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 a method for providing information for selecting a customized material for alleviating irritation caused by vitamin C, the method comprising a step of identifying bases at polymorphic sites of genetic polymorphism markers that have significant correlation with the degree of irritation caused by vitamin C. The present invention also relates to: a composition for diagnosing skin type susceptible to skin irritation caused by vitamin C, the composition comprising a probe for detecting genetic polymorphism markers that have significant correlation with skin irritation caused by vitamin C or an agent for amplifying the markers; a kit or a microarray, each comprising the composition; and a method for providing information about skin type susceptible to skin irritation caused by vitamin C, by using the genetic polymorphism markers or a combination of the markers.

Description

Composition for alleviating skin irritation caused by vitamin C

[0001] The present invention relates to a composition for alleviating irritation caused by vitamin C and a method for providing information for selecting a customized substance for alleviating irritation caused by vitamin C, which comprises a step of identifying a base at a polymorphic site of a genetic polymorphism marker having a significant correlation with the degree of irritation caused by vitamin C. The customized substance of the present invention relates to a cosmetic composition for alleviating irritation caused by vitamin C, preventing discoloration in a formulation through improving the stability of vitamin C, and enhancing efficacy. 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 an agent capable of amplifying a genetic polymorphism marker having a significant correlation with the degree of irritation caused by vitamin C, a kit or microarray comprising the composition, and a method for providing information for determining the degree of irritation caused by vitamin C using the genetic polymorphism marker or a combination of markers.

[0002]

[0003] The skin is the outermost organ of the body, covering a vast area (1.5-2 m2). Its role is to defend against external factors such as UV rays and foreign substances. Exposure to various external factors and the aging process causes the skin to synthesize and release melanin, which can lead to cosmetic problems and diminish the quality of life. Vitamin C is a well-known whitening agent that prevents pigmentation caused by melanin.

[0004] Vitamin C, also known as ascorbic acid, is an essential nutrient that cannot be synthesized within the body and must be supplemented through diet. Furthermore, it alleviates skin pigmentation by inhibiting tyrosinase activity, which is involved in melanin production. It also promotes collagen protein synthesis and, with its powerful antioxidant activity, reduces free radicals in the body, delaying skin aging. This makes it a key whitening and anti-aging ingredient in cosmetics.

[0005] Although vitamin C exhibits excellent anti-aging and skin beautifying effects due to its high antioxidant capacity, it is also unstable and sensitive to external environments such as air, especially oxygen, heat, and light, easily decomposing through oxidation and causing discoloration within the formulation. Therefore, to complement these problems of vitamin C and improve stability, vitamin C derivatives (ascorbic acid derivatives) with a modified vitamin C structure are widely used. However, vitamin C derivatives have the disadvantage of significantly lower antioxidant and pigmentation inhibition effects compared to pure vitamin C.

[0006] Meanwhile, growing consumer interest in vitamin C and research showing that concentrations between 10% and 20% can be more effective are driving demand for high-concentration products. However, higher concentrations not only increase formulation instability but also lead to skin irritation caused by vitamin C. However, because the exact mechanism of this irritation remains unknown, it is currently common to assess skin sensitivity after using the product. Therefore, to minimize side effects and ensure more effective application of vitamin C to the skin, prior research into the causes of vitamin C irritation and the development of diagnostic methods are necessary. Some studies have shown that topical vitamin C application increases gene expression in dermal collagen and the synthesis of MMP-1 inhibitors (pmid 11407971). Other disease studies have linked mutations in the vitamin C transporter gene to common chronic diseases (pmid 26980812). Therefore, skin reactivity to vitamin C is expected to vary depending on genotype.

[0007] However, specific genetic correlations between skin reactivity to vitamin C, particularly its level of irritation, are scarce. Therefore, identifying genetic variants and genes associated with vitamin C skin irritation is expected to provide valuable information for understanding the biological mechanisms and predicting individual vitamin C use and its associated irritation.

[0008]

[0009] In this invention, candidate gene analysis was used to identify genetic variants and genes associated with vitamin C skin irritation in Koreans. Furthermore, a model for predicting vitamin C skin irritation was developed using these genetic variants. Furthermore, based on genetic and skin measurement data, an effective material capable of alleviating vitamin C skin irritation in Koreans was developed and applied to products.

[0010]

[0011] One object of the present invention is to provide a composition for alleviating skin irritation by vitamin C.

[0012] Another object of the present invention is to provide a cosmetic composition or a quasi-drug composition containing the composition as an active ingredient.

[0013] 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 its efficacy.

[0014] Another object of the present invention is to provide an information-providing method for selecting a customized substance for relieving irritation by vitamin C, which comprises a step of identifying a base at a polymorphic site of a significant single nucleotide polymorphism marker associated with skin irritation by vitamin C.

[0015] Another object of the present invention is to provide a single nucleotide polymorphism (SNP) marker for diagnosing the degree of stimulation by vitamin C.

[0016] Another aspect of the present invention provides a composition for diagnosing the degree of irritation caused by vitamin C, comprising a probe or an agent capable of detecting or amplifying a single nucleotide polymorphism (SNP) marker for diagnosing the degree of irritation caused by vitamin C.

