Gene marker for predicting retinol efficacy, composition for alleviating wrinkles, and method for predicting retinol efficacy

Abstract

The present invention relates to: a genetic polymorphic marker significantly correlated with the degree of retinol efficacy; a composition for determining retinol efficacy, the composition comprising a probe capable of detecting the genetic polymorphic marker or an agent capable of amplifying the genetic polymorphic marker; and a kit or microarray comprising the composition. In addition, the present invention relates to: a composition for determining retinol efficacy; an information providing method for selecting a customized material for improving and enhancing retinol efficacy, the method comprising a step for identifying a base at a polymorphic site of a genetic polymorphic marker significantly correlated with the degree of retinol efficacy; and a method and system for predicting efficacy information including the degree of wrinkle improvement when retinol is applied.

Description

Genetic markers predicting the efficacy of retinol, compositions for improving wrinkles, and methods for predicting the efficacy of retinol

[0001] The present invention relates to a composition for determining retinol efficacy, comprising a genetic polymorphism marker having a significant correlation with the degree of retinol efficacy, a probe capable of detecting the same, or an agent capable of amplifying the same, and a kit or microarray comprising the composition. In addition, the present invention relates to a composition for determining retinol efficacy, a method for providing information for selecting a customized substance for improving retinol efficacy, the method comprising a step of identifying bases of a polymorphic region of a genetic polymorphism marker having a significant correlation with the degree of retinol efficacy, and a method and system for predicting efficacy information, including the degree of wrinkle improvement upon application of retinol.

[0002]

[0003] Retinol, a form of vitamin A, has been scientifically proven and approved for its wrinkle-improving and anti-aging effects and is widely used in anti-aging functional cosmetics. Retinol is known to increase collagen synthesis in fibroblasts, inhibit the expression of matrix metalloproteinases (MMPs), and promote skin turnover in the dermis and epidermis. This results in a thickening of the dermis and epidermis, which reduces wrinkles. Furthermore, rapid skin turnover removes dead skin cells, improving pigmentation and skin texture. However, the irritation and efficacy of retinol use vary significantly among individuals, and the exact mechanism of this variation remains unknown. Therefore, it is generally recommended to assess retinol sensitivity after using the product. Some studies have shown that individual differences in retinoid reactions may be due to genetic differences. Furthermore, some genome-wide association studies (GWAS) have revealed that the efficacy of substances with specific effects is significantly associated with genetic variants located within multiple genes. Furthermore, individual gene studies have confirmed that the efficacy of the same substance can vary depending on genotype. Therefore, retinol's skin efficacy is expected to exhibit similar characteristics.

[0004] However, specific genetic correlations with retinol's skin benefits are rare. Therefore, identifying genetic variants and genes associated with retinol's skin benefits is expected to provide valuable information for understanding the biological mechanisms and predicting individual retinol skin benefits.

[0005]

[0006] The present invention utilizes candidate gene analysis to identify genetic variants and causative genes associated with retinol skin efficacy in Koreans. Furthermore, the present invention develops a model for predicting retinol skin efficacy using these genetic variants. Furthermore, based on big data from genetic and skin measurement data, the goal is to develop and apply effective ingredients that can enhance retinol responsiveness and efficacy in Koreans.

[0007]

[0008] One object of the present invention is to provide a single nucleotide polymorphism (SNP) marker for determining the efficacy of retinol.

[0009] Another object of the present invention is to provide a composition for determining the efficacy of retinol, comprising a probe capable of detecting or amplifying a single nucleotide polymorphism (SNP) marker for determining the efficacy of retinol.

[0010] Another object of the present invention is to provide a kit or microarray for determining the efficacy of retinol comprising the composition.

[0011] Another object of the present invention is to provide a method and system for predicting efficacy information including the degree of wrinkle improvement when applying retinol.

[0012] Another object of the present invention is to provide single nucleotide polymorphism (SNP) markers that affect skin efficacy when applying a substance containing retinol.

[0013] Another object of the present invention is to provide a composition for improving the efficacy of retinol.

[0014] Another object of the present invention is to provide a cosmetic composition or a quasi-drug composition containing the composition as an active ingredient.

[0015] Another object of the present invention is to provide a method for providing information for selecting a customized substance for improving the efficacy of retinol, which includes a step of identifying a base at a polymorphic site of a single nucleotide polymorphism marker for determining the efficacy of retinol.

[0016]

[0017] The present invention can provide information on the degree of retinol efficacy of an individual through genetic polymorphism markers having a correlation significance with the degree of retinol efficacy, and furthermore, can develop customized ingredients or products that can improve the efficacy of retinol based on the information on genetic polymorphism markers observed in an individual.

[0018] In addition, through the prediction method and system of the present invention, it is possible to provide efficacy information on the degree of wrinkle improvement when applying a substance containing retinol.

[0019] Additionally, the accuracy of prediction is high because the results of machine learning are used in the process of predicting efficacy.

[0020]

[0021] Figures 1 and 2 are schematic block diagrams illustrating a retinol efficacy prediction system according to the present invention.

[0022] Figure 3 is a flowchart illustrating an embodiment of a method for predicting retinol efficacy according to the present invention.

[0023] Figure 4 shows the promotion of RARB expression by hesperidin of the present invention.

[0024] Figure 5 shows the inhibition of CYP2C9 expression by baicalin of the present invention.

[0025] Figure 6 shows the promotion of LAMC2 and / or ITGB8 expression by sitosterol of the present invention.

[0026] Figure 7 shows the promotion of EGFR expression by troxerutine of the present invention.

[0027] Figure 8 shows the results of treatment with a substance for improving the efficacy of retinol of the present invention, segmentation of the low-sensitivity group and the high-sensitivity group, and the rate of change in wrinkles between the groups.

[0028] Figure 9 shows the correlation between the retinol efficacy-associated significant polymorphism markers and the retinol efficacy booster.

[0029] Figure 10 is a diagram showing the distribution of efficacy scores when the value derived through the mathematical formula is used as is (left diagram) and when the value obtained by taking -Log (specifically -Log 2) of the value derived through the mathematical formula is used (right diagram).

[0030]

[0031] This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention can also be applied to each other description and embodiment. In other words, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Furthermore, the scope of the present invention should not be considered limited by the specific descriptions described below.

[0032] Furthermore, those skilled in the art will recognize or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific embodiments of the invention described herein. Furthermore, such equivalents are intended to be encompassed by the present invention.

[0033]

[0034] As one aspect for achieving the purpose of the present invention, the present invention provides a single nucleotide polymorphism (SNP) marker for determining the efficacy of retinol.

[0035] In another aspect, the present invention provides a composition for determining retinol efficacy, comprising a probe capable of detecting or amplifying a single nucleotide polymorphism (SNP) marker for determining retinol efficacy.

[0036] Depending on the degree of retinol efficacy, an individual's skin type can be distinguished.

[0037] The term "retinol" used in the present invention refers to a type of vitamin A, also known as pure vitamin A. It exists in the mucosal cells of the intestines of animals and is abundant in green-yellow plants. It is also transformed into the active form, retinoic acid, and is known to play an important role in maintaining the original function of epidermal cells. In addition, it is known to have the effect of promoting cell differentiation through the regulation of skin cell differentiation genes and promoting the biosynthesis of collagen, a fibrous solid protein existing between animal cells, and elastin, a fibrous fiber-like protein, thereby reducing wrinkles and increasing skin elasticity.

[0038] The term "retinol efficacy" in the present invention encompasses all effects of retinol, including wrinkle improvement, exfoliation, and anti-pigmentation. Specifically, the term may refer to wrinkle improvement due to retinol, but is not limited thereto. Furthermore, the term may be used interchangeably with "wrinkle improvement due to retinol."

[0039] For the purposes of the present invention, the term "skin wrinkle" refers to the observed appearance of skin folds due to loss of elasticity in a certain area of ​​an individual's skin. Due to the skin folding, the wrinkled area is shaded compared to the surrounding skin, and thus appears relatively darker than the surrounding skin. Furthermore, since wrinkles are mostly linear, they can be defined as wrinkles when the shaded areas are connected and observed to extend beyond a certain length, but are not limited thereto.

