Composition for improving skin wrinkles

Abstract

The present invention relates to a composition for improving skin wrinkles and an information providing method for selecting a custom-made substance for improving wrinkles, the method comprising a step of identifying a base of a polymorphic site of a gene polymorphic marker that is significant in association with the degree of skin wrinkles. Furthermore, the present invention relates to: a composition for diagnosing the type of skin in which wrinkles are likely to be generated, the composition comprising a probe capable of detecting a gene polymorphism marker having significance associated with the degree of wrinkles in the skin or a preparation capable of amplifying the gene polymorphism marker; a kit or microarray comprising the composition; and a method for providing information on the type of skin on which wrinkles are likely to be generated, using the gene polymorphism marker or the combination of markers. The wrinkles relate to skin fine wrinkles, drooping pore wrinkles or nasolabial wrinkles.

Description

Technical Field

[0001] The present invention relates to a composition for improving skin wrinkles and a method for providing information for selecting a customized substance for improving wrinkles, the method including a step of identifying the base at a polymorphic site of a gene polymorphism marker having an association significance with the degree of skin wrinkles. Further, the present invention relates to a composition for diagnosing whether a skin type prone to wrinkle formation is present or not, which includes a probe capable of detecting a gene polymorphism marker having an association significance with the degree of skin wrinkles or a preparation capable of amplifying the gene polymorphism marker, a kit or a microarray including the above composition, and a method for providing information on whether a skin type prone to wrinkle formation is present or not by using the above gene polymorphism marker or a combination of markers. The above wrinkles refer to skin fine lines, sagging pore wrinkles or nasolabial folds. Background Art

[0002] The skin is a protective film having various physiological functions and is also an important organ for obtaining aesthetic favor from others. Skin aging is generally caused by two processes. The changes that occur to anyone over time without special environmental factors are called intrinsic aging, and the changes that occur on the face, neck, hands, etc. due to long-term exposure to environmental factors such as sunlight are called photoaging, that is, extrinsic aging. Among the two aging processes, the reduction of collagen in the dermis is most closely related to wrinkle formation. In intrinsic aging, the aging of skin fibroblasts leading to a decrease in the ability to generate new collagen fibers is one of the main reasons. In extrinsic aging, damage to skin fibroblasts by ultraviolet rays, reactive oxygen species, etc. leading to a decrease in the ability to generate new collagen fibers, or an increase in collagen-degrading enzymes leading to a decrease in the total amount of dermal collagen is one of the reasons for wrinkle formation.

[0003] As people age, skin aging, reduced elasticity, pigmentation, and other signs of skin aging occur naturally. To date, the trend has been to care for these signs of aging after they have occurred. However, recently, with the active research on genes, if one can understand an individual's genetic constitution related to the skin, it is possible to maintain beautiful skin for a longer time through more proactive management (Kemp et al., Molecules. 2017 Feb 26; 22(3), 2017). In particular, through services such as DTC testing (Direct To Consumer, a genetic testing service where consumers directly commission a genetic testing agency), various companies are providing genetic testing services that analyze SNPs related to an individual's DNA to understand each characteristic. Skin genetic testing is a type of genetic testing that analyzes SNPs (Single Nucleotide Polymorphisms) related to skin pigmentation, elasticity, aging, and vitamin C concentration in an individual's unique DNA, helping to prevent skin aging and effectively manage the skin.

[0004] Currently, a very diverse range of gene polymorphism markers have been revealed. However, most of them are markers related to skin color, whitening, moisture, ultraviolet rays, etc. In fact, the situation where they are accompanied by mechanism research is not common. In particular, it can be considered that there are few cases where mechanism research has been conducted or customized materials have been developed for polymorphism markers related to wrinkles, specifically, polymorphism markers related to fine lines on the skin, polymorphism markers related to sagging pores and wrinkles, and polymorphism markers related to nasolabial folds. Summary of the Invention

[0005] Technical Problem

[0006] The inventors of the present invention aimed to construct a skin characteristic classification system for Koreans by building a big data of genetic information and skin type information, to construct a scientific skin classification standard by understanding the genetic characteristics that determine an individual's skin characteristics, and to develop personalized active ingredients based on this, and through the subdivision of various products, contribute to the development of customized cosmetics for different skin characteristics. After unremitting efforts, it was confirmed that specific single nucleotide polymorphism (SNP) markers significantly related to skin fine lines, sagging pores and wrinkles, and nasolabial folds can be screened, and personalized substances that can help improve fine lines, improve sagging pores and wrinkles, and improve nasolabial folds are provided based on genetic information.

[0007] Means for Solving the Problem

[0008] One object of the present invention is to provide a composition for improving skin fine lines. Another object of the present invention is to provide a cosmetic composition or a pharmaceutical external composition containing the above composition as an active ingredient.

[0009] Another object of the present invention is to provide a method for providing information on a customized substance for improving fine lines, which includes the step of confirming the bases at the polymorphic sites of single nucleotide polymorphism markers associated with the degree of skin fine lines.

[0010] One object of the present invention is to provide a single nucleotide polymorphism (SNP) marker for diagnosing whether a skin type is prone to generating fine lines.

[0011] Another object of the present invention is to provide a composition for diagnosing whether a skin type is prone to generating fine lines, which contains a probe capable of detecting a single nucleotide polymorphism (SNP) marker for diagnosing whether a skin type is prone to generating fine lines or a preparation capable of amplifying the single nucleotide polymorphism (SNP) marker.

[0012] Another object of the present invention is to provide a kit or microarray for diagnosing whether a skin type is prone to generating fine lines, which contains the above composition.

[0013] Another object of the present invention is to provide a method for providing information on whether a skin type is prone to generating fine lines, which includes the step of confirming the polymorphic sites of the above single nucleotide polymorphism markers.

[0014] One object of the present invention is to provide a composition for improving sagging pores and wrinkles.

[0015] Another object of the present invention is to provide a cosmetic composition or a pharmaceutical external composition containing the above composition as an active ingredient.

[0016] Another object of the present invention is to provide a method for providing information on a customized substance for improving sagging pores and wrinkles, which includes the step of confirming the bases at the polymorphic sites of single nucleotide polymorphism markers associated with the degree of sagging pores and wrinkles.

[0017] One object of the present invention is to provide a single nucleotide polymorphism (SNP) marker for diagnosing whether a skin type is prone to generating sagging pores and wrinkles.

[0018] Another object of the present invention is to provide a composition for diagnosing whether a skin type is prone to generating sagging pores and wrinkles, which contains a probe capable of detecting a single nucleotide polymorphism (SNP) marker for diagnosing whether a skin type is prone to generating sagging pores and wrinkles or a preparation capable of amplifying the single nucleotide polymorphism (SNP) marker.

[0019] Another object of the present invention is to provide a kit or microarray for diagnosing whether a skin type is prone to generating sagging pores and wrinkles, which contains the above composition.

[0020] Another object of the present invention is to provide a method for providing information on whether a skin type is prone to forming drooping pore wrinkles, which includes the step of confirming the polymorphic sites of the above-mentioned single nucleotide polymorphism markers.

[0021] One object of the present invention is to provide a composition for improving nasolabial folds.

[0022] 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.

[0023] Another object of the present invention is to provide a method for providing information for selecting a customized substance for improving nasolabial folds, which includes the step of confirming the bases of the polymorphic sites of the single nucleotide polymorphism markers significantly associated with the degree of nasolabial folds.

[0024] One object of the present invention is to provide a single nucleotide polymorphism (SNP) marker for diagnosing whether a skin type is prone to forming nasolabial folds.

[0025] Another object of the present invention is to provide a composition for diagnosing whether a skin type is prone to forming nasolabial folds, which contains a probe capable of detecting the single nucleotide polymorphism (SNP) marker for diagnosing whether a skin type is prone to forming nasolabial folds or a preparation capable of amplifying the single nucleotide polymorphism (SNP) marker.

[0026] Another object of the present invention is to provide a kit or microarray for diagnosing whether a skin type is prone to forming nasolabial folds, which contains the above composition.

[0027] Another object of the present invention is to provide a method for providing information on whether a skin type is prone to forming nasolabial folds, which includes the step of confirming the polymorphic sites of the above-mentioned single nucleotide polymorphism markers.

[0028] Advantages of the Invention

[0029] Predictive information on the degree of skin wrinkle occurrence of an individual can be provided through the gene polymorphism markers significantly associated with the degree of skin wrinkles of the present invention. Furthermore, customized ingredients or products capable of alleviating the degree of skin wrinkles can be developed based on the information of the gene polymorphism markers observed in the individual. The above wrinkles include skin fine lines, drooping pore wrinkles, or nasolabial folds. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 It shows the promotion of collagen expression brought about by the treatment with phloretin and / or oryzanol.

[0031] Figure 2 It shows the promotion of collagen expression brought about by the treatment with lupeol and / or sucralfate.

[0032] Figure 3 Shows Antera 3D and the analysis positions of crow's feet.

[0033] Figure 4 Shows the reference images for expert visual evaluation.

[0034] Figure 5 Shows the improvement rate of crow's feet relative to retinol when applying a cream containing phloretin, oryzanol, lupeol, and sucralfate.

[0035] Figure 6 Shows the inhibition of BNC2 expression brought about by the treatment with phloretin and / or oryzanol.

[0036] Figure 7 Shows the promotion of EDAR expression brought about by the treatment with lupeol and / or sucralfate.

[0037] Figure 8 And Figure 9 Shows the improvement rate of crow's feet relative to retinol when applying a cream containing phloretin, oryzanol, lupeol, and sucralfate to the high-risk group.

[0038] Figure 10 Shows the promotion of collagen expression brought about by the treatment with fucoidan and / or N-feruloylserotonin.

[0039] Figure 11 Shows the promotion of collagen expression brought about by the treatment with emodin and / or formononetin.

[0040] Figure 12 Shows Antera 3D and the analysis positions of the cheek area.

[0041] Figure 13 Shows the reference images for expert visual evaluation.

[0042] Figure 14 Shows the improvement rate of sagging pores relative to retinol when applying a cream containing fucoidan, N-feruloylserotonin, emodin, and formononetin.

[0043] Figure 15 Shows the inhibition of SGPP2 expression brought about by the treatment with fucoidan and / or N-feruloylserotonin.

[0044] Figure 16 Shows the promotion of ADIPOQ expression brought about by the treatment with emodin and / or formononetin.

[0045] Figure 17 And Figure 18Shows the improvement rate of drooping pore wrinkles relative to retinol when applying a cream containing fucoidan, N-feruloylserotonin, emodin, and formononetin to the high-risk group.

[0046] Figure 19 and Figure 20 Shows the promotion of collagen expression brought about by the treatment with the substance of the present invention.

[0047] Figure 21 Shows Antera 3D and the analysis positions of nasolabial folds.

[0048] Figure 22 Shows the reference images for expert visual evaluation.

[0049] Figure 23 Shows the improvement rate of nasolabial folds relative to retinol when applying a cream containing the substance of the present invention.

[0050] Figure 24 Shows the improvement rate of nasolabial folds relative to retinol when applying a cream containing the substance of the present invention to the high-risk group. Detailed Description

[0051] The specific description is as follows. On the other hand, each description and embodiment disclosed in the present invention can also be applied to each other description and embodiment. That is, all combinations of various elements disclosed in the present invention belong to the scope of the present invention. In addition, it should not be considered that the scope of the present invention is limited by the specific descriptions described below.

[0052] In addition, those of ordinary skill in the art can recognize or confirm multiple equivalents to specific aspects of the present invention described in the present invention only by using conventional experiments. In addition, such equivalents are intended to be included in the present invention.

[0053] As one aspect for achieving the object of the present invention, the present invention provides a composition for improving skin fine lines. As an example, the above composition contains one or more substances selected from the group consisting of phloretin, oryzanol, lupeol, and sucralfate as active ingredients. As another example, the above composition may contain (i) phloretin and oryzanol; (ii) lupeol and sucralfate; or (iii) phloretin, oryzanol, lupeol, and sucralfate as active ingredients, but is not limited thereto.

[0054] The above-mentioned term "phloretin" is a dihydrochalcone plant-derived phenol with a C6-C3-C6 skeleton, which is rich in apples (mainly the peel) and apple leaves and exists in the form of free or glucosidic phloridzin (phloretin 2'-O-glucose). Phloretin has various biological functions including antioxidant and anti-cancer activities and is related to the prevention of heart disease. The molecular formula of phloretin is C 15 H 14 O5, with a molecular weight of 274.272, and can be represented by the following Chemical Formula 1. The above-mentioned phloretin is not limited to its obtaining method and can be chemically synthesized by methods well-known in the art or commercially available substances can be used.

[0055] [Chemical Formula 1]

[0056]

[0057] The above-mentioned term "oryzanol" is a substance extracted from rice bran, corn, barley, etc. It is a compound in which ferulic acid is used as the parent nucleus and sterols such as campesterol and β-sitosterol are ester-bonded with triterpene alcohols. It is reported that this oryzanol is the main component of drugs for treating menopausal disorders and autonomic nerve dysfunction abnormalities. The above-mentioned oryzanol is not limited to its obtaining method and can be chemically synthesized by methods well-known in the art or commercially available substances can be used. According to an embodiment of the present invention, the above-mentioned oryzanol can be γ-oryzanol (HMARU PHAROUS CO.), but is not limited thereto.

[0058] The above-mentioned term "lupeol" is a fat-soluble component of plant origin, specifically of leguminous plant origin. It can directly dissolve sebum, enter pores to kill acne bacteria, and play a role in inhibiting various causes of acne such as sebum production, inflammation, and keratinization, which are the causes of acne. The molecular formula of lupeol is C 30 H 50 O, with a molecular weight of 426.729, and can be represented by the following Chemical Formula 2. The above-mentioned lupeol is not limited to its obtaining method and can be chemically synthesized by methods well-known in the art or commercially available substances can be used.

[0059] [Chemical Formula 2]

[0060]

[0061] The above-mentioned term "sucralfate" is a drug used to treat gastric ulcers, gastroesophageal reflux disease, gastritis, etc. and to prevent stress ulcers. The salt part of sucralfate binds to fibroin on the gastric wall to form a membrane, and the above-mentioned membrane coats the gastric wall to protect the stomach from gastric acid, bile, etc., thereby relieving the pain of the stomach and enabling sucralfate to function as an antacid. The above-mentioned sucralfate is not limited to its method of obtaining and can be chemically synthesized by methods known in the art or commercially available substances can be used.

[0062] Based on the total weight of the composition, the content of the substance of the present invention can be about 0.0001 to 10% by weight, but is not limited thereto. As an example, based on the total weight of the composition, the skin fine line improvement material of the present invention can be 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.

[0063] For the purpose of the present invention, the above-mentioned substance can impart an effect of promoting the improvement of skin fine lines.

[0064] For the purpose of the present invention, the above-mentioned term "skin wrinkle" refers to the appearance of the skin being folded due to a decrease in the elasticity of a part of an individual's skin. Due to the folding of the skin, a shadow is formed at the wrinkle part compared to the surrounding skin, and therefore, it is observed to be relatively dark compared to the surrounding skin. In addition, most wrinkles have a linear appearance, and therefore, when the shadow parts are connected and observed to be longer than a certain length, they can be defined as wrinkles, but are not limited thereto.

[0065] The above-mentioned skin wrinkles are related to human aging and can form relatively shallow fine lines around the corners of the eyes or mouth. The term called skin fine lines can be used interchangeably with terms such as "relief", "furrow", "wrinkle", "fine wrinkle", "line", etc. In addition, the above-mentioned skin fine lines can be fine lines under the eyes or fine lines at the corners of the mouth, but are not limited thereto.

[0066] For the purpose of the present invention, the above-mentioned substance can be characterized by promoting collagen synthesis. Specifically, when phloretin and / or oryzanol in the above-mentioned substance are treated, the synthesis of type I collagen or type IV collagen can be promoted, and when lupeol and / or sucralfate are treated, the synthesis of type III collagen or type IV collagen can be promoted, but is not limited thereto.

[0067] As an example, the cream containing the substance of the present invention which is effective in promoting collagen synthesis was applied to the corners of the eyes, and as a result, it was confirmed that it showed an excellent wrinkle improvement rate compared to the application of retinol as a control group.

[0068] In fact, many wrinkle improvement raw materials for collagen have been launched on the market. However, it is understood that the actual perceived improvement rate by consumers is quite low. In addition, in fact, non-retinol raw materials that exceed retinol, which is the raw material with the most excellent known wrinkle improvement effect among existing raw materials, are not common. However, in the present invention, it was confirmed that non-retinol substances have an excellent effect on promoting collagen synthesis, and from this point of view, it can be considered significant.

[0069] For the purpose of the present invention, the above-mentioned substance may be characterized by promoting the expression of EDAR or inhibiting the expression of BNC2. Specifically, when treating phloretin and / or oryzanol in the above-mentioned substance, the expression of BNC2 can be inhibited, and when treating lupeol and / or sucralfate, the expression of EDAR can be promoted, but it is not limited thereto. It can be seen that the reactivity to the substance of the present invention varies according to the type of specific gene polymorphism, which indicates that the efficiency of fine wrinkle improvement can be increased by providing a customized formulation for high-risk groups with a high level of risk of fine wrinkle occurrence.