[0017] Another object of the present invention is to provide a kit or microarray for diagnosing the degree of stimulation by vitamin C comprising the above composition.

[0018] Another object of the present invention is to provide a method for providing information on the degree of stimulation by vitamin C, which includes a step of identifying a polymorphic site of the single nucleotide polymorphism marker.

[0019] Another object of the present invention is to provide an information providing method for prescribing a customized substance targeting a high-risk group susceptible to stimulation by vitamin C, including a step of identifying a polymorphic site of the single nucleotide polymorphism marker.

[0020]

[0021] The present invention can provide information on the degree of stimulation by vitamin C of an individual through genetic polymorphism markers having a correlation with the degree of stimulation by vitamin C, and furthermore, it will be possible to develop customized ingredients or products that can alleviate the degree of stimulation by vitamin C based on the information on genetic polymorphism markers observed in an individual.

[0022]

[0023] Figure 1 illustrates a vitamin C-specific skin stimulation recognition gene expression control material of the present invention.

[0024] Figure 2 shows the results of a synergy analysis of ASIC2 expression reduction by quercetin, tannic acid or quercetin, tannic acid, and resveratrol of the present invention.

[0025] Figure 3 shows an example of rs10512451 (17:32014117:A:G), a single nucleotide polymorphism marker within ASIC2, which differs in whether stimulation is perceived depending on the sub-allele type when applying vitamin C of the present invention.

[0026] Figure 4 shows the results of an analysis of the degree of improvement in irritation according to the addition of the vitamin C skin irritation relief material of the present invention.

[0027] Figure 5 shows the difference in improvement in vitamin C stimulation in low-risk and high-risk groups when the personalized vitamin C stimulation relief composition of the present invention is applied.

[0028] Figure 6 shows a visual evaluation of the degree of discoloration due to heat treatment of pure vitamin C.

[0029] Figures 7 and 8 illustrate the discovery and evaluation of a material for recovering discoloration and antioxidant activity of pure vitamin C at high temperatures.

[0030] Figure 9 shows the results of whitening and antioxidant activity analysis of tannic acid and tocopherol, which are vitamin C stimulation relief materials of the present invention.

[0031]

[0032] This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention can also be applied to each other description and embodiment. In other words, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Furthermore, the scope of the present invention should not be considered limited by the specific descriptions described below.

[0033] Furthermore, those skilled in the art will recognize or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific embodiments of the invention described herein. Furthermore, such equivalents are intended to be encompassed by the present invention.

[0034]

[0035] In one embodiment to achieve the object of the present invention, the present invention provides a composition for alleviating skin irritation by vitamin C. For example, the composition comprises, as an active ingredient, at least one substance selected from the group consisting of quercetin, phloretin, genistein, tannic acid, zinc PCA, madecassoside, resveratrol, tocopherol, asiaticoside, and green citrus extract. In another example, the composition comprises

[0036] (i) quercetin, phloretin, genistein, tannic acid, zinc PCA, madecassoside and resveratrol; or (ii) madecassoside, tannic acid, tocopherol, genistein, asiaticoside and citrus extract; but is not limited thereto.

[0037]

[0038] The term "quercetin" refers to a plant-based flavonoid compound derived from the flavonoid group of polyphenols. Quercetin is found in many fruits, vegetables, leaves, seeds, and grains. It has a bitter taste and is used as an ingredient in health supplements, beverages, and foods. Quercetin is also called quercetin. Quercetin is not limited by its method of acquisition, and can be chemically synthesized using methods known in the art or commercially available materials.

[0039] [Chemical Formula 1]

[0040]

[0041]

[0042] The term 'phloretin' above refers to a dihydrochalcone plant-derived phenol with a C6-C3-C6 skeleton, which is abundant in apples (mainly peels) and apple leaves, and exists as phloridzin (phloretin 2'-O-glucose) in free or glucosidic form. Apple peels contain 0.2 to 5 mg / g of phloretin. Phloretin has many biological functions, including antioxidant and anticancer activities, and is associated with the prevention of heart disease. This component is particularly superior to other antioxidant components in its ability to penetrate the skin barrier. Phloretin is not limited by its acquisition method, and can be chemically synthesized by a method known in the art, or a commercially available substance can be used.

[0043] [Chemical Formula 2]

[0044]

[0045]

[0046] The term "Genistein" above refers to one of the isoflavones isolated from soybeans, and is known to have various effects, including anti-tumor, anti-osteoporosis, and antioxidant properties. Because it contains phytoestrogen activity, it is a substance widely used as an ingredient in pharmaceuticals and health functional foods, especially for alleviating various menopausal symptoms in postmenopausal women. The method of obtaining genistein is not limited, and it can be chemically synthesized by a method known in the art, or a commercially available substance can be used.

[0047] [Chemical Formula 3]

[0048]

[0049]

[0050] The term 'tannic acid' above is one of the most abundant polyphenols found in plants such as fruits, vegetables, olives, and cacao. Recently, tannic acid has been used as a multifunctional coating molecule. It is also known as a molecule with excellent affinity for 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 moieties rich in phenolic hydroxyl groups (five galol groups (three -OH groups connected to an aromatic ring) and five catechol groups (two -OH groups covalently connected to an aromatic ring)). The tannic acid is not limited to the method of obtaining it, and can be chemically synthesized by a method known in the art, or a commercially available material can be used. The INCI name of the substance is tannic acid, and it can be used interchangeably with tannic acid.