[0040]

[0041] In the present invention, the term "polymorphism" refers to a case where two or more alleles exist at a single genetic locus, and among the polymorphic sites, a single nucleotide polymorphism (SNP) in which only a single base differs from person to person is called a single nucleotide polymorphism. A desirable polymorphic marker has two or more alleles that exhibit an occurrence frequency of 1% or more, more specifically 10% or 20% or more, in a selected population. A 'genetic polymorphic marker' generally refers to a case in which two or more alleles are observed at the same genetic location (base), and generally, depending on the individual, there are cases of major allele / major allele, major allele / minor allele, and minor allele / minor allele. In the present invention, it may be used interchangeably with “polymorphic marker” and may mean a base and base portion of a lower allele, or may be defined together with the number and base position of a chromosome, but is not limited thereto.

[0042] In the present invention, the term "allele" refers to multiple types of a gene existing at the same genetic locus of a homologous chromosome. Alleles are also used to indicate polymorphism, and for example, SNPs have two types of alleles. In addition, it refers to a combination of two or more bases having the same number and base position on a chromosome, 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.

[0043]

[0044] Specifically, the genetic polymorphism markers of the present invention are significantly associated with the efficacy of retinol, and when one or more minor alleles are present among two alleles, the efficacy of retinol is significantly higher than that of an individual having the major allele / major allele. That is, in the case of the major allele / minor allele and the minor allele / minor allele, it can be seen that the skin characteristics have a higher or lower degree of efficacy by retinol compared to the case of possessing the major allele / major allele. Specifically, it can be seen that the skin characteristics have a higher or lower degree of improvement in skin wrinkles by retinol.

[0045] Since the single nucleotide polymorphism marker of the present invention can predict an individual's unique retinol efficacy characteristics, it can also provide information on effective ingredients that effectively affect changes in efficacy due to retinol, and thus can provide information on customized skin care products and lifestyle patterns for individuals, but is not limited thereto.

[0046] In the present invention, the term "rs_id" refers to an rs-ID, an independent marker assigned to all SNPs initially registered by NCBI, which began accumulating SNP information in 1998. The rs_id described in this table refers to an SNP marker, which is a polymorphic marker of the present invention.

[0047]

[0048] The above 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 correlation with the degree of retinol efficacy.

[0049] Specifically, the single nucleotide polymorphism marker may be one or more single nucleotide polymorphism markers selected from Table 1 related to the degree of skin efficacy by retinol.

[0050] The association significance of the single nucleotide polymorphism marker of the present invention with skin efficacy by retinol is characterized by a p-value such as, but not limited to, 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 may be less than 0.01, and more specifically, the p-value may be less than 0.1, but is not limited thereto.

[0051]

[0052] The single nucleotide polymorphism (SNP) marker of the present invention may be one or more of the markers shown in Table 1, but is not limited thereto. The single nucleotide polymorphism (SNP) marker may be one or more, and may be used in combinations of two or more, three or more, four or more, or any number of markers capable of determining the efficacy of retinol, but is not limited thereto.

[0053] The marker may be, but is not limited to, a SNP itself, a polynucleotide consisting of 5-100 contiguous DNA sequences including the SNP position, or a polynucleotide consisting of a complementary sequence thereof.

[0054] As a specific example, the single nucleotide polymorphism marker may be, but is not limited to, any one or more of the markers listed in Table 1 associated with efficacy by retinol.

[0055] The markers selected from among those shown in Table 1 may be described as follows.

[0056]

[0057] For example, if the SNP ID is rs10494560, Chr.Position (GRCh ver. 37) is described as "1: 183172548" and the Allele is set to C > T, this indicates that the 183172548th base on human chromosome 1 is C or T, and the base located to the left of the ">" of the allele may indicate the major allele and the base located to the right may indicate the minor allele.

[0058] As a specific example, the marker selected from Table 1 may be, but is not limited to, one or more polynucleotides selected from the group consisting of a polynucleotide comprising 5 to 100 contiguous DNA sequences including the 183172548th base of human chromosome 1, wherein the 183172548th base is C or T (rs10494560); and complementary polynucleotides thereof. The markers described above are only some of the examples described in Table 1, and markers at other locations in the chromosome may be selected in the same manner as described above.

[0059]

[0060] The above alleles of the present invention have the same number of chromosomes in each individual, and among them, there are major alleles and minor alleles of SNPs, and as the base of the polymorphic site of the polymorphic marker increases by one to the minor allele, the major alleles may decrease by one, and as the base of the polymorphic site of the polymorphic marker increases by one to the major allele, the minor alleles may decrease by one. However, the ranges in which the minor alleles and major alleles can increase and decrease can be within three types: i) major allele / major allele, ii) major allele / minor allele, iii) minor allele / minor allele, and alleles can decrease or increase within the ranges of the three types, but are not limited thereto.

[0061] In addition, in the present invention, the marker is a marker that can determine the efficacy of retinol as the base of the polymorphic site of the polymorphic marker of the individual increases by one minor allele. Specifically, an individual having (1) a major allele / minor allele, (2) a minor allele / minor allele) that has at least one minor allele among two alleles can be determined to have skin characteristics with a higher or lower degree of skin efficacy by retinol compared to a person having a major allele / major allele, which is a general individual.

[0062] More specifically, the degree of increase or decrease in skin efficacy by retinol can be determined as the minor allele increases by one among the markers shown in Table 1. For example, if the major allele is C and the minor allele is T at the 100335586th base of the 4th chromosome of the individual among the markers shown in Table 1 (rs284785), it can be determined that the skin efficacy by retinol increases or decreases in the case of having C / T or T / T compared to a person having C / C, but is not limited thereto.

[0063]

[0064] The above bases are only examples of a portion of Table 1 and are not specifically described, but can be interpreted and derived as explained above.

[0065]

[0066] In the present invention, the term "probe capable of detecting a marker for judging the efficacy of retinol" refers to a composition capable of diagnosing the degree of skin efficacy of retinol by specifically hybridizing with a polymorphic region of the gene as described above, and the specific method of such gene analysis is not particularly limited and may be any gene detection method known in the technical field to which this invention pertains. In addition, the term may be used interchangeably with the terms "for judging the degree of skin efficacy of retinol" and "for judging improvement of skin wrinkles by retinol."

[0067] In the present invention, the term "agent capable of amplifying a marker for judging retinol efficacy" refers to a composition capable of diagnosing the degree of skin efficacy by retinol by confirming the polymorphic site of the gene as described above through amplification, and specifically refers to a primer capable of specifically amplifying the polynucleotide of the marker for judging retinol efficacy. In addition, the term may be used interchangeably with the terms "for diagnosing retinol efficacy," "for diagnosing the degree of skin efficacy by retinol," "for judging the degree of skin efficacy by retinol," "for diagnosing skin wrinkle improvement by retinol," and "for judging skin wrinkle improvement by retinol."

[0068] The primer used for the amplification of the above polymorphic marker refers to a single-stranded oligonucleotide that can act as an initiator of template-directed DNA synthesis under appropriate conditions (e.g., four different nucleoside triphosphates and a polymerizing agent such as DNA, RNA polymerase, or reverse transcriptase) in an appropriate buffer and at an appropriate temperature. The appropriate length of the primer may vary depending on the intended use, but is typically 15 to 30 nucleotides. Shorter primer molecules generally require lower temperatures to form stable hybrids with the template. The primer sequence need not be perfectly complementary to the template, but should be sufficiently complementary to hybridize with the template.

[0069] As used herein, the term "primer" refers to a short sequence of bases having a short free 3'-terminal hydroxyl group, which can form base pairs with a complementary template and serves as a starting point for copying the template strand. The primer can initiate DNA synthesis in the presence of a reagent for polymerization (i.e., DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates in an appropriate buffer and temperature. Skin type can be predicted by performing polymerase chain reaction (PCR) amplification and the degree of production of the desired product. PCR conditions and the lengths of the sense and antisense primers can be modified based on those known in the art.

[0070] The probes or primers of the present invention can be chemically synthesized using the phosphoramidite solid support method or other well-known methods. These nucleic acid sequences can also be modified using many means known in the art. Non-limiting examples of such modifications include methylation, "capping," substitution with one or more homologs of a natural nucleotide, and modifications between nucleotides, such as modification with uncharged linkers (e.g., methyl phosphonate, phosphotriester, phosphoroamidate, carbamate, etc.) or charged linkers (e.g., phosphorothioate, phosphorodithioate, etc.).