[0070] In the present invention, the term "about" may be given before a specific numerical value. The term "about" used in this application includes not only the exact number described after the term, but also a range that is almost or close to that number. Considering the context in which the number is given, it can be determined whether it is close to the specific number mentioned or almost the number. As an example, the term "about" may refer to a range of -10% to +10% of the numerical value. As another example, the term "about" may refer to a range of -5% to +5% of the given numerical value. However, it is not limited thereto.

[0071] The above-mentioned composition is characterized in that it is a personalized customized type. Specifically, a personalized customized substance can be provided for an individual having a single nucleotide polymorphism (SNP) marker significantly associated with a specific degree of skin fine wrinkles.

[0072] As an example, the single nucleotide polymorphism (SNP) marker significantly associated with the degree of skin fine wrinkles may be any one or more selected from Table 1, but it is not limited thereto.

[0073] In the present invention, the term "polymorphism" refers to a situation where there are more than two alleles at a locus, and among polymorphic sites, those with only a single base difference according to different individuals are called single nucleotide polymorphisms (SNPs). Preferred polymorphic markers have more than two alleles with an occurrence frequency of 1% or more, more specifically 10% or 20% or more, in a selected population. A "gene polymorphism marker" generally refers to a situation where more than two alleles are observed at the same gene locus (base), and usually, according to different individuals, there are cases of major allele / major allele, major allele / minor allele, and minor allele / minor allele. In the present invention, it can be used interchangeably with "polymorphism marker", and refers to the base and base locus of the minor allele, or can be defined together with the chromosome number and base position, but is not limited thereto.

[0074] In the present invention, the term "allele" refers to multiple types of a gene existing at the same locus on homologous chromosomes. Alleles are also used to represent polymorphisms. For example, SNPs have two types of bialleles. Additionally, it refers to a combination of more than two bases with the same chromosome number and base position, and the above bases include the major allele with a high occurrence frequency in individuals of a specific population and the minor allele with an occurrence frequency lower than that of the major allele.

[0075] Specifically, the gene polymorphism marker of the present invention has a significant association with skin fine lines, can have two alleles, and can be classified into cases of major allele / minor allele, minor allele / minor allele, and major allele / major allele.

[0076] When having an allele of a skin fine line-associated significant polymorphism marker, depending on the allele the individual has, the effect of improving skin fine lines through the reaction caused by the substance of the present invention may vary. As an example, when holding a major allele / major allele, major allele / minor allele, or minor allele / minor allele, the reactivity caused by the substance of the present invention can be measured, that is, the skin characteristic with a high or low degree of fine line improvement. Therefore, it can provide information for selecting a customized substance for individuals with 4 or more minor alleles to increase the improvement of fine lines.

[0077] In the present invention, the term "rs_id" refers to the independent identifier rs-ID assigned to all SNPs initially registered by NCBI that have accumulated SNP information since 1998. The rs_id recorded in this table refers to the polymorphism marker of the present invention, that is, the SNP marker.

[0078] The above single nucleotide polymorphism markers can be one or more single nucleotide polymorphism markers selected from the single nucleotide polymorphism markers shown in Table 1 and Table 4. Whether the single nucleotide polymorphism markers shown in Table 1 and Table 4 are related to the degree of skin fine lines can be determined.

[0079] The degree of skin wrinkles of the single nucleotide polymorphism marker of the present invention is judged by measuring the frequency number of each marker. The significance of this is characterized by but not limited to the p-value, such as a p-value 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 (p-value). Specifically, the p-value can be less than 0.01, more specifically, the p-value can be less than 0.001, and more specifically can be less than 0.0001, but not limited to this.

[0080] The single nucleotide polymorphism (SNP) marker of the present invention can be any one or more selected from the markers shown in Table 1 or Table 4, but not limited to this. The above single nucleotide polymorphism (SNP) markers can be one or more, and can be used in combinations of 2 or more, 3 or more, 4 or more, etc. that can determine the number of wrinkles, but not limited to this.

[0081] The above marker can be the SNP itself, or a polynucleotide composed of 5 - 100 consecutive DNA sequences containing the above SNP site, or a polynucleotide composed of its complementary sequence, but not limited to this.

[0082] As an example, the above-mentioned fine wrinkle degree-related significant single nucleotide polymorphism markers may be one or more single nucleotide polymorphism markers present in the genes encoding proteins of EDAR or BNC2. Specifically, they may be any one or more selected from the group consisting of rs10865025, rs10756807, rs1952688, rs10116469, and rs12338608, but are not limited thereto.

[0083] As an example, when the SNP ID is rs10865025, the Chr.Position (GRCh ver.37) is recorded as "2:109511765". If the allele is disclosed as G>A, this means that the 109511765th base of human chromosome 2 is G or A, and the base on the left side of the ">" in the allele may mean the major allele, and the base on the right side may mean the minor allele.

[0084] As a specific example, the markers selected from Table 1 may be composed of one or more polynucleotides selected from the group consisting of the following polynucleotides; and their complementary polynucleotides, but are not limited thereto: a polynucleotide composed of 5-100 consecutive DNA sequences containing the 109511765th base of human chromosome 2, wherein the 109511765th base is G or A (rs10865025); a polynucleotide composed of 5-100 consecutive DNA sequences containing the 16756041st base of human chromosome 9, wherein the 16756041st base is G or A (rs10756807); a polynucleotide composed of 5-100 consecutive DNA sequences containing the 16782156th base of human chromosome 9, wherein the 16782156th base is A or G (rs1952688); a polynucleotide composed of 5-100 consecutive DNA sequences containing the 16783095th base of human chromosome 9, wherein the 16783095th base is A or T (rs10116469); a polynucleotide composed of 5-100 consecutive DNA sequences containing the 16800470th base of human chromosome 9, wherein the 16800470th base is C or T (rs12338608).

[0085] As another embodiment, one or more of the single nucleotide polymorphism (SNP) markers shown in Table 4 in the same manner as Table 1 may be selected, but are not limited thereto.

[0086] The composition of the present invention can be 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 solutions, topical ointments, creams, foams, nutritive lotions, softening lotions, face masks, softening waters, emulsions, makeup bases, serums, soaps, liquid cleansers, bath agents, sunscreen creams, sunscreen oils, suspensions, emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleaning agents, oils, powder foundations, emulsion foundations, wax foundations, patches, and sprays, but is not limited thereto.

[0087] In addition, the cosmetic composition of the present invention may further contain one or more cosmetically acceptable carriers incorporated into general skin cosmetics. As conventional ingredients, for example, oils, water, surfactants, humectants, lower alcohols, thickeners, chelating agents, pigments, preservatives, fragrances, etc. can be appropriately incorporated, but are not limited thereto.

[0088] The cosmetically acceptable carriers contained in the cosmetic composition of the present invention vary depending on the dosage form.

[0089] When the dosage form of the present invention is an ointment, paste, cream or gel, as carrier components, animal oils, vegetable oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silica, talc, zinc oxide or mixtures thereof can be used.

[0090] When the dosage form of the present invention is a powder or spray, as carrier components, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or mixtures thereof can be used. In particular, when it is a spray, a propellant such as chlorofluorocarbon, propane / butane or dimethyl ether can be additionally contained.

[0091] When the dosage form of the present invention is a solution or emulsion, as carrier components, solvents, solubilizers or emulsifiers can be used. For example, water, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol oil can be used. In particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol fatty acid esters, polyethylene glycol or fatty acid esters of sorbitan can be used.

[0092] When the dosage form of the present invention is a suspension, as carrier components, liquid diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitan esters and polyoxyethylene sorbitan anhydride esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth can be used.

[0093] When the dosage form of the present invention is soap, as carrier components, alkali metal salts of fatty acids, fatty acid semi-ester salts, fatty acid protein hydrolysates, hydroxyethyl sulfonates, lanolin derivatives, fatty alcohols, vegetable oils, glycerol, sugars, etc. can be used.

[0094] On the other hand, the above composition provides a pharmaceutical external preparation composition for alleviating or improving skin fine lines.

[0095] In the present invention, the term "pharmaceutical external preparation" refers to an article with a milder effect than pharmaceuticals among articles used for the purpose of diagnosing, treating, improving, alleviating, disposing of, or preventing diseases of humans or animals. For example, according to the Korean Pharmaceutical Affairs Act, pharmaceutical external preparations do not include articles used for the purposes of pharmaceuticals, and include products for the treatment or prevention of diseases of humans and animals, products with a mild or indirect effect on the human body, etc.

[0096] As an example, the pharmaceutical external preparation composition of the present invention can be prepared in a form selected from the group consisting of body wash, shampoo, conditioner, foam, soap, facial mask, ointment, cream, lotion, essence, and spray, but is not limited thereto.

[0097] On the other hand, the present invention provides a method for providing information for selecting a customized substance for improving fine lines, which includes the following steps: (a) obtaining a biological sample from an individual; (b) amplifying or hybridizing with a probe a polymorphism site of any one or more single nucleotide polymorphism markers selected from the group consisting of rs10865025, rs10756807, rs1952688, rs10116469, and rs12338608 from the above biological sample; and (c) confirming the base of the polymorphism site amplified or hybridized in the above step (b).

[0098] Skin fine lines can be improved by treating the selected customized substance to an individual with a specific polymorphism site.

[0099] The term "individual" in the present invention refers to a subject for diagnosing the degree of skin fine lines. In the above test sample, DNA can be obtained from samples such as hair, urine, blood, various body fluids, isolated tissues, isolated cells, or saliva, but is not limited thereto.

[0100] The method for obtaining genomic DNA in the above step (a) can use any method known to those skilled in the art.

[0101] The step of amplifying the polymorphic site of the above single nucleotide polymorphism marker or hybridizing it with a probe from the DNA obtained in the above step (a) can be carried out using any method known to those skilled in the art. For example, it can be obtained by PCR amplifying the target nucleic acid and purifying it. 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-sustained sequence replication (Guatelli et al., Proc. Natl. Acad. Sci. USA 87, 1874 (1990)), and nucleic acid sequence-based amplification (NASBA) can be used.

[0102] In the above method, the determination of the base of the polymorphic site in step (b) includes sequencing analysis, hybridization by microarray, allele specific PCR, dynamic allele hybridization technology (dynamic allele-specific hybridization, DASH), PCR extension analysis, SSCP, PCR-RFLP analysis, or TaqMan technology, SNPlex platform (Applied Biosystems), mass spectrometry (such as the MassARRAY system of Sequenom), mini-sequencing method, Bio-Plex system (BioRad), CEQ and SNPstream system (Beckman), Molecular Inversion Probe array technology (such as Affymetrix GeneChip), and BeadArray Technologies (such as Illumina GoldenGate and Infinium assays), but is not limited thereto. By the above method or other methods available to those skilled in the art to which the present invention pertains, one or more alleles in polymorphic markers including microsatellite markers, SNPs, or other types of polymorphic markers can be confirmed. The determination of the base of such a polymorphic site can specifically be carried out using a SNP chip.

[0103] The above substance can be one or more selected from the group consisting of phloretin, oryzanol, lupeol, and sucralfate, but is not limited thereto.

[0104] For the purpose of the present invention, the above-mentioned fine line degree-related significant genes can provide information capable of selecting specific substances for each individual. As an example, for an individual having one or more single nucleotide polymorphism markers in the gene encoding the protein of EDAR or BNC2, when treating one or more substances selected from the group consisting of lupeol, phloretin, oryzanol, lupeol, and sucralfate, the improvement of fine lines can be enhanced, but the optimal substance combination can be provided according to the gene characteristics of each individual.

[0105] This indicates that information can be provided to enable screening of specific single nucleotide polymorphism (SNP) markers significantly related to skin fine lines and selection of personalized substances capable of maximizing the effect of improving fine lines based on genetic information.

[0106] As an example, an information-providing method can be provided to select phloretin and oryzanol when the bases of rs10865025, rs10756807, rs1952688, rs10116469, and rs12338608 contain 4 or more minor alleles; or select lupeol and sucralfate; or select phloretin, oryzanol, lupeol, and sucralfate, but not limited thereto.

[0107] In the present invention, the term "SNP chip" refers to one of the DNA microarrays capable of simultaneously identifying the bases of hundreds of thousands of SNPs.

[0108] The TaqMan method includes: (1) the step of designing and manufacturing primers and TaqMan probes to enable amplification of the required DNA fragment; (2) the step of labeling the probes of different alleles with FAM dye and VIC dye (Applied Biosystems); (3) the step of performing PCR using the above DNA as a template with the above primers and probes; (4) the step of analyzing and confirming the TaqMan analysis plate with a nucleic acid analyzer after the completion of the above PCR reaction; and (5) the step of determining the genotype of the polynucleotide in step (1) according to the above analysis results.

[0109] In the above, sequencing analysis can be performed using conventional methods for determining base sequences and can be carried out using an automated gene analyzer. In addition, allele-specific PCR refers to a PCR method in which a primer set including a primer designed with the base where the SNP is located as the 3'-end is used to amplify the DNA fragment where the above SNP is located. The principle of the above method utilizes the following: for example, when a specific base is substituted from A to G, when a PCR reaction is carried out by designing a primer containing the above A as the 3'-end base and a reverse primer capable of amplifying a DNA fragment of an appropriate size, if the base at the above SNP site is A, a band at the expected position is observed due to the normal progress of the amplification reaction. If the above base is substituted with G, the primer can bind complementarily to the template DNA, but since the 3'-end does not undergo complementary binding, the amplification reaction cannot proceed normally. DASH can be carried out by conventional methods, specifically by the method of Prince et al.

[0110] On the other hand, PCR extension analysis is achieved through the following process: First, a DNA fragment containing the base where the single nucleotide polymorphism is located is amplified with a primer pair, then all nucleotides added to the reaction are inactivated by dephosphorylation, and a SNP-specific extension primer, a dNTP mixture, a dideoxynucleotide, a reaction buffer, and a DNA polymerase are added thereto to carry out a primer extension reaction. At this time, the extension primer has the base immediately adjacent to the 5'-direction of the base where the SNP is located as the 3'-end, the dNTP mixture does not contain a nucleic acid having the same base as the dideoxynucleotide, and the above dideoxynucleotide is selected from one of the base types showing the SNP. For example, in the case of substitution from A to G, when a mixture of dGTP, dCTP, and dTTP and ddATP are added to the reaction, in the base where the above substitution occurs, the primer is extended by DNA polymerase, and after several 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 this position, so the base type showing the SNP can be judged by comparing the lengths of the above extended primers.

[0111] At this time, as a detection method, when the extension primer or the dideoxynucleotide is fluorescently labeled, the above SNP can be detected by detecting fluorescence using a conventional gene analyzer for determining base sequences (for example, Model 3700 of ABI Company, etc.), and when an unlabeled extension primer and dideoxynucleotide are used, the above SNP can be detected by measuring the molecular weight using MALDI-TOF (matrix assisted laser desorption ionization-time of flight) technology.

[0112] As one aspect for achieving the object of the present invention, the present invention provides a single nucleotide polymorphism (SNP) marker for diagnosing the degree of skin fine lines.

[0113] As another aspect, the present invention provides a composition for diagnosing the degree of skin fine lines, which comprises a probe capable of detecting a single nucleotide polymorphism (SNP) marker for diagnosing the degree of skin fine lines or a preparation capable of amplifying the single nucleotide polymorphism (SNP) marker.

[0114] For the object of the present invention, the above term "degree of skin fine lines" refers to the result of the detection ratio of the relative total detection site obtained by adding all the target areas when the linear part of a certain length or more due to skin wrinkles is observed to be relatively darker than the surrounding skin, but is not limited thereto. Specifically, it refers to judging the ease of fine line formation by using one or more single nucleotide polymorphisms (SNP) markers selected from Table 1 or Table 4. More specifically, the skin type with easy fine line formation refers to the skin type with a high proportion of more wrinkles generated per skin area due to a fast aging process.

[0115] For the above alleles of the present invention, the number of chromosomes in each individual is the same, and there are a major allele and a minor allele of the SNP. As the base at the polymorphic site of the polymorphism marker increases one by one to the minor allele, the major allele can decrease one by one, and as the base increases one by one to the major allele, the minor allele can decrease one by one. However, the range in which the minor allele and the major allele can increase or decrease can be within three types: i) major allele / major allele, ii) major allele / minor allele, iii) minor allele / minor allele, and within the range of the above three types, the alleles can decrease or increase, but are not limited thereto.

[0116] In the present invention, the term "probe capable of detecting a marker for diagnosing the degree of skin fine lines" refers to a composition capable of diagnosing the degree of skin fine lines by confirming through a specific hybridization reaction with the polymorphic site of the above gene, and there is no particular limitation on the specific method of such gene analysis, and it can be carried out according to all gene detection methods known in the technical field to which the present invention belongs. In addition, the above term can be used interchangeably with the term "for diagnosing whether it is a skin type prone to fine line formation".