[0051] [Chemical Formula 4]

[0052]

[0053]

[0054] The above term 'zinc PCA (ZnPCA)' is a raw material synthesized using glutamic acid obtained from sugar beets. Zinc is a mineral essential for physiological activities in the body and is known to have various functions (cell differentiation protein synthesis, antibacterial action). In particular, zinc inhibits 5-α reductase to reduce sebum secretion, and the above zinc PCA can be used interchangeably with zinc PCA and zinc pyrrolidone carboxylic acid. The above zinc PCA is not limited to the method of obtaining it, and can be synthesized by a method known in the art or a commercially available material can be used.

[0055]

[0056] The above term 'Madecassoside' refers to one of the four main components of the extract of Centella asiatica, a plant that grows widely in the subtropical regions along the Indian Ocean coast. The extract of this plant has been used as a folk medicine by the indigenous people of the region since ancient times to treat skin diseases and leprosy (especially the ulcer area of ​​leprosy). In particular, madecassoside is known to not only provide a protective effect by suppressing inflammation, preventing hyperkeratinization, and inhibiting the penetration of external environmental pollutants, but also promote collagen production to quickly regenerate new skin, thereby minimizing scarring. The above madecassoside is not limited to the method of obtaining it, and may be chemically synthesized by a method known in the art, or a commercially available substance may be used.

[0057]

[0058] The term 'resveratrol' above 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 resveratrol is C14H12O3 and the molecular weight is 228.25. It is a type of stilbene and is produced in plants by the enzyme stilbene synthase (STS), and exists in two structural isomers. It is found in many plants, including mulberries, peanuts, grapes, berries such as raspberries, and cranberries. It is found in more quantities than other plants because grapes secrete a powerful antibacterial substance called resveratrol as a defense substance to protect themselves when attacked by fungi, as a phytoalexin secreted when plants are stressed. It can be obtained by extracting natural products by conventional methods or by synthesizing them through a compound synthesis process.

[0059] Non-limiting examples of the above extraction methods include hot water extraction, ultrasonic extraction, filtration, and reflux extraction, and these may be performed alone or in combination of two or more methods.

[0060] In the present invention, the type of solvent used for the extraction is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the extraction solvent include water, alcohol, or a mixture thereof, which may be used alone or in combination of one or more types. Specifically, water may be used. When alcohol is used as the solvent, an alcohol having 1 to 4 carbon atoms may be specifically used.

[0061]

[0062] The above term 'tocopherol' refers to 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 may be used, and any of those isolated from extracts, synthesized, or commercially available products may be used without limitation.

[0063]

[0064] The term "asiaticoside" refers to a natural organic compound that reduces skin irritation, improves elasticity, and possesses antioxidant properties. Asiaticoside, a triterpene saponin compound, can exhibit skin moisturizing and antioxidant effects. Asiaticoside is not limited by its method of acquisition, and can be synthesized using methods known in the art or using commercially available materials.

[0065]

[0066] The above term 'green tangerine extract' refers to an extract extracted from the fruit of green tangerine (Citrus nippokoreana Tanaka). It is also called 'tangerine peel extract' by its INCI name. The green tangerine is different from the commonly called tangerine (Citrus unshiu Marcov.) by its specific name. The green tangerine is an evergreen tree of the Rutaceae family that mainly grows in Jeju City. The genus name is derived from kitron, an old Greek name for an African fragrant tree. The green tangerine grows to a height of about 7m, and the branches are thin and densely arranged and have small thorns. The leaves of the green tangerine are lanceolate, 65mm long, and 26mm wide. The petiole is about 10 mm, the wing leaves are small, the flowers bloom in June, and the fruit of the above-mentioned green tangerine is spherical, with a longitudinal diameter of 3 to 4 cm and a transverse diameter of 4 to 5 cm. The method for obtaining the above-mentioned green tangerine extract is not particularly limited, and can be extracted according to a method commonly used in the relevant technical field.

[0067] Non-limiting examples of the above extraction methods include hot water extraction, ultrasonic extraction, filtration, and reflux extraction, and these may be performed alone or in combination of two or more methods.

[0068] In the present invention, the type of solvent used for the extraction is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the extraction solvent include water, alcohol, or a mixture thereof, which may be used alone or in combination of one or more types. Specifically, water may be used. When alcohol is used as the solvent, an alcohol having 1 to 4 carbon atoms may be specifically used.

[0069]

[0070] The material of the present invention may be contained in an amount of about 0.0001 to 10 wt% based on the total weight of the composition, but is not limited thereto. For example, the irritation-relieving material by vitamin C of the present invention may be contained in an amount of about 0.0001 to 10 wt%, about 0.0005 to 5 wt%, about 0.001 to 5 wt%, about 0.01 to 5 wt%, about 0.0001 to 3 wt%, about 0.0005 to 3 wt%, about 0.001 to 3 wt%, about 0.01 to 3 wt%, about 0.0001 to 1 wt%, about 0.0005 to 1 wt%, about 0.001 to 1 wt%, or about 0.01 to 1 wt% based on the total weight of the composition.