[0071]

[0072] Another aspect of the present invention provides a kit for assessing retinol efficacy, comprising the composition for assessing retinol efficacy. The kit may be, but is not limited to, an RT-PCR kit or a DNA chip kit.

[0073] The kit of the present invention can diagnose skin type by amplifying a SNP polymorphism marker, which is a marker for determining retinol efficacy, or by determining the mRNA expression level of the SNP polymorphism marker. As a specific example, the kit for measuring the mRNA expression level of the marker for determining retinol efficacy in the present invention may be a kit including the essential elements required for performing RT-PCR. In addition to each primer pair specific to the gene of the marker for determining retinol efficacy, the RT-PCR kit may include a test tube or other appropriate container, a reaction buffer (with various pH and magnesium concentrations), deoxynucleotides (dNTPs), an enzyme such as Taq polymerase and reverse transcriptase, DNase, RNAse inhibitor, DEPC-water, sterile water, etc. In addition, it may include a primer pair specific to a gene used as a quantitative control. In addition, specifically, the kit of the present invention may be a kit for determining retinol efficacy that includes the essential elements required for performing a DNA chip. A DNA chip kit is a tool that attaches nucleic acid species in a gridded array to a flat solid support, typically a glass surface no larger than a microscope slide, so that the nucleic acids are arranged uniformly on the chip surface, allowing multiple hybridization reactions to occur between the nucleic acids on the DNA chip and complementary nucleic acids contained in a solution treated on the chip surface, enabling massive parallel analysis.

[0074]

[0075] Another aspect of the present invention provides a microarray for determining retinol efficacy comprising the composition for determining retinol efficacy.

[0076] The above microarray may comprise DNA or RNA polynucleotides. The above microarray is composed of a conventional microarray except that the probe polynucleotides comprise the polynucleotides of the present invention.

[0077] Methods for manufacturing microarrays by immobilizing probe polynucleotides on a substrate are well known in the art. The probe polynucleotide refers to a hybridizable polynucleotide, and refers to an oligonucleotide capable of sequence-specific binding to complementary strands of nucleic acids. The probe of the present invention is an allele-specific probe, which hybridizes to a DNA fragment derived from one member of the same species but not to a fragment derived from the other member due to the presence of a polymorphic site in the nucleic acid fragments derived from two members of the same species. In this case, hybridization conditions must be sufficiently stringent to ensure that only one of the alleles hybridizes, as there is a significant difference in hybridization intensity between alleles. This can induce a favorable hybridization difference between different allele forms. The probe of the present invention can be used in methods for diagnosing skin types, etc. by detecting alleles. These diagnostic methods include detection methods based on nucleic acid hybridization, such as Southern blot, and in methods using DNA chips, the probe may be provided in a form pre-bound to the substrate of the DNA chip. The above hybridization can usually be performed under stringent conditions, for example, a salt concentration of 1 M or less and a temperature of 25°C or higher. For example, conditions of 5x SSPE (750 mM NaCl, 50 mM Na Phosphate, 5 mM EDTA, pH 7.4) and 25-30°C may be suitable for allele-specific probe hybridization.

[0078] The process of immobilizing the probe polynucleotide associated with the skin diagnosis of the present invention onto a substrate can also be easily manufactured using these conventional techniques. Furthermore, hybridization of nucleic acids on microarrays and detection of hybridization results are well known in the art. For example, the detection can be accomplished by labeling a nucleic acid sample with a label capable of generating a detectable signal, such as a fluorescent substance such as Cy3 or Cy5, followed by hybridization on a microarray and detection of the signal generated from the label.

[0079]

[0080] In another aspect, the present invention provides a method for providing information for selecting a customized substance for improving the efficacy of retinol, comprising the steps of (a) amplifying or hybridizing a polymorphic site of the single nucleotide polymorphism marker with a probe in DNA obtained from a sample isolated from an individual; and (b) identifying a base of the amplified or hybridized polymorphic site in step (a).

[0081] In the present invention, the term "subject" refers to a subject for diagnosing skin wrinkles. DNA may be obtained from, but is not limited to, samples such as hair, urine, blood, various bodily fluids, isolated tissue, isolated cells, or saliva.

[0082] Any method known to those skilled in the art can be used to obtain genomic DNA in the above step (a).

[0083] Any method known to those skilled in the art can be used to amplify the polymorphic site of the single nucleotide polymorphism marker from the DNA obtained in step (a) or to hybridize it with a probe. For example, the target nucleic acid can be amplified by PCR and purified. In addition, ligase chain reaction (LCR) (Wu and Wallace, Genomics 4, 560 (1989), Landegren et al., Science 241, 1077 (1988)), transcription amplification (Kwoh et al., Proc. Natl. Acad. Sci. USA 86, 1173 (1989)), self-sustaining sequence replication (Guatelli et al., Proc. Natl. Acad. Sci. USA 87, 1874 (1990)), and nucleic acid-based sequence amplification (NASBA) can be used.

[0084] Among the above methods, determining the base of the polymorphic site in step (b) includes, but is not limited to, sequencing analysis, hybridization by microarray, allele specific PCR, dynamic allele-specific hybridization (DASH), PCR extension analysis, SSCP, PCR-RFLP analysis or TaqMan technique, SNPlex platform (Applied Biosystems), mass spectrometry (e.g., Sequenom's MassARRAY system), mini-sequencing method, 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). One or more alleles at a polymorphic marker, including a microsatellite, SNP, or other type of polymorphic marker, can be identified using the above methods or other methods available to those skilled in the art to which the present invention pertains. Determining the base pairs at such polymorphic sites can be specifically performed using a SNP chip.

[0085] The method may further determine whether wrinkle improvement according to retinol efficacy is superior or inferior if (c) the base of the amplified or hybridized polymorphic site includes at least one base that is a minor allele according to the single nucleotide polymorphism marker, but is not limited thereto. In addition, the method may further determine whether wrinkle improvement according to retinol efficacy is superior or inferior if (c) the base of the amplified or hybridized polymorphic site includes at least one base that is a major allele according to the single nucleotide polymorphism marker, but is not limited thereto.

[0086] In the present invention, the term "SNP chip" means one of the DNA microarrays capable of identifying each base of hundreds of thousands of SNPs at once.

[0087] The TaqMan method comprises the steps of (1) designing and manufacturing primers and TaqMan probes to amplify a desired DNA fragment; (2) labeling probes of different alleles with FAM dye and VIC dye (Applied Biosystems); (3) performing PCR using the DNA as a template and the primers and probes; (4) analyzing and confirming the TaqMan assay plate with a nucleic acid analyzer after the PCR reaction is completed; and (5) determining the genotype of the polynucleotides of step (1) from the analysis results.

[0088] In the above, the sequencing analysis can use a conventional method for determining the base sequence, and can be performed using an automated genetic analyzer. In addition, allele-specific PCR refers to a PCR method that amplifies a DNA fragment at which the SNP is located using a primer set including a primer designed with the base at which the SNP is located as the 3' end. The principle of the method is that, for example, when a specific base is substituted from A to G, if a primer including the A as the 3'-terminal base and an opposite primer capable of amplifying 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 can complementarily bind to the template DNA, but the 3'-end side cannot complementarily bind, so the amplification reaction is not performed properly. This utilizes the fact that DASH can be performed using a conventional method, and specifically, can be performed using the method of Prince et al.

[0089] Meanwhile, PCR extension analysis is performed by first amplifying a DNA fragment containing the base where the single nucleotide polymorphism is located using a primer pair, then inactivating all nucleotides added to the reaction by dephosphorylation, and then adding a SNP-specific extension primer, a dNTP mixture, a dideoxynucleotide, a reaction buffer, and DNA polymerase to perform a primer extension reaction. At this time, the extension primer uses the base immediately 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 base types representing the SNP. For example, when there is an A to G substitution, 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 occurred, and after a few bases, the primer extension reaction is terminated by ddATP at the position where the A base first appears. If the above substitution does not occur, the extension reaction is terminated at that position, so that the type of base representing the SNP can be determined by comparing the length of the extended primer.