[0117] In the present invention, the term "agent capable of amplifying a marker for diagnosing the degree of fine lines of the skin" refers to a composition capable of diagnosing the degree of fine lines of the skin by amplifying and confirming the polymorphic sites of the above genes. Specifically, it refers to a primer capable of specifically amplifying the polynucleotide of the marker for diagnosing the degree of fine lines of the skin. In addition, the above term can be used interchangeably with the term "for diagnosing whether the skin type is prone to generating fine lines".

[0118] The primer for amplifying the above polymorphic marker refers to a single-stranded oligonucleotide that can function as a starting point for template-directed DNA synthesis under appropriate conditions (such as four different nucleoside triphosphates and a polymerase such as DNA, RNA polymerase or reverse transcriptase) in an appropriate buffer and at an appropriate temperature. The appropriate length of the above primer may vary according to the purpose of use, but is usually 15 to 30 nucleotides. Short primer molecules usually require a lower temperature to form a stable hybrid with the template. The primer sequence does not have to be completely complementary to the template, but must be complementary enough to hybridize with the template.

[0119] In the present invention, the term "primer", as a short base sequence with a free 3'-terminal hydroxyl group (free 3'-hydroxyl group), refers to a short sequence that can form a base pair with a complementary template and serve as a starting point for template strand replication. The primer can initiate DNA synthesis in the presence of a reagent for a polymerization reaction (i.e., DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates in an appropriate buffer solution and at an appropriate temperature. By performing PCR amplification, the skin type can be predicted based on the degree of generation of the desired product. The PCR conditions and the lengths of the sense and antisense primers can be modified based on those well known in the art.

[0120] The probes or primers of the present invention can be chemically synthesized using the phosphoramidite solid support method or other widely known methods. Such nucleic acid sequences can also be modified using many means well known in the art. Non-limiting examples of such modifications include methylation, "capping", substitution of one or more natural nucleotides with homologs, and modifications between nucleotides, for example, modification to an uncharged linker (such as methyl phosphonate, phosphotriester, phosphoramidate, carbamate, etc.) or a charged linker (such as phosphorothioate, phosphorodithioate, etc.).

[0121] As another aspect, the present invention provides a kit for diagnosing the degree of fine lines of the skin, which comprises the above-mentioned composition for diagnosing the degree of fine lines of the skin. The above-mentioned kit may be an RT-PCR kit or a DNA chip kit, but is not limited thereto.

[0122] The kit of the present invention can diagnose the degree of fine lines of the skin by amplifying and confirming the SNP polymorphism marker, which is a marker for diagnosing the degree of fine lines of the skin, or by confirming the expression level of the SNP polymorphism marker and the expression level of mRNA. As a specific example, in the present invention, the kit for measuring the mRNA expression level of the marker for diagnosing the degree of fine lines of the skin may be a kit comprising the necessary elements required for performing RT-PCR. In addition to the respective primer pairs specific to the gene of the marker for diagnosing the degree of fine lines of the skin, the RT-PCR kit may further include test tubes or other suitable containers, reaction buffers (with various pH and magnesium concentrations), deoxynucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, RNase inhibitor, DEPC-water, sterile water, etc. Additionally, it may include primer pairs specific to the gene used as a quantitative control group. Specifically, the kit of the present invention may be a kit for diagnosing the degree of fine lines of the skin comprising the necessary elements required for performing a DNA chip. The DNA chip kit is generally a DNA chip kit in which nucleic acid species are attached to a flat solid support plate, typically a glass surface not larger than a microscope slide, in a gridded array, and is a tool for performing a large number of parallel analyses by allowing multiple hybridization reactions to occur between the nucleic acids on the DNA chip and the complementary nucleic acids contained in the solution processed on the chip surface, with the nucleic acids being uniformly arranged on the chip surface.

[0123] As another aspect, the present invention provides a microarray for diagnosing the degree of fine lines of the skin, which comprises the above-mentioned composition for diagnosing the degree of fine lines of the skin.

[0124] The above-mentioned microarray may contain DNA or RNA polynucleotides. Except for containing the polynucleotide of the present invention in the probe polynucleotide, the above-mentioned microarray is composed of a conventional microarray.

[0125] Methods for preparing microarrays by immobilizing probe polynucleotides on a substrate are well known in the art. The above-mentioned probe polynucleotides refer to hybridizable polynucleotides, which are oligonucleotides capable of sequence-specific binding to complementary strands of nucleic acids. The probes of the present invention are allele-specific probes. The polymorphic sites are present in nucleic acid fragments from two members of the same species, hybridize with the DNA fragment from one member, but do not hybridize with the fragment from the other member. In this case, the hybridization conditions show significant differences in the hybridization intensity between alleles, so they must be stringent enough to hybridize only with one of the alleles. By doing so, good hybridization differences between different allelic forms can be induced. The above-mentioned probes of the present invention can be used in methods for diagnosing skin types by detecting alleles, etc. The above-mentioned diagnostic methods include detection methods based on nucleic acid hybridization, such as Southern blot hybridization, etc., and in the method using a DNA chip, they can also be provided in a form pre-bound to the substrate of the DNA chip. The above-mentioned hybridization can usually be carried out under stringent conditions, such as a salt concentration of less than 1M and a temperature of more than 25°C. For example, the conditions of 5x SSPE (750 mM NaCl, 50 mM sodium phosphate, 5 mM EDTA, pH 7.4) and 25 - 30°C are suitable for allele-specific probe hybridization.

[0126] The process of immobilizing the probe polynucleotides related to the diagnosis of skin fine line degree of the present invention on a substrate can also be easily prepared using this prior art. In addition, the hybridization of nucleic acids on a microarray and the detection of hybridization results are well known in the art. The above-mentioned detection can detect the hybridization results through the following process: for example, labeling a nucleic acid sample with a labeling material capable of generating a detectable signal (including fluorescent materials, such as materials like Cy3 and Cy5), then hybridizing on the microarray and detecting the signal generated by the above-mentioned labeling material.

[0127] As another aspect, the present invention provides a method for providing information about the degree of skin fine lines, which includes the following steps; (a) amplifying or hybridizing the polymorphic sites of the above-mentioned single nucleotide polymorphism markers in the DNA obtained from a sample isolated from an individual; and (b) confirming the bases of the polymorphic sites amplified or hybridized in the above step (a).

[0128] The above terms "individual, step (a), step (b)" are as described above.

[0129] In the above method, additionally, when (c) the bases of the amplified or hybridized polymorphic sites contain 4 or more minor alleles (i.e., bases) according to the above-mentioned single nucleotide polymorphism markers, it can be judged as a high-risk group with more fine lines, but it is not limited to this.

[0130] As an aspect for achieving the object of the present invention, the present invention provides a composition for improving drooping pores and wrinkles. As an example, the above composition contains one or more substances selected from the group consisting of fucoidan, N-feruloylserotonin, emodin, and formononetin as active ingredients. As another example, the above composition may contain (i) fucoidan and N-feruloylserotonin; (ii) emodin and formononetin; or (iii) fucoidan, N-feruloylserotonin, emodin, and formononetin as active ingredients, but is not limited thereto.

[0131] The above term "fucoidan" is a component in the slimy mucilage contained in brown algae, secreted by the mucosal tubes in the leaves, and thus plays a role in resisting the invasion of bacteria when the leaves and stems of seaweeds are damaged by the flow of tides and sand. Fucoidan has similar physiological properties to the sulfuric acid polysaccharide present in the blood, i.e., heparin, and various physiological functions have been revealed, and various physiological active functions such as anticancer, antiviral, anti-allergic, and immune enhancement have been reported, and it is made by extracting brown algae such as Undaria pinnatifida, Laminaria japonica, Potamogetonaceae, and Sargassum fusiforme. The above fucoidan is not limited to its acquisition method and can be chemically synthesized by methods known in the art or commercially available substances can be used.

[0132] The above term "N-feruloylserotonin", as a derivative of serotonin, is an alkaloid and polyphenol found in safflower seeds. It is known to be an amide formed chemically between serotonin and ferulic acid and has anti-atherosclerotic activity. The molecular formula of N-feruloylserotonin is C 20 H 20 N2O4, with a molecular weight of 352.390, and can be represented by the following chemical formula 3. The above N-feruloylserotonin is not limited to its acquisition method and can be chemically synthesized by methods known in the art or commercially available substances can be used.

[0133] [Chemical formula 3]

[0134]

[0135] The above term "emodin", as a compound of the anthraquinone series, can be isolated from rhubarb, buckthorn, etc. The molecular formula of emodin is C 15 H 10O5, with a molecular weight of 270.24, can be represented by the following Chemical Formula 4. The above emodin is not limited to its obtaining method and can be chemically synthesized by methods well-known in the art or using commercially available substances.

[0136] [Chemical Formula 4]

[0137]

[0138] The above term "Formononetin", as an O-methylated isoflavone, is found in numerous plants and herbs. It is reported to promote angiogenesis. The molecular formula of formononetin is C 16 H 12 O4, with a molecular weight of 284.26, can be represented by the following Chemical Formula 5. The above formononetin is not limited to its obtaining method and can be chemically synthesized by methods well-known in the art or using commercially available substances.

[0139] [Chemical Formula 5]

[0140]

[0141] Based on the total weight of the composition, the content of the substance of the present invention can be about 0.0001 to 10% by weight, but is not limited thereto. As an example, based on the total weight of the composition, the retinol booster of the present invention can be 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.

[0142] For the purpose of the present invention, the above substance can impart an effect of promoting the improvement of sagging pore wrinkles.

[0143] For the purpose of the present invention, the above term "sagging pore wrinkles" refers to pores that spread and sag like ellipses due to aging, and these sagging pores are connected to appear like wrinkles. The pores that spread into ellipses mainly from the side of the nose wing towards the cheek are called sagging pores. Due to the folding of the skin, the wrinkle part forms a shadow compared with the surrounding skin, so it is observed to be relatively dark compared with the surrounding skin. In addition, most wrinkles have a linear appearance, so when the shadow parts are connected and observed to be longer than a certain length, they can be defined as wrinkles, but are not limited thereto.

[0144] For the purpose of the present invention, the above-mentioned substances may be characterized by promoting collagen synthesis. Specifically, when treating fucoidan sulfate and / or N-feruloylserotonin in the above-mentioned substances, the synthesis of type IV collagen can be promoted, and when treating emodin and / or formononetin, the synthesis of type I collagen or type IV collagen can be promoted, but not limited thereto.

[0145] As an example, as a result of applying a cream containing the substances of the present invention that are effective in promoting collagen synthesis to the cheek area, it was confirmed that compared with the application of retinol as a control group, an excellent wrinkle improvement rate was exhibited.

[0146] In fact, many wrinkle improvement raw materials for collagen have been launched on the market, but it is understood that the actual perception rate of consumers for wrinkle improvement is quite low. In addition, in fact, non-retinol raw materials that exceed retinol, which is the raw material with the most excellent known wrinkle improvement effect among existing raw materials, are not common. However, in the present invention, it was confirmed that non-retinol substances have an excellent effect on promoting collagen synthesis, and from this point of view, it can be considered significant.

[0147] For the purpose of the present invention, the above-mentioned substances may be characterized by promoting the expression of ADIPOQ or inhibiting the expression of SGPP2. Specifically, when treating emodin and / or formononetin in the above-mentioned substances, the expression of ADIPOQ can be promoted, and when treating fucoidan sulfate and / or N-feruloylserotonin, the expression of SGPP2 can be inhibited, but not limited thereto. It can be seen that the reactivity to the substances of the present invention varies according to the presence or absence of specific genes, which indicates that the improvement efficiency of sagging pores and wrinkles can be increased by providing a customized formula for high-risk groups with a high risk level of sagging pores and wrinkles.

[0148] In the present invention, the term "about" is as described above.

[0149] The above-mentioned composition is characterized in that it is a personalized customization type. Specifically, substances for personal customization can be provided for individuals with single nucleotide polymorphism (SNP) markers significantly associated with a specific degree of sagging pores and wrinkles.

[0150] As an example, the single nucleotide polymorphism (SNP) markers significantly associated with the degree of sagging pores and wrinkles may be any one or more selected from Table 2 or Table 5, but not limited thereto.

[0151] In the present invention, the terms "polymorphism", "allele" and "rs_id" are as described above.

[0152] Specifically, the gene polymorphism marker of the present invention has a significant association with drooping pores and wrinkles, can have two alleles, and can be divided into cases of having a major allele / minor allele, minor allele / minor allele, and major allele / major allele.

[0153] When having the allele of the polymorphism marker with significant association with drooping pores and wrinkles, depending on the allele possessed by the individual, the effect of improving drooping pores and wrinkles through the reaction caused by the substance of the present invention may be different. As an example, when having major allele / major allele, major allele / minor allele, or minor allele / minor allele, the reactivity caused by the substance of the present invention can be measured, that is, the skin characteristics with a high or low degree of improvement in drooping pores and wrinkles, so it can provide information for selecting customized substances for individuals containing one or more minor alleles to increase the improvement of drooping pores and wrinkles.

[0154] The above single nucleotide polymorphism marker can be one or more single nucleotide polymorphism markers selected from the single nucleotide polymorphism markers shown in Table 2 and Table 5. The single nucleotide polymorphism markers shown in Table 2 and Table 5 can determine whether there is an association with the degree of drooping pores and wrinkles.

[0155] The single nucleotide polymorphism (SNP) marker of the present invention can be any one or more of the markers shown in Table 2 or Table 5, but is not limited thereto. The above single nucleotide polymorphism (SNP) marker can be one or more, and can be used in combinations such as two or more that can determine the number of wrinkles, but is not limited thereto. The above marker can be the SNP itself, or a polynucleotide composed of 5 - 100 consecutive DNA sequences containing the above SNP site, or a polynucleotide composed of its complementary sequence, but is not limited thereto.

[0156] As an example, the single nucleotide polymorphism marker with significant association with the degree of drooping pores and wrinkles can be one or more single nucleotide polymorphism markers present in the gene encoding the protein of ADIPOQ or SGPP2. Specifically, it can be any one or more selected from the group consisting of rs10203329, rs4592839, rs76071583, and rs2036373, but is not limited thereto.

[0157] As an example, when the SNP ID is rs10203329, the Chr.Position (GRCh ver.37) is recorded as "2:223350001". If the allele is publicly available as A>G, this means that the 223,350,001st base of human chromosome 2 is A or G, and the base on the left side of the ">" in the allele may mean the major allele, and the base on the right side may mean the minor allele.

[0158] As a specific example, the markers selected from Table 2 may be composed of one or more polynucleotides selected from the group consisting of the following polynucleotides; and their complementary polynucleotides, but are not limited thereto: a polynucleotide composed of 5 to 100 consecutive DNA sequences containing the 223,350,001st base of human chromosome 2, wherein the 223,350,001st base is A or G (rs10203329); a polynucleotide composed of 5 to 100 consecutive DNA sequences containing the 223,412,813th base of human chromosome 2, wherein the 223,412,813th base is T or C (rs4592839); a polynucleotide composed of 5 to 100 consecutive DNA sequences containing the 186,557,872nd base of human chromosome 3, wherein the 186,557,872nd base is A or G (rs76071583); a polynucleotide composed of 5 to 100 consecutive DNA sequences containing the 186,570,191st base of human chromosome 3, wherein the 186,570,191st base is T or G (rs2036373).

[0159] As another embodiment, one or more of the single nucleotide polymorphism (SNP) markers shown in Table 5 in the same manner as Table 2 may be selected, but are not limited thereto.

[0160] The composition of the present invention can be used as a cosmetic composition as described above.

[0161] On the other hand, the above composition provides a pharmaceutical external preparation composition for relieving or improving sagging pore wrinkles. In the present invention, the term "pharmaceutical external preparation" is as described above.

[0162] As another aspect, the present invention provides a method for providing information for selecting a customized substance for improving sagging pores and wrinkles, comprising the following steps: (a) obtaining a biological sample from an individual; (b) amplifying or hybridizing with a probe a polymorphism site of any one or more single nucleotide polymorphism markers selected from the group consisting of rs4592839 and rs76071583 from the above biological sample; and (c) confirming the base of the amplified or hybridized polymorphism site in step (b).

[0163] The sagging pores and wrinkles can be improved by treating the selected customized substance to an individual having a specific polymorphism site.

[0164] The term "individual" in the present invention refers to a subject for diagnosing the degree of sagging pores and wrinkles. In the above test sample, DNA can be obtained from samples such as hair, urine, blood, various body fluids, isolated tissues, isolated cells or saliva, etc., but not limited thereto. Steps (a) and (b) are the same as described above.

[0165] The above substance can be one or more selected from the group consisting of fucoidan, N-feruloylserotonin, emodin and formononetin, but not limited thereto.

[0166] For the purpose of the present invention, the above-mentioned gene significantly associated with the degree of sagging pores and wrinkles can provide information capable of selecting a specific substance for each individual. As an example, for an individual having one or more single nucleotide polymorphism markers present in the gene encoding the protein of ADIPOQ or SGPP2, when treating with one or more substances selected from the group consisting of emodin, fucoidan, N-feruloylserotonin, emodin and formononetin, the improvement of sagging pores and wrinkles can be enhanced, but according to the gene characteristics of each individual, an optimal substance combination can be provided.

