on lifestyle habits (drinking, smoking, eating habits, sleeping habits, etc.) and used in the analysis (for example, linear regression analysis was performed).

[0090] Representative examples of SNP markers significantly associated with skin wrinkles are shown in Table 1.

[0091] [Table 1] EXAMPLES

[0092] A test to confirm retinol responsiveness to improving skin wrinkles Based on the polymorphic markers showing an association with the degree of skin wrinkles obtained in Example 1, an attempt was made to find a substance that can maximize the effect of retinol in improving skin wrinkles and to formulate a customized retinol booster.

[0093] 2-1: Selection of subjects With approval from the Institutional Review Board (IRB, Korea) (LG-Wr_Healing-2020-1124), and in accordance with the standard operating guidelines and clinical trial standards of the LG Household & Healthcare Institutions Bioethics Committee, 18 healthy subjects were selected who were adults in their 20s to 50s with many wrinkles around the eyes and under the eyes, had no major abnormalities in the appearance of their skin, and had not undergone any dermatological treatment within one month prior to the start of the study.

[0094] Among the subjects, the following were excluded: 1) those with eczema / infectious skin diseases, 2) those with specific allergic constitutions or hypersensitivity, 3) those who were pregnant or breastfeeding, 4) those with severe nutritional disorders, 5) those with drug or alcohol addiction, 6) those with moles, acne, tattoos, erythema, burn scars, etc. at the test site, 7) those who had participated in the same test less than one month ago, and 8) those with high blood pressure or diabetes.

[0095] 2-2: Exam information An essence containing 2500 IU of retinol was applied under both eyes and to the corners of the eyes (every other day for the first two weeks, and every day from the third week onwards).

[0096] After that, Essence A-1 (Lupeol 0.015%, Sucralfate 0.015%) or Essence A-2 (Glutathione 0.01%) was applied to the entire right side of the face, and Essence B (vehicle formulation) was applied to the entire left side of the face twice a day (morning and evening). Specific information on Essence B (vehicle formulation) is shown in Table 2.

[0097] The test was conducted for a total of 8 weeks. Wrinkles were measured every 4 weeks using the Antera 3D, and the overall size of fine wrinkles under the eyes was then calculated. The overall size of fine wrinkles refers to the total volume of the area that is lower than the surface area when measured using the Antera divided by the total area.

[0098] [Table 2]

[0099] 2-3: Calculation of wrinkle improvement rate The wrinkle improvement rate (%) was calculated using the following formula.

[0100]

number

[0101] Statistical significance was determined using the t-test in the Excel statistical data analysis program. The significance test result was *p<0.05. EXAMPLES

[0102] Confirmation of the correlation between polymorphic markers with significance related to the degree of skin wrinkles and retinol treatment Based on the polymorphic markers showing an association with the degree of skin wrinkles obtained in Example 1, the degree of wrinkle improvement by retinol was confirmed. As a result, as shown in FIG. 1, it was confirmed that the responsiveness to retinol differs depending on whether or not one possesses an allele of a specific gene SNP.

[0103] For example, in the case of EDAR (rs3827760) among the genes in Table 1, it was confirmed that retinol responsiveness was high in the group that did not have the minor allele (A), and retinol responsiveness was low in the group that had at least one minor allele.

[0104] In addition, in the case of BNC2 (rs16935073), it was confirmed that retinol responsiveness was high in the group carrying two minor alleles (C), and retinol responsiveness was low in the group carrying at least one major allele (A).

[0105] Therefore, by informing the group who are highly responsive to retinol and are expected to see improvement in wrinkles of this fact before prescribing retinol alone, it is possible to reduce resistance to various side effects that appear during the adaptation period and to increase the degree to which the efficacy is felt, leading to continued use of retinol products. EXAMPLES

[0106] Identification of polymorphic markers associated with the degree of skin wrinkles and the effect of retinol responsiveness booster Based on the correlation between the polymorphic markers having relative significance with the degree of skin wrinkles obtained in Example 3 and retinol treatment, an attempt was made to provide a substance that can improve the wrinkle improvement efficiency in a group with low retinol responsiveness.