[0090] At this time, in the case of fluorescently labeled extension primers or dideoxynucleotides, the SNP can be detected by detecting fluorescence using a genetic analyzer (e.g., ABI's Model 3700) used for general base sequence determination, and in the case of using non-labeled extension primers and dideoxynucleotides, the SNP can be detected by measuring the molecular weight using the MALDI-TOF (matrix-assisted laser desorption ionization-time of flight) technique.

[0091]

[0092] A method for predicting retinol efficacy according to one aspect of the present invention includes a step of inputting genetic information of a prediction target individual into a prediction terminal, a step of transmitting the input genetic information to a server, a step of calculating efficacy information according to a result of comparing a threshold efficacy score predetermined in the server with an efficacy score calculated according to the input genetic information, and a step of transmitting and outputting the calculated efficacy information to the prediction terminal, wherein the efficacy score may be determined according to the number and type of one or more single nucleotide polymorphism markers selected from Table 1 while existing in the genetic information of a specific individual.

[0093] In another aspect of the present invention, the single nucleotide polymorphism marker type may include a major allele-major allele type, a major allele-minor allele type, and a minor allele-minor allele type.

[0094] In another aspect of the present invention, the efficacy score may be lower the more bases have an OR value lower than 1 for a specific single nucleotide polymorphism marker in the genetic information, and may be higher the more bases have an OR value higher than 1 for a specific single nucleotide polymorphism marker in the genetic information.

[0095] In another aspect of the present invention, the efficacy score can be determined according to the following mathematical formula.

[0096]

[0097] Here, is the efficacy score, and OR k is the efficacy weight of the kth single nucleotide polymorphism marker, Xk is a number determined according to the single nucleotide polymorphism marker type of the kth single nucleotide polymorphism marker, k may be an integer greater than or equal to 1, and m may be the number of one or more single nucleotide polymorphism markers that may be utilized in calculating the efficacy score (for example, it may be 45 with reference to Table 1).

[0098]

[0099] In another aspect of the present invention, the efficacy information may include the degree of wrinkle improvement when retinol is applied to a predicted target subject.

[0100] In addition, a retinol efficacy prediction system according to one aspect of the present invention includes a prediction terminal that receives genetic information of a prediction target individual and outputs efficacy information calculated by a server according to the input genetic information, and a server that transmits the genetic information input to the prediction terminal and calculates efficacy information according to a comparison result of a predetermined threshold efficacy score and the efficacy score calculated according to the input genetic information, wherein the efficacy score can be determined according to the number and type of one or more single nucleotide polymorphism markers selected from Table 1 while existing in the genetic information of a specific individual.

[0101] In addition, a computer program according to one aspect of the present invention may be stored in a computer-readable recording medium to execute the above-described method for predicting retinol efficacy.

[0102]

[0103] Another aspect of the present invention provides a method and system for predicting the efficacy of retinol. This will be described in detail with reference to FIGS. 1 to 3.

[0104] The retinol efficacy prediction system includes a prediction terminal (100) and a server (200), and may optionally further include a data provision server (300). Here, the prediction terminal (100) and the server (200) may be separate components, or may be integrated into a single component. In another aspect of the present invention, functions (storage, calculation, output, etc.) performed by the prediction terminal (100) may also be performed by the server (200), and conversely, functions (storage, calculation, output, etc.) performed by the server (200) may also be performed by the prediction terminal (100).

[0105]

[0106] The prediction terminal (100) predicts efficacy information expected to appear in a prediction target object when a substance containing retinol is applied to the prediction target object. Here, the efficacy information may refer to whether / to what extent wrinkles are improved when a substance containing retinol is applied to the prediction target object.

[0107] Referring to FIG. 2, the prediction terminal (100) includes a communication unit (110), an input unit (120), a memory (130), a processor (140), an output unit (150), a control unit (160), a power supply unit (170), and an interface unit (180).

[0108] The communication unit (110) is configured for communication with an external device, and data transmission and reception with the external device can be possible through the communication unit (110).

[0109] The input unit (120) can receive commands or data to be used in a component (e.g., a processor) of the prediction terminal (100) from an external source (e.g., a user) of the server (100). The input unit (120) can include, for example, a microphone, a mouse, a keyboard, a key (e.g., a button), or a digital pen (e.g., a stylus pen).

[0110] The memory (130) can store various data used in the prediction terminal (100). The data stored in the memory (130) may be, for example, a critical efficacy score, and may also include input data or output data for software and commands related thereto.

[0111] Additionally, the memory (130) can store both information input to the prediction terminal (100) and information generated from the prediction terminal (100).

[0112] The processor (140) may perform a data processing or calculation function, and as at least part of the data processing or calculation function, may store commands or data received from other components in volatile memory, process the commands or data stored in the volatile memory, and store result data in non-volatile memory. The processor (140) may include an efficacy score calculation unit and an efficacy information calculation unit.

[0113] The efficacy score calculation unit calculates the efficacy score by a preset method. Specifically, the efficacy score can be calculated by multiplying the assigned scores of each of one or more single polymorphisms (SNPs) selected by the SNP marker selection device described below. The assigned scores can be calculated using a first value indicating the degree to which each single polymorphism affects wrinkle improvement, and a second value determined according to the number of major alleles and minor alleles included in each single polymorphism, and more specifically, the score can be calculated by multiplying the first value by the second value.

[0114] Specifically, the efficacy score calculation unit calculates the efficacy score of the prediction target object using the following mathematical formula.

[0115]

[0116] Here, is the efficacy score, and OR kis the efficacy weight of the kth single nucleotide polymorphism marker, Xk is a number determined according to the single nucleotide polymorphism marker type of the kth single nucleotide polymorphism marker, and k is an integer greater than or equal to 1. In addition, m may correspond to the number of single nucleotide polymorphism markers included in [Table 1] and may be 45.

[0117] The efficacy information calculation unit is configured to compare an efficacy score calculated according to genetic information input into the prediction terminal (100) with a predetermined threshold efficacy score and to calculate efficacy information according to the comparison result. In one embodiment of the present invention, the threshold efficacy score may be a numerical value that distinguishes groups in which the wrinkle improvement effect is prominent or not prominent when applying retinol. Meanwhile, the efficacy score according to an embodiment of the present invention may be applied with a value calculated using the above mathematical formula and taking the -Log. When the value calculated using the above mathematical formula is used as the efficacy score, a problem occurs in which the distribution of the efficacy score is biased toward some areas because single nucleotide polymorphism markers having an OR value less than 1 are multiplied multiple times (see the left drawing of FIG. 10). Therefore, in one embodiment of the present invention, by taking the -Log of the value calculated using the above mathematical formula, the efficacy score calculated as a number less than 1 is converted into a positive number, thereby solving the problem in which the distribution of the efficacy score is biased toward some areas. More specifically, a value calculated using the above mathematical formula and taking -Log 2 is used as the efficacy score (see the right drawing of FIG. 10). Meanwhile, the critical efficacy score according to one aspect of the present invention may have a value of 8.5 to 9.5 based on -Log 2 (value) obtained by taking -Log 2 from the value calculated through the above mathematical formula, and more specifically, may be 9.

[0118] The efficacy information calculated in the efficacy information calculation unit can be, for example, a 'high improvement group' or a 'low improvement group', and through this, it is possible to determine in advance whether a significant wrinkle improvement effect will be produced when a substance containing retinol is applied to the skin of the prediction target individual simply by inputting his or her genetic information.

[0119]

[0120] As one aspect for achieving the purpose of the present invention, the present invention provides a composition for improving the efficacy of retinol.

[0121] Specifically, the present invention aims to provide a composition that maximizes the effects of retinol by using various substances to enhance its efficacy. For the purposes of the present invention, the above substances may be referred to as "retinol boosters."