[0071]

[0072] In the present invention, the term "about" may be presented before a specific numerical value. As used herein, the term "about" encompasses not only the exact number described after the term, but also a range that is or is nearly that number. Whether a number is or is nearly the specific number described can be determined based on the context in which it is presented. For example, the term "about" may refer to a range of -10% to +10% of a numerical value. For another example, the term "about" may refer to a range of -5% to +5% of a given numerical value. However, this is not a limitation.

[0073]

[0074] For the purpose of the present invention, the above material has a skin irritation relief effect due to vitamin C.

[0075] For the purpose of the present invention, the term 'skin irritation by vitamin C' means an abnormal reaction such as stinging, burning, itching, or other reactions (erythema, rash, allergic reaction, swelling, or nervousness) that occurs when an external substance comes into contact with the skin.

[0076]

[0077] For the purposes of the present invention, the above-mentioned substance may be characterized by a reduction in melanin production. Specifically, treatment with tannic acid, tocopherol, and a combination thereof among the above-mentioned substances may promote a reduction in melanin.

[0078] For example, when tannic acid and / or tocopherol, which are effective in reducing melanin production, were treated simultaneously with vitamin C, it was confirmed that there was a synergistic effect in reducing melanin when treated simultaneously with vitamin C compared to when vitamin C, tannic acid, or tocopherol were treated individually.

[0079]

[0080] For the purposes of the present invention, the above-mentioned substance may be characterized by improved antioxidant capacity, and in particular, may be capable of restoring antioxidant capacity that has been reduced by high temperatures. Specifically, when any one or more selected from the group consisting of resveratrol, tannic acid, quercetin, madecassoside, genistein, and zinc PCA is treated, the improvement and / or recovery of antioxidant capacity may be promoted.

[0081] For example, when resveratrol, tannic acid, quercetin, madecassoside, genistein, and / or zinc PCA were treated simultaneously with vitamin C, it was confirmed that there was a synergistic effect in improving antioxidant capacity when vitamin C, resveratrol, tannic acid, quercetin, madecassoside, genistein, or zinc PCA were treated simultaneously with vitamin C, compared to when each of them was treated separately.

[0082]

[0083] For the purpose of the present invention, the above material may be characterized by improving the stability of vitamin C, and specifically, may be characterized by preventing discoloration due to heat by improving the stability of vitamin C. More specifically, when any one or more selected from the group consisting of resveratrol, tannic acid, madecassoside, genistein, and zinc PCA is treated among the above materials, the stability of vitamin C can be improved and discoloration due to heat can be prevented.

[0084] For example, when resveratrol, tannic acid, madecassoside, genistein, and / or zinc PCA were treated simultaneously with vitamin C, it was confirmed that when vitamin C, resveratrol, tannic acid, madecassoside, genistein, or zinc PCA were treated simultaneously with vitamin C, there was a synergistic effect in preventing heat-induced discoloration of vitamin C compared to when vitamin C, resveratrol, tannic acid, madecassoside, genistein, or zinc PCA were treated alone.

[0085]

[0086] For the purpose of the present invention, the substance may be characterized by inhibiting the expression of ASIC2 or PRKCE. Specifically, when treating with at least one selected from the group consisting of quercetin, phloretin, genistein, tannic acid, zinc PCA, madecassoside, and resveratrol, the expression of ASIC2 may be inhibited, and when treating with at least one selected from the group consisting of madecassoside, tannic acid, tocopherol, genistein, asiaticoside, and green citrus extract, the expression of PRKCE may be inhibited, but is not limited thereto. It can be seen that the responsiveness to the substance of the present invention varies depending on a specific genetic polymorphism type, and it is suggested that the efficiency of alleviating irritation by vitamin C may be increased by providing a customized prescription for alleviating irritation by vitamin C.

[0087]

[0088] The above composition is characterized by being personalized. Specifically, it can provide a personalized substance targeting an individual having a significant single nucleotide polymorphism (SNP) marker associated with a specific level of stimulation by vitamin C.

[0089] For example, the significant single nucleotide polymorphism (SNP) marker associated with the degree of stimulation by vitamin C may be, but is not limited to, one or more selected from Table 1.

[0090] In the present invention, the term "polymorphism" refers to a case where two or more alleles exist at a single genetic locus, and among the polymorphic sites, a polymorphism in which only a single base differs from person to person is called a single nucleotide polymorphism (SNP). A desirable polymorphic marker has two or more alleles that exhibit an occurrence frequency of 1% or more, more specifically 10% or 20% or more, in a selected population. A 'genetic polymorphic marker' generally refers to a case where two or more alleles are observed at the same genetic location (base), and generally, depending on the individual, there are cases of major allele / major allele, major allele / minor allele, and minor allele / minor allele. In the present invention, it may be used interchangeably with "polymorphic marker" and may mean a base and base portion of a lower allele, or may be defined together with the number and base position of a chromosome, but is not limited thereto.

[0091] In the present invention, the term "allele" refers to multiple types of a gene existing at the same genetic locus of a homologous chromosome. Alleles are also used to indicate polymorphism, and for example, SNPs have two types of alleles. In addition, it refers to a combination of two or more bases having the same number and base position on a chromosome, and the bases include a major allele that occurs frequently in individuals of a specific population, and a minor allele that occurs less frequently than the major allele.