[0167] This indicates that information can be provided such that specific single nucleotide polymorphism (SNP) markers significantly associated with sagging pores and wrinkles can be screened and personalized customized substances capable of maximizing the effect of improving sagging pores and wrinkles can be selected based on genetic information.

[0168] As an example, a method for providing information can be provided so that when one or more minor alleles are included in the bases of rs4592839 and rs76071583, fucoidan and N-feruloylserotonin are selected; or emodin and formononetin are selected; or fucoidan, N-feruloylserotonin, emodin and formononetin are selected, but not limited thereto.

[0169] In the present invention, the term "SNP chip" is as described above.

[0170] As one aspect for achieving the object of the present invention, the present invention provides a single nucleotide polymorphism (SNP) marker for diagnosing the degree of sagging pore wrinkles.

[0171] As another aspect, the present invention provides a composition for diagnosing the degree of sagging pore wrinkles, which comprises a probe capable of detecting a single nucleotide polymorphism (SNP) marker for diagnosing the degree of sagging pore wrinkles or a preparation capable of amplifying the single nucleotide polymorphism (SNP) marker.

[0172] For the object of the present invention, the above-mentioned term "degree of sagging pore wrinkles" means that due to aging, pores spread and sag like an ellipse, and these sagging pores are connected to look like wrinkles, but are not limited thereto. The pores that spread in an ellipse mainly from the side of the nose wing towards the cheek are called sagging pores. Fibrous proteins such as collagen and elastin decrease with age, so while the skin sags, the pores also become larger and present a wrinkle-like appearance. It refers to the result of the detection ratio of the relative total of all object areas when a linear part of a certain length or more due to sagging pore wrinkles is observed to be relatively darker than the surrounding skin, but is not limited thereto. Specifically, it refers to judging the ease of generation of sagging pore wrinkles by using one or more single nucleotide polymorphism (SNP) markers selected from Table 2 or Table 5. More specifically, the skin type with an easy generation of sagging pore wrinkles refers to a skin type with a high proportion of generating more sagging pores per skin area due to a fast aging process.

[0173] In the present invention, the term "probe capable of detecting a marker for diagnosing the degree of sagging pore wrinkles of the skin" refers to a composition capable of diagnosing the degree of sagging pore wrinkles by confirming through a specific hybridization reaction with the polymorphic site of the above gene, and there is no particular limitation on the specific method of this gene analysis, and it can be carried out according to all gene detection methods known in the technical field to which the present invention belongs. In addition, the above term can be used interchangeably with the term "for diagnosing whether it is a skin type prone to generating sagging pore wrinkles".

[0174] In the present invention, the term "preparation capable of amplifying a marker for diagnosing the degree of sagging pore wrinkles of the skin" refers to a composition capable of diagnosing the degree of sagging pore wrinkles by amplifying and confirming the polymorphic site of the above gene, specifically, it refers to a primer capable of specifically amplifying the polynucleotide of the marker for diagnosing the degree of sagging pore wrinkles of the skin. In addition, the above term can be used interchangeably with the term "for diagnosing whether it is a skin type prone to generating sagging pore wrinkles".

[0175] The primers for amplifying the above-mentioned polymorphism markers refer to single-stranded oligonucleotides that can function as the starting point of template-directed DNA synthesis under appropriate conditions in an appropriate buffer (for example, 4 different nucleoside triphosphates and polymerizing agents such as DNA, RNA polymerase, or reverse transcriptase), and at an appropriate temperature. The appropriate length of the above primers may vary according to the purpose of use, but is usually 15 to 30 nucleotides. Short primer molecules usually require a lower temperature to form a stable hybrid with the template. The primer sequence does not have to be completely complementary to the template, but must be complementary enough to hybridize with the template.

[0176] In the present invention, the term "primer" is as described above.

[0177] As another aspect, the present invention provides a kit for diagnosing the degree of sagging pores and wrinkles of the skin, which contains the above-mentioned composition for diagnosing the degree of sagging pores and wrinkles. The above kit may be an RT-PCR kit or a DNA chip kit, but is not limited thereto.

[0178] As another aspect, the present invention provides a microarray for diagnosing the degree of sagging pores and wrinkles of the skin, which contains the above-mentioned composition for diagnosing the degree of sagging pores and wrinkles.

[0179] The process of immobilizing the probe polynucleotide related to the diagnosis of the degree of sagging pores and wrinkles of the present invention on the substrate can also be easily prepared using such prior art. In addition, the hybridization of nucleic acids on the microarray and the detection of the hybridization results are well known in the art. The above detection can detect the hybridization results through the following process: for example, labeling the nucleic acid sample with a labeling material (including fluorescent materials such as Cy3 and Cy5 and other materials) that can generate a detectable signal, and then hybridizing on the microarray and detecting the signal generated by the above labeling material.

[0180] As another aspect, the present invention provides a method for providing information about the degree of sagging pores and wrinkles, which includes the following steps; (a) amplifying the polymorphic sites of the above-mentioned single nucleotide polymorphism markers or hybridizing with a probe in the DNA obtained from a sample isolated from an individual; and (b) confirming the bases of the polymorphic sites amplified or hybridized in the above step (a).

[0181] The above terms "individual, step (a), step (b)" are as described above.

[0182] In the above method, additionally, when (c) the bases of the amplified or hybridized polymorphic sites contain 4 or more bases according to the minor allele of the above-mentioned single nucleotide polymorphism marker, it can be determined as a high-risk group with more sagging pores and wrinkles, but is not limited thereto.

[0183] As an aspect for achieving the object of the present invention, the present invention provides a composition for improving nasolabial folds. As an example, the above composition contains one or more substances selected from the group consisting of niacinamide, tranexamic acid, pyridoxine HCl, camphor, cordyceps extract, sodium mannose phosphate, and retinol as active ingredients. As another example, the above composition may contain (i) camphor, niacinamide, and sodium mannose phosphate; (ii) tranexamic acid and pyridoxine HCl; or (iii) retinol, camphor, niacinamide, sodium mannose phosphate, tranexamic acid, pyridoxine HCl, and cordyceps extract as active ingredients, but not limited thereto.

[0184] The above term "niacinamide" is also called nicotinamide. As a kind of vitamin B3 derivative, it is widely distributed in the plant kingdom together with nicotinic acid. It is known that it is widely distributed as a component of NAD (nicotinamide adenine dinucleotide) and NADP (nicotinamide adenine dinucleotide phosphate) in living organisms. The molecular formula of niacinamide is C 6 H 6 N 2 O , which can be represented by the following Chemical Formula 6. The above niacinamide is not limited to its acquisition method and can be chemically synthesized by methods known in the art or commercially available substances can be used.

[0185] [Chemical Formula 6]

[0186]

[0187] The above term "tranexamic acid" is a kind of amino acid. It is known that in pharmaceuticals, it is used as a coagulant, anti-allergic agent, anti-inflammatory agent, etc. In the cosmetic field, it inhibits the production of melanin by regulating melanocyte-stimulating substances. The molecular formula of tranexamic acid is C8H 15 NO2, and the molecular weight is 157.21, which can be represented by the following Chemical Formula 7. The above tranexamic acid is not limited to its acquisition method and can be chemically synthesized by methods known in the art or commercially available substances can be used.

[0188] [Chemical Formula 7]

[0189]

[0190] The above-mentioned term "Pyridoxine HCl" is a water-soluble vitamin that, as vitamin B6, is a component of enzymes important for protein metabolism and is involved in the production of neurotransmitters and the secretion of serotonin (hormone). When proteins are digested, homocysteine is produced, and when the concentration of homocysteine in the blood becomes high, the likelihood of developing cardiovascular diseases and dementia increases. Vitamin B6 combines with folic acid and vitamin B12 to break down homocysteine, thus significantly reducing the risk of heart disease. In addition, it is also required when tryptophan is converted to vitamin B3 and when vitamin B12 is absorbed. The above-mentioned Pyridoxine HCl is not limited to its method of obtaining and can be chemically synthesized by methods well-known in the art or commercially available substances can be used.

[0191] The above-mentioned term "Camphor" is a monoterpene-based compound with the chemical formula C 10 H 16 O, with a molecular weight of 156.2, and can be represented by the following Chemical Formula 8. The above-mentioned Camphor is not limited to its method of obtaining and can be chemically synthesized by methods well-known in the art or commercially available substances can be used.

[0192] [Chemical Formula 8]

[0193]

[0194] The above-mentioned term "Cordyceps sinensis" is a small mushroom belonging to the order Clavicipitales, family Cordycipitaceae, class Ascomycetes. It parasitizes insects in winter and grows like a plant in summer, which is the origin of its name. Cordyceps sinensis varies somewhat in its composition depending on its species, but generally consists of water, fat, crude protein, carbohydrates, and ash, and the protein contains about 18 kinds of essential amino acids for the human body. In addition, the following facts are known: Cordyceps sinensis contains special components such as Cordyceps Polysaccharides and Cordycepic Acid, and thus has been used not only as a therapeutic agent for tuberculosis, asthma, jaundice, etc. since ancient times, but also has the functions of enhancing immunity and anti-cancer effects. The extract of the above-mentioned Cordyceps sinensis is not limited to its method of obtaining, and all those extracted by methods well-known in the art are included, and commercially available substances can be used.

[0195] The molecular formula of the above-mentioned term "Sodium mannose phosphate" is C6H 12O9PNa, with a molecular weight of 282.12. Specifically, the above-mentioned sodium mannan phosphate can be sodium mannose 6-phosphate or sodium D-mannose-6-phosphate (D-Mannose-6-phosphate sodium salt), but is not limited thereto, and can be represented by the following Chemical Formula 9. The above-mentioned sodium mannan phosphate is not limited to its acquisition method and can be chemically synthesized by methods well-known in the art or commercially available substances can be used.

[0196] [Chemical Formula 9]

[0197]

[0198] As non-retinol substances, the above 6 substances can act alone, thereby having a wrinkle improvement effect superior to that of retinol, which is the raw material with the most excellent known wrinkle improvement effect, and at the same time act together with retinol, thereby acting as a retinol enhancer that increases the wrinkle improvement effect. The substances of the present invention are not particularly limited as long as they act alone or act together with retinol to increase the reactivity caused by retinol, thereby contributing to maximizing the skin wrinkle improvement effect.

[0199] The above term "retinol" is a type of vitamin A and is 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 exists in the mucosal cells of the animal intestine and is also abundantly contained in green and yellow plants. It is reported that it will also be converted into the active form of retinoic acid and plays an important role in maintaining the original functions of epidermal cells. In addition, it is reported that in the cell nucleus existing in skin cells, DNA expresses RNA, promotes cell differentiation, and promotes the biosynthesis of collagen (a hard protein existing in the form of fibrous solids between animal cells) and elastin (composed of elastic fibers), etc., thereby having the effects of reducing wrinkles and increasing skin elasticity.

[0200] Based on the total weight of the composition, the content of the substance of the present invention can be about 0.0001 to 10% by weight, but is not limited thereto. As an example, based on the total weight of the composition, the retinol enhancer of the present invention can be 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.

[0201] For the purpose of the present invention, the above substance can impart an effect that promotes the improvement of nasolabial folds.

[0202] For the purposes of the present invention, the above-mentioned term "nasolabial fold", as an expression wrinkle, refers to the shape of an inverted V formed by the depression around the mouth when smiling or speaking. Deeper nasolabial folds are caused by cheek flesh that has lost elasticity and sagged due to aging. Due to the folding of the skin, a shadow is formed at the wrinkle area compared to the surrounding skin, so it is observed as relatively dark compared to the surrounding skin. In addition, most of the wrinkles have a linear shape, so when the shadow areas are connected and observed to be longer than a certain length, they can be defined as wrinkles, but it is not limited to this. In addition, nasolabial folds can present in various forms, including drooping nasolabial folds that sag downward due to gravity, sunken nasolabial folds that are only deeply sunken without sagging, and wrinkles that combine drooping nasolabial folds and sunken nasolabial folds, but it is not limited to this.

[0203] For the purposes of the present invention, the above-mentioned substance may be characterized by promoting collagen synthesis. Specifically, when treating camphor, niacinamide, and sodium mannosephosphate; tranexamic acid and pyridoxine hydrochloride; or retinol, camphor, niacinamide, sodium mannosephosphate, tranexamic acid, pyridoxine hydrochloride, and Cordyceps sinensis extract in the above-mentioned substance, it can promote the synthesis of type I collagen, but it is not limited to this.

[0204] In fact, many wrinkle-improving raw materials targeting collagen have been launched on the market, but it is understood that the actual perception rate of consumers for wrinkle improvement is quite low. In addition, in fact, non-retinol raw materials that exceed retinol, which is known to have the most excellent wrinkle-improving effect among existing raw materials, are not common. However, in the present invention, it has been confirmed that non-retinol substances have excellent effects on promoting collagen synthesis, and from this point of view, it can be considered significant.

[0205] For the purposes of the present invention, the above-mentioned substance may be characterized by promoting the expression of PPARGC1B, SPATA5, or EN1. Specifically, when treating niacinamide, camphor, and / or sodium mannosephosphate in the above-mentioned substance, it can promote the expression of PPARGC1B. When treating tranexamic acid and / or pyridoxine hydrochloride, it can promote the expression of SPATA5. When treating Cordyceps sinensis extract, it can promote the expression of EN1, but it is not limited to this. It can be seen that the reactivity to the substance of the present invention varies according to the presence or absence of specific genes, which indicates that the efficiency of nasolabial fold improvement can be increased by providing a customized formula for high-risk groups with a high risk level of nasolabial fold occurrence.

[0206] In the present invention, the term "about" is as described above.

[0207] The above-mentioned composition is characterized in that it is a personalized customized type. Specifically, a personalized customized substance can be provided for individuals with single nucleotide polymorphism (SNP) markers significantly associated with a specific degree of nasolabial folds.

[0208] As an example, the single nucleotide polymorphism (SNP) marker significantly associated with the degree of nasolabial folds may be any one or more selected from Table 3 or Table 6, but is not limited thereto.

[0209] In the present invention, the terms "polymorphism", "allele", and "rs_id" are as described above.

[0210] Specifically, the gene polymorphism marker of the present invention is significantly associated with nasolabial folds, can have two alleles, and can be divided into cases of having a major allele / minor allele, minor allele / minor allele, or major allele / major allele.

[0211] When having an allele of the polymorphism marker significantly associated with nasolabial folds, depending on the allele possessed by an individual, the effect of improving nasolabial folds through the reaction caused by the substance of the present invention may be different. As an example, when having a major allele / major allele, major allele / minor allele, or minor allele / minor allele, the reactivity caused by the substance of the present invention, that is, the skin property of having a high or low degree of nasolabial fold improvement, can be measured. Therefore, it can provide information for selecting a customized substance for an individual containing one or more minor alleles to increase the improvement of nasolabial folds.

[0212] 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 3 and Table 6. Whether the single nucleotide polymorphism markers shown in Table 3 and Table 6 are associated with the degree of nasolabial folds can be determined.

[0213] The single nucleotide polymorphism (SNP) marker of the present invention may be any one or more selected from the markers shown in Table 3 or Table 6, but is not limited thereto. The above single nucleotide polymorphism (SNP) marker may be one or more, and may be used in combinations such as two or more that can determine the number of wrinkles, but is not limited thereto.

[0214] As an example, the above-mentioned nasolabial fold degree-related significant single nucleotide polymorphism markers may be one or more single nucleotide polymorphism markers present in the genes encoding the proteins of PPARGC1B, SPATA5 or EN1. Specifically, it may be any one or more selected from the group consisting of rs76148197, rs2166027 and rs41444645, but is not limited thereto.

[0215] As an example, when the SNP ID is rs76148197, the Chr.Position (GRCh ver.37) is recorded as "2:119602515". If the allele is disclosed as A>G, this means that the 119602515th base of human chromosome 2 is A or G. The base on the left side of the ">" in the allele may mean the major allele, and the base on the right side may mean the minor allele.

[0216] As a specific example, the markers selected from Table 3 may be composed of one or more polynucleotides selected from the group consisting of the following polynucleotides; and their complementary polynucleotides, but are not limited thereto: a polynucleotide composed of 5-100 consecutive DNA sequences containing the 119602515th base of human chromosome 2, wherein the 119602515th base is A or G (rs76148197); a polynucleotide composed of 5-100 consecutive DNA sequences containing the 124171618th base of human chromosome 4, wherein the 124171618th base is T or C (rs2166027); a polynucleotide composed of 5-100 consecutive DNA sequences containing the 149131368th base of human chromosome 5, wherein the 149131368th base is C or G (rs41444645).

[0217] As another embodiment, any one or more of the single nucleotide polymorphism (SNP) markers shown in Table 6 in the same manner as Table 3 may be selected, but are not limited thereto.

[0218] The composition of the present invention can be used as a cosmetic composition as described above.

[0219] On the other hand, the above composition provides a pharmaceutical external composition for relieving or improving nasolabial folds. In the present invention, the term "pharmaceutical external product" is as described above.

[0220] As another aspect, the present invention provides a method for providing information for selecting a customized substance for improving nasolabial folds, which includes the following steps: (a) obtaining a biological sample from an individual; (b) amplifying or hybridizing with a probe a polymorphism site of any one or more single nucleotide polymorphism markers selected from the group consisting of rs76148197, rs2166027, and rs41444645 from the above biological sample; and (c) confirming the base of the polymorphism site amplified or hybridized in the above step (b).