[0122] In the present invention, a 'retinol booster' refers to a substance that works together with retinol to amplify the effect of improving wrinkles. Retinol has the effect of reducing skin wrinkles and increasing skin elasticity, but the reactivity to retinol may vary depending on the individual. Therefore, a retinol booster is not particularly limited as long as it works together with retinol to increase the reactivity to retinol and help maximize the effect of improving skin wrinkles. In addition, it refers to a substance that has a synergistic effect with respect to a positive correlation index that increases when retinol is treated compared to the control group in a cell experiment on wrinkle-related gene markers, and has an effect of restoring to the level of the control group with respect to a negative correlation index that decreases when retinol is treated compared to the control group. In the present invention, a 'retinol booster' may be used interchangeably with a 'retinol efficacy booster', a 'composition for improving retinol efficacy', or a 'retinol efficacy enhancing improvement', but is not limited thereto. For example, the retinol booster may include, but is not limited to, one or more substances selected from the group consisting of beta-sitosterol, hesperidin, baicalin, and troxerutin as an active ingredient.

[0123]

[0124] The above term 'beta-sitosterol' is a type of phytosterol that exists in plants with a structure similar to cholesterol, and is found in large quantities in plants such as rice bran, peanuts, beans, pumpkin seeds, corn oil, radish, cabbage, and wolfberry. In particular, beta-sitosterol is known to have blood cholesterol reduction, blood sugar reduction, anticancer, immune-enhancing, antibacterial, antioxidant, and anti-inflammatory effects. The above beta-sitosterol is not limited to the method of obtaining it, and can be chemically synthesized by a method known in the art, or a commercially available substance can be used. The IUPAC name of the above substance is Stigmast-5-en-3β-ol.

[0125] [Chemical Formula 1]

[0126]

[0127] The above term 'hesperidin' is a type of polyphenol that is abundantly found in citrus fruits, etc., and is also known as vitamin P along with rutin and quercetin as a vitamin-like substance. It is known to help collagen production and has anti-inflammatory effects and is effective for diseases related to aging, and to affect joints and cartilage. It is also known that if it is deficient in the human body, it can cause decreased absorption of vitamin C, arteriosclerosis, and decreased immunity. The above hesperidin is not limited to the method of obtaining it, and can be chemically synthesized by a method known in the art, or a commercially available substance can be used. The IUPAC name of the above substance is (2S)-3', 5-Dihydroxy-4'-methoxy-7-[α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranosyloxy]flavan-4-one.

[0128] [Chemical Formula 2]

[0129]

[0130] The above term 'baicalin' is a type of flavonoid, which is mainly contained in the roots of Scutellaria baicalensisGeorgi, a plant of the Lamiaceae family, and is known to have pharmacological effects such as anti-infection, anti-allergy, and anti-hypertensive effects. The above baicalin can be chemically synthesized and obtained using various extraction methods known in the art, and if necessary, can be obtained through an additional purification process. In addition, the baicalin of the present invention can be used as the component itself or as a gold extract containing baicalin. The IUPAC name of the above substance is 5,6-Dihydroxy-4-oxoflav-2-en-7-ylβ-D-glucopyranosiduronic acid.

[0131] [Chemical Formula 3]

[0132]

[0133] The term "Troxerutin" refers to a natural flavonoid derived from the Japanese maple tree (Sophora Japonica). One of the most stable bioflavonoids, troxerutin is known to inhibit lipoxygenase, which causes inflammation and allergies, and to suppress the formation of prostaglandins, which are inflammatory mediators. Furthermore, troxerutin stabilizes the skin by regulating capillary resistance and promoting healthy blood and lymphatic microcirculation. Furthermore, troxerutin protects skin cells from UV-induced cell death, restricted cell migration, growth arrest, and DNA damage. The method of obtaining troxerutin is not limited, and can be chemically synthesized using methods known in the art or using commercially available substances.

[0134] [Chemical Formula 4]

[0135]

[0136]

[0137] The material of the present invention may be contained in an amount of about 0.0001 to 10 wt% based on the total weight of the composition, but is not limited thereto. For example, the retinol booster material of the present invention may be contained in an amount of about 0.0001 to 10 wt%, about 0.0005 to 5 wt%, about 0.001 to 5 wt%, about 0.01 to 5 wt%, about 0.0001 to 3 wt%, about 0.0005 to 3 wt%, about 0.001 to 3 wt%, about 0.01 to 3 wt%, about 0.0001 to 1 wt%, about 0.0005 to 1 wt%, about 0.001 to 1 wt%, or about 0.01 to 1 wt% based on the total weight of the composition.

[0138]

[0139] For the purpose of the present invention, the above substance has an effect of improving the efficacy of retinol.

[0140] The term 'retinol efficacy' is as described above.

[0141]

[0142] For the purpose of the present invention, the substance may be characterized by promoting the expression of RARB, LAMC2, ITGB8 or EGFR; or inhibiting the expression of CYP2C9. Specifically, among the substances, when hesperidin is treated, the expression of RARB is promoted, when beta-sitosterol is treated, the expression of LAMC2 or ITGB8 is promoted, when troxerutine is treated, the expression of EGFR is promoted, and when baicalin is treated, the expression of CYP2C9 is inhibited, but is not limited thereto. It can be seen that the responsiveness to the substance of the present invention varies depending on a specific genetic polymorphism type, and it is suggested that the wrinkle improvement efficiency by retinol can be increased by providing a customized prescription according to the efficacy information related to the efficacy of retinol.

[0143]

[0144] In the present invention, the term "about" may be presented before a specific numerical value. As used herein, the term "about" encompasses not only the exact number described after the term, but also a range that is or is nearly that number. Whether a number is or is nearly the specific number described can be determined based on the context in which it is presented. For example, the term "about" may refer to a range of -10% to +10% of a numerical value. For another example, the term "about" may refer to a range of -5% to +5% of a given numerical value. However, this is not a limitation.

[0145]

[0146] The above composition is characterized by being personalized. Specifically, it can provide a personalized substance targeting an individual having a significant single nucleotide polymorphism (SNP) marker associated with a specific degree of retinol efficacy.

[0147] For example, the significant single nucleotide polymorphism (SNP) marker associated with the degree of retinol efficacy may be, but is not limited to, one or more selected from Table 1.

[0148]

[0149] The composition of the present invention can be used as a cosmetic composition for improving the efficacy of retinol, and can be formulated in various forms. The cosmetic composition according to the present invention can be prepared in a formulation selected from the group consisting of a solution, an external ointment, a cream, a foam, a nourishing toner, an emollient toner, a pack, an emollient, an emulsion, a makeup base, an essence, a soap, a liquid cleanser, a bath agent, a sunscreen cream, a sun oil, a suspension, an emulsion, a paste, a gel, a lotion, a powder, a soap, 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.

[0150] In addition, the cosmetic composition of the present invention may additionally include one or more cosmetically acceptable carriers that are blended with general skin cosmetics, and may appropriately blend conventional ingredients such as oil, water, surfactants, moisturizers, lower alcohols, thickeners, chelating agents, pigments, preservatives, fragrances, etc., but is not limited thereto.

[0151] The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation.

[0152] When the formulation of the present invention is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or a mixture thereof may be used as a carrier component.

[0153] When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or a mixture thereof may be used as a carrier component, and particularly in the case of a spray, a propellant such as chlorofluorohydrocarbon, propane / butane or dimethyl ether may be additionally included.

[0154] When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, and for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil can be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol fatty acid ester, polyethylene glycol or fatty acid ester of sorbitan can be used.

[0155] When the formulation of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth, etc. can be used as a carrier component.

[0156] When the formulation of the present invention is soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolyzates, isethionates, lanolin derivatives, fatty alcohols, vegetable oils, glycerol, sugars, etc. can be used as carrier components.

[0157]

[0158] In another aspect, the composition provides an over-the-counter drug composition for improving the efficacy of retinol.

[0159] In the present invention, the term "quasi-drug" means a product that has a milder effect than a pharmaceutical product among products used for the purpose of diagnosing, treating, improving, alleviating, managing or preventing diseases of humans or animals. For example, according to the Pharmaceutical Affairs Act of the Republic of Korea, a quasi-drug is a product excluding products used for the purpose of pharmaceutical products, and includes products used for treating or preventing diseases of humans or animals, products that have a mild effect on the human body or do not act directly, etc.

[0160] In one embodiment, the pharmaceutical composition of the present invention can be manufactured in a form selected from the group consisting of, but not limited to, body cleanser, shampoo, conditioner, foam, soap, mask, ointment, cream, lotion, essence, and spray.

[0161]

[0162] Hereinafter, the present invention will be described in more detail through examples. These examples are intended merely to illustrate the present invention and are not to be construed as limiting the scope of the present invention.