[0092] Specifically, the genetic polymorphism markers of the present invention are significantly associated with skin irritation by vitamin C, and can have two alleles, and can be divided into cases of having a major allele / minor allele, a minor allele / minor allele, and a major allele / major allele.

[0093] In the case where an individual has an allele of a polymorphic marker with a significant correlation with the degree of irritation by vitamin C, the effect of relieving irritation by vitamin C may vary depending on the reactivity to the substance of the present invention depending on the allele that the individual has. For example, in the case where the individual has a major allele / major allele, a major allele / minor allele, or a minor allele / minor allele, the reactivity to the substance of the present invention, that is, the skin characteristics of high or low degree of irritation by vitamin C, can be measured, and thus, information can be provided for selecting a customized substance for increasing the degree of irritation relief by vitamin C in an individual that includes one or more of the minor alleles.

[0094]

[0095] In the present invention, the term "rs_id" refers to an rs-ID, an independent marker assigned to all SNPs initially registered by NCBI, which began accumulating SNP information in 1998. The rs_id described in this table refers to an SNP marker, which is a polymorphic marker of the present invention.

[0096] The above single nucleotide polymorphism marker may be one or more single nucleotide polymorphism markers selected from the single nucleotide polymorphism markers shown in Table 1. The single nucleotide polymorphism marker shown in Table 1 may be used to determine the degree of correlation with the degree of stimulation by vitamin C.

[0097] The degree of stimulation of the single nucleotide polymorphism marker of the present invention by vitamin C was determined by measuring the frequency of each marker. Such significance is characterized by a p-value such as, but not limited to, a p-value of less than 0.05, less than 0.01, less than 0.001, less than 0.0001, less than 0.00001, less than 0.000001, less than 0.00000001, less than 0.00000001, or less than 0.000000001. Specifically, the p-value may be less than 0.01, more specifically, the p-value may be less than 0.001, and even more specifically, may be less than 0.0001, but is not limited thereto.

[0098]

[0099] The single nucleotide polymorphism (SNP) marker of the present invention may be one or more of the markers shown in Table 1, but is not limited thereto. The single nucleotide polymorphism (SNP) marker may be one or more, and may be used in combinations of two or more, three or more, or any number that can determine the degree of stimulation by vitamin C, but is not limited thereto.

[0100] The marker may be, but is not limited to, a SNP itself, a polynucleotide consisting of 5-100 contiguous DNA sequences including the SNP position, or a polynucleotide consisting of a complementary sequence thereof.

[0101] For example, the significant single nucleotide polymorphism marker associated with the degree of stimulation by the vitamin C may be one or more single nucleotide polymorphism markers present in a gene encoding a protein of ASIC2, PRKCE, TRPM3, KCND3 or KLK8, and specifically, may 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.

[0102] For example, if the SNP ID is rs940052, Chr.Position (GRCh ver. 37) is described as "2:45981142", and the Allele is initiated as A>G, this indicates that the 45981142nd base on the human chromosome 2 is A or G, and the base located to the left of the ">" of the allele may indicate the major allele, and the base located to the right may indicate the minor allele.

[0103] In one specific example, the marker selected from Table 1 is a polynucleotide comprising 5-100 contiguous DNA sequences including base 45981142 of human chromosome 2, wherein base 45981142 is A or G (rs940052); a polynucleotide comprising 5-100 contiguous DNA sequences including base 32250107 of human chromosome 17, wherein base 32250107 is C or T (rs902557); a polynucleotide comprising 5-100 contiguous DNA sequences including base 31833531 of human chromosome 17, wherein base 31833531 is C or T (rs12602965); A polynucleotide comprising 5-100 contiguous DNA sequences including base 73437824 of human chromosome 9, wherein base 73437824 is G or A (rs7038646); A polynucleotide comprising 5-100 contiguous DNA sequences including base 46040237 of human chromosome 2, wherein base 46040237 is G or A (rs79738529); A polynucleotide comprising 5-100 contiguous DNA sequences including base 112464376 of human chromosome 1, wherein base 112464376 is T or C (rs17029069); A polynucleotide comprising 5-100 contiguous DNA sequences including base 46078964 of human chromosome 2, wherein base 46078964 is G or A (rs7581914); A polynucleotide comprising 5-100 contiguous DNA sequences including base 32297419 of human chromosome 17, wherein base 32297419 is A or G (rs9902778); A polynucleotide comprising 5-100 contiguous DNA sequences including base 31677062 of human chromosome 17, wherein base 31677062 is C or T (rs452439);A polynucleotide comprising 5-100 contiguous DNA sequences including base 45982030 of human chromosome 2, wherein base 45982030 is G or A (rs867286); A polynucleotide comprising 5-100 contiguous DNA sequences including base 51500192 of human chromosome 19, wherein base 51500192 is A or C (rs1722562); A polynucleotide comprising 5-100 contiguous DNA sequences including base 112356676 of human chromosome 1, wherein base 112356676 is A or G (rs641724); A polynucleotide comprising 5 to 100 contiguous DNA sequences including the 32266753rd base of human chromosome 17, wherein the 32266753rd base is A or G (rs1490895); A polynucleotide comprising 5 to 100 contiguous DNA sequences including the 32014117th base of human chromosome 17, wherein the 32014117th base is A or G (rs10512451); and a complementary polynucleotide thereof, but is not limited thereto.