[0221] The nasolabial folds can be improved by treating the selected customized substance to an individual having a specific polymorphism site.

[0222] The term "individual" in the present invention refers to a subject for diagnosing the degree of nasolabial folds. In the above test sample, DNA can be obtained from samples such as hair, urine, blood, various body fluids, isolated tissues, isolated cells, or saliva, etc., but is not limited thereto. The above steps (a) and (b) are the same as described above.

[0223] The above substance can be one or more selected from the group consisting of N-nicotinamide, tranexamic acid, pyridoxine hydrochloride, camphor, Cordyceps sinensis extract, sodium mannosephosphate, and retinol, but is not limited thereto.

[0224] For the purpose of the present invention, the above nasolabial fold degree-related significant gene can provide information capable of selecting a specific substance for each individual. As an example, for an individual having one or more single nucleotide polymorphism markers present in the gene encoding PPARGC1B, SPATA5, or EN1 protein, when treating one or more substances selected from the group consisting of nicotinamide, tranexamic acid, pyridoxine hydrochloride, camphor, Cordyceps sinensis extract, sodium mannosephosphate, and retinol, the improvement of nasolabial folds can be enhanced, but according to the gene characteristics of each individual, an optimal substance combination can be provided.

[0225] This indicates that information can be provided such that specific single nucleotide polymorphism (SNP) markers significantly related to nasolabial folds can be screened and personalized customized substances capable of maximizing the effect of improving nasolabial folds can be selected based on genetic information.

[0226] As an example, a method for providing information can be provided so that when one or more minor alleles are included in the bases of rs76148197, rs2166027, and rs41444645, camphor, nicotinamide, and sodium mannosephosphate are selected; or tranexamic acid and pyridoxine hydrochloride are selected; or retinol, camphor, nicotinamide, sodium mannosephosphate, tranexamic acid, pyridoxine hydrochloride, and Cordyceps sinensis extract are selected, but is not limited thereto.

[0227] In the present invention, the term "SNP chip" is as described above.

[0228] As one aspect for achieving the object of the present invention, the present invention provides a single nucleotide polymorphism (SNP) marker for diagnosing the extent of nasolabial folds.

[0229] As another aspect, the present invention provides a composition for diagnosing the extent of nasolabial folds, comprising a probe capable of detecting a single nucleotide polymorphism (SNP) marker for diagnosing the extent of nasolabial folds or an agent capable of amplifying the single nucleotide polymorphism (SNP) marker.

[0230] For the purpose of the present invention, the above-mentioned term "degree of nasolabial folds" as expression lines refers to the depression of the mouth area into an eight-shaped shape when smiling or talking, but is not limited to this. Due to aging, the cheek flesh loses elasticity and sags, and at the same time, deeper wrinkles are formed from the sides of the nose around the mouth to the cheeks. Fibrous proteins such as collagen and elastin decrease with age, so the skin sags and presents a wrinkle-like appearance. It refers to the result of adding all object areas and the detection ratio relative to all detection areas when the linear area with a certain length or more is observed to be relatively dark compared to the surrounding skin due to nasolabial folds, but is not limited to this. Specifically, it refers to the use of any one or more single nucleotide polymorphism (SNP) markers selected from Table 3 or Table 6 to determine whether nasolabial folds are easy to form. More specifically, the skin type that is easy to form nasolabial folds refers to the skin type with a high proportion of more pores generated per skin area due to the rapid aging process.

[0231] In the present invention, the term "a probe capable of detecting a marker for diagnosing the extent of nasolabial folds of the skin" refers to a composition capable of diagnosing the extent of nasolabial folds by confirming a specific hybridization reaction with the polymorphic site of the above-mentioned gene, and the specific method of such gene analysis is not particularly limited, and can be performed according to all gene detection methods known in the technical field to which the present invention belongs. In addition, the above term can be used interchangeably with the term "for diagnosing whether the skin type is prone to the formation of nasolabial folds".

[0232] In the present invention, the term "a preparation capable of amplifying a marker for diagnosing the extent of nasolabial folds of the skin" refers to a composition capable of diagnosing the extent of nasolabial folds by amplifying and confirming the polymorphic sites of the above-mentioned gene, and specifically refers to a primer capable of specifically amplifying the polynucleotide of the marker for diagnosing the extent of nasolabial folds of the skin. In addition, the above term can be used interchangeably with the term "for diagnosing whether the skin type is prone to the formation of nasolabial folds".

[0233] The primers for amplifying the above-mentioned polymorphism markers refer to single-stranded oligonucleotides that can function as the starting points of template-directed DNA synthesis under appropriate conditions in an appropriate buffer (e.g., 4 different nucleoside triphosphates and polymerizing agents such as DNA, RNA polymerase, or reverse transcriptase), and at an appropriate temperature. The appropriate length of the above-mentioned primers may vary depending on the purpose of use, but is usually 15 to 30 nucleotides. Short primer molecules usually require a lower temperature to form a stable hybrid with the template. The primer sequence does not have to be completely complementary to the template, but must be complementary enough to hybridize with the template.

[0234] In the present invention, the term "primer" is as described above.

[0235] As another aspect, the present invention provides a kit for diagnosing the degree of nasolabial folds of the skin, which contains the above-mentioned composition for diagnosing the degree of nasolabial folds. The above-mentioned kit may be an RT-PCR kit or a DNA chip kit, but is not limited thereto.

[0236] As another aspect, the present invention provides a microarray for diagnosing the degree of nasolabial folds of the skin, which contains the above-mentioned composition for diagnosing the degree of nasolabial folds.

[0237] The process of immobilizing the probe polynucleotide related to the diagnosis of the degree of nasolabial folds of the present invention on the substrate can also be easily prepared using such prior art. In addition, the hybridization of nucleic acids on the microarray and the detection of hybridization results are well known in the art. The above-mentioned detection can detect the hybridization results through the following process: for example, labeling a nucleic acid sample with a labeling material (including fluorescent materials such as Cy3 and Cy5 and other materials) capable of generating a detectable signal, and then hybridizing on the microarray and detecting the signal generated by the above-mentioned labeling material.

[0238] As another aspect, the present invention provides a method for providing information about the degree of nasolabial folds, which includes the following steps; (a) amplifying the polymorphic sites of the above-mentioned single nucleotide polymorphism markers in the DNA obtained from a sample isolated from an individual or hybridizing with a probe; and (b) confirming the bases of the polymorphic sites amplified or hybridized in the above step (a).

[0239] The above terms "individual, step (a), step (b)" are as described above.

[0240] In the above method, additionally, when (c) the bases of the amplified or hybridized polymorphic sites contain 4 or more bases according to the minor allele of the above-mentioned single nucleotide polymorphism marker, it can be determined as a high-risk group with more nasolabial folds, but is not limited thereto.

[0241] Embodiments of the invention

[0242] Hereinafter, the present invention will be described in more detail by way of examples. These examples are only for illustrating the present invention, and the scope of the present invention should not be construed as being limited by these examples.

[0243] Example 1: Derivation of significantly associated gene polymorphisms for fine lines, sagging pores, wrinkles, and nasolabial folds

[0244] An attempt was made to identify genomic loci (gene variations) that show differences in the presence or absence and degree (numerical value) of facial skin wrinkles among Koreans (females) based on genetic information. In order to identify genomic loci (gene variations) related to wrinkles, the present invention used a microarray genotyping chip (product of Illumina) that can screen the entire genome level without pre-screening candidate genes.

[0245] Specifically, in the present invention, a professional instrument for image-based skin diagnosis (Janus3 of PIE) was used to evaluate the presence or absence and degree of fine lines, the presence or absence and degree of drooping pore wrinkles, and / or the presence or absence and degree of nasolabial folds. In order to minimize the external effects that may affect the presence or absence and degree (numerical value) of skin fine lines, the presence or absence and degree (numerical value) of drooping pore wrinkles, and / or the presence or absence and degree (numerical value) of nasolabial folds, it was used after correction with age, BMI, and cluster principal component values.

[0246] In order to confirm the association between genomic loci (gene variations) and the presence or absence and degree (numerical value) of skin fine lines, the presence or absence and degree (numerical value) of drooping pore wrinkles, and / or the presence or absence and degree (numerical value) of nasolabial folds, linear regression analysis was used to quantify the correlation significance and genetic effects.

[0247] 1-1: Method for evaluating the degree of skin fine lines, the degree of drooping pore wrinkles, and the degree of nasolabial folds

[0248] To derive gene polymorphism markers that can explain the degree of fine lines under the eyes, the degree of sagging pores, and / or the degree of nasolabial folds, healthy Korean women in their 20s to 70s were recruited. In addition, to measure the skin, all subjects washed their faces with facial cleanser or soap, waited for 30 minutes without applying any products to allow the skin to adapt to the measurement environment, and then the degree of skin wrinkles was evaluated. A professional image-based skin diagnosis instrument (Janus3 from PIE Company) was used in the evaluation (measurement and analysis were performed according to the instrument manufacturer's manual). Specifically, the subject wore a hair band and wrapped a black cloth around the clothes, then fixed the face on the forehead support and chin support built into the instrument, maintained the posture for detection with eyes closed, and a high-resolution image of the subject's face was taken through the camera built into the Janus 3 instrument. Only the bilateral lower eye parts as the evaluation target areas were extracted from the captured high-resolution image, de-identified, and then, one or more trained skin experts judged the presence or absence of fine lines under the eyes, or evaluated and quantified the grade according to the fine line grade criteria under the eyes. In addition, one or more trained skin experts judged the presence or absence of sagging pore wrinkles, or evaluated and quantified the grade according to the sagging pore wrinkle grade criteria. In addition, one or more trained skin experts judged the presence or absence of nasolabial folds, or evaluated and quantified the grade according to the nasolabial fold grade criteria.

[0249] For visual evaluation, a high-resolution image of the subject's face was taken through the camera built into the Janus 3 instrument to obtain a facial image. Then, using facial landmark information, images of the designated lower eye area, the central cheek area, and the facial image were extracted (de-identified) to extract the evaluation target areas. One or more trained skin experts evaluated the presence or absence and grade of fine lines under the eyes, the presence or absence and grade of sagging pore wrinkles, and the presence or absence and grade of nasolabial folds. The reference images for evaluating the grade were respectively as Figure 4 , Figure 13 , Figure 23 shown, Figure 4 , Figure 13 From the upper left corner, it represents grades 0 to 4, Figure 23 From the upper end, it represents grades 0 to 5.

[0250] Gene collection was achieved through saliva collection. For effective gene collection, all subjects were prohibited from consuming any food, including water, starting 30 minutes before collection.

[0251] Among the above-mentioned subjects, the following cases are excluded from the subjects: ① pregnancy, lactation, or planning to be pregnant within 6 months; ② using topical steroid skin preparations for more than 1 month for the treatment of skin diseases; ③ less than 6 months have passed after participating in the same trial; ④ having sensitive or allergic skin; ⑤ having skin abnormalities such as moles, acne, erythema, telangiectasia, etc. at the test site; ⑥ using the same or similar cosmetics or pharmaceuticals at the test site within 3 months before the start of the trial; ⑦ having received or planning to have surgery (skin peeling, Botulinum toxin (Botox), other skin care) at the test site within 6 months; ⑧ having chronic wasting diseases (such as asthma, diabetes, hypertension, etc.); ⑨ having atopic dermatitis; ⑩ in addition, according to the judgment of the principal investigator, cases judged to be difficult to conduct the trial are excluded from the subjects.

[0252] 1-2: Gene Collection and Analysis Methods

[0253] For gene analysis by extracting genes from saliva, human genomic DNA is extracted using the QIAamp mini prep kit (QIAGEN). Its quality is confirmed by testing the absorbance (OD 260 / 280) or 1.7, concentration 50 ng / ul, and the band of 1x TAE 1% agarose gel, and gene analysis is performed only on genes that pass the quality test.

[0254] Gene analysis is performed using the microarray genotyping chip of Illumina. Specifically, the genes of the analyzed subjects are analyzed using the global screening array product of the same company.

[0255] The gene analysis experiment of the Illumina microarray genotyping chip is carried out according to the provided manual, using the provided reagents, and performing the processes of genomic DNA amplification, DNA fragmentation, precipitation, hybridization, staining, washing, coating, and scanning.

[0256] The completed microarray genotyping chip was scanned using the iScan Control Software (Illumina). After the scanning was completed, idat files were automatically generated and the data quality management (sample call rate 98%, marker call rate 98%) and gene information confirmation were performed using the GenomeStudio (Illumina) program.

[0257] In this experiment, only the data that passed the data quality management after gene analysis was used.

[0258] 1 - 3: Export gene polymorphism markers related to the presence or absence of fine lines on the skin, sagging pores and wrinkles, and nasolabial folds

[0259] To export gene polymorphism markers that are significantly associated with the presence or absence of fine lines on the skin, sagging pores and wrinkles, and nasolabial folds, a correlation analysis was performed between the presence or absence of fine lines under the eyes, sagging pores and wrinkles, nasolabial folds, and gene polymorphism markers. Specifically, for more than 1,000 Korean women, the phenotypic data was the expert visual evaluation data for judging the presence or absence of fine lines under the eyes, sagging pores and wrinkles, and nasolabial folds.

[0260] For the genotype, genes with a high likelihood of functionally affecting the occurrence of wrinkles were selected, and the data of gene polymorphism markers located on these genes was used.

[0261] However, for the quality management of the target gene polymorphism markers to be analyzed, it was limited to those exceeding the benchmarks of minor allele frequency > 0.01 and Hardy - Weinberg equilibrium > 0.000001 for each gene polymorphism marker for analysis. In addition, the significance of the correlation between the presence or absence of fine lines under the eyes, sagging pores and wrinkles, nasolabial folds, and gene polymorphism markers was evaluated using the F - statistics of logistic regression analysis, and the benchmark was set as P - value < 0.05. The analysis used the PLINK v 1.90 and SNP&Variation suite (Golden Helix, Inc., Bozeman, Montana, USA) programs.

[0262] In order to minimize external effects that may affect the degree of skin fine lines, the degree of sagging pores and wrinkles, and the degree of nasolabial folds, and to derive effects based on genetic information, the presence or absence of fine lines under the eyes, sagging pores and wrinkles, and nasolabial folds can be corrected using information such as age, menopause status, or BMI (body mass index) information or lifestyle habits (alcohol consumption, smoking, eating habits, sleep habits, etc.), and used for analysis.

[0263] First, Table 1 below shows representative examples of SNP markers that are significantly associated with the presence or absence of skin fine lines, especially fine lines under the eyes.

[0264] [Table 1]

[0265] Gene polymorphism markers associated with the presence or absence of fine lines under the eyes (P<0.05)

[0266] Number Phenotype of analysis subject <![CDATA[SNP 1) > <![CDATA[Gene 1) > <![CDATA[Chromosome: Location 1) > <![CDATA[Allele 2) > <![CDATA[MAF 3) > <![CDATA[P-value 4) > <![CDATA[Effect size (β) 5) > 1 Fine lines under eyes rs10865025 EDAR 2:109511765 G>A 0.08 0.04 0.40 2 Fine lines under eyes rs10756807 BNC2 9:16756041 G>A 0.25 0.02 0.33 3 Fine lines under eyes rs1952688 BNC2 9:16782156 A>G 0.33 0.03 0.28 4 Fine lines under eyes rs10116469 BNC2 9:16783095 A>T 0.34 0.04 0.27 5 Fine lines under eyes rs12338608 BNC2 9:16800470 C>T 0.20 0.01 0.39

[0267] 1) National Institutes of Health (NIH) ID, the sequence can be confirmed on the relevant website

[0268] 2) (Major allele) > (Minor allele) meaning

[0269] 3) Minor allele frequency = (2mm + Mm) / 2(MM + Mm + mm)

[0270] 4) Statistical significance of phenotypic differences among the three genotypes (M / M, M / m, m / m) (M: major allele, m: minor allele)

[0271] 5) Degree of change in phenotypic risk as the minor allele increases one by one (-: risk of fine lines under the eyes decreases, +: risk of fine lines under the eyes increases)

[0272] Presence or absence of fine lines under the eyes = Observation of fine lines under the eyes (expert visual evaluation)

[0273] Table 2 below shows representative examples of SNP markers that are significantly associated with the presence or absence of sagging pores.