[0163]

[0164] Example 1: Derivation of genetic polymorphism markers associated with retinol efficacy.

[0165]

[0166] Thirty-two subjects used a 0.1% retinol cream for four weeks, and the same expert classified wrinkle length improvement rates of 5% or more as “improved”. After dichotomizing the presence or absence of retinol efficacy in 30 subjects, logistic regression analysis was performed to derive genetic polymorphism markers that showed a significant association with wrinkle improvement.

[0167] To detect genetic polymorphism markers, human genomic DNA was extracted from the saliva of the subjects using the QIAamp mini prep kit (QIAGEN), and its quality was confirmed by an absorbance (0D 260 / 280) of 1.7, a concentration of 50 ng / μl, and a band test using 1X TAE 1% agarose gel. Only for the cases that passed the quality, the genetic polymorphism markers of the subjects were detected using the Illumina microarray genotyping chip, specifically, the global screening array product of the same company. The Illumina microarray genotyping chip genetic polymorphism marker detection experiment was performed using the provided manual and reagents, and the completed microarray genotyping chip was scanned using iScan Control Software (Illumina) to confirm the data quality (sample call rate 98%, marker call rate 98%) and genetic polymorphism marker information.

[0168] Prior to proceeding with regression analysis, human genomic DNA was extracted from the saliva of the subjects using a QIAamp mini prep kit (QIAGEN) to detect genetic polymorphism markers in the subjects. The quality was confirmed through a band test using a 1X TAE 1% agarose gel with an absorbance (0D 260 / 280) of 1.7 and a concentration of 50 ng / μl. Detection of genetic polymorphism markers was attempted only for those that passed the quality test.

[0169] The Illumina microarray genotyping chip genetic analysis experiment was conducted according to the provided manual, and the processes of genomic DNA amplification, DNA fragmentation, precipitation, hybridization, staining, washing, coating, and scanning were performed using the provided reagents.

[0170] The microarray genotyping chip for which the experiment was completed was scanned using iScan Control Software (Illumina). Upon completion of scanning, an idat file was automatically generated, and data quality control (sample call rate 98%, marker call rate 98%) and genetic information confirmation were performed using the GenomeStudio (Illumina) program.

[0171] Each genetic polymorphism marker was used only if it exceeded the criteria of a minor allele frequency of 1% or more and a Hardy-Weinberg equilibrium of 0.0001% or more for quality control.

[0172] The genetic polymorphism markers analyzed are a set of genetic polymorphism markers that are thought to be associated with retinol metabolism, and are markers located within genes selected through a literature review related to retinol for candidate gene analysis.

[0173] In the candidate gene analysis, the significance of genetic polymorphism markers showing association with the efficacy (wrinkle improvement) of retinol was evaluated through logistic regression analysis Z-statistics, and the criterion was set at P-value < 0.1.

[0174] As a result, 45 polymorphic markers with significant association with retinol efficacy were discovered (Table 1).

[0175]

[0176] Significant polymorphism markers associated with retinol efficacy (P < 0.1) SNPGENEP 1) OR 2) CHR 3) BP 4) A1 5) A2 6)<h2 style=";text-align:left;direction:ltr">1rs10494560LAMC2 0.0031020.052081183172548TC2rs3823974ITGB80.00830810.89720442796CT3rs16844790ITGB60.013450.052082160968628GA4rs75951061 ITGB80.015227.606720383888GA5rs3738824LAMC20.01870.12241183192496GA6rs1023522ITGB80.022965.243720413590AG7rs8832IL4R0.02 580.28151627375787GA8rs3735619ITGB80.02665.3720418678TC9rs35769460RARB0.028580.1874325290986GT10rs17016738RARB0.037415.565325600646TC11rs6673920LAMC20.040470.17581183189331AG12rs76795558RARB0.040470.1758324880096AG13rs2189300ITGB80.042334.4 42720429735AG14rs8099763LAMA30.045110.18891821409064AG15rs10486391ITGB80.047584.527720376018GA16rs6730023ITGB60.049510.21522161019869AG17rs2290706ITGB60.049510.21522161026336GA18rs2069837IL60.049830.1786722768027GA19rs13306267COL3A10.05784 0.162189860630CA20rs12339187RXRA0.057840.169137229327GA21rs11185659RXRA0.057840.169137243383TC22rs2860905CYP2C90.057840.161096702295AG23rs4086116CYP2C90.057840.161096707202TC24rs4917639CYP2C90.057840.161096725535CA25rs9332172CYP2C90.057840.161096731788GA26rs1934963CYP2C90.057840.161096734676CT27rs9849459RARB0.059074.028325009827TC28rs22949 88COL9A30.060840.23642061453549AG29rs13074694RARRES10.063160.30353158424331AG30rs6964705EGFR0.063160. 3035755209637CA31rs11976696EGFR0.067770.3275755232333GA32rs2271680COL3A10.068630.17142189862352AG33rs 284785ADH70.076390.29414100335586TC34rs7535205DHRS30.078065.062112650069CT35rs1914037COL3A10.079470.27 872189845515GA36rs12607787LAMA30.079470.27871821496166CT37rs1286772RARB0.085560.3572325580776CG38rs13 403826ITGB60.087783.3022160963372CT39rs76077040RARB0.087783.302325007603GT40rs1864909RARB0.09311732552 2717AC41rs58312942RARB0.093117325553634CA42rs2878906LAMA30.097130.26241821364642AG43rs62094786LAMA30. 097130.26241821375683TG44rs8098244LAMA30.097130.26241821405553AG45rs7616062RARB0.09893.751325578892TC.

[0177] 1) P: P value calculated from Z-statistics of logistic regression analysis 2) OR: Odds ratio, the odds ratio of genetic mutation information, which is the proportion of people with retinol efficacy and those without retinol efficacy who have a specific mutation type

[0178] 3) CHR: Chromosome

[0179] 4) BP: Basepair

[0180] 5) A1: Minor allele, the frequency of the minor allele among the frequency of occurrence of the variant type in the population within the data set

[0181] 6) A2: Major allele, the frequency of the major allele among the frequency of occurrence of the variant type in the population within the data set

[0182]

[0183] Example 2: Establishment of a polygenic effector score model to predict retinol efficacy.

[0184]

[0185] In the results of the logistic regression model analysis using the 'retinol efficacy' phenotype and the available genotype data, some of the genetic polymorphism markers (P-value < 0.1) showing association significance and the odds ratios of those markers were nonlinearly combined to define a polygenic risk score formula representing retinol efficacy, as shown in the following formula.

[0186]

[0187] i: ith subject

[0188] k: kth SNP (1 ≤ k ≤ m)

[0189] OR k : OR value of the kth SNP

[0190] X k : Number of minor alleles of the kth SNP

[0191] OR snp1 : OR value of SNP1

[0192] n1-i: Number of SNP1s in subject i

[0193] n2-i: Number of SNP2 in subject i

[0194]

[0195] The above equation calculates the efficacy score E for subject i using a combination of m SNPs, where OR (the first weight) represents the effect size (odds ratio) of each SNP, and X (the second weight) represents the number of minor alleles for each SNP (n=0,1,2). Since the effect size (odds ratio) of each SNP is a ratio or probability scale with a value between 0 and 1, the efficacy score E for subject i is calculated by starting from βO (=1) and sequentially multiplying the effect sizes (odds ratios) of a total of m SNPs by the number of minor alleles.

[0196] In the above formula, X k Or, the minor allele that constitutes n1-i, n2-i refers to the minor allele among the major allele and the minor allele, and the number can be higher as the number of minor alleles increases.

[0197]

[0198] When applying the above formula, 27 of the 45 markers in Table 1 were selected and used. The 27 markers are as shown in Table 2 below.