[0104]

[0105] The composition of the present invention 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 according to the present invention can be prepared in a formulation selected from the group consisting of a solution, an external ointment, a cream, a foam, a nourishing toner, an emollient toner, a pack, an emollient, an emulsion, a makeup base, an essence, a soap, a liquid cleanser, a bath agent, a sunscreen cream, a sun oil, a suspension, an emulsion, a paste, a gel, a lotion, a powder, a soap, a surfactant-containing cleansing, an oil, a powder foundation, an emulsion foundation, a wax foundation, a patch, and a spray, but is not limited thereto.

[0106] In addition, the cosmetic composition of the present invention may additionally include one or more cosmetically acceptable carriers that are blended with general skin cosmetics, and may appropriately blend conventional ingredients such as oil, water, surfactants, moisturizers, lower alcohols, thickeners, chelating agents, pigments, preservatives, fragrances, etc., but is not limited thereto.

[0107] The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation.

[0108] When the formulation of the present invention is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or a mixture thereof may be used as a carrier component.

[0109] When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or a mixture thereof may be used as a carrier component, and particularly in the case of a spray, a propellant such as chlorofluorohydrocarbon, propane / butane or dimethyl ether may be additionally included.

[0110] When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizer or an emulsifier is used as a carrier component, and for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil can be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan can be used.

[0111] When the formulation of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth, etc. can be used as a carrier component.

[0112] When the formulation of the present invention is soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolyzates, isethionates, lanolin derivatives, fatty alcohols, vegetable oils, glycerol, sugars, etc. can be used as carrier components.

[0113] All ingredients described in the present invention preferably do not exceed the maximum usage levels specified in the Regulations on Cosmetic Safety Standards, etc. (Korean Regulations), Cosmetic Safety Technical Standards (Chinese Regulations), etc.

[0114]

[0115] In another aspect, the composition provides an over-the-counter drug composition for alleviating or improving irritation by vitamin C.

[0116] In the present invention, the term "quasi-drug" means a product that has a milder effect than a pharmaceutical product among products used for the purpose of diagnosing, treating, improving, alleviating, managing or preventing diseases of humans or animals. For example, according to the Pharmaceutical Affairs Act of the Republic of Korea, a quasi-drug is a product excluding products used for the purpose of pharmaceutical products, and includes products used for treating or preventing diseases of humans or animals, products that have a mild effect on the human body or do not act directly, etc.

[0117] In one embodiment, the pharmaceutical composition of the present invention can be manufactured in a form selected from the group consisting of, but not limited to, body cleanser, shampoo, conditioner, foam, soap, mask, ointment, cream, lotion, essence, and spray.

[0118] All ingredients described in the present invention preferably do not exceed the maximum usage levels specified in the Regulations on Cosmetic Safety Standards, etc. (Korean Regulations), Cosmetic Safety Technical Standards (Chinese Regulations), etc.

[0119]

[0120] In another aspect, the present invention provides a method for providing information for selecting a customized substance for relieving irritation by vitamin C, comprising the steps of: (a) obtaining a biological sample from an individual; (b) amplifying or hybridizing a polymorphic site of a single nucleotide polymorphism marker consisting of rs10512451 from the biological sample with a probe; and (c) identifying a base of the amplified or hybridized polymorphic site of step (b).

[0121] The above-mentioned selected customized material can be applied to an individual having a specific polymorphic site to improve skin irritation caused by vitamin C.

[0122]

[0123] In the present invention, the term "subject" refers to a subject for diagnosing the degree of skin irritation caused by vitamin C. DNA may be obtained from samples such as, but not limited to, hair, urine, blood, various body fluids, isolated tissue, isolated cells, or saliva.

[0124] Any method known to those skilled in the art can be used to obtain genomic DNA in the above step (a).

[0125] Any method known to those skilled in the art can be used to amplify the polymorphic site of the single nucleotide polymorphism marker from the DNA obtained in step (a) or to hybridize it with a probe. For example, the target nucleic acid can be amplified by PCR and purified to obtain the result. 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-based sequence amplification (NASBA) can be used.

[0126] Determining the base of the polymorphic site in step (b) of the above method includes, but is not limited to, sequencing analysis, hybridization by microarray, allele specific PCR, dynamic allele-specific hybridization (DASH), PCR extension analysis, SSCP, PCR-RFLP analysis or TaqMan technique, SNPlex platform (Applied Biosystems), mass spectrometry (e.g., Sequenom's MassARRAY system), mini-sequencing method, Bio-Plex system (BioRad), CEQ and SNPstream system (Beckman), Molecular Inversion Probe array technology (e.g., Affymetrix GeneChip), and BeadArray Technologies (e.g., Illumina GoldenGate and Infinium assays). One or more alleles at a polymorphic marker, including a microsatellite, SNP, or other type of polymorphic marker, can be identified using the above methods or other methods available to those skilled in the art to which the present invention pertains. Determining the base pairs at such polymorphic sites can be specifically performed using a SNP chip.