[0107] 4-1: Confirmation of the effect of retinol booster on gene expression related to skin wrinkles based on EDAR analysis data Since the minor allele (A) carrier group of the skin regeneration protein EDAR (rs3827760) shows low reactivity to retinol, we attempted to customize a treatment for them.

[0108] Specifically, 2500IU of retinol was combined with 0.015% lupeol and 0.015% sucralfate (Essence A-1).As a result, it was confirmed that the wrinkle improvement efficacy of the group was improved by approximately 3.7 times, from 3.3% to 12.3% (Figure 2).

[0109] 4-2: Confirmation of the effect of retinol booster on gene expression related to skin wrinkles based on BNC2 analysis data Since patients with the major allele (A) of the cutaneous photosensitive protein BNC2 (rs1693073) have low retinol sensitivity, we attempted to tailor a treatment for them.

[0110] Specifically, 2500IU of retinol was combined with 0.01% glutathione (Essence A-2). As a result, wrinkles were observed in the group treated with retinol alone, whereas the combined treatment with the said formula improved wrinkles by 12.2% (Figure 3).

[0111] Therefore, it can be seen that for groups with low responsiveness to retinol, the wrinkle improvement efficiency can be improved by providing a customized prescription for each group.

[0112] From the above description, a person skilled in the art to which the present invention pertains will understand that the present invention can be implemented in other specific forms without changing its technical ideas or essential features. It should be understood that the above examples are merely illustrative and not limiting. The present invention should be interpreted as including all modifications or alterations derived from the meaning and scope of the claims and their equivalent concepts, rather than the specification.

Claims

1. A composition for improving wrinkles, comprising as active ingredients retinol and at least one retinol booster selected from the group consisting of lupeol, sucralfate and glutathione.

2. The composition according to claim 1, wherein the retinol booster is contained in an amount of 0.0001 to 10% by weight based on the total weight of the composition.

3. 2. The composition according to claim 1, wherein the retinol is contained in an amount of 2000 to 6000 IU.

4. The composition according to claim 1 , wherein the wrinkle improvement is targeted to an individual having at least one single nucleotide polymorphism marker present in a gene encoding an EDAR or BNC2 protein.

5. The composition of claim 4, wherein the single nucleotide polymorphism marker is rs3827760 or rs16935073.

6. The single nucleotide polymorphism marker is A polynucleotide consisting of a continuous DNA sequence of 5 to 100 bases including the 109513601st base of human chromosome 2, in which the 109513601st base is G or A (rs3827760); A polynucleotide consisting of a continuous DNA sequence of 5 to 100 bases including the 16795790th base of human chromosome 9, in which the 16795790th base is A or C (rs16935073); 5. The composition of claim 4, comprising at least one polynucleotide selected from the group consisting of:

7. (a) obtaining a biological sample from an individual; (b) amplifying or hybridizing with a probe a polymorphic site of a single nucleotide polymorphism marker that is rs3827760 or rs16935073 from the biological sample; (c) a step of confirming the base of the amplified or hybridized polymorphic site in the (b) step.

8. The method for providing information according to claim 7, wherein the retinol booster is at least one selected from the group consisting of lupeol, sucralfate and glutathione.

9. The method for providing information according to claim 7, wherein, when the base of rs3827760 contains at least one minor allele, lupeol and sucralfate are selected as customized retinol boosters for improving wrinkles.

10. The method for providing information according to claim 7, wherein a customized retinol booster for improving wrinkles is not selected when the base of rs3827760 contains two major alleles.

11. The method for providing information according to claim 7, wherein glutathione is selected as a customized retinol booster for improving wrinkles when the base of rs16935073 contains at least one major allele.

12. The method for providing information according to claim 7, wherein a customized retinol booster for improving wrinkles is not selected when the base of rs16935073 contains two minor alleles.

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