[0199]

[0200] Significant polymorphism markers associated with retinol efficacy (P < 0.1) Gene OR (32 people) SNPA1 A21 ADH70.2941 rs284785 TC2 COL9 A30.2364 rs2294988 GA3 CYP2 C90.16 rs1934963 CT4 DHRS35.062 rs7535205 CT5 DHRS30.3168 rs4846127 CT6 EGFR0.3275 rs11976696 GA7 EGFR0.3035 rs6964 705CA8IL4R0.299rs3024530AG9IL4R0.2815rs8832GA10IL60.1786rs2069837GA11ITGB63.3 02rs13403826CT12ITGB60.2152rs6730023AG13ITGB60.2152rs2290706GA14ITGB85.3rs373 5619TC15ITGB84.527rs10486391GA16LAMA30.2787rs12607787CT17LAMA30.1889rs8099763 AG18RARB5.557rs1021702GA19RARB4.028rs9849459CT20RARB3.302rs76077040GT21RARB3. 059rs4568101CA22RARB0.2491rs4280597GA23RARB0.2491rs1021701GA24RARB0.1874rs321 526AC25RARB0.1874rs35769460GT26RXRA0.16rs12339187GA27LAMC20.05208rs10494560TC

[0201]

[0202] Example 3: Development of a personalized prescription

[0203]

[0204] For some of the genes in Table 1, we examined the level of gene expression during retinol treatment and selected materials capable of modulating this expression. Through the following experiments, we aimed to develop a retinol formulation containing beta-sitosterol, hesperidin, baicalin, and troxerutin.

[0205]

[0206] 3-1: Discovery of Materials Regulating RARB Gene Expression

[0207]

[0208] The present invention was applied to various materials to confirm the correlation between RARB expression level and collagen synthesis ability.

[0209] Specifically, fibroblasts were seeded in 12-well plates and cultured for 24 hours. Dulbecco's Modified Eagle Medium (DMEM) was used as the medium during culture, and an incubator maintained at 37°C and CO2 was used. After removing the medium, the cells were washed once with phosphate-buffered saline (PBS), and 500 μl of PBS was added. Subsequently, 25 mJ / cm2 of UVB was irradiated using a UV irradiator. After removing the PBS, DMEM containing the test substance of the present invention was added to the cells. After 24 hours, RNA was extracted, and cDNA was synthesized using this RNA. Using the cDNA, the expression levels of collagen (COL1A1) and RARB genes were analyzed by RT-qPCR.

[0210] As a result, it was confirmed that there was a correlation between RARB expression levels and collagen synthesis ability when various substances were treated in the present invention (Fig. 4a). In addition, among the various substances, it was confirmed that hesperidin increased RARB expression by 180% and COL1A expression by 120% (Fig. 4b).

[0211]

[0212] 3-2: Discovery of Materials Regulating CYP2C9 Gene Expression

[0213] Exogenous retinoids are degraded and inactivated by various CYP family enzymes in the body. In the skin, it has been shown that exogenous retinoids are rapidly hydroxylated and degraded by CYP family enzymes in epidermal keratinocytes (Elizabeth Pavez Lorie et al., The involvement of cytochrome p450 (CYP) 26 in the retinoic acid metabolism of human epidermal keratinocytes, Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, 2009 Mar;1791(3):220-8). The role of CYP2C9 in skin tissue has not been clearly elucidated.

[0214] Specifically, keratinocytes (HaCat) were seeded in 12-well plates and cultured for 24 hours. Dulbecco's Modified Eagle Medium (DMEM) was used as the culture medium, and an incubator maintained at 37°C and CO2 was used. After removing the medium, the cells were washed once with PBS (Phosphate Buffered Saline). After removing the PBS, DMEM containing the test substance of the present invention was added to the cells. After 24 hours, RNA was extracted, and cDNA was synthesized using this RNA. Using the cDNA, CYP2C9 gene expression was analyzed by RT-qPCR.

[0215] As a result, it was confirmed that CYP2C9 was overexpressed when keratinocytes were treated with retinol (Fig. 5). Among the various substances of the present invention, baicalin was confirmed to effectively suppress the overexpression of CYP2C9 induced by retinol. This suggests that it can increase the bioactivity of retinol.

[0216]

[0217] 3-3: Discovery of materials that regulate the expression of skin barrier-related genes (LAMC2, ITGB8, EGFR)

[0218] The LAMC2 gene encodes the Laminin Gamma-2 (Laminin γ2) protein, which is a component of the skin's basement membrane and plays a role in maintaining cell-to-cell adhesion and basement membrane integrity. It is also known to regulate cell migration and differentiation within skin tissues and participate in wound healing and the recovery of damaged tissue, thereby contributing to maintaining the structure of the skin and recovering from damage (Tracy L Adair-Kirk et al., Keratinocyte-Targeted Expression of Human Laminin γ2 Rescues Skin Blistering and Early Lethality of Laminin γ2 Deficient Mice, PLoS One. 2012;7(9):e45546). It has been reported that when LAMC2 is underexpressed, epidermal detachment is observed due to basement membrane collapse.

[0219] In addition, Integrin Beta 8, encoded by the gene ITGB8, is known to exist on the cell surface, interact with and bind to the surrounding extracellular matrix, and affect cell migration and differentiation within skin tissues, and is known to control various inflammatory responses.

[0220] Epidermal Growth Factor Receptor protein encoded by the EGFR gene is known to play a role in regulating the growth, proliferation, and differentiation of skin cells, and is known to be involved in epithelial regeneration and healing, skin barrier formation, etc. It has been reported that low expression of EGFR is related to abnormal skin tissue formation such as delayed wound healing (Hong Fang et al., EGFR inhibitor gefitinib regulates barrier function in human epidermal keratinocytes via the modulation of the expression of claudins, International Journal of Molecular Medicine, 2019 Mar;43(3):1522-1530.; H Shiraha et al., Aging fibroblasts present reduced epidermal growth factor (EGF) responsiveness due to preferential loss of EGF receptors Journal of Biological Chemistry, 2000 Jun 23;275(25):19343-51.).

[0221] Accordingly, the present invention was applied to various materials to determine whether LAMC2, ITGB8, and EGFR genes were expressed.

[0222] Specifically, keratinocytes (HaCat) were seeded in 12-well plates and cultured for 24 hours. Dulbecco's Modified Eagle Medium (DMEM) was used as the culture medium, and an incubator maintained at 37°C and CO2 was used. After removing the medium, the cells were washed once with phosphate-buffered saline (PBS). After removing the PBS, DMEM containing the test substance of the present invention was added to the cells. After 24 hours, RNA was extracted, and cDNA was synthesized using this RNA. Using the cDNA, the expression levels of LAMC2, ITGB8, and EGFR genes were analyzed by RT-qPCR.

[0223]

[0224] As a result, it was confirmed that the expression of LAMC2 and ITGB8 decreased when retinol was treated in keratinocytes corresponding to the epidermis (Fig. 6a). This indicates the collapse of the basement membrane and skin barrier, and it was confirmed that the expression of LAMC2 and ITGB8 genes, which had been reduced by retinol treatment, was restored when sitosterol was treated (Fig. 6b).

[0225] In addition, when retinol is treated in keratinocytes corresponding to the epidermis, EGFR expression is reduced, which induces incomplete differentiation of epidermal keratinocytes, which causes skin barrier defects (Nicolas Joly-Tonetti et al., EGFR inhibitors switch keratinocytes from a proliferative to a differentiative phenotype affecting epidermal development and barrier function, BMC Cancer, 2021 Jan 5;21(1):5; Jerome F Sah et al., Retinoids suppress epidermal growth factor-associated cell proliferation by inhibiting epidermal growth factor receptor-dependent ERK1 / 2 activation J Biol Chem, 2002 Mar 22;277(12):9728-35. Epub 2002 Jan 11.). It was confirmed that EGFR expression reduced by retinol can be restored when troxerutine is treated (Fig. 7).

[0226]

[0227] Example 4: Validation of a Polygenic Effector Score Model for Efficacy Evaluation of Newly Developed Prescriptions and Prediction of Prescription Efficacy

[0228]

[0229] Ten subjects aged between 40 and 60 were recruited, and the number of risk alleles carried by each subject is shown in Table 3.