[0127] The above substance may be one or more selected from the group consisting of quercetin, phloretin, genistein, tannic acid, zinc PCA, madecassoside, resveratrol, tocopherol, asiaticoside, and citrus extract, but is not limited thereto.

[0128] For the purpose of the present invention, the significant genes associated with the degree of stimulation by vitamin C can provide information for selecting a specific substance for each individual. For example, in the case of an individual having one or more single nucleotide polymorphism markers present in a gene encoding ASIC2 or PRKCE protein, when treating one or more substances selected from the group consisting of quercetin, phloretin, genistein, tannic acid, zinc PCA, madecassoside, resveratrol, tocopherol, asiaticoside, and citrus extract, the relief of stimulation by vitamin C can be increased, but the optimal combination of substances can be provided according to the genetic characteristics of each individual.

[0129] This suggests that it may be possible to identify specific single nucleotide polymorphism (SNP) markers that are significantly correlated with skin irritation caused by vitamin C and provide information to select personalized substances that can maximize the effect of alleviating irritation caused by vitamin C based on genetic information.

[0130] Specifically, if the risk score obtained by multiplying the number of sub-alleles 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 skin irritation caused by vitamin C may be classified as a low-risk group, and if the risk score is greater than 1, the risk of skin irritation caused by vitamin C may be classified as a high-risk group. For example, in a case where the risk of irritation caused by the above vitamin C is high-risk, an information provision method may be provided that allows selection of one or more selected from the group consisting of quercetin, phloretin, genistein, tannic acid, zinc PCA, madecassoside, resveratrol, tocopherol, asiaticoside, and green citrus extract as a customized substance, but is not limited thereto.

[0131]

[0132] In the present invention, the term "SNP chip" means one of the DNA microarrays capable of identifying each base of hundreds of thousands of SNPs at once.

[0133] The TaqMan method comprises the steps of (1) designing and manufacturing primers and TaqMan probes to amplify a desired DNA fragment; (2) labeling probes of different alleles with FAM dye and VIC dye (Applied Biosystems); (3) performing PCR using the DNA as a template and the primers and probes; (4) analyzing and confirming the TaqMan assay plate with a nucleic acid analyzer after the PCR reaction is completed; and (5) determining the genotype of the polynucleotide of step (1) from the analysis results.

[0134] In the above, the sequencing analysis can use a conventional method for determining the base sequence, and can be performed using an automated genetic analyzer. In addition, allele-specific PCR refers to a PCR method that amplifies a DNA fragment at which the SNP is located using a primer set including a primer designed with the base at which the SNP is located as the 3' end. The principle of the method is that, for example, when a specific base is substituted from A to G, if a primer including the A as the 3'-terminal base and an opposite primer capable of amplifying a DNA fragment of an appropriate size are designed and a PCR reaction is performed, if the base at the SNP position is A, the amplification reaction is performed normally and a band at the desired position is observed, and if the base is substituted with G, the primer can complementarily bind to the template DNA, but the 3'-end side cannot complementarily bind, so the amplification reaction is not performed properly. This utilizes the fact that DASH can be performed using a conventional method, and specifically, can be performed using the method of Prince et al.

[0135] Meanwhile, PCR extension analysis is performed by first amplifying a DNA fragment containing the base where the single nucleotide polymorphism is located using a primer pair, then inactivating all nucleotides added to the reaction by dephosphorylation, and then adding a SNP-specific extension primer, a dNTP mixture, a dideoxynucleotide, a reaction buffer, and DNA polymerase to perform a primer extension reaction. At this time, the extension primer uses the base immediately adjacent to the 5' side of the base where the SNP is located as the 3' end, and the dNTP mixture excludes nucleic acids having the same base as the dideoxynucleotide, and the dideoxynucleotide is selected from one of the base types representing the SNP. For example, when there is an A to G substitution, if a mixture of dGTP, dCTP, and dTTP and ddATP are added to the reaction, the primer is extended by DNA polymerase at the base where the substitution occurred, and after a few bases, the primer extension reaction is terminated by ddATP at the position where the A base first appears. If the above substitution does not occur, the extension reaction is terminated at that position, so that the type of base representing the SNP can be determined by comparing the length of the extended primer.

[0136] At this time, in the case of fluorescently labeled extension primers or dideoxynucleotides, the SNP can be detected by detecting fluorescence using a genetic analyzer (e.g., ABI's Model 3700) used for general base sequence determination, and in the case of using non-labeled extension primers and dideoxynucleotides, the SNP can be detected by measuring the molecular weight using the MALDI-TOF (matrix-assisted laser desorption ionization-time of flight) technique.

[0137]

[0138] As one aspect for achieving the purpose of the present invention, the present invention provides a single nucleotide polymorphism (SNP) marker for diagnosing the degree of stimulation by vitamin C.

[0139] In another aspect, the present invention provides a composition for diagnosing the degree of irritation by vitamin C, comprising a probe or an agent capable of detecting or amplifying a single nucleotide polymorphism (SNP) marker for diagnosing the degree of irritation by vitamin C.