[0274] [Table 2]

[0275] Gene polymorphism markers associated with the presence or absence of sagging pores and wrinkles (P<01)

[0276] Number Phenotype of analysis subject <![CDATA[SNP 1) > <![CDATA[Gene 1) > <![CDATA[Chromosome: Location 1) > <![CDATA[Allele 2) > <![CDATA[MAF 3) > <![CDATA[P-value 4) > <![CDATA[Effect size (β) 5) > 1 Sagging pores and wrinkles rs10203329 SGPP2 2:223350001 A>G 0.34 0.07 -0.17 2 Sagging pores and wrinkles rs4592839 SGPP2 2:223412813 T>C 0.11 0.08 0.25 3 Sagging pores and wrinkles rs76071583 ADIPOQ 3:186557872 A>G 0.02 0.09 0.57 4 Sagging pores and wrinkles rs2036373 ADIPOQ 3:186570191 T>G 0.03 0.04 -0.56

[0277] 1) National Institutes of Health (NIH) ID, the sequence can be confirmed on the relevant website

[0278] 2) (Major allele, major allele) > (Minor allele, minor allele) meaning

[0279] 3) Minor allele frequency = (2mm + Mm) / 2(MM + Mm + mm)

[0280] 4) Statistical significance of phenotypic differences among the three genotypes (M / M, M / m, m / m) (M: major allele, m: minor allele)

[0281] 5) Degree of change in phenotypic risk as the minor allele increases one by one (-: Decrease in the risk of sagging pores, +: Increase in the risk of sagging pores)

[0282] Presence or absence of sagging pore wrinkles = Observation of pore sagging in the cheek area (Expert visual evaluation)

[0283] Table 3 below shows representative examples of SNP markers significantly associated with the presence or absence of nasolabial folds.

[0284] [Table 3]

[0285] Gene polymorphism markers associated with the presence or absence of nasolabial folds (P < 0.05)

[0286] Number Phenotype of analysis subject <![CDATA[SNP 1) > <![CDATA[Gene 1) > <![CDATA[Chromosome: Location 1) > <![CDATA[Allele 2) > <![CDATA[MAF 3) > <![CDATA[P-value 4) > <![CDATA[Effect size (β) 5) > 1 Nasolabial folds rs76148197 EN1 2:119602515 A>G 0.14 0.047 0.27 2 Nasolabial folds rs2166027 SPATA5 4:124171618 T>C 0.43 0.01 -0.23 3 Nasolabial folds rs41444645 PPARGC1B 5:149131368 C>G 0.04 0.02 0.57

[0287] 1) National Institutes of Health (NIH) ID, the sequence can be confirmed on the relevant webpage

[0288] 2) (Major allele, major allele) > (Minor allele, minor allele) meaning

[0289] 3) Minor allele frequency = (2mm + Mm) / 2(MM + Mm + mm)

[0290] 4) Statistical significance of phenotypic differences among the three genotypes (M / M, M / m, m / m) (M: major allele, m: minor allele)

[0291] 5) Degree of change in phenotypic risk as the minor allele increases one by one (-: Decrease in the risk of nasolabial folds, +: Increase in the risk of nasolabial folds)

[0292] Presence or absence of nasolabial folds = Observation of nasolabial folds (Expert visual evaluation)

[0293] 1 - 4: Derivation of gene polymorphism markers associated with the degree of skin fine lines, the degree of sagging pore wrinkles, and the degree of nasolabial folds

[0294] In order to derive gene polymorphism markers that are significantly associated with the degree of fine lines on the skin, the degree of sagging pores and wrinkles, and the degree of nasolabial folds, an association analysis was performed between the degree of fine lines under the eyes, the degree of sagging pores and wrinkles, the degree of nasolabial folds, and the gene polymorphism markers. Specifically, for more than 2,000 Korean women, the phenotypes used the expert visual evaluation data for judging the presence or absence of fine lines under the eyes, the presence or absence of sagging pores and wrinkles, and the presence or absence of nasolabial folds. For the visual evaluation, high-resolution images of the faces of the subjects to be analyzed were taken through the camera built into the Janus 3 instrument to obtain facial images. Then, using the facial landmark information, the images of the designated areas under the eyes, the central part of the cheeks, and the nasolabial fold area were extracted (de-identified) to extract the evaluation target areas. One or more trained skin experts evaluated the presence or absence and grades of fine lines under the eyes, the presence or absence and grades of sagging pores and wrinkles, and the presence or absence and grades of nasolabial folds. The reference images for the evaluation grades are as Figure 4 , Figure 13 shown, starting from the upper left end of Figure 4 , Figure 13 it indicates grades 0 to 4. In addition, the reference images for evaluating the presence or absence and grades of nasolabial folds are as Figure 23 shown, starting from the upper left end of Figure 23 it indicates grades 0 to 5.

[0295] For the genotypes, all gene polymorphism marker data observed at the full-length genome level generated by the microarray chip were used. However, for the quality control of the target gene polymorphism markers for analysis, it was limited to those exceeding the minor allele frequency > 0.01 and the Hardy-Weinberg equilibrium > 0.000001 benchmarks for each gene polymorphism marker for analysis. In addition, the significance of the association between the degree of fine lines under the eyes and the gene polymorphism markers was evaluated through the F-statistics of the logistic regression analysis, and the benchmark was set as the P-value < 0.0001. The analysis used the PLINK v 1.90 and SNP&Variationsuite (Golden Helix, Inc., Bozeman, Montana, USA) programs.

[0296] To minimize the external effects that may affect the degree of fine lines under the eyes, the degree of drooping pore wrinkles, and the degree of nasolabial folds and to derive effects based on genetic information, the degree of fine lines under the eyes, the degree of drooping pore wrinkles, and the degree of nasolabial folds can be corrected using information on age, menopause status, or BMI (body mass index) information or lifestyle habits (alcohol consumption, smoking, eating habits, sleep habits, etc.) and used for analysis.

[0297] First, Table 4 below shows representative examples of SNP markers that are significantly associated with skin fine lines, especially the degree of fine lines under the eyes.

[0298] [Table 4]

[0299] Gene polymorphism markers associated with the degree of fine lines under the eyes (P<0.0001)

[0300]

[0301]

[0302]

[0303]

[0304]

[0305]

[0306]

[0307]

[0308]

[0309]

[0310]

[0311]

[0312]

[0313]

[0314]

[0315]

[0316]

[0317]

[0318]

[0319]

[0320]

[0321] 1) National Institutes of Health (NIH) ID, the sequence can be confirmed on the relevant website

[0322] 2) Meaning of (major allele) > (minor allele)

[0323] 3) Minor allele frequency = (2mm + Mm) / 2(MM + Mm + mm)

[0324] 4) Statistical significance of the phenotypic differences among the three genotypes (M / M, M / m, m / m) (M: major allele, m: minor allele)

[0325] 5) Degree of increase or decrease in phenotype with the gradual increase of the minor allele (-: reduction in the risk of fine lines under the eyes, +: increase in the risk of fine lines under the eyes)

[0326] Presence or absence of fine lines under the eyes = severity level of fine lines under the eyes (evaluated by experts with the naked eye)

[0327] Table 5 below shows representative examples of SNP markers significantly associated with sagging pore wrinkles, especially the degree of sagging pore wrinkles.

[0328] [Table 5]

[0329] Gene polymorphism markers associated with the degree of sagging pore wrinkles (P < 0.0001)

[0330]

[0331]

[0332]

[0333]

[0334]

[0335]

[0336]

[0337]

[0338]

[0339]

[0340]

[0341]

[0342]

[0343]

[0344]

[0345]

[0346]

[0347]

[0348]

[0349]

[0350]

[0351] 1) The ID of the National Institutes of Health (NIH) of the United States can confirm the sequence on the relevant webpage

[0352] 2) The meaning of (major allele) > (minor allele)

[0353] 3) Minor allele frequency = (2mm + Mm) / 2(MM + Mm + mm)

[0354] 4) The statistical significance of the phenotypic differences among the three genotypes (M / M, M / m, m / m) (M: major allele, 5m: minor allele)

[0355] 5) As the minor allele increases one by one, the degree of increase or decrease in phenotype (-: the risk of sagging pores and wrinkles decreases,

[0356] +: the risk of sagging pores and wrinkles increases)

[0357] The degree of sagging pores and wrinkles = the severity level of pore sagging in the cheek area (evaluated by experts with the naked eye)

[0358] Table 6 below shows representative examples of SNP markers significantly associated with the degree of nasolabial folds.

[0359] 10[Table 6]

[0360] Genetic polymorphism markers associated with the degree of nasolabial folds (P<0.0001)

[0361]

[0362]

[0363]

[0364]

[0365]

[0366]

[0367]

[0368]

[0369]

[0370]

[0371]

[0372]

[0373]

[0374]

[0375]

[0376]

[0377]

[0378]

[0379]

[0380]

[0381]

[0382]

[0383]

[0384]

[0385]

[0386]

[0387]

[0388]

[0389]

[0390]

[0391]

[0392]

[0393]

[0394]

[0395]

[0396]

[0397]

[0398]

[0399]

[0400]

[0401]

[0402]

[0403]

[0404]

[0405]

[0406]

[0407]

[0408]

[0409]

[0410]

[0411]

[0412]

[0413]

[0414]

[0415]

[0416]

[0417]

[0418]

[0419]

[0420]

[0421]

[0422]

[0423]

[0424]

[0425]

[0426]

[0427]

[0428]

[0429]

[0430]

[0431]

[0432] 1) The ID of the National Institutes of Health (NIH) of the United States can confirm the sequence on the relevant webpage.

[0433] 2) The meaning of (major allele) > (minor allele)

[0434] 3) Minor allele frequency = (2mm + Mm) / 2(MM + Mm + mm)

[0435] 4) Statistical significance of phenotypic differences among the three genotypes (M / M, M / m, m / m) (M: major allele, m: minor allele)

[0436] 5) Degree of increase or decrease in phenotype with the gradual increase of the minor allele (-: reduction in nasolabial fold risk, +: increase in nasolabial fold risk)

[0437] Nasolabial fold degree = severity grade of wrinkles at the nasolabial fold area of the face (evaluated by experts with the naked eye)

[0438] 6) Degree of sagging: Degree of wrinkles sagging downward due to gravity, referring to Figure 4 the respective grade images and grade criteria for the degree of sagging shown

[0439] 7) Degree of depression: Degree of depression on the skin surface without sagging, referring to Figure 4 the respective grade images and grade criteria for the degree of depression shown

[0440] Example 2: Analysis results of the effect of various substances on promoting collagen synthesis

[0441] 2-1. Analysis results of the collagen synthesis promoting effects of various substances for improving skin fine lines

[0442] Attempts were made to confirm the effectiveness of promoting collagen synthesis by treating various substances capable of promoting collagen synthesis on the corresponding genes and related mechanisms.

[0443] Specifically, phloretin, oryzanol, lupeol, and sucralfate were used as the above substances.

[0444] Human skin fibroblasts (NHDF-Neo-Human Dermal Fibroblasts, Neonatal, Lonza, CC-2509) were purchased from Lonza and used. The purchased cells were cultured in a culture medium prepared by mixing 10% FBS (Fetal Bovine Serum) and 1% antibiotics (Penicillin streptomycin) in DMEM (Dulbecco Modified Eagle Medium - Thermo Fisher Scientific) in a 37°C, 5% CO2 incubator.

[0445] To analyze the efficacy of promoting collagen mRNA synthesis, human skin fibroblasts were dispensed at 1.5×10^5 cells / well onto a 6-well plate and cultured in a 37°C, 5% CO2 incubator for 24 hours. The substances were added to serum-free media at appropriate concentrations / combinations and then cultured again under the same conditions for 24 hours. Subsequently, mRNA was obtained using an RNA-spin total RNA extraction kit (RNA-spin total RNA extraction kit) (iNtRON Biotechnology, 17211), and cDNA was synthesized using a cDNA synthesis kit (cDNA synthesis kit) (PhileKorea, ET21100). Quantitative PCR (qPCR) was performed using the synthesized cDNA and taqman probes (taqman probe) (assay IDs: COL1A1-Hs00164004_m1, COL3A1-Hs00943809_m1, COL4A1-Hs00266237_m1, BNC2-Hs00417700_m1, EDAR-Hs00223468_m1) to analyze the expression levels of each gene.

[0446] As a result, as Figure 1 shown, it was confirmed that when phloretin or oryzanol was treated alone, the expression of type I collagen (COL1A1), type III collagen (COL3A1), and type IV collagen (COL4A1) was promoted, and when phloretin and oryzanol were treated in combination, a synergistic effect on the gene expression of type I collagen (COL1A1) and type IV collagen (COL4A1) was shown.

[0447] In addition, as Figure 2 shown, it was confirmed that when lupeol or sucralfate was treated alone, the expression of type III collagen (COL3A1) was promoted, and when lupeol and sucralfate were treated in combination, a synergistic effect on the gene expression of type III collagen (COL3A1) and type IV collagen (COL4A1) was shown.

[0448] 2-2. Analysis results of the effect of various substances for improving sagging pores and wrinkles on promoting collagen synthesis

[0449] Attempts were made to confirm whether there was an effect on promoting collagen synthesis by treating various substances that could promote collagen synthesis on the corresponding genes and related mechanisms.

[0450] Specifically, the above substances used fucoidan, N-feruloylserotonin, emodin, and formononetin.

[0451] Human skin fibroblasts (NHDF-Neo-Human Dermal Fibroblasts, Neonatal, Lonza, CC-2509) were purchased from Lonza and used. The purchased cells were cultured in a medium prepared by mixing 10% FBS (Fetal Bovine Serum) and 1% antibiotics (Penicillin streptomycin) in DMEM (Dulbecco Modified Eagle Medium - Thermo Fisher Scientific) in a 37°C, 5% CO2 incubator.

[0452] To analyze the efficacy of promoting collagen mRNA synthesis, human skin fibroblasts were dispensed at 1.5X10^5 cells / well onto a 6-well plate and cultured in a 37°C, 5% CO2 incubator for 24 hours. The substances were added to the serum-free medium at appropriate concentrations / combinations, and then cultured again under the same conditions for 24 hours. Subsequently, mRNA was obtained using the RNA-spin total RNA extraction kit (iNtRON Biotechnology, 17211), and cDNA was synthesized using the cDNA synthesis kit (Phile Korea, ET21100). qPCR was performed using the synthesized cDNA and taqman probes (detection IDs: COL1A1-Hs00164004_m1, COL4A1-Hs00266237_m1, sgpp2-Hs00544786_m1), ADIPOQ F primer (5'-CAGGCCGTGATGGCAGAGATG-3'), and ADIPOQ R primer (5'-GGTTTCACCGATGTCTCCCTTAG-3') to analyze the expression levels of each gene.

[0453] As a result, as Figure 10 shown, it was confirmed that when fucoidan or N-feruloylserotonin was treated alone, the expression of type IV collagen (COL4A1) was promoted, and when fucoidan and N-feruloylserotonin were treated in combination, a synergistic effect on the gene expression of type IV collagen (COL4A1) was shown.

[0454] In addition, as Figure 11 shown, it was confirmed that when emodin or formononetin was treated alone, the expression of type I collagen (COL1A1) and type IV collagen (COL4A1) was promoted, and when emodin and formononetin were treated in combination, a synergistic effect on the gene expression of type I collagen (COL1A1) and type IV collagen (COL4A1) was shown.

[0455] 2-3. Analysis results of the collagen synthesis promoting effects of various substances for improving nasolabial folds

[0456] An attempt was made to confirm whether there was an effect on promoting collagen synthesis by treating various substances capable of promoting collagen synthesis on the corresponding genes and related mechanisms.

[0457] Specifically, the above substances used were niacinamide, tranexamic acid, pyridoxine HCl, camphor, Cordyceps sinensis extract, and sodium mannose phosphate.

[0458] Human skin fibroblasts (NHDF-Neo-Human Dermal Fibroblasts, Neonatal, Lonza, CC-2509) were purchased from Lonza and used. The purchased cells were cultured in a medium prepared by mixing 10% FBS (fetal bovine serum) and 1% antibiotics (penicillin streptomycin) in DMEM (Dulbecco's Modified Eagle Medium - Thermo Fisher Scientific) in a 37°C, 5% CO2 incubator.

[0459] To analyze the efficacy of promoting collagen mRNA synthesis, human skin fibroblasts were dispensed into a 6-well plate at 1.5X10^5 cells / well and cultured in a 37°C, 5% CO2 incubator for 24 hours. The substance was added to the serum-free medium at an appropriate concentration / combination, and then cultured again under the same conditions for 24 hours. Subsequently, mRNA was obtained using the RNA-spin total RNA extraction kit (iNtRON Biotechnology, 17211), and cDNA was synthesized using the cDNA synthesis kit (PhileKorea, ET21100). Quantitative PCR (qPCR) was performed using the synthesized cDNA and taqman probes (detection IDs: COL1A1-Hs00164004_m1, COL3A1-Hs00943809_m1, PPARGC1B-Hs00993805_m1, SPATA5-Hs01047535_m1, EN1-Hs00154977_m1) to analyze the expression levels of each gene.

[0460] As a result, as Figure 19 shown, it was confirmed that when camphor, niacinamide, or sodium mannosephosphate was treated alone, the expression of type I collagen (COL1A1) was promoted, and when camphor, niacinamide, and sodium mannosephosphate were treated in combination, a synergistic effect on the gene expression of type I collagen (COL1A1) was shown.

[0461] At the same time, it was confirmed that when tranexamic acid or pyridoxine hydrochloride was treated alone, the expression of type I collagen (COL1A1) was promoted, and when tranexamic acid and pyridoxine hydrochloride were treated in combination, a synergistic effect on the gene expression of type I collagen (COL1A1) was shown.