[0230] Significant polymorphism markers associated with retinol efficacy in 10 subjects Gene OR (32 subjects) SNPA1 A2 Subject 1 Subject 2 Subject 3 Subject 4 Subject 5 Subject 6 Subject 7 Subject 8 Subject 9 Subject 10 ADH70.2941 rs284785 TC2100000110 COL9A30.2364 rs2294988 GA0102121112 CYP2C90.16 rs1934963 CT0101000000 DHRS35.062 rs7535205 CT1011010111 DHRS30.3168 rs4846127 CT0010111011 EGFR0.3275 rs11976696 GA1201100111EGFR0.3035rs6964705CA1201100100IL4R0.299rs3024530AG1110110102IL4R0.2815rs8832GA0110111100IL60.1786 rs2069837GA1010000011ITGB63.302rs13403826CT0001111100ITGB60.2152rs6730023AG2201110111ITGB60.2152rs2290706GA2201 110111ITGB85.3rs3735619TC1011201111ITGB84.527rs10486391GA1011100111LAMA30.2787rs12607787CT1101110100LAMA30.188 9rs8099763AG1001100100RARB5.557rs1021702TGA0011110100RARB4.028rs9849459CT1101120102RARB3.302rs76077040GT1121102 110RARB3.059rs4568101CA1100120211RARB0.2491rs4280597GA0010100110RARB0.2491rs1021701GA0010100110RARB0.1874rs321 526AC1000110101RARB0.1874rs35769460GT1000010011RXRA0.16rs12339187GA1000001001LAMC20.05208rs10494560TC0011000000

[0231] The subjects used a cream containing 0.1% retinol, 0.1% hesperidin, 0.002% baicalin, 0.1% beta-sitosterol, and 0.1% troxerutin for 8 weeks. Afterwards, the subjects' wrinkles around the eyes were analyzed using Antera 3D equipment, and the wrinkle improvement rate compared to week 0 was measured.

[0232] A cream formulation was prepared with 0.1% retinol, 0.1% hesperidin, 0.002% baicalin, 0.1% beta-sitosterol, and 0.1% troxerutin at appropriate concentrations.

[0233] After applying the polygenic effector score model of Experimental Example 2 to the data in Table 3, -Log2 was taken and the data were divided into two groups, High and Low, based on values ​​around 9. The average wrinkle improvement rate of the groups was calculated and the results are shown in the graph in Figure 8. As a result, a significant difference in efficacy was observed between the two groups.

[0234]

[0235] Based on the calculated efficacy scores, the entire study population was divided into 1:1 groups. This was then applied to a set of subjects divided into 1:1 groups based on actual wrinkle improvement rates (rates of change). This confirmed the accuracy, sensitivity, and specificity of predicting retinol efficacy (Table 4). This demonstrates that this model can demonstrate excellent predictive performance.

[0236]

[0237]

[0238] Example 5: Correlation between retinol efficacy significant polymorphism markers and new prescriptions (boosters)

[0239]

[0240] Among the genes in Table 1, among the genes closely related to efficacy, when one or more minor alleles are possessed among the alleles, the efficacy of retinol was observed to be lower when the efficacy booster of the present invention was prescribed compared to the control group (Fig. 9). Specifically, for the SNPs in Table 1, the OR (odds ratio) value was less than 1, confirming that the degree of efficacy decreased as the number of minor alleles increased. In other words, it was confirmed that the efficacy of retinol increased as the number of major alleles increased, resulting in the observation of reduced wrinkles.

[0241]

[0242] This suggests that the efficacy booster of the present invention can be tailored to individuals with genes closely related to efficacy.

[0243]

[0244] From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without altering its technical spirit or essential characteristics. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as encompassing all changes or modifications derived from the meaning and scope of the following claims and their equivalent concepts, rather than the detailed description above.

Claims

1. A composition for determining the efficacy of retinol, comprising a probe or an agent capable of detecting or amplifying one or more single nucleotide polymorphism (SNP) markers for determining the efficacy of retinol selected from Table 1.

2. A composition according to claim 1, wherein the single nucleotide polymorphism marker for determining the retinol efficacy further comprises a polynucleotide corresponding to one or more single nucleotide polymorphism markers selected from any one of Table 1; and one or more polynucleotides selected from the group consisting of complementary polynucleotides thereof.

3. A kit for assessing the efficacy of retinol comprising the composition of claim 1 or 2.

4. In the third paragraph, the kit is a kit for determining the efficacy of retinol, which is an RT-PCR kit or a DNA chip kit.

5. A microarray for assessing retinol efficacy, comprising a single nucleotide polymorphism (SNP) marker for assessing retinol efficacy according to Article 1. 6.(a) A step of amplifying or hybridizing with a probe a polymorphic site of a single nucleotide polymorphism marker for determining retinol efficacy according to Article 1 in DNA obtained from a sample separated from an individual; and (b) A method for providing information for selecting a customized substance for improving the efficacy of retinol, comprising the step of identifying the base of the amplified or hybridized polymorphic site of step (a).

7. A method for providing information in paragraph 6, wherein the sample is hair, urine, blood, various body fluids, isolated tissue, isolated cells, or saliva.

8. A method for providing information in paragraph 6, wherein the amplification and confirmation of the polymorphic site uses a SNP chip.

9. In paragraph 6, A method for providing information, wherein the substance is at least one selected from the group consisting of beta-sitosterol, hesperidin, baicalin, and troxerutin.

10. A step in which genetic information of a prediction target entity is input into the prediction terminal; and A step in which the input genetic information is transmitted to the server, the server calculates efficacy information based on a comparison result of a threshold efficacy score determined in advance in the server and an efficacy score calculated based on the input genetic information, and the calculated efficacy information is transmitted and output to the prediction terminal; The above efficacy scores are, The number of single nucleotide polymorphism markers selected from Table 1 and the type of single nucleotide polymorphism markers that are present in the genetic information of a specific individual are determined according to the How to predict retinol efficacy.

11. In paragraph 10, The above single nucleotide polymorphism marker type is, Major allele-major allele type; Superordinate allele - minor allele type; and Suballelic - a suballelic type; including; How to predict retinol efficacy.

12. In paragraph 10, The above efficacy scores are, In the above genetic information, the OR value of a specific single nucleotide polymorphism marker according to a combination of one or more of the above single nucleotide polymorphism markers is lower the more bases there are lower than 1, and higher the OR value of a specific single nucleotide polymorphism marker is higher the more bases there are higher than 1. How to predict retinol efficacy.

13. In paragraph 12, The above efficacy score is determined according to the following mathematical formula: Here, is the efficacy score, and OR k is the efficacy weight of the kth single nucleotide polymorphism marker, Xk is a number determined according to the single nucleotide polymorphism marker type of the kth single nucleotide polymorphism marker, and k is an integer greater than or equal to 1. How to predict retinol efficacy.

14. In paragraph 13, The above efficacy information is, Including the degree of wrinkle improvement when applying retinol to the predicted target object, How to predict retinol efficacy.

15. A prediction terminal that receives genetic information of a prediction target object and outputs efficacy information calculated by the server based on the input genetic information; and A server is included, which transmits genetic information input to the above prediction terminal and calculates efficacy information based on a comparison result between a predetermined threshold efficacy score and an efficacy score calculated based on the input genetic information; The above efficacy scores are, The number of single nucleotide polymorphism markers selected from Table 1 and the type of single nucleotide polymorphism markers that are present in the genetic information of a specific individual are determined according to the Retinol efficacy prediction system.

16. A computer program stored in a computer-readable recording medium to execute a method according to any one of claims 10 to 15.

17. A composition for improving the efficacy of retinol, comprising as an active ingredient at least one substance selected from the group consisting of beta-sitosterol, hesperidin, baicalin, and troxerutin.

18. In paragraph 17, A composition characterized in that the above material is contained in an amount of 0.0001 to 10 wt% based on the total weight of the composition.

19. In Article 17, A composition characterized in that the substance promotes the expression of RARB, LAMC2, ITGB8 or EGFR; or inhibits the expression of CYP2C9.

20. In paragraph 19, A composition wherein the substance for promoting the expression of RARB is hesperidin.

21. In paragraph 19, A composition wherein the substance for promoting the expression of LAMC2 or ITGB8 is beta-sitosterol.

22. In paragraph 19, A composition wherein the substance for promoting the expression of EGFR is troxerutine.

23. In paragraph 19, A composition wherein the substance for inhibiting the expression of CYP2C9 is baicalin.

24. In paragraph 17, The composition is a customized composition targeting an individual having one or more single nucleotide polymorphism markers present in a gene encoding a protein of RARB, LAMC2, ITGB8, EGFR or CYP2C9.

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