Composition for alleviating skin irritation caused by vitamin C

Abstract

The present invention relates to a composition for alleviating skin irritation caused by vitamin C, and an information providing method for selecting a custom-made substance for alleviating irritation caused by vitamin C, comprising a step of confirming a base of a polymorphic site of a gene polymorphic marker which is significant in relation to the degree of irritation caused by vitamin C. The invention also relates to a composition for alleviating skin irritation caused by vitamin C and an information providing method for selecting the custom-made substance for alleviating irritation caused by vitamin C. Furthermore, the present invention relates to: a composition for diagnosing whether or not a skin type in which skin irritation caused by vitamin C is likely to occur, the composition comprising a probe capable of detecting a gene polymorphism marker having significance in association with skin irritation caused by vitamin C or a preparation capable of amplifying the gene polymorphism marker; a kit or a microarray comprising the composition; and a method for providing information on the type of skin, which is susceptible to skin irritation caused by vitamin C, using the gene polymorphism marker or the combination of markers.

Description

TECHNICAL FIELD

[0001] The present invention relates to a composition for alleviating irritation caused by vitamin C and an information providing method for selecting a customized material for alleviating irritation caused by vitamin C, which includes a step of confirming the base of a polymorphic site of a genetic polymorphic marker associated with the degree of irritation caused by vitamin C. The customized material of the present invention relates to a cosmetic composition for preventing discoloration in a dosage form and improving efficacy by alleviating irritation induced by vitamin C, improving stability of vitamin C. In addition, the present invention relates to a composition for diagnosing the degree of irritation caused by vitamin C, which comprises a probe capable of detecting or a preparation capable of amplifying a genetic polymorphic marker associated with the degree of irritation caused by vitamin C, a kit or a microarray comprising the above composition, and a method for providing information on judging the degree of irritation caused by vitamin C using the above genetic polymorphic marker or a combination of markers. BACKGROUND

[0002] Skin, which is located at the outermost part of our body, is an organ occupying the largest area (1.5-2 m 2 ) and is responsible for the defense function against external environments such as ultraviolet rays and foreign substances. In the process of exposure to various external environments and aging, melanin is synthesized and released in the skin, which causes an adverse cosmetic result, thereby becoming a factor for reducing the quality of life. Vitamin C is well known as one of representative whitening raw materials for preventing such pigmentation caused by melanin.

[0003] Vitamin C, also known as ascorbic acid, is one of essential nutrients that cannot be synthesized in the body itself and thus must be supplemented through food. In addition, it has a function of not only alleviating pigmentation of the skin by inhibiting tyrosinase activity involved in the melanin production process, but also delaying skin aging by promoting collagen synthesis in the body and reducing active oxygen in the body with excellent antioxidant activity, and thus is one of representative active ingredients for whitening and preventing aging in cosmetics.

[0004] As such, vitamin C exhibits excellent anti-aging and skin cosmetic effects due to its high antioxidant capacity, but correspondingly has the following problems: it is unstable, sensitive to air, in particular, oxygen and heat, external environments such as light, and easily oxidized and decomposed, inducing discoloration in a dosage form. Therefore, in order to compensate for these problems of vitamin C and improve stability, vitamin C derivatives (ascorbic acid derivatives) that change the structure of vitamin C are widely used. However, vitamin C derivatives have the disadvantage that the effects such as antioxidant capacity, pigmentation inhibition capacity, etc. are significantly decreased compared to pure vitamin C.

[0005] On the other hand, the demand for high content products is increasing due to the increased consumer interest in vitamin C and research showing higher efficacy at concentrations of 10-20%. However, as the content increases, not only does the formulation become unstable, but irritation caused by vitamin C is induced on the skin. However, the mechanism of its occurrence is not fully understood, and thus, currently, the sensitivity to vitamin C products is generally judged after direct use of the product. Therefore, in order to minimize side effects and more effectively apply vitamin C to the skin, it is necessary to first conduct research on the causes of vitamin C-induced irritation and develop a diagnostic method. According to some research, it was confirmed that when vitamin C is applied topically to the skin, the gene expression of collagen in the dermis layer and the synthesis of inhibitors of MMP-1 are increased (pmid 11407971), and in other disease research, it was confirmed that there is a correlation between variations in the vitamin C transporter gene and general chronic diseases (pmid 26980812). Therefore, it is expected that the skin reactivity of vitamin C will also vary depending on the genotype.

[0006] However, it is difficult to find examples of specific genetic correlation analysis of skin reactivity to vitamin C, particularly the degree of irritation. Therefore, the discovery of genetic variations and genes associated with vitamin C skin irritation is expected to provide important information for understanding the biological mechanism and understanding and predicting the application of vitamin C to individuals and irritation thereof. SUMMARY

[0007] PROBLEMS TO BE SOLVED BY THE INVENTION

[0008] In the present invention, genetic variations and genes associated with vitamin C skin irritation in Koreans were discovered using candidate gene analysis, and a model for predicting vitamin C skin irritation using genetic variations was developed. In addition, it was aimed to develop an efficacy material that can alleviate vitamin C skin irritation in Koreans based on data on genetic information and skin measurement information and apply it to products.

[0009] MEANS FOR SOLVING THE PROBLEMS

[0010] An object of the present invention is to provide a composition for alleviating skin irritation caused by vitamin C.

[0011] Another object of the present invention is to provide a cosmetic composition or a quasi-drug composition comprising the above composition as an active ingredient.

[0012] Another object of the present invention is to provide a composition having the effect of not only alleviating irritation caused by vitamin C but also improving the stability of vitamin C and enhancing the efficacy of vitamin C.

[0013] Another object of the present application is to provide an information providing method for selecting a customized material for relieving irritation caused by vitamin C, which comprises a step of confirming a base of a polymorphic site of a single nucleotide polymorphism marker associated with significance of skin irritation caused by vitamin C.

[0014] Another object of the present application is to provide a single nucleotide polymorphism (SNP) marker for diagnosing a degree of irritation caused by vitamin C.

[0015] Another object of the present application is to provide a composition for diagnosing a degree of irritation caused by vitamin C, which comprises a probe capable of detecting a single nucleotide polymorphism (SNP) marker for diagnosing a degree of irritation caused by vitamin C or a preparation capable of amplification.

[0016] Another object of the present application is to provide a kit or a microarray for diagnosing a degree of irritation caused by vitamin C, which comprises the above composition.

[0017] Another object of the present application is to provide a method of providing information on a degree of irritation caused by vitamin C, which comprises a step of confirming a polymorphic site of the above single nucleotide polymorphism marker.

[0018] Another object of the present application is to provide an information providing method of prescribing a customized material for a high-risk group susceptible to irritation caused by vitamin C, which comprises a step of confirming a polymorphic site of the above single nucleotide polymorphism marker.

[0019] Inventive Effects

[0020] Information on a degree of irritation caused by vitamin C of an individual can be provided by a genetic polymorphism marker associated with significance of a degree of irritation caused by vitamin C according to the present application, and further, a customized ingredient or product capable of relieving a degree of irritation caused by vitamin C can be developed according to information on a genetic polymorphism marker observed in an individual. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 A vitamin C-specific skin irritation recognition gene expression modulation material of the present application is shown.

[0022] Figure 2 An example of rs10512451 (17:32014117:A:G) in a single nucleotide polymorphism marker in ASIC2 in which there is a difference in irritation recognition according to a minor allele type upon application of vitamin C is shown.

[0023] Figure 3 An example of rs10512451 (17:32014117:A:G) in a single nucleotide polymorphism marker in ASIC2 in which there is a difference in irritation recognition according to a minor allele type upon application of vitamin C is shown.

[0024] Figure 4 The degree of improvement of irritation by adding the vitamin C skin irritation alleviating material of the present application is shown.

[0025] Figure 5 The difference in the improvement of vitamin C irritation between the low risk group and the high risk group when applying the personal customized vitamin C irritation alleviating composition of the present application is shown.

[0026] Figure 6 The degree of discoloration of pure vitamin C caused by heat treatment is shown by naked eye evaluation.

[0027] Figure 7 And Figure 8 The discoloration of pure vitamin C caused by high temperature and the discovery and evaluation of the antioxidant ability recovery material are shown.

[0028] Figure 9 The analysis results of the whitening and antioxidant ability of tannic acid, tocopherol as the vitamin C irritation alleviating material of the present application are shown. DETAILED DESCRIPTION

[0029] The specific description is as follows. On the other hand, each of the explanations and embodiments disclosed in the present application can be applied to each of the other explanations and embodiments. That is, all combinations of the various elements disclosed in the present application belong to the scope of the present application. In addition, it cannot be considered that the scope of the present application is limited by the specific description described below.

[0030] In addition, those skilled in the art can recognize or confirm, only using conventional experiments, that there are a plurality of equivalents to the specific aspects of the present application described in the present application. In addition, such equivalents are intended to be included in the present application.