[0462] In addition, as Figure 20 shown, it was confirmed that when a mixture of 7 substances including 6 substances of the present invention (camphor, niacinamide, sodium mannosephosphate, tranexamic acid, pyridoxine hydrochloride, and Cordyceps sinensis extract) and retinol was treated, the most excellent synergistic effect on the gene expression of type I collagen (COL1A1) was shown.

[0463] Example 3: Human applicability test for confirming the improvement degree of fine lines, sagging pores, wrinkles, and nasolabial folds relative to retinol

[0464] An attempt was made to discover substances that can maximize the effects of improving skin fine lines, sagging pores and wrinkles, and nasolabial folds based on the polymorphic markers obtained in Example 1 that are associated with the presence or absence of skin fine lines, sagging pores and wrinkles, nasolabial folds, and their degrees, and to formulate a customized substance formula.

[0465] 3-1: Selection of subjects

[0466] After approval by the Institutional Review Board (IRB, South Korea) (LG-Wr_Fine-2021-1013), the study was carried out in accordance with the standard operating guidelines and clinical trial standards of the LG Household & Healthcare Institute's Bioethics Committee. In order to confirm the degree of improvement in skin fine lines, 11 healthy subjects with no major abnormalities in skin appearance and no dermatological surgery experience within 3 months before the start of the trial were selected from adults in their 20s to 50s with many wrinkles at the corners of the eyes and under the eyes.

[0467] In addition, in order to confirm the degree of improvement in sagging pores and wrinkles, 8 healthy subjects with no major abnormalities in skin appearance and no dermatological surgery within 3 months before the start of the trial were selected from adults in their 20s to 50s who had many sagging pores and wrinkles near the cheeks.

[0468] In order to confirm the degree of improvement in nasolabial folds, 10 healthy subjects were selected from adults in their 20s to 50s who had no major abnormalities in skin appearance and had no experience of dermatological surgery within 3 months before the start of the trial.

[0469] 3-2: Subject Exclusion Criteria

[0470] The above subjects were excluded if 1) they had eczematous / infectious skin diseases, 2) they had allergic constitutions or allergies, 3) they were pregnant or breastfeeding, 4) they had obvious nutritional disorders, 5) they were drug or alcohol intoxicated, 6) they had moles, acne, tattoos, erythema, burn marks, etc. on the test site, 7) it had not been 1 month since they participated in the same test, and 8) they had hypertension or diabetes.

[0471] 3-3: Test information

[0472] 3-3-1: Trial information on the degree of improvement of skin fine lines

[0473] Control group cream: 0 to 2 weeks is the skin adaptation period, during which retinol 0.1% (3300 IU) cream is applied to the left canthus every other night. From 2 weeks to the end of the experiment, it is applied to the left canthus every night.

[0474] Experimental group cream: Phloretin 0.05%, oryzanol 0.1%, lupeol (raw material name: collageneer) 2%, sucralfate (raw material name: Sucralfatesome) 2% were applied to the right eye corner, twice a day in the morning and evening.

[0475] It was used for 4 weeks in total. Wrinkles before and after use were measured using Antera 3D, and then the overall size (texture overall size) (Ra value) of the fine lines under the eyes was calculated.

[0476] The overall size of fine lines - Ra value refers to the value obtained by dividing the total volume of the higher or lower parts compared to the surface area by the total area when measured by Antera.

[0477] 3 - 3 - 2: Test information for the improvement degree of skin sagging, pores, and wrinkles

[0478] Control group cream: From week 0 to week 2 was the skin adaptation period. During this period, a 0.1% (3300 IU) retinol cream was applied to the left cheek of the face every other evening, and from week 2 to week 4, it was applied to the left cheek of the face every evening.

[0479] Experimental group cream: Containing 0.3% N - Feruloylserotonin, 2% Fucoidan, 0.5% Emodin, and 1% Formononetin, it was applied to the right cheek of the face 2 times a day, in the morning and evening.

[0480] It was used for 4 weeks in total and measured using Antera 3D. Before the experiment (week 0) and after the experiment (week 4), wrinkles were measured using Antera 3D respectively. Then, the volume of sagging pores in the butterfly area of the cheek was selected, and then ImageJ analysis was performed.

[0481] 3 - 3 - 3: Test information for the improvement degree of nasolabial folds

[0482] Control group cream: From week 0 to week 2 was the skin adaptation period. During this period, a 0.1% (2500 IU) retinol cream was applied to the left nasolabial fold area every other day, and from week 3, it was applied to the left nasolabial fold area every evening.

[0483] Experimental group cream: 2% niacinamide, 2% tranexamic acid, 0.1% pyridoxine hydrochloride, 0.03% camphor, 1% Cordyceps sinensis extract, and 0.1% sodium mannitol phosphate were applied to the right nasolabial fold area of the face 2 times a day, in the morning and evening.

[0484] It was used for 8 weeks in total. Every 4 weeks, wrinkles were measured using Antera 3D, and then the fold wrinkle length and depth of the concave wrinkles in the nasolabial fold area were measured.

[0485] 3 - 4: Wrinkles measuring instruments and measuring / analyzing methods

[0486] 3-4-1: Under-eye Fine Line Measuring Instrument and Measuring / Analyzing Method

[0487] Use Antera 3D to take photos of the area around the eyes (take photos of the left and right eyes separately) to obtain 3D images (3D image) (take photos 3 times repeatedly) ( Figure 3 ).

[0488] 1) Analysis item: Texture

[0489] 2) Select stripe from the area designation tool → Designate the under-eye skin directly below the eyelashes (designate according to the shape of the under-eye patch, width 9 mm), and automatically select the same area of the 0-week and 4-week images

[0490] 3) When analyzing the texture, use the Ra value among the parameters of score, Ra, Rq, and Rmax for analysis, calculate the average of the values taken 3 times repeatedly and compare them

[0491] 3-4-2: Sagging Pore and Wrinkle Measuring Instrument and Measuring / Analyzing Method

[0492] Use Antera 3D to take photos of the pores near the cheeks between the nose and cheekbones (take photos of the left and right cheeks separately) to obtain 3D images (take photos 3 times repeatedly) ( Figure 12 ).

[0493] 1) Analysis item: Volume

[0494] 2) Select stripe from the area designation tool → Measure the prominent part of the pores in the diagonal direction next to the nose (width 19 mm), and automatically select the same area of the 0-week and 4-week images

[0495] 3) When analyzing the volume

[0496] Threshold: After analyzing with a width of 2 mm, in Image J, in Adjust>Color Threshold, in the Filtered Analyze Particles mode with Hue 90 - 140, after setting 0 - 1000 size, use the average of the results corresponding to Major (the long axis length of the particles) for analysis, calculate the average of the values taken 3 times repeatedly and compare them

[0497] 3-4-3: Nasolabial Fold Measuring Instrument and Measuring / Analyzing Method

[0498] Use Antera 3D to take photos of the nasolabial fold area (take photos of the left and right separately) to obtain 3D images (take photos 3 times repeatedly) ( Figure 22 ).

[0499] 1) Analysis item: Fold (the maximum width of the detected wrinkle is fixed at 3 mm)

[0500] 2) Select the polygon shape from the area designation tool → Designate the nasolabial fold area extending from beside the nose wing to beside the mouth corner, and automatically select the same area of the 0-week and 4-week images

[0501] 3) When analyzing the fold numerical values, use the length value to analyze the length of the sunken nasolabial fold, and use the maximum depth value to analyze the maximum depth of the sunken nasolabial fold. Calculate the average of the values taken 3 times repeatedly and make a comparison

[0502] 3 - 5: Confirm the improvement degree of skin wrinkles relative to retinol

[0503] 3 - 5 - 1: Improvement degree of skin fine lines

[0504] In Example 2, four substances (phloretin, oryzanol, lupeol, and sucralfate) of the present invention that are effective in promoting collagen synthesis are mixed and applied to the right eye corner, while the control group cream (retinol) is applied to the left eye corner

[0505] As a result, as Figure 5 shown, it was confirmed that after 4 weeks of application, when applying the cream containing phloretin, oryzanol, lupeol, and sucralfate, an excellent wrinkle improvement rate was shown, and the fine line improvement rate was 2.1 times that relative to retinol

[0506] 3 - 5 - 2: Confirm the improvement degree of sagging pore wrinkles

[0507] In Example 2, four substances (fucoidan, N-feruloylserotonin, emodin, and formononetin) of the present invention that are effective in promoting collagen synthesis are mixed and applied to the right cheek, while the control group cream (retinol) is applied to the left cheek

[0508] As a result, as Figure 14 shown, it was confirmed that after 4 weeks of application, when applying the cream containing fucoidan, N-feruloylserotonin, emodin, and formononetin, an excellent wrinkle improvement rate was shown, and the sagging pore wrinkle improvement rate was 1.8 times that relative to retinol

[0509] Example 4: Confirmation of the correlation between significantly associated gene polymorphisms related to skin wrinkle degree and treatment with wrinkle-improving substances Relationship

[0510] 4 - 1: Confirm the correlation between the polymorphic markers related to the degree of skin fine lines and the treatment with fine line improvement substances

[0511] An attempt was made to confirm the degree of improvement in fine lines of the substance of the present invention based on the polymorphic markers showing a correlation with the degree of skin fine lines obtained in Example 1.

[0512] As an example, it has been reported that the more the expression level of BNC2, one of the transcription factors of skin cells (especially, keratinocytes), the darker the skin color, and the mutations occurring in the BNC2 gene are associated with pigmented spots on the skin. Therefore, reducing the expression level of BNC2 is to reduce the reactivity to reactive oxygen species while preventing cell aging or death. That is, it can be seen that when the substance of the present invention is treated to reduce the expression level of BNC2, it is effective in improving wrinkles.

[0513] As Figure 6 shown, it was confirmed that when phloretin and oryzanol were treated alone, the expression of BNC2 was inhibited, and when phloretin and oryzanol were treated in combination, due to the synergistic effect, the expression of BNC2 was more effectively inhibited compared to the single treatment.

[0514] As another example, it has been reported that EDAR, an EDA receptor known to be crucial for ectodermal development during the ontogeny of organisms, the more its expression level decreases, the more the NF-kB signal is inhibited, the skin becomes thinner and drier. In particular, it will induce hypohidrotic ectodermal dysplasia (HED) with wrinkles and pigmentation around the eyes. Therefore, increasing the expression level of EDAR is to increase chemokines through NF-Kb activity and enhance the ability to repair stimuli such as photoaging. That is, it can be seen that when the substance of the present invention is treated to increase the expression level of EDAR, it is effective in improving wrinkles.

[0515] As Figure 7 shown, it was confirmed that when lupeol and sucralfate were treated alone, the expression of EDAR increased, and when lupeol and sucralfate were treated in combination, due to the synergistic effect, the expression of EDAR was more effectively promoted compared to the single treatment.

[0516] Thus, it can be seen that by treating the substance of the present invention, which has a higher reactivity to the improvement of fine lines than retinol, customized formulations can be prescribed according to the types of polymorphic markers related to the presence or absence of fine lines.

[0517] 4-2: Confirming the correlation between the polymorphic markers showing a significant association with the degree of sagging pores and wrinkles and the treatment with the substance for improving sagging pores and wrinkles

[0518] An attempt was made to confirm the degree of improvement in sagging pores and wrinkles of the substance of the present invention based on the polymorphic markers showing a correlation with the degree of sagging pores and wrinkles obtained in Example 1.

[0519] As an example, SGPP2 is a gene that expresses an enzyme that can play a major role in inflammatory conditions. When SGPP2 is overactive, it may be more vulnerable to inflammatory signals. In the case of skin aging, inflammatory responses gradually accumulate. Therefore, when the activity of SGPP2 is high, it may be vulnerable to skin inflammatory signals, and inflammatory responses are likely to accumulate, easily leading to skin aging. Thus, reducing the expression level of SGPP2 is to reduce the reactivity to inflammation and at the same time prevent the progression of skin aging. That is, it can be seen that when the substance of the present invention is treated to reduce the expression level of SGPP2, it is effective in improving wrinkles.

[0520] As Figure 15 shown, it was confirmed that when fucoidan and N-feruloylserotonin were treated alone, the expression of SGPP2 was inhibited, and it was confirmed that when fucoidan and N-feruloylserotonin were treated in combination, due to the synergistic effect, compared with the single treatment, the expression of SGPP2 was more effectively inhibited.

[0521] As another example, ADIPOQ is a gene that expresses adiponectin, a substance mainly secreted in adipose tissue, and in the skin, ADIPOQ is mainly known for its anti-inflammatory effect. The lower the ADIPOQ reactivity in the skin, the more the anti-inflammatory effect in the skin is reduced, easily inducing an inflammatory environment. In addition, it can lead to the formation of neutral fat, the reduction of dermal elasticity, and the formation of sagging. Therefore, increasing the expression level of ADIPOQ is to reduce neutral fat and increase dermal elasticity, reducing skin sagging. That is, it can be seen that when the substance of the present invention is treated to increase the expression level of ADIPOQ, it is effective in improving wrinkles.

[0522] As Figure 16 shown, it was confirmed that when emodin and formononetin were treated alone, the expression of ADIPOQ increased, and it was confirmed that when emodin and formononetin were treated in combination, due to the synergistic effect, compared with the single treatment, the expression of ADIPOQ was more effectively promoted.

[0523] Thus, it can be seen that by treating the substance of the present invention, which has a higher reactivity to improving sagging pores and wrinkles than retinol, customized formulations can be prescribed according to the types of polymorphism markers related to the presence or absence of sagging pores and wrinkles.

[0524] 4-3: Confirmation of the correlation between significantly associated gene polymorphisms related to nasolabial fold degree and treatment with nasolabial fold-improving substances

[0525] An attempt was made to confirm the degree of nasolabial fold improvement of the substance of the present invention based on the polymorphism markers obtained in Example 1 that showed a correlation with the degree of nasolabial folds.

[0526] As an example, PPARGC1B is one of the co-activators of peroxisome proliferator-activated receptor-γ (PPARγ), and it is known to be involved in melanogenesis in the skin. Regarding PPARGC1B, it has been reported that a decrease in mitochondrial function leads to the occurrence of wrinkles and that restoring the decreased mitochondrial function reduces wrinkles (Cell Death & Disease volume 9, Article number: 735 (2018)). Therefore, increasing the expression level of PPARGC1B plays a role in restoring the function of mitochondria, which are the cell's energy-producing organs, and thus can prevent and restore deep-set wrinkles, namely nasolabial folds, that have accumulated over a relatively long period. That is, it can be seen that when the substance of the present invention is treated to increase the expression level of PPARGC1B, it is effective in improving wrinkles.

[0527] As shown in Table 7 below, it was confirmed that when treated with niacinamide, camphor, or sodium mannitol phosphate, the expression of PPARGC1B increased.

[0528] [Table 7]

[0529] Substance Gene expression level (PPARGC1B) No treatment 1.00 Niacinamide 1.52 Camphor 2.63 Sodium mannose phosphate 1.53

[0530] As another example, SPATA5 is related to spermatogenesis and is a gene involved in mitochondrial morphogenesis during early spermatogenesis. It has been revealed that SPATA5 is involved in mitochondrial remodeling and ATP production regulation during brain development (Neurology Apr 2019, 92(15 Supplement) P4.6-062;). Although its role in the skin has not been studied, based on the above research, it can play a role in mitochondrial function and ATP production. Therefore, increasing the expression level of SPATA5 plays a role in restoring the function of mitochondria, which are the cell's energy-producing organs, and thus can prevent and restore deep-set wrinkles, namely nasolabial folds, that have accumulated over a relatively long period. That is, when the substance of the present invention is treated to increase the expression level of SPATA5, it is effective in improving wrinkles.

[0531] As shown in Table 8 below, it was confirmed that when treated with tranexamic acid or pyridoxine hydrochloride, the expression of SPATA5 increased.

[0532] [Table 8]

[0533] Substance Gene expression level (SPATA5) No treatment 1.00 Tranexamic acid 1.21 Pyridoxine HCl 1.36

[0534] As another example, EN1 is a transcription factor gene that is known to play a role in the wound healing process of the skin. When EN1 expression increases, the rapid increase of extracellular matrix (ECM) components will accelerate wound healing, thus contributing to the reduction of wrinkles. That is, when the substance of the present invention is treated to increase the expression of EN1, it has an effect on improving wrinkles.

[0535] As shown in Table 9 below, it was confirmed that the expression of EN1 increased when the Cordyceps sinensis extract was treated.

[0536] [Table 9]

[0537] Substance Gene expression level (EN1) No treatment 1.00 Cordyceps sinensis extract 2.49

[0538] It can be seen from this that the substance of the present invention, which has a higher responsiveness to the improvement of nasolabial folds than retinol, can be prescribed in a customized formulation according to the type of polymorphic marker associated with the presence or absence of nasolabial folds.

[0539] Example 5: Human applicability test for developing a customized wrinkle-improving formula for high-risk groups of wrinkles

[0540] An attempt is made to provide a substance that can enhance the efficiency of improving fine lines, sagging pores wrinkles, and / or nasolabial folds based on the correlation between the significant polymorphism markers associated with the degree of skin fine lines, sagging pores wrinkles, and / or nasolabial folds obtained in Example 3 and the treatment with the substance of the present invention, according to the risk level of wrinkle occurrence.