[0140]

[0141] 'Skin irritation due to vitamin C' refers to, but is not limited to, abnormal reactions such as stinging, burning, itching, and other reactions (erythema, rash, allergic reaction, swelling, and nervousness) that occur when an external substance comes into contact with the skin. Specifically, it refers to determining whether skin irritation due to vitamin C is likely to occur using one or more single nucleotide polymorphism (SNP) markers selected from Table 1.

[0142] The above alleles of the present invention have the same number of chromosomes in each individual, and among them, there are major alleles and minor alleles of SNPs, and as the base of the polymorphic site of the polymorphic marker increases by one to the minor allele, the major alleles may decrease by one, and as the base of the polymorphic site of the polymorphic marker increases by one to the major allele, the minor alleles may decrease by one. However, the ranges in which the minor alleles and major alleles can increase and decrease can be within three types: i) major allele / major allele, ii) major allele / minor allele, iii) minor allele / minor allele, and alleles can decrease or increase within the ranges of the three types, but are not limited thereto.

[0143]

[0144] In the present invention, 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 specifically hybridizing with a polymorphic region of the gene as described above, and the specific method of such gene analysis is not particularly limited and may be any gene detection method known in the technical field to which this invention belongs. In addition, the term may be used interchangeably with the terms "for diagnosing whether a skin type is prone to skin irritation caused by vitamin C" or "for diagnosing whether a high-risk skin type is prone to skin irritation caused by vitamin C."

[0145] In the present invention, the term "agent capable of amplifying 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 polymorphic site of the gene as described above through amplification, and specifically means a primer capable of specifically amplifying a polynucleotide of the marker for diagnosing the degree of irritation caused by vitamin C. In addition, the term may be used interchangeably with the terms "for diagnosing whether a skin type is prone to skin irritation caused by vitamin C" or "for diagnosing whether a high-risk skin type is prone to skin irritation caused by vitamin C."

[0146] The primer used for amplifying the above polymorphic marker refers to a single-stranded oligonucleotide that can act as an initiator of template-directed DNA synthesis under appropriate conditions (e.g., four different nucleoside triphosphates and a polymerizing agent such as DNA, RNA polymerase, or reverse transcriptase) in an appropriate buffer and at an appropriate temperature. The appropriate length of the primer may vary depending on the intended use, but is typically 15 to 30 nucleotides. Shorter primer molecules generally require lower temperatures to form stable hybrids with the template. The primer sequence need not be perfectly complementary to the template, but should be sufficiently complementary to hybridize with the template.

[0147] As used herein, the term "primer" refers to a short sequence of bases having a short free 3'-terminal hydroxyl group, capable of forming base pairs with a complementary template, and serving as a starting point for copying the template strand. The primer can initiate DNA synthesis in the presence of a reagent for polymerization (i.e., DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates in an appropriate buffer and temperature. Skin type can be predicted by performing PCR amplification and determining the degree of production of the desired product. PCR conditions and the lengths of the sense and antisense primers can be modified based on those known in the art.

[0148] The probes or primers of the present invention can be chemically synthesized using the phosphoramidite solid support method or other well-known methods. These nucleic acid sequences can also be modified using many means known in the art. Non-limiting examples of such modifications include methylation, "capping," substitution with one or more homologs of a natural nucleotide, and modifications between nucleotides, such as modification with uncharged linkers (e.g., methyl phosphonate, phosphotriester, phosphoramidate, carbamate, etc.) or charged linkers (e.g., phosphorothioate, phosphorodithioate, etc.).

[0149]

[0150] In another aspect, the present invention provides a kit for diagnosing the degree of irritation caused by vitamin C, comprising the composition for diagnosing the degree of irritation caused by vitamin C. The kit may be, but is not limited to, an RT-PCR kit or a DNA chip kit.

[0151] The kit of the present invention can diagnose the degree of stimulation by vitamin C by amplifying a SNP polymorphism marker, which is a marker for diagnosing the degree of stimulation by vitamin C, or by confirming the expression level of mRNA of the SNP polymorphism marker. As a specific example, the kit for measuring the mRNA expression level of the marker for diagnosing the degree of stimulation by vitamin C in the present invention may be a kit including essential elements required for performing RT-PCR. In addition to each primer pair specific for the gene of the marker for diagnosing the degree of stimulation by vitamin C, the RT-PCR kit may include a test tube or other appropriate container, a reaction buffer (with various pH and magnesium concentrations), deoxynucleotides (dNTPs), an enzyme such as Taq polymerase and reverse transcriptase, DNase, RNase inhibitor, DEPC-water, sterile water, etc. In addition, it may include a primer pair specific for a gene used as a quantitative control. Also specifically, the kit of the present invention may be a kit for diagnosing the degree of stimulation by vitamin C, which includes essential elements necessary for performing a DNA chip. A DNA chip kit is a tool in which nucleic acid species are attached in a gridded array to a flat solid support plate, typically a glass surface no larger than a microscope slide, so that the nucleic acids are uniformly arranged on the chip surface, thereby allowing multiple hybridization reactions to occur between the nucleic acids on the DNA chip and complementary nucleic acids contained in a solution treated on the chip surface, thereby enabling massive parallel analysis.

[0152]

[0153] In another aspect, the present invention provides a microarray for diagnosing