[0031] As one aspect for achieving the object of the present application, the present application provides a composition for alleviating skin irritation caused by vitamin C. As an example, the above composition contains one or more substances selected from the group consisting of Quercetin, Phloretin, Genistein, Tannic acid, ZnPCA, Madecassoside, Resveratrol, Tocopherol, Asiaticoside, and Ponciri extract as an effective ingredient. As another example, the above composition can contain (i) Quercetin, Phloretin, Genistein, Tannic acid, ZnPCA, Madecassoside, and Resveratrol; or (ii) Madecassoside, Tannic acid, Tocopherol, Genistein, Asiaticoside, and Ponciri extract as an effective ingredient, but is not limited thereto.

[0032] The above term "Quercetin" is a plant flavonoid (polyphenolic flavonoid) compound extracted from the flavonoid group of polyphenols. The above Quercetin is found in many fruits, vegetables, leaves, seeds, and grains. It has a bitter taste and can be used as an ingredient of health supplements, beverages, and foods. The above Quercetin is also called Quercetin. The above Quercetin is not limited to its obtaining method, and can be chemically synthesized by a method known in the art, or using a commercially available substance.

[0033] [Chemical Formula 1]

[0034]

[0035] The above term "Phloretin" is a dihydrochalcone plant-derived phenol having a C6-C3-C6 skeleton, which is abundant in apples (mainly the skin) and apple leaves, and exists as phloridzin (phloretin 2'-O-glucose) in a free form or a glucosidic form. Apple skin contains 0.2 to 5 mg / g of phloretin. Phloretin has many biological functions including antioxidant, anticancer activity, and is related to the prevention of heart disease. This ingredient is particularly superior to other antioxidant ingredients in the ability to pass through the skin barrier. Phloretin is not limited to its obtaining method, and can be chemically synthesized by a method known in the art, or using a commercially available substance.

[0036] [Chemical Formula 2]

[0037]

[0038] The above term "Genistein" is one of isoflavones isolated from soybeans, which is known to have various effects such as anti-tumor, anti-osteoporosis, antioxidant, etc. Since it contains phytoestrogen activity, it is widely used as a substance for medical products or health functional foods, etc. as an ingredient for alleviating various menopausal symptoms, particularly in women in the menopausal period. The above Genistein is not limited to its obtaining method, and can be chemically synthesized by a method known in the art, or using a commercially available substance.

[0039] [Chemical Formula 3]

[0040]

[0041] The above term "tannic acid" is one of the most abundant polyphenols found in plants such as fruits, vegetables, olives, cacao, etc., and recently, tannic acid has been used as a multifunctional coating molecule. In addition, it is known to be an excellent molecule in affinity with biomacromolecules including DNA and proline-rich proteins such as thrombin, gelatin, collagen, and mucin. Tannic acid can bind to proteins through multiple hydrogen bonds and hydrophobic interactions with the phenolic hydroxyl-rich portion (5 gallol groups (3 -OH groups attached to an aromatic ring) and 5 catechol groups (2 -OH groups attached to an aromatic ring through a covalent bond)) of the target protein. The above tannic acid is not limited to its method of obtaining, and can be chemically synthesized by a method known in the art, or using a commercially available material. The above material has an INCI name of tannic acid, and can be used in combination with tannic acid.

[0042] [Chemical Formula 4]

[0043]

[0044] The above term "ZnPCA" is a raw material synthesized using glutamic acid obtained from sugar beet, and zinc, which is a mineral necessary for physiological activities in the body, is known to have various effects (cell differentiation protein synthesis, antibacterial effect). In particular, zinc reduces sebum secretion by inhibiting 5-alpha reductase, and the above ZnPCA can be used in combination with ZnPCA, zinc pyrrolidone carboxylic acid. The above ZnPCA is not limited to its method of obtaining, and can be synthesized by a method known in the art, or using a commercially available material.

[0045] The above term "madecassoside" is one of four main components of an extract of a plant named "Centella asiatica" widely grown in subtropical regions along the coast of the Indian Ocean, and the extract of the plant has been used as a folk medicine to treat skin diseases or leprosy (particularly, ulcer sites of leprosy) by local indigenous people since ancient times. In particular, it is known that madecassoside not only provides a protective effect of inhibiting inflammation, preventing excessive keratinization, and inhibiting penetration of external environmental pollutants, but also promotes collagen production, allowing rapid regeneration of new flesh, thereby leaving as few scars as possible. The above madecassoside is not limited to its method of obtaining, and can be chemically synthesized by a method known in the art, or using a commercially available material.

[0046] The above term "Resveratrol" can be isolated from one or more extracts selected from the group consisting of grape skin, grape seed, peanut, mulberry and cocoa powder. The chemical formula of the above Resveratrol is C14H12O3, and the molecular weight is 228.25. As a stilbene, it is produced in plants by the action of an enzyme, stilbene synthase (STS), and exists in two structural isomers. It is found in many plants including berries such as mulberry, peanut, grape, raspberry, cranberry, etc., and is a phytoalexin secreted when plants are stimulated (stressed), and when grapes are attacked by mold, a powerful antibacterial substance called Resveratrol is secreted as a defense substance to protect itself, and thus is found more than other plants. It can be obtained by extracting natural products by conventional methods, or can be synthesized by a compound synthesis process.

[0047] As non-limiting examples of the above extraction method, hot water extraction, ultrasonic extraction, filtration, reflux extraction, etc. can be exemplified, which can be performed alone or in combination with two or more methods.

[0048] In the present application, the type of solvent used for the above extraction is not particularly limited, and any solvent known in the related art can be used. As non-limiting examples of the above extraction solvent, water, alcohol or a mixed solvent thereof, etc. can be exemplified, which can be used alone or in combination with one or more, and specifically, water can be used. When alcohol is used as a solvent, specifically, an alcohol having a carbon number of 1 to 4 can be used.

[0049] The above term "Tocopherol" is a group of derivatives of fat-soluble vitamin E, and any one or more of α-tocopherol, β-tocopherol, γ-tocopherol and δ-tocopherol or a mixture thereof can be used, and any of those isolated from an extract, synthesized or commercially available products can be used without limitation.

[0050] The above term "Asiaticoside" is a natural organic compound that reduces skin irritation, improves elasticity, and has an oxidation prevention effect. The above Asiaticoside is a triterpene saponin compound, and can exhibit skin moisturizing and oxidation prevention effects. The above Asiaticoside is not limited to its method of obtaining, and can be synthesized by a method known in the art, or a commercially available material is used.

[0051] The above term "yuzu extract" refers to an extract obtained from the fruit of yuzu (Citrus nippokoreana Tanaka). In addition, it is also known as the INCI name "orange peel extract". The above yuzu is different from the commonly known orange (Citrus unshiu Marcov.) in the species name. The above yuzu is an evergreen shrub of the Rutaceae family, and is mainly grown in the city of Jeju. The genus name is derived from the ancient Greek name kitron for a juniper tree grown in Africa. The above yuzu grows to a height of about 7 m, has thin and densely grown branches, and has small thorns. The leaves of the above yuzu are lanceolate, 65 mm long and 26 mm wide. The petiole is about 10 mm, and the alar lobe is small. The flowers bloom in June, and the fruit of the above yuzu is oblate, 3 to 4 cm in vertical diameter, and 4 to 5 cm in horizontal diameter. The method of obtaining the above yuzu extract is not particularly limited, and can be extracted according to the method commonly used in the related art.

[0052] As non-limiting examples of the above extraction method, hot water extraction, ultrasonic extraction, filtration, reflux extraction, etc. can be exemplified, which can be performed alone or in combination with two or more methods.

[0053] In the present application, the type of solvent used for the above extraction is not particularly limited, and any solvent known in the related art can be used. As non-limiting examples of the above extraction solvent, water, alcohol, or a mixed solvent thereof, etc. can be exemplified, which can be used alone or in combination with one or more, and specifically water can be used. When alcohol is used as the solvent, specifically, an alcohol having a carbon number of 1 to 4 can be used.

[0054] The content of the substance of the present application can be about 0.0001 to 10% by weight, based on the total weight of the composition, but is not limited thereto. For example, the irritation relieving material caused by vitamin C of the present application 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, based on the total weight of the composition.

[0055] In the present application, the term "about" can be given before a specific numerical value. The term "about" used in the present application includes not only the exact number written after the term but also a range almost the number or close to the number. Whether close to the specific number or almost the number can be determined in consideration of the context in which the number is given. As an example, the term "about" can refer to a range of -10% to +10% of the numerical value. As another example, the term "about" can refer to a range of -5% to +5% of the given numerical value. However, it is not limited thereto.

[0056] In the present application, the above-mentioned substance has a skin irritation alleviating effect caused by vitamin C.

[0057] In the present application, the above-mentioned term "skin irritation caused by vitamin C" refers to abnormal reactions such as stinging, burning, itching, other reactions (erythema, rash, allergic reaction, swelling, discomfort), etc. that occur when an external substance contacts the skin.