[0541] 5-1: Selecting the subjects

[0542] 5-1-1: Select the subjects with the degree of fine lines

[0543] After approval by the Institutional Review Board (IRB, South Korea) (LG-Wr_Fine-2021-1013), the study was carried out in accordance with the standard operating guidelines and clinical trial standards of the LG Household & Healthcare Institute's Bioethics Committee. Ten healthy subjects with no major abnormalities in skin appearance and no dermatological surgery experience within 3 months before the start of the trial were selected from adults in their 20s to 50s with many wrinkles at the corners and under the eyes.

[0544] As follows, based on the gene polymorphism markers associated with the presence or absence of fine lines under the eyes in Table 1, the subjects were classified into a low-risk group and a high-risk group according to the risk level of fine line occurrence.

[0545] Among the 5 markers, including 3 or less minor alleles > low risk group (n = 5)

[0546] Among the 5 markers, including more than 4 minor alleles > high-risk group (n = 5)

[0547] 5-1-2: Selecting subjects with drooping pore and wrinkle degrees

[0548] After approval by the Institutional Review Board (IRB, Korea) (LG-Wr_Fine-2021-1013), and in accordance with the standard operating guidelines and clinical trial standards of the LG Life Sciences Institutional Review Board, 8 healthy subjects aged 20 to 50 with more drooping pores and wrinkles near the cheeks were selected. These subjects had no major abnormalities in skin appearance and no dermatological surgery experience within 1 month before the start of the trial.

[0549] As follows, according to the risk levels of drooping pore and wrinkle occurrence for 2 markers SGPP2 (rs4592839) and ADIPOQ (rs76071583) with positive effect sizes among the 4 markers related to the presence or absence of drooping pores and wrinkles in Table 2, the subjects were classified into low-risk and high-risk groups.

[0550] Among the 2 markers, including 0 or fewer minor alleles > low-risk group (n = 4)

[0551] Among the 2 markers, including 1 or more minor alleles > high-risk group (n = 4)

[0552] 5-1-3: Selecting subjects with nasolabial fold degrees

[0553] After approval by the Institutional Review Board (IRB, Korea) (LG-Wr_Deep-2021-1018), and in accordance with the standard operating guidelines and clinical trial standards of the LG Life Sciences Institutional Review Board, 10 healthy subjects aged 20 to 50 with more nasolabial folds near the cheeks were selected. These subjects had no major abnormalities in skin appearance and no dermatological surgery experience within 1 month before the start of the trial.

[0554] As follows, according to the risk levels of nasolabial fold depression occurrence for 3 markers related to nasolabial fold depression (listed in Annex 2) in Table 3, the subjects were classified into low-risk and high-risk groups.

[0555] The sum of the number of minor alleles * effect size (β) for each marker is 0.08 or more > high-risk group (n = 5)

[0556] The sum of the number of minor alleles of each marker * effect size (β) is less than 0.08 > low-risk group (n = 5)

[0557] * Example 1: When the number of minor alleles of the markers for Subject 1 is PPARGC1B = 1, SPATA5 = 1, and EN1 = 2, the risk level = 1 * 0.57 + 1 * (-0.23) + 2 * 0.27 = 0.88 high-risk group

[0558] * Example 2: When the number of minor alleles of the markers for Subject 2 is PPARGC1B = 0, SPATA5 = 2, and EN1 = 0, the risk level = 2 * (-0.23) = -0.46 low-risk group

[0559] 5-2: Subject exclusion criteria

[0560] Same as Example 3-2

[0561] 5-3: Test information

[0562] Same as Example 3-3

[0563] 5-4: Wrinkles measuring instrument and measuring / analysis method

[0564] Same as Example 3-4

[0565] 5-5: Confirming the wrinkle improvement rate brought by the treatment of the substance of the present invention

[0566] 5-5-1: Confirming the improvement rate of fine lines

[0567] It was confirmed that when applying a cream containing 4 substances (phloretin, oryzanol, lupeol, and sucralfate) of the present invention, as Figure 8 and Figure 9 shown, in the low-risk group, when applying the customized formula compared with retinol alone, the length of the fine lines under the eyes improved to a similar level, but in the high-risk group, when applying the customized formula compared with retinol alone, the improvement rate of the length of the fine lines under the eyes increased by about 3.6 times.

[0568] It can be seen that according to the presence or absence of alleles of a specific gene SNP, the reactivity to substances will be different, and it can be seen that providing a customized formula for the high-risk group with a high risk level of fine line occurrence can improve the fine line improvement efficiency.

[0569] 5-5-2: Confirming the improvement rate of sagging pore wrinkles

[0570] Retinol formulations and customized formulations (0.3% N-Feruloylserotonin, 2% Fucoidan, 0.5% Emodin, 1% Formononetin) were formulated for the low-risk group and the high-risk group, and it was confirmed that the efficacy of improving drooping pore wrinkles of the customized formulation in the high-risk group increased by 2.26 times compared with the low-risk group.

[0571] It was confirmed that in the low-risk group, when applying the customized formulation, the average length of drooping pore wrinkles was improved to a similar level compared with retinol alone, while in the high-risk group, when applying the customized formulation, the average length of drooping pore wrinkles was 2.7 times better than retinol alone.

[0572] From this, it can be seen that the reactivity to substances varies depending on whether or not specific gene SNPs carry alleles, and it can be seen that providing a customized formulation for the high-risk group with a high risk level of drooping pore wrinkles can improve the efficiency of improving drooping pore wrinkles.

[0573] 5-5-3: Confirm the improvement rate of nasolabial folds

[0574] It was confirmed that in the low-risk group, when applying the customized formulation, the length of the sunken nasolabial folds was improved to a similar level compared with retinol, and the maximum depth was significantly improved by about 44% ( Figure 21 ).

[0575] It was confirmed that in the high-risk group, when applying the customized formulation, compared with retinol, not only was the length of the sunken nasolabial folds significantly improved by more than about 58.2%, but also the maximum depth was significantly improved by about 173.9% ( Figure 21 ).

[0576] From this, it can be seen that the reactivity to substances varies depending on whether or not specific gene SNPs carry alleles, and it can be seen that providing a customized formulation for the high-risk group with a high risk level of nasolabial folds can improve the efficiency of improving nasolabial folds.

[0577] Based on the above description, those skilled in the art to which the present invention pertains can understand that the present invention can be implemented in other specific forms without changing the technical idea or essential features of the present invention. In this regard, it should be understood that the above-described embodiments are exemplary in all respects and not restrictive. The scope of the present invention should be construed to include the meaning and scope of the claims described below and all modifications or variations derived from its equivalent concepts, and should not be construed to include only the above detailed description.

Claims

1. A composition for improving skin fine lines, which contains one or more substances selected from the group consisting of phloretin, oryzanol, lupeol, and sucralfate as active ingredients.

2. The composition according to claim 1, wherein, The above composition contains (i) phloretin and oryzanol; (ii) lupeol and sucralfate; or (iii) phloretin, oryzanol, lupeol, and sucralfate as active ingredients.

3. The composition according to claim 1, wherein Based on the total weight of the composition, the content of the above substances is 0.0001 to 10% by weight.

4. The composition according to claim 1, wherein, The above skin fine lines are fine lines under the eyes or fine lines at the corners of the mouth.

5. The composition according to claim 1, characterized in that, The above composition promotes collagen synthesis.

6. The composition according to claim 5, wherein, The substances for promoting the synthesis of type I collagen or type IV collagen are phloretin and oryzanol.

7. The composition according to claim 5, wherein, The substances for promoting the synthesis of type III collagen or type IV collagen are lupeol and sucralfate.

8. The composition according to claim 1, wherein The above substances promote the expression of EDAR or inhibit the expression of BNC2.

9. The composition according to claim 8, wherein The substances for inhibiting the expression of BNC2 are phloretin or oryzanol.

10. The composition according to claim 8, wherein, The substances for promoting the expression of EDAR are lupeol or sucralfate.

11. The composition according to claim 1, wherein, The above improvement of wrinkles is customized for individuals having one or more single nucleotide polymorphism markers in the gene encoding the protein of EDAR or BNC2.

12. The composition according to claim 11, wherein, The above single nucleotide polymorphism markers are any one or more selected from the group consisting of rs10865025, rs10756807, rs1952688, rs10116469, and rs12338608.

13. The composition according to claim 11, wherein, The above single nucleotide polymorphism markers are composed of one or more polynucleotides selected from the group consisting of the following polynucleotides; and their complementary polynucleotides: A polynucleotide composed of 5 - 100 consecutive DNA sequences containing the 109511765th base of chromosome 2 of humans, wherein the 109511765th base is G or A (rs10865025); A polynucleotide composed of 5 - 100 consecutive DNA sequences containing the 16756041st base of chromosome 9 of humans, wherein the 16756041st base is G or A (rs10756807); A polynucleotide composed of 5 - 100 consecutive DNA sequences containing the 16782156th base of chromosome 9 of humans, wherein the 16782156th base is A or G (rs1952688); A polynucleotide composed of 5 - 100 consecutive DNA sequences containing the 16783095th base of chromosome 9 of humans, wherein the 16783095th base is A or T (rs10116469); A polynucleotide composed of 5 - 100 consecutive DNA sequences containing the 16800470th base of chromosome 9 of humans, wherein the 16800470th base is C or T (rs12338608).

14. An information providing method for selecting customized substances for improving fine lines, which includes the following steps: (a) Obtaining a biological sample from an individual; (b) Amplifying or hybridizing with a probe a polymorphic site of any one or more single nucleotide polymorphism markers selected from the group consisting of rs10865025, rs10756807, rs1952688, rs10116469, and rs12338608 from the above biological sample; and (c) Identifying the bases of the polymorphic site amplified or hybridized in step (b) above.

15. The information providing method according to claim 14, wherein, The above substance is one or more selected from the group consisting of phloretin, oryzanol, lupeol, and sucralfate.

16. The information providing method according to claim 14, wherein, When there are 4 or more minor alleles among the bases of rs10865025, rs10756807, rs1952688, rs10116469, and rs12338608, the customized substances for improving fine lines are selected from phloretin and oryzanol; lupeol and sucralfate; or phloretin, oryzanol, lupeol, and sucralfate.

17. A composition for improving drooping pores and wrinkles, which contains any one or more substances selected from the group consisting of fucoidan, N-feruloylserotonin, emodin, and formononetin as an active ingredient.

18. The composition according to claim 17, wherein, The above composition contains (i) fucoidan and N-feruloylserotonin; (ii) emodin and formononetin; or (iii) fucoidan, N-feruloylserotonin, emodin, and formononetin as an active ingredient.

19. The composition according to claim 17, wherein Based on the total weight of the composition, the content of the above substance is 0.0001 to 10% by weight.

20. The composition according to claim 17, characterized in that, The above composition promotes collagen synthesis.

21. The composition according to claim 20, wherein, The substances for promoting type IV collagen synthesis are fucoidan and N-feruloylserotonin.

22. The composition according to claim 20, wherein, The substances for promoting type I collagen or type IV collagen synthesis are emodin and formononetin.

23. The composition according to claim 17, wherein The above substances promote the expression of ADIPOQ or inhibit the expression of SGPP2.

24. The composition according to claim 23, wherein, The substances for promoting ADIPOQ expression are emodin or formononetin.

25. The composition according to claim 23, wherein, The substances for inhibiting SGPP2 expression are fucoidan or N-feruloylserotonin.

26. The composition according to claim 17, wherein, The above wrinkle improvement is a customized type for individuals having one or more single nucleotide polymorphism markers present in the gene encoding the protein of ADIPOQ or SGPP2.

27. The composition according to claim 26, wherein, The above single nucleotide polymorphism markers are any one or more selected from the group consisting of rs10203329, rs4592839, rs76071583, and rs2036373.

28. The composition according to claim 26, wherein The above single nucleotide polymorphism markers are composed of one or more polynucleotides selected from the group consisting of the following polynucleotides; and their complementary polynucleotides: A polynucleotide composed of 5 - 100 consecutive DNA sequences containing the 223350001st base of human chromosome 2, wherein the 223350001st base is A or G (rs10203329); A polynucleotide consisting of 5 - 100 consecutive DNA sequences of the 223412813th base of chromosome 2 containing a human, wherein the 223412813th base is T or C (rs4592839); A polynucleotide consisting of 5 - 100 consecutive DNA sequences of the 186557872th base of chromosome 3 containing a human, wherein the 186557872th base is A or G (rs76071583); A polynucleotide consisting of 5 - 100 consecutive DNA sequences of the 186570191th base of chromosome 3 containing a human, wherein the 186570191th base is T or G (rs2036373).

29. An information - providing method for selecting a customized substance for improving sagging pore wrinkles, comprising the following steps: (a) Obtaining a biological sample from an individual; (b) Amplifying or hybridizing with a probe a polymorphic site of one or more single - nucleotide polymorphism markers selected from the group consisting of rs4592839 and rs76071583 from the biological sample; and (c) Confirming the base of the polymorphic site amplified or hybridized in step (b).

30. The information providing method according to claim 29, wherein, The above - mentioned substance is one or more selected from the group consisting of fucoidan, N - feruloylserotonin, emodin, and formononetin.

31. The information providing method according to claim 30, wherein, When one or more minor alleles are included in the bases of rs4592839 and rs76071583, the customized substance for improving sagging pore wrinkles selects fucoidan and N - feruloylserotonin; emodin and formononetin; or fucoidan, N - feruloylserotonin, emodin, and formononetin.

32. A composition for improving nasolabial folds, which contains one or more substances selected from the group consisting of niacinamide, tranexamic acid, pyridoxine HCl, camphor, Cordyceps sinensis extract, sodium mannose phosphate, and retinol as active ingredients.

33. The composition according to claim 32, wherein, The above - mentioned composition contains (i) camphor, niacinamide, and sodium mannose phosphate; (ii) tranexamic acid and pyridoxine HCl; or (iii) retinol, camphor, niacinamide, sodium mannose phosphate, tranexamic acid, pyridoxine HCl, and Cordyceps sinensis extract as active ingredients.

34. The composition according to claim 32, wherein, Based on the total weight of the composition, the content of the above - mentioned substances is 0.0001 to 10% by weight.

35. The composition according to claim 32, wherein The above - mentioned composition promotes collagen synthesis.

36. The composition according to claim 35, wherein, The above - mentioned collagen is type I collagen.

37. The composition according to claim 32, wherein The above - mentioned substances promote the expression of PPARGC1B, SPATA5, or EN1.

38. The composition according to claim 37, wherein, The substances for promoting the expression of PPARGC1B are niacinamide, camphor, or sodium mannose phosphate.

39. The composition according to claim 37, wherein, The substances for promoting the expression of SPATA5 are tranexamic acid or pyridoxine HCl.

40. The composition according to claim 37, wherein, The substance for promoting the expression of EN1 is Cordyceps sinensis extract.

41. The composition according to claim 32, wherein, The above-mentioned wrinkle improvement is customized for individuals having one or more single nucleotide polymorphism markers in the genes encoding the proteins of PPARGC1B, SPATA5, or EN1.

42. The composition according to claim 41, wherein, The above-mentioned single nucleotide polymorphism markers are any one or more selected from the group consisting of rs76148197, rs2166027, and rs41444645.

43. The composition according to claim 10, wherein The above-mentioned single nucleotide polymorphism markers are composed of one or more polynucleotides selected from the group consisting of the following polynucleotides; and their complementary polynucleotides: A polynucleotide composed of 5-100 consecutive DNA sequences containing the 119,602,515th base of chromosome 2 of a human, wherein the 119,602,515th base is A or G (rs76148197); A polynucleotide composed of 5-100 consecutive DNA sequences containing the 124,171,618th base of chromosome 4 of a human, wherein the 124,171,618th base is T or C (rs2166027); A polynucleotide composed of 5-100 consecutive DNA sequences containing the 149,131,368th base of chromosome 5 of a human, wherein the 149,131,368th base is C or G (rs41444645).

44. An information providing method for selecting a customized substance for improving nasolabial folds, comprising the following steps: (a) Obtaining a biological sample from an individual; (b) Amplifying or hybridizing with a probe a polymorphic site of one or more single nucleotide polymorphism markers selected from the group consisting of rs76148197, rs2166027, and rs41444645 from the above-mentioned biological sample; and (c) Confirming the base of the polymorphic site amplified or hybridized in the step (b).

45. The information providing method according to claim 44, wherein, The above-mentioned substance is one or more selected from the group consisting of niacinamide, tranexamic acid, pyridoxine hydrochloride, camphor, Cordyceps sinensis extract, sodium mannitol phosphate, and retinol.

46. The information providing method according to claim 44, wherein, When one or more minor alleles are included in the bases of rs76148197, rs2166027, and rs41444645, the customized substances for improving nasolabial folds are selected as camphor, niacinamide, and sodium mannitol phosphate; tranexamic acid and pyridoxine hydrochloride; or retinol, camphor, niacinamide, sodium mannitol phosphate, tranexamic acid, pyridoxine hydrochloride, and Cordyceps sinensis extract.

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