[0058] In the present application, the above-mentioned substance can be characterized by reducing melanin production. Specifically, when tannic acid, tocopherol, and combinations thereof in the above-mentioned substance are treated, melanin reduction can be promoted.

[0059] As an example, as a result of simultaneously treating tannic acid and / or tocopherol, which are effective in reducing melanin production, with vitamin C, it was confirmed that there was a synergistic effect on reducing melanin when treated with vitamin C simultaneously compared to when treated with vitamin C, tannic acid, or tocopherol alone.

[0060] In the present application, the above-mentioned substance can be characterized by improving antioxidant capacity, and in particular, can restore antioxidant capacity that has decreased due to high temperature. Specifically, when any one or more selected from the group consisting of resveratrol, tannic acid, quercetin, hydroxyasiatic acid, genistein, and zinc PCA in the above-mentioned substance are treated, improvement and / or restoration of antioxidant capacity can be promoted.

[0061] As an example, as a result of simultaneously treating resveratrol, tannic acid, quercetin, hydroxyasiatic acid, genistein, and / or zinc PCA with vitamin C, it was confirmed that there was a synergistic effect on improving antioxidant capacity when treated with vitamin C simultaneously compared to when treated with vitamin C, resveratrol, tannic acid, quercetin, hydroxyasiatic acid, genistein, or zinc PCA alone.

[0062] On the purpose of the present application, the above-mentioned substance can be characterized by improving the stability of vitamin C, and specifically, the characteristic can be characterized by preventing discoloration due to heat by improving the stability of vitamin C. More specifically, when treating any one or more selected from the group consisting of resveratrol, tannic acid, hydroxyasiatic acid, genistein, and zinc PCA among the above-mentioned substances, discoloration due to heat can be prevented by improving the stability of vitamin C.

[0063] As an example, as a result of simultaneously treating resveratrol, tannic acid, hydroxyasiatic acid, genistein, and / or zinc PCA with vitamin C, it was confirmed that, when treated simultaneously with vitamin C, there was a synergistic effect on preventing discoloration of vitamin C due to heat, compared to when treated individually with vitamin C, resveratrol, tannic acid, hydroxyasiatic acid, genistein, or zinc PCA.

[0064] On the purpose of the present application, the above-mentioned substance can be characterized by inhibiting the expression of ASIC2 or PRKCE. Specifically, when treating any one or more selected from the group consisting of quercetin, phloretin, genistein, tannic acid, zinc PCA, hydroxyasiatic acid, and resveratrol, the expression of ASIC2 can be inhibited, and when treating any one or more selected from the group consisting of hydroxyasiatic acid, tannic acid, tocopherol, genistein, asiatic acid, and ponciri extract, the expression of PRKCE can be inhibited, but is not limited thereto. It can be seen that the reactivity to the substance of the present application differs depending on the type of specific genetic polymorphism, which indicates that the efficiency of alleviating irritation caused by vitamin C can be improved by providing a customized prescription for alleviating irritation caused by vitamin C.

[0065] The above-mentioned composition is characterized in that it is a personal customization. Specifically, a substance for personal customization can be provided for an individual having a single nucleotide polymorphism (SNP) marker of significant relevance to the degree of irritation caused by a specific vitamin C.

[0066] As an example, the single nucleotide polymorphism (SNP) marker of significant relevance to the degree of irritation caused by vitamin C can be any one or more selected from Table 1, but is not limited thereto.

[0067] In the present application, the term "polymorphism" refers to a case where two or more alleles exist at one locus, and in the polymorphic site, only a single base different according to the person is called a single nucleotide polymorphism (SNP). A preferred polymorphic marker has two or more alleles showing a frequency of occurrence of 1% or more, more specifically 10% or 20% or more in a selected population. A "genetic polymorphic marker" generally refers to a case where two or more alleles are observed at the same genetic position (base), and there are generally a major allele / major allele, a major allele / minor allele, a minor allele / minor allele according to different individuals. In the present application, it can be used interchangeably with "polymorphic marker", and refers to the base and base site of the minor allele, or can be defined together with the number of chromosomes and the base position, but is not limited thereto.

[0068] In the present application, the term "allele" refers to a plurality of types of one gene present at the same locus of homologous chromosomes. The allele is also used to indicate polymorphism, for example, a SNP has two types of biallele. In addition, it refers to a combination of two or more bases having the same number of chromosomes and base positions, and the above-mentioned bases include a major allele having a high frequency of occurrence in individuals of a specific population and a minor allele having a lower frequency of occurrence than the major allele.

[0069] Specifically, the genetic polymorphic marker of the present application is significantly associated with skin irritation caused by vitamin C, can have two alleles, and can be classified into a case having a major allele / minor allele, a minor allele / minor allele, a major allele / major allele.

[0070] When the individual has an allele of the marker of the degree of irritation caused by vitamin C-associated significant polymorphism, the reactivity caused by the substance of the present application can make the effect of alleviating the irritation caused by vitamin C different depending on the allele the individual has, and as an example, when the individual has major allele / major allele, major allele / minor allele, or minor allele / minor allele, the skin property of having high or low reactivity caused by the substance of the present application, i.e., the degree of irritation caused by vitamin C can be determined, and thus information can be provided in selecting a customized substance for enhancing the alleviation of the irritation caused by vitamin C for an individual having one or more minor alleles.

[0071] In the present application, the term "rs_id" refers to an independent identifier rs-ID given to all SNPs initially registered by NCBI which has accumulated SNP information since 1998. The rs_id described in this table refers to the polymorphic marker of the present application, i.e., the SNP marker.

[0072] The above-mentioned single nucleotide polymorphism marker can be one or more selected from the single nucleotide polymorphism markers shown in Table 1. The single nucleotide polymorphism marker shown in the above-mentioned Table 1 can determine whether it is associated with the degree of irritation caused by vitamin C.

[0073] The degree of irritation caused by vitamin C of the single nucleotide polymorphism marker of the present application is determined by measuring the frequency of each marker. The significance is characterized by, but not limited to, a p-value, for example, less than 0.05, less than 0.01, less than 0.001, less than 0.0001, less than 0.00001, less than 0.000001, less than 0.0000001, less than 0.00000001, or less than 0.000000001. Specifically, the p-value can be less than 0.01, more specifically, the p-value can be less than 0.001, and more specifically, the p-value can be less than 0.0001, but is not limited thereto.

[0074] The single nucleotide polymorphism (SNP) marker of the present application can be one or more selected from the markers shown in Table 1, but is not limited thereto. The above-mentioned single nucleotide polymorphism (SNP) marker can be one or more, and can be used in a combination of two or more, three or more, etc., the number of which can determine the degree of irritation caused by vitamin C, but is not limited thereto.

[0075] The marker can be the SNP itself, or a polynucleotide consisting of 5 to 100 continuous DNA sequences including the SNP position, or a polynucleotide consisting of a complementary sequence thereof, but is not limited thereto.

[0076] As an example, the marker of the degree of stimulation associated significance caused by vitamin C can be one or more single nucleotide polymorphism markers present in a gene encoding a protein of ASIC2, PRKCE, TRPM3, KCND3, or KLK8, specifically, can be one or more selected from the group consisting of rs940052, rs902557, rs12602965, rs7038646, rs79738529, rs17029069, rs7581914, rs9902778, rs452439, rs867286, rs1722562, rs641724, rs1490895, and rs10512451, but is not limited thereto.

[0077] As an example, for SNP ID rs940052, if Chr. Position (GRCh ver. 37) is recorded as "2:45981142" and the allele (Allele) is disclosed as A>G, this indicates that the 45981142th base of the human chromosome 2 is A or G, and the base located on the left side of the allele (allele) ">" can represent the major allele, and the base located on the right side can represent the minor allele.

[0078] As a specific example, the marker selected from Table 1 can consist of one or more polynucleotides selected from the group consisting of: a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 45981142th base, wherein the above-mentioned 45981142th base of chromosome 2 of human is A or G (rs940052); a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 32250107th base, wherein the above-mentioned 32250107th base of chromosome 17 of human is C or T (rs902557); a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 31833531th base, wherein the above-mentioned 31833531th base of chromosome 17 of human is C or T (rs12602965); a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 73437824th base, wherein the above-mentioned 73437824th base of chromosome 9 of human is G or A (rs7038646); a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 46040237th base, wherein the above-mentioned 46040237th base of chromosome 2 of human is G or A (rs79738529); a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 112464376th base, wherein the above-mentioned 112464376th base of chromosome 1 of human is T or C (rs17029069); a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 46078964th base, wherein the above-mentioned 46078964th base of chromosome 2 of human is G or A (rs7581914); a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 32297419th base, wherein the above-mentioned 32297419th base of chromosome 17 of human is A or G (rs9902778); a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 31677062th base, wherein the above-mentioned 31677062th base of chromosome 17 of human is C or T (rs452439); a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 45982030th base, wherein the above-mentioned 45982030th base of chromosome 2 of human is G or A (rs867286); a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 51500192th base, wherein the above-mentioned 51500192th base of chromosome 19 of human is A or C (rs1722562); and complements thereof, but are not limited thereto.A polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 112356676th base, wherein the above 112356676th base of the human chromosome 1 is A or G (rs641724); a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 32266753rd base, wherein the above 32266753rd base of the human chromosome 17 is A or G (rs1490895); a polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 32014117th base, wherein the above 32014117th base of the human chromosome 17 is A or G (rs10512451).

[0079] The composition of the present application can be used as a cosmetic composition for alleviating or improving skin irritation caused by vitamin C and can be formulated in various forms. The cosmetic composition of the present application can be prepared in a dosage form selected from the group consisting of a solution, an external ointment, a cream, a foam, a nourishing cosmetic water, a skin softener, a pack, a softener, a lotion, a primer, an essence, a soap, a liquid cleanser, a bath agent, a sunscreen, a sunscreen oil, a suspension, an emulsion, a paste, a gel, a serum, a powder, a soap, a surfactant-containing cleanser, an oil, a powder foundation, an emulsion foundation, a wax foundation, a patch, and a spray, but is not limited thereto.

[0080] In addition, the cosmetic composition of the present application can further include one or more than one cosmetically acceptable carrier used in general skin cosmetics, and as a conventional ingredient, for example, oil, water, a surfactant, a humectant, a lower alcohol, a thickening agent, a chelating agent, a pigment, a preservative, a perfume, etc. can be appropriately included, but is not limited thereto.

[0081] The cosmetically acceptable carrier included in the cosmetic composition of the present application is various depending on the dosage form.

[0082] When the dosage form of the present application is an ointment, a paste, a cream, or a gel, as a carrier ingredient, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, a cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, or a mixture thereof can be used.

[0083] When the dosage form of the present application is a powder or a spray, as a carrier ingredient, lactose, talc, silica, aluminum hydroxide, calcium silicate, a polyamide powder, or a mixture thereof can be used, and in particular, when it is a spray, a propellant such as chlorofluorocarbon, propane / butane, or dimethyl ether can be additionally included.

[0084] When the dosage form of the present application is a solution or emulsion, as a carrier component, solvents, solubilizers or emulsifiers, for example, water, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, in particular, cottonseed oil, peanut oil, corn embryo oil, olive oil, castor oil and sesame oil, glycerol fatty ester, polyethylene glycol or fatty acid ester of sorbitan can be used.

[0085] When the dosage form of the present application is a suspension, as a carrier component, liquid diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth gum, etc. can be used.

[0086] When the dosage form of the present application is a soap, as a carrier component, alkali metal salt of fatty acid, fatty acid half ester salt, fatty acid protein hydrolysate, isethionate, lanolin derivative, fatty alcohol, vegetable oil fat, glycerol, sugar, etc. can be used.

[0087] All the components described in the present application are preferably not more than the maximum use value prescribed in the relevant regulations such as the cosmetic safety standards (Korean regulations), the cosmetic safety technical standards (Chinese regulations), etc.

[0088] As another aspect, the above composition provides a quasi-drug composition for relieving or improving irritation caused by vitamin C.

[0089] In the present application, the term "quasi-drug" refers to an item whose action is lighter than that of a pharmaceutical product among items used for the purpose of diagnosis, treatment, improvement, alleviation, management or prevention of diseases of humans or animals, for example, according to the Korean Pharmaceutical Affairs Act, a quasi-drug is an item that does not include an item used for the purpose of a pharmaceutical product, including a product for treatment or prevention of diseases of humans, animals, a product whose action on the human body is light or does not directly act, etc.

[0090] As a specific example, the quasi-drug composition of the present application can be prepared in a form selected from the group consisting of shower gel, shampoo, hair conditioner, foam, soap, pack, ointment, cream, lotion, serum and spray, but is not limited thereto.

[0091] All the components described in the present application are preferably not more than the maximum use value prescribed in the relevant regulations such as the cosmetic safety standards (Korean regulations), the cosmetic safety technical standards (Chinese regulations), etc.

[0092] As another aspect, the present application provides an information providing method for selecting a customized material for relieving irritation caused by vitamin C, comprising: (a) a step of obtaining a biological specimen from an individual; (b) a step of amplifying or hybridizing a polymorphic site of a single nucleotide polymorphism marker consisting of rs 10512451 from the above biological specimen; and (c) a step of confirming the base of the amplified or hybridized polymorphic site of the above (b) step.

[0093] By treating the individual having the specific polymorphic site with the selected customized material, the skin irritation caused by vitamin C can be improved.

[0094] The term "individual" of the present application refers to a subject for diagnosing the degree of irritation of the skin caused by vitamin C. In the above test sample, DNA can be obtained from a specimen such as hair, urine, blood, various body fluids, isolated tissues, isolated cells, or saliva, etc., but is not limited thereto.

[0095] The method for obtaining genome DNA of the above (a) step can use any method known to those skilled in the art.

[0096] The step of amplifying or hybridizing the polymorphic site of the single nucleotide polymorphism marker from the DNA obtained in the above (a) step can use any method known to those skilled in the art. For example, it can be obtained by amplifying the target nucleic acid by PCR and purifying it. In addition, it can use 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).

[0097] 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-specific 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 Sequenom's MassARRAY system), 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 analysis method), but is not limited to these. The above methods or other methods available to those skilled in the art can be used to identify one or more alleles of polymorphic markers, including microsatellites, SNPs, or other types of polymorphic markers. The bases at such polymorphic sites can be determined specifically using a SNP chip.

[0098] The substance may be one or more selected from the group consisting of quercetin, phloretin, genistein, tannic acid, PCA zinc, madecassoside, resveratrol, tocopherol, asiaticoside and green tangerine extract, but is not limited thereto.

[0099] For the purposes of the present invention, genes significantly associated with the degree of irritation caused by vitamin C can provide information that enables the selection of specific substances for each individual. For example, for individuals who have one or more single nucleotide polymorphism markers present in genes encoding ASIC2 or PRKCE proteins, treatment with one or more substances selected from the group consisting of quercetin, phloretin, genistein, tannic acid, PCA zinc, madecassoside, resveratrol, tocopherol, asiaticoside, and green tangerine extract can enhance the alleviation of irritation caused by vitamin C. However, an optimal combination of substances can be provided based on the genetic characteristics of each individual.

[0100] This indicates that specific single nucleotide polymorphism (SNP) markers having a significant correlation with skin irritation caused by vitamin C can be screened and information can be provided based on genetic information so that a personal customized substance that can maximize the effect of alleviating irritation caused by vitamin C can be selected.

[0101] Specifically, when the risk score is 1 or less, which is obtained by multiplying the number of minor alleles of rs940052, rs902557, rs12602965, rs7038646, rs79738529, rs17029069, rs7581914, rs9902778, rs452439, rs867286, rs1722562, rs641724, and rs1490895 by the effect size (β) and adding them, the risk of skin irritation caused by vitamin C can be classified as a low risk group, and when the risk score exceeds 1, the risk of irritation caused by vitamin C can be classified as a high risk group. As an example, when the risk of irritation caused by vitamin C is a high risk group, an information providing method can be provided so that one or more selected from the group consisting of quercetin, phloretin, genistein, tannic acid, zinc PCA, hydroxyasiaticoside, resveratrol, tocopherol, asiaticoside, and ponciri extract is selected as a customized substance, but is not limited thereto.

[0102] In the present application, the term "SNP chip" refers to one of DNA microarrays that can confirm several tens of thousands of SNPs in each base at a time.

[0103] The TaqMan method includes: (1) a step of designing and manufacturing primers and TaqMan probes so as to enable amplification of a desired DNA fragment; (2) a step of labeling probes for different alleles with FAM dye and VIC dye (Applied Biosystems); (3) a step of performing PCR using the above-described DNA as a template, using the above-described primers and probes; (4) a step of analyzing and confirming a TaqMan analysis plate using a nucleic acid analyzer after the above-described PCR reaction is completed; and (5) a step of determining the genotype of the polynucleotide of step (1) according to the above-described analysis result.

[0104] In the above, the sequencing analysis can use a conventional method for determining a base sequence, and can be performed using an automated gene analyzer. In addition, the allele-specific PCR refers to a PCR method in which a primer set including a primer designed to have a 3' end at a base where a SNP is located is used to amplify a DNA fragment where the above SNP is located. The principle of the above method utilizes the fact that, for example, when a specific base is substituted from A to G, if a primer containing the above A as a 3' end base and a reverse primer capable of amplifying a DNA fragment of an appropriate size are designed to perform a PCR reaction, if the base at the SNP position is A, a band at the desired position is observed due to the normal progress of the amplification reaction, and if the base is substituted with G, the primer can be complementarily bound to the template DNA, but since the 3' end does not complementarily bind, the amplification reaction does not progress normally. The DASH can be performed by a conventional method, and specifically can be performed by the method of Prince et al.

[0105] Meanwhile, the PCR extension analysis is achieved by first amplifying a DNA fragment containing a base where a single nucleotide polymorphism is located with a primer pair, then dephosphorylating all nucleotides added to the reaction to inactivate them, and adding a SNP-specific extension primer, a dNTP mixture, a dideoxynucleotide, a reaction buffer, and a DNA polymerase thereto to perform a primer extension reaction. At this time, the extension primer has a 3' end at a base directly adjacent in the 5' direction of the base where the SNP is located, the dNTP mixture does not contain a nucleic acid having the same base as the dideoxynucleotide, and the above dideoxynucleotide is selected from one showing one of the base types of the SNP. For example, in the case of substitution of A with 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 the DNA polymerase, and after a few bases, the primer extension reaction is terminated by the ddATP at the position where the A base first appears. If the above substitution does not occur, the extension reaction is terminated at that position, so the base type showing the SNP can be judged by comparing the lengths of the above extended primers.

[0106] 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 a base sequence (for example, Model 3700 of ABI, etc.), and when unlabeled extension primers and dideoxynucleotides 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.

[0107] As one aspect for achieving the object of the present application, the present application provides a single nucleotide polymorphism (SNP) marker for diagnosing the degree of irritation caused by vitamin C.

[0108] As another aspect, the present application provides a composition for diagnosing the degree of irritation caused by vitamin C, which comprises a probe capable of detecting or a preparation capable of amplifying a single nucleotide polymorphism (SNP) marker for diagnosing the degree of irritation caused by vitamin C.

[0109] The "skin irritation caused by vitamin C" means abnormal reactions such as stinging, burning, itching, other reactions (erythema, rash, allergic reaction, swelling, discomfort), etc. that occur when an external substance contacts the skin, but is not limited thereto. Specifically, it means judging whether or not the skin type is susceptible to the skin irritation caused by vitamin C using any one or more of the single nucleotide polymorphism (SNP) markers selected from Table 1.

[0110] As to the above alleles of the present application, the number of chromosomes is the same in each individual, in which the major allele and the minor allele of the SNP exist, and the major allele can be decreased one by one as the base of the polymorphic site of the polymorphic marker is increased to the minor allele, and the minor allele can be decreased one by one as the base is increased to the major allele. However, the range in which the minor allele and the major allele can be increased or decreased can be within three types of i) major allele / major allele, ii) major allele / minor allele, iii) minor allele / minor allele, and the alleles can be decreased or increased within the range of the above three types, but is not limited thereto.

[0111] In the present application, the term "probe capable of detecting a marker for diagnosing the degree of irritation caused by vitamin C" means a composition capable of diagnosing the degree of irritation caused by vitamin C by confirming the specific hybridization reaction with the polymorphic site of the above gene, and the specific method of such gene analysis is not particularly limited and can be performed according to all the gene detection methods known in the technical field to which the present application pertains. In addition, the above term can be used interchangeably with the term "for diagnosing whether or not the skin type is susceptible to the skin irritation caused by vitamin C" or "for diagnosing whether or not the skin type is a high-risk group susceptible to the skin irritation caused by vitamin C".

[0112] In the present invention, the term "agent capable of amplifying a marker for diagnosing the degree of irritation caused by vitamin C" refers to a composition capable of diagnosing the degree of irritation caused by vitamin C by confirming the polymorphic site of the above-mentioned gene through amplification, and specifically, refers to a primer capable of specifically amplifying the above-mentioned polynucleotide for diagnosing the degree of irritation caused by vitamin C. In addition, the above-mentioned term can be used interchangeably with the term "for diagnosing whether a skin type is susceptible to the generation of skin irritation caused by vitamin C" or "for diagnosing whether a skin type is a high-risk group susceptible to the generation of skin irritation caused by vitamin C".

[0113] The primer for amplifying the above-mentioned polymorphic marker refers to a single-stranded oligonucleotide that can function as a starting point for template-directed DNA synthesis in an appropriate buffer under appropriate conditions (e.g., 4 different nucleoside triphosphates and DNA, RNA polymerase or reverse transcriptase, etc. polymerase) and at an appropriate temperature. The appropriate length of the above-mentioned primer can vary depending on the purpose of use, but is generally 15 to 30 nucleotides. Short primer molecules generally require lower temperatures to form stable hybrids with templates. The primer sequence does not need to be completely complementary to the template, but must be sufficiently complementary to the extent of hybridization with the template.

[0114] In the present invention, the term "primer" as a base sequence having a short free 3' end hydroxyl group (free 3' hydroxyl group) refers to a short sequence that can form base pairs with a complementary template and serve as a starting point for template strand replication. The primer can initiate DNA synthesis in the presence of reagents for polymerization reaction (i.e., DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates in an appropriate buffer solution and temperature. The degree of generation of the desired product can be predicted by performing PCR amplification to predict the skin type. The length of the sense and antisense primers can be modified based on what is well known in the art.

[0115] The probes or primers of the present invention can be chemically synthesized using a phosphoramidite solid support method or other widely known methods. Such nucleic acid sequences can also be modified using a number of means well known in the art. As non-limiting examples of such modifications, there are methylation, "capping", substitution with one or more homologues of natural nucleotides, and modification between nucleotides, for example, modification of uncharged linkers (e.g., methyl phosphonate, phosphotriester, phosphoroamidate, carbamate, etc.) or charged linkers (e.g., phosphorothioate, phosphorodithioate, etc.).

[0116] As another aspect, the present application provides a kit for diagnosing the degree of stimulation caused by vitamin C, comprising the above-described composition for diagnosing the degree of stimulation caused by vitamin C. The above-described kit can be an RT-PCR kit or a DNA chip kit, but is not limited thereto.

[0117] The kit of the present application can diagnose the degree of stimulation caused by vitamin C by amplifying the marker for diagnosing the degree of stimulation caused by vitamin C, i.e., the SNP polymorphism marker, or confirming the expression level of the SNP polymorphism marker and the expression level of mRNA. As a specific example, in the present application, the kit for measuring the mRNA expression level of the marker for diagnosing the degree of stimulation caused by vitamin C can be a kit comprising essential elements necessary for performing RT-PCR. In addition to each primer pair specific to the gene of the marker for diagnosing the degree of stimulation caused by vitamin C, the RT-PCR kit can include a test tube or other appropriate container, a reaction buffer (pH and magnesium concentration are various), deoxynucleotides (dNTPs), Taq-polymerase and other enzymes such as reverse transcriptase, a DNase inhibitor, an RNase inhibitor, DEPC-water, sterile water, etc. In addition, a primer pair specific to a gene used as a quantitative control group can be included. In addition, specifically, the kit of the present application can be a kit for diagnosing the degree of stimulation caused by vitamin C comprising essential elements necessary for performing a DNA chip. The DNA chip kit generally attaches nucleic acid species in a gridded array to a flat solid support plate, typically a glass surface no larger than a microscope slide, and is a tool in which nucleic acids are uniformly arranged on the surface of a chip to enable multiple hybridization reactions between nucleic acids on a DNA chip and complementary nucleic acids contained in a solution treated on the surface of the chip, thereby performing a large number of parallel analyses.

[0118] As another aspect, the present application provides a microarray for diagnosing the degree of stimulation caused by vitamin C, comprising the above-described composition for diagnosing the degree of stimulation caused by vitamin C.

[0119] The above-described microarray can comprise DNA or RNA polynucleotides. The above-described microarray consists of a conventional microarray, except that the polynucleotide of the present application is included in the probe polynucleotide.

[0120] The method of preparing a microarray by immobilizing a probe polynucleotide on a substrate is well known in the art. The above-mentioned probe polynucleotide refers to a hybridizable polynucleotide, which refers to an oligonucleotide capable of binding to a complementary strand of a nucleic acid in a sequence-specific manner. The probe of the present application is an allele-specific probe, and the polymorphic site exists in a nucleic acid fragment derived from two members of the same species, hybridizes with a DNA fragment derived from one member, but does not hybridize with a fragment derived from the other member. In this case, the hybridization conditions show a significant difference in the strength of hybridization between alleles, and thus must be stringent enough to hybridize with only one of the alleles. By doing so, a good hybridization difference between different allelic forms can be induced. The above-mentioned probe of the present application can detect an allele and is used in a diagnostic method for alleviation of skin irritation caused by vitamin C, etc. The above-mentioned diagnostic method includes a detection method based on nucleic acid hybridization, such as Southern blot hybridization, etc., and can be provided in a form pre-bound on a substrate of a DNA chip in a method using a DNA chip. The above-mentioned hybridization can be performed under stringent conditions, such as a salt concentration of 1 M or less and a temperature of 25°C or more. For example, conditions of 5x SSPE (750 mM NaCl, 50 mM sodium phosphate, 5 mM EDTA, pH 7.4) and 25 to 30°C can be suitable for allele-specific probe hybridization.

[0121] The process of immobilizing the probe polynucleotide of the present application associated with the degree of irritation caused by vitamin C on a substrate can also be easily prepared using this prior art. In addition, hybridization of nucleic acids on a microarray and detection of hybridization results are well known in the art. The above-mentioned detection can detect hybridization results by labeling a nucleic acid sample with a labeling material capable of generating a detectable signal, including a fluorescent material such as Cy3 and Cy5, etc., and then hybridizing on a microarray and detecting a signal generated from the above-mentioned labeling material.

[0122] As another aspect, the present application provides a method of providing information on the degree of irritation caused by vitamin C, comprising: (a) a step of amplifying or hybridizing with a probe a polymorphic site of the above-mentioned single nucleotide polymorphism marker in DNA obtained from a sample isolated from an individual; and (b) a step of confirming the base of the amplified or hybridized polymorphic site of the above-mentioned (a) step.

[0123] The above-mentioned terms "individual", (a) step and (b) step are as previously described.

[0124] The above-mentioned method further, (c) when the hazard score obtained by multiplying the effect amount (β) of the amplified or hybridized single nucleotide polymorphism marker and adding is more than 1, it can be judged that the high-risk group having a high risk of occurrence of skin irritation caused by vitamin C, but is not limited thereto.

[0125]

[0126] Hereinafter, the present application will be explained in more detail by examples. These examples are only for illustrating the present application, and the scope of the present application should not be construed as being limited by these examples.

[0127] Example 1: Derivation of vitamin C skin irritation-associated significant gene polymorphism markers

[0128] Genomic loci (genetic variations) that appear differences in vitamin C stimulation (questionnaire answers) according to genetic information were intended to be explored. In order to explore genomic loci (genetic variations) associated with vitamin C stimulation, candidate genes presumed to be associated with vitamin C stimulation, particularly genes associated with burning, itching, stinging, keratin barrier function, vitamin C metabolism, redness, rash, etc. were screened and used for analysis. As a device for screening the genome, a microarray genotyping chip (Illumina product) was used.

[0129] In the present application, in order to evaluate the presence or absence and degree of vitamin C stimulation, the subjects were asked to apply 20% vitamin directly to the facial skin (cheeks) after cleansing at night for 2 days, and a phenotypic questionnaire answer was collected at an interval of 1 point from no stimulation (0 points) to stimulation (maximum 6 points). The collected vitamin C skin stimulation scale was used as a phenotype.

[0130] In order to confirm the association of genomic loci (genetic variations) with the degree of skin stimulation (scale), the correlation significance and genetic effect were numerically evaluated using linear regression analysis.

[0131] 1-1. Vitamin C stimulation degree evaluation method

[0132] In order to derive a genetic polymorphism marker that can explain the degree of vitamin C stimulation, a total of 84 healthy Koreans in their 20s to 70s were recruited. As the first evaluation, 51 subjects were asked to apply the distributed vitamin C preparation in the first step of skin management after cleansing at night, and then whether or not stimulation was collected in the form of a scale (0 points ~ 6 points), and as detailed stimulation, burning, stinging, itching were distinguished and collected.

[0133] 1-2. Gene collection and analysis method

[0134] ​For gene analysis from the extracted genes from saliva, human genomic DNA was extracted using QIAmp mini prep kit (QIAGEN), the quality of which was confirmed by absorbance (OD 260 / 280) > 1.7, concentration 50 ng / ul, band test of 1x TAE 1% agarose gel, and only the genes passing the quality were subjected to gene analysis.

[0135] Gene analysis was performed using Illumina's microarray genotyping chip, and specifically, the genes of the analysis subjects were analyzed using the same company's global screening array product.

[0136] The microarray genotyping chip gene analysis experiment of Illumina was performed according to the provided manual, using the provided reagents, and the processes of genomic DNA amplification, DNA fragmentation, precipitation, hybridization, staining, washing, coating, and scanning were performed.

[0137] The microarray genotyping chip experiment was scanned using iScan Control Software (Illumina), and after scanning was completed, idat files were automatically generated and data quality management (sample call rate > 98%, marker call rate > 98%) and gene information confirmation were performed using the GenomeStudio (Illumina) program.

[0138] In the present application, only data passing the data quality management after gene analysis was used.

[0139] 1-3. Vitamin C stimulation degree-related gene polymorphism marker derivation

[0140] In order to derive gene polymorphism markers related to the degree of vitamin C stimulation, linear regression analysis was performed using the gene polymorphism markers of the analysis subjects. Linear regression analysis of the gene markers of the subjects was performed only for gene markers having a frequency of 2% or more of the minor allele in the subject group.

[0141] The significance of the gene polymorphism markers associated with the degree of vitamin C stimulation was evaluated by the T statistic of the linear regression analysis result, and the standard was set to P-value < 0.1. Many significant gene polymorphism markers associated with the degree of vitamin C stimulation were derived (Table 1).

[0142] [Table 1]

[0143] Vitamin C stimulation-associated gene polymorphism markers

[0144]

[0145]

[0146] 1) SNP: National Institutes of Health (NIH) ID, the sequence can be confirmed on the relevant website

[0147] 2) rsID: Reference SNP

[0148] 3) Beta: Linear regression coefficient

[0149] 4) MAF (Minor allele frequency): Frequency of the minor allele in the subject group

[0150] 5) Minor allele: Minor allele

[0151] 6) Major allele: Major allele

[0152] Example 2: Analysis results of vitamin C skin irritation-associated gene expression regulation effects of various substances

[0153] Among the various substances having skin-improving effects, a screening work of the substance affecting the expression of the vitamin C skin stimulation-associated gene was performed.

[0154] Human keratinocytes (HaCaT) were purchased from ATCC and used. The purchased cells were cultured in a medium in which 10% FBS (fetal bovine serum) and 1% penicillin / streptomycin were put into DMEM (Dulbecco Modified Eagle Medium-GIBCO) in a 37°C, 5% CO2 incubator.

[0155] In order to analyze the mRNA expression change effect of the vitamin C skin stimulation-associated gene, human keratinocytes were cultured at 1 x 10 5Cells / well were dispensed into 12 wells and cultured in a 37°C, 5% CO2 incubator for 1 day. Then, these substances were added to the culture medium at appropriate concentrations and cultured under the same conditions for two days. Total RNA was obtained using the AccuPrep Universal RNA Extraction Kit (Bioneer, K-3140) and used RocketScript TM cDNA was synthesized by Cycle (Bioneer, K-2202) and the expression level of each gene was analyzed by qPCR using the synthesized cDNA and Taqman probes (detection ID: human ASIC2-Hs00153756_m1, human PRKCE-Hs00942886_m1).

[0156] The results, such as Figure 1 As shown, the material confirmed to reduce the expression of human ASIC2 is quercetin, phloretin, genistein, tannic acid, PCA zinc, madecassoside or resveratrol, and the material confirmed to reduce the expression of human PRKCE is madecassoside, tannic acid, tocopherol, genistein, asiaticoside or green orange extract.

[0157] Example 3: Confirmation of synergistic effects of pure vitamin C irritation-improving materials

[0158] The purpose was to confirm whether the combination of substances screened in Example 2 has a synergistic effect in inhibiting the expression of human ASIC2. Human keratinocytes were cultured at 1×10 5 Cells were plated at 100 cells / well in a 12-well plate and cultured in a 37°C, 5% CO2 incubator for one day. The plates were then placed in a culture medium supplemented with the relevant substances at appropriate concentrations and combinations and cultured again in the incubator for one day. RNA was then extracted and cDNA synthesized in the same manner as in Example 2. qPCR was then performed to confirm whether the expression of the relevant genes was synergistically inhibited.

[0159] It was confirmed that when treated with 15 ppm of quercetin and 15 ppm of tannic acid, ASIC2 expression decreased by approximately 10% (X) and 36% (Y), respectively. When quercetin and tannic acid were treated simultaneously at corresponding concentrations, gene expression decreased by 94% (T, effect measurement value). This is 47% greater than the Colby-predicted formula (E1) for the combination of the two substances, and therefore it was determined that there was a synergistic effect in reducing ASIC2 expression ( Figure 2 ), as a specific example, even for rs10512451 (17:32014117:A:G) in the ASIC2 gene, which may affect the skin irritation of vitamin C depending on the type of the minor allele ( Figure 3 ), but is not limited to this single nucleotide polymorphism marker.

[0160] Colby formula E1=X+Y-(XY / 100)

[0161] Furthermore, when treated with 15 ppm quercetin, 15 ppm tannic acid, or 25 ppm resveratrol as other combinations, ASIC2 expression decreased by 10%, 36%, and 31% respectively. However, when quercetin, tannic acid, and resveratrol were treated simultaneously at corresponding concentrations, ASIC2 expression decreased by 94% (T, effect measurement value). This is greater than the Colby-predicted value (E2) of 77% for the three-substance combination, indicating a synergistic effect in reducing ASIC2 expression. Figure 2 )

[0162] Colby formula E2 = (X + Y + Z) - (XY + XZ + YZ / 100) + XYZ / 10000

[0163] Example 4: Human application test for developing pure vitamin C skin irritation-recognizing high-risk group customized, irritation-relief prescription materials Figure 4

[0164] In Example 2, substances that regulate the expression of a significant polymorphic marker associated with pure vitamin C-specific skin irritation were screened. Thus, the aim is to provide a substance that can improve the efficiency of irritation improvement according to the risk level of vitamin C irritation.

[0165] 4-1. Selection of subjects

[0166] After approval from the Institutional Review Board (IRB, Korea) (LGHH-20220324-AB-04-03), the study was conducted according to the standard operating guidelines and clinical trial standards of the LG Household & Health Care Corporation's Bioethics Committee. 84 healthy subjects aged 20 to 50 years with no major abnormalities in skin appearance were selected.

[0167] 4-2. Subject selection criteria

[0168] Based on the [Human Derived Material Research Consent Form (IRB No.LGHH-20180727-AA-03, LGHH-20201210-AB-03)] already held by the responsible research institution (LG Household & Health Care), the research subjects do not meet the corresponding conditions

[0169] i) Those who agree to the content item "Shelf Life of Human Origin Materials" but disagree with the item "1. Permanent Storage"

[0170] ii) Those who agree to "3. Disagree" in the "Provision for Secondary Use During the Retention Period" section of the Contents section

[0171] 4-3. Subject Exclusion Criteria

[0172] Among the above-mentioned subjects, 1) those who are also stimulated or allergic to general salt water, or 2) those who are judged by the investigator to be unsuitable for the test in addition, were excluded from the subjects.

[0173] 4-4. Vitamin C skin irritation alleviating material irritation evaluation test / analysis method

[0174] Among the total of 84 subjects, 33 of the second test group, except for the first test group, were divided into 20% vitamin C serum as a control group and serum added with vitamin C skin irritation alleviating material as an experimental group, and applied to the right cheek and left cheek, respectively. The total evaluation was performed for 2 days, on the 1st day and the 3rd day, while on the 2nd day, considering that repeated irritation exposure can cause a feeling of numbness, it was suggested to rest. For the same reason, on the 1st day and the 3rd day, the cheeks to which the control group and the experimental group serum were applied were exchanged.

[0175] The evaluation scale was designated as 0 to 6, 1-2 was slight (slightly uncomfortable, ambiguous degree), 3-4 was moderate (degree of feeling irritation), and 5-6 was strong (degree of wanting to wash off due to strong irritation). The irritation category was divided into burning, stinging, itching, and others (erythema, urticaria, etc.). The irritation values thus evaluated on the 1st day and the 3rd day were averaged according to the irritation category to calculate the "irritation improvement degree". As a result, it was confirmed that for burning, it was significantly decreased from 1.82 of the control group to 1.32 (p<0.05), stinging was alleviated from 2.06 to 1.64, and itching was alleviated from 1.58 to 1.40( Figure 4 ).

[0176] For the irritation improvement sensation rate, the number of people who felt improvement among the people who felt irritation was calculated in percentage according to each irritation category. As a result, it showed an improvement sensation rate of 64% for burning, 62% for stinging, and 55% for itching. Finally, the degree of improvement of the sensation of the irritation alleviating component according to the degree of existing vitamin C irritation was expressed using a color chart, and as can be seen from the figure, the more strongly the existing irritation was felt by the subject, the more effective the improvement of irritation was( Figure 5 ).

[0177] 4-4. Difference in irritation improvement according to irritation gene score when a prescription customized vitamin C irritation alleviating composition

[0178] The vitamin C irritation gene score (PRS) was calculated according to the Beta value of the skin irritation-associated gene polymorphism marker of Table 1 (below). According to the calculated score, 32 of the 33 second test subjects were divided into a low-risk group and a high-risk group as follows. (1 was an evaluation dropout)

[0179]

[0180] I = subject

[0181] J = jth SNP

[0182] SNP j = number of minor alleles of jth SNP

[0183] m = total number of SNPs (here, 13)

[0184] Sum of effect size (β) of 13 genetic markers exceeds 1 → high risk group (n = 19)

[0185] Sum of effect size (β) of 13 genetic markers is 1 or less → low risk group (n = 13)

[0186] (As a reference, the above 13 genetic markers are skin irritation-associated genetic polymorphism markers consisting of rs940052, rs902557, rs12602965, rs7038646, rs79738529, rs17029069, rs7581914, rs9902778, rs452439, rs867286, rs1722562, rs641724, and rs1490895 in Table 1.)

[0187] When the vitamin C irritation gene score is divided into a low risk group and a high risk group with 1 as a reference, the more the vitamin C irritation high risk group, the more the burning irritation is reduced significantly in the application of a vitamin C prescription including a vitamin C irritation relieving material, compared to the low risk group Example 5: Analysis of vitamin C irritation-relief materials for improving vitamin C discoloration and antioxidant ability caused by high temperature (whether or not is determined with respect to the burning irritation felt by the subject when a control prescription is applied). This means that the vitamin C irritation relieving material prescription is more effective for a subject group predicted to have a higher vitamin C irritation gene score, and thus, it can be seen that the prescription is expected to be more effective for a subject group having a higher possibility of feeling vitamin C irritation.

[0188] Figure 6 Figure 7

[0189] For various materials considered to have an effect of improving vitamin C irritation, improvement in discoloration and antioxidant ability of vitamin C was confirmed.

[0190] First, when a solution in which 10% and 15% of vitamin C was dissolved in water was heated at 80 degrees for 8 hours, it was confirmed that the color turned yellow and the antioxidant ability decreased Figure 8 ).

[0191] At this time, the discoloration was compared by the degree of yellowing when observed by the naked eye and the absorbance at 390 nm, and the antioxidant capacity was measured by the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging reaction. Also, the solution of the candidate substance was mixed in a 10%, 15% vitamin C solution and heated at 80 degrees for 8 hours to measure the change in discoloration and antioxidant capacity Example 6: Confirmation of pigment deposition / antioxidant-enhancing effects of pure vitamin C nerve irritation-relief and stability-improving materials and Figure 9 ).

[0192] As a result, it was confirmed that when vitamin C and various substances were treated together, the discoloration of vitamin C was improved in some substances, and the decreased antioxidant capacity was also improved to the level of the unheated vitamin C solution. Specifically, in 500 ppm of resveratrol, the discoloration of 10% and 15% of vitamin C was improved by 57%, 72%, respectively, and for the antioxidant capacity, 27% of the antioxidant capacity was exhibited in 10% of vitamin C alone, and 45% of the antioxidant capacity was exhibited when 500 ppm of resveratrol was added to vitamin C, thereby not only improving the antioxidant capacity, but also confirming the synergistic effect of the combination compared to resveratrol alone (1%).

[0193] Also, 30%, 33% levels of improvement in discoloration were exhibited in 500 ppm of PCA zinc, the antioxidant capacity of vitamin C was 21%, PCA zinc was 10%, the combination was 55%, and the improvement in antioxidant capacity and the synergistic effect of the combination were confirmed.

[0194] 62%, 73% of improvement in discoloration were exhibited in 200 ppm of hydroxyaspidin, the antioxidant capacity of vitamin C was 27%, hydroxyaspidin was 5%, the combination was 43%, and the improvement in antioxidant capacity and the synergistic effect of the combination were confirmed.

[0195] No improvement in discoloration was confirmed in 200 ppm of quercetin, the antioxidant capacity of 10% of vitamin C was 27%, quercetin was 3%, the combination was 48%, and the improvement in antioxidant capacity and the synergistic effect were confirmed.

[0196] 50%, 60% of improvement in discoloration were exhibited in 100 ppm of genistein, the antioxidant capacity of vitamin C was 37%, genistein was 3%, the combination was 54%, and the improvement in antioxidant capacity and the synergistic effect were confirmed.

[0197] 19%, 25% of improvement in discoloration were exhibited in 100 ppm of tannic acid, the antioxidant capacity of vitamin C was 21%, tannic acid was 5%, the combination was 52%, and the improvement in antioxidant capacity and the synergistic effect were confirmed.

[0198] Figure 9 ​

[0199] To confirm whether the two materials (tannin, tocopherol) screened in Example 2 can enhance the whitening effect and antioxidant effect of vitamin C at the cellular level.

[0200] First, for whitening evaluation, mouse melanoma cells (B16F10) were dispensed at 1 x 10 5 cells / well in a 12-well plate and cultured for one day in a 37°C, 5% CO2 incubator, and then treated with 10 nM of a-MSH. The medium was placed with the appropriate concentration and combination of substances and cultured again for one day in a 37°C, 5% CO2 incubator to confirm whether the substances inhibit melanin production increased by a-MSH. The cells attached to the plate were recovered in PBS and the cell pellet was obtained by centrifugation. The cell pellet was photographed after macroscopic determination. A 1N NaOH solution containing 10% DMSO (dimethyl sulfoxide) was placed in the cell pellet and dissolved at 80°C for 10 minutes, and then placed in a 96-well plate, and the 405 nm absorbance was measured on an Epoch (BioteK) instrument. The melanin was quantified using the measured absorbance and normalized with the protein concentration of the test material, and thus the melanin concentration was measured. For the melanin production inhibition rate of the test material, the ratio of the melanin concentration of the test material-treated group to the melanin concentration increased by a-MSH treatment was calculated.

[0201] As a result, it was confirmed that when tannin or tocopherol was simultaneously treated with vitamin C, melanin production was more effectively inhibited than in the vitamin C-only treated group ​ ).

[0202] Next, for antioxidant efficacy evaluation, mouse melanoma cells (B16F10) were dispensed at 1 x 10 4Cells / well were dispensed in a black clear bottom 96 well plate and cultured in a CO2 incubator for one day, then on the second day, the medium was replaced with the medium added with the substance, and cultured again, and on the third day of culture, DCF-DA assay was performed. Specifically, first, the cell culture solution was removed, then washed with PBS, then DCF-DA stain was mixed with serum-free medium and incubated with the cells for 30 minutes. The medium was removed again, washed with PBS, then replaced with HBSS (Hank's Balanced Salt Solution), and then measured under the condition of plate reader excitation 529 nm, emission 529 nm.

[0203] As a result, it was confirmed that when tannic acid or tocopherol was simultaneously treated with vitamin C, the antioxidant ability was more excellent than that of the group treated with vitamin C alone ​ ).

[0204] Based on the above description, those skilled in the art to which the present application pertains can understand that the present application can be implemented in other specific forms without changing the technical idea or essential characteristics of the present application. In this regard, it should be understood that the above-described embodiments are exemplary in all aspects and are not limiting. The scope of the present application should be interpreted as including the meaning and scope of the claims described below and all modifications or variations derived from the equivalent concept thereof, not just the above detailed description.

Claims

1. A composition for alleviating skin irritation caused by vitamin C, comprising one or more substances selected from the group consisting of quercetin, phloretin, genistein, tannic acid, zinc PCA (ZnPCA), madecassoside, resveratrol, tocopherol, asiaticoside and green tangerine extract as active ingredients.

2. The composition of claim 1, wherein, The above composition contains the following as active ingredients: (i) quercetin, phloretin, genistein, tannic acid, PCA zinc, madecassoside, and resveratrol; or (ii) Madecassoside, tannic acid, tocopherol, genistein, asiaticoside and green mandarin orange extract.

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

4. The composition of claim 1, wherein, The above composition reduces melanin production.

5. The composition of claim 4, wherein, The substances used to reduce the above-mentioned melanin production are tannic acid, tocopherol and a combination thereof.

6. The composition of claim 1, wherein, The above composition improves antioxidant capacity.

7. The composition of claim 6, wherein, The substance for improving the antioxidant capacity is at least one selected from the group consisting of resveratrol, tannic acid, quercetin, madecassoside, genistein, and PCA zinc.

8. The composition of claim 1, wherein, The above composition improves the stability of vitamin C.

9. The composition of claim 8, wherein, The ingredient for improving the stability of vitamin C is at least one selected from the group consisting of resveratrol, tannic acid, madecassoside, genistein, and PCA zinc.

10. The composition of claim 1, wherein, The aforementioned alleviation of skin irritation caused by vitamin C is for an individual comprising any one or more single nucleotide polymorphism (SNP) markers for diagnosis associated with vitamin C irritation selected from Table 1.

11. The composition of claim 10, wherein, The single nucleotide polymorphism marker for diagnosis associated with vitamin C stimulation is one or more single nucleotide polymorphism markers present in genes encoding one or more proteins selected from the group consisting of ASIC2, PRKCE, TRPM3, KCND3, and KLK8.

12. The composition of claim 10, wherein, The single nucleotide polymorphism marker for diagnosis associated with vitamin C stimulation is one or more selected from the group consisting of rs940052, rs902557, rs12602965, rs7038646, rs79738529, rs17029069, rs7581914, rs9902778, rs452439, rs867286, rs1722562, rs641724, rs1490895 and rs10512451.

13. The composition of claim 1, wherein, The above substances inhibit the expression of ASIC2 or PRKCE.

14. The composition of claim 13, wherein, The substance for inhibiting ASIC2 expression is at least one selected from the group consisting of quercetin, phloretin, genistein, tannic acid, PCA zinc, madecassoside, and resveratrol.

15. The composition of claim 13, wherein, The substance for inhibiting PRKCE expression is at least one selected from the group consisting of madecassoside, tannic acid, tocopherol, genistein, asiaticoside, and citrus fruit extract.

16. The composition of claim 12, wherein, The SNP marker for diagnosis of the irritation caused by vitamin C is selected from the group consisting of the following polynucleotides: one or more polynucleotides selected from the group consisting of: a polynucleotide consisting of 5-100 consecutive DNA sequences comprising base number 45981142, wherein the aforementioned base number 45981142 of chromosome 2 of humans is A or G (rs940052); a polynucleotide consisting of 5-100 consecutive DNA sequences comprising base number 32250107, wherein the aforementioned base number 32250107 of chromosome 17 of humans is C or T (rs902557); a polynucleotide consisting of 5-100 consecutive DNA sequences comprising base number 31833531, wherein the aforementioned base number 31833531 of chromosome 17 of humans is C or T (rs12602965); a polynucleotide consisting of 5-100 consecutive DNA sequences comprising base number 73437824, wherein the aforementioned base number 73437824 of chromosome 9 of humans is G or A (rs7038646); a polynucleotide consisting of 5-100 consecutive DNA sequences comprising base number 46040237, wherein the aforementioned base number 46040237 of chromosome 2 of humans is G or A (rs79738529); a polynucleotide consisting of 5-100 consecutive DNA sequences comprising base number 112464376, wherein the aforementioned base number 112464376 of chromosome 1 of humans is T or C (rs17029069); a polynucleotide consisting of 5-100 consecutive DNA sequences comprising base number 46078964, wherein the aforementioned base number 46078964 of chromosome 2 of humans is G or A (rs7581914); a polynucleotide consisting of 5-100 consecutive DNA sequences comprising base number 32297419, wherein the aforementioned base number 32297419 of chromosome 17 of humans is A or G (rs9902778); a polynucleotide consisting of 5-100 consecutive DNA sequences comprising base number 31677062, wherein the aforementioned base number 31677062 of chromosome 17 of humans is C or T (rs452439); a polynucleotide consisting of 5-100 consecutive DNA sequences comprising base number 45982030, wherein the aforementioned base number 45982030 of chromosome 2 of humans is G or A (rs867286); a polynucleotide consisting of 5-100 consecutive DNA sequences comprising base number 51500192, wherein the aforementioned base number 51500192 of chromosome 19 of humans is A or C (rs1722562); a polynucleotide consisting of 5-100 consecutive DNA sequences comprising base number 112356676, wherein the aforementioned base number 112356676 of chromosome 1 of humans is A or G (rs641724);polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 322,667,53rd base wherein the above-mentioned 322,667,53rd base of human chromosome 17 is A or G (rs 149,0895); a polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 320,141,17th base wherein the above-mentioned 320,141,17th base of human chromosome 17 is A or G (rs 105,12451).

17. An information providing method for selecting a customized substance for relieving irritation caused by vitamin C, comprising: (a) a step of obtaining a biological sample from an individual; (b) a step of amplifying or hybridizing with a probe a polymorphic site of the SNP marker consisting of rs940052, rs902557, rs12602965, rs7038646, rs79738529, rs17029069, rs7581914, rs9902778, rs452439, rs867286, rs1722562, rs641724, rs1490895 and rs10512451 from the above biological sample; and (c) a step of confirming the base of the amplified or hybridized polymorphic site of the above (b).

18. The information providing method according to Claim 17, wherein The substance is one or more selected from the group consisting of quercetin, phloretin, genistein, tannic acid, zinc PCA, hydroxyasiaticoside, resveratrol, tocopherol, asiaticoside and chinpi extract.

19. The information providing method according to Claim 17, wherein When the risk score obtained by multiplying the genotype of rs940052, rs902557, rs12602965, rs7038646, rs79738529, rs17029069, rs7581914, rs9902778, rs452439, rs867286, rs1722562, rs641724 and rs1490895 by the effect size (β) and adding them is 1 or less, the risk of the irritation caused by vitamin C is classified as a low risk group, and when the risk score exceeds 1, the risk of the irritation caused by vitamin C is classified as a high risk group.

20. The information providing method according to Claim 19, wherein The prescription customized substance when the risk of the skin irritation caused by vitamin C is a high risk group.

21. The information providing method according to Claim 17, wherein The sample is hair, urine, blood, various body fluids, isolated tissues, isolated cells or saliva.

22. The information providing method according to Claim 17, wherein The amplification and confirmation of the polymorphic site use an SNP chip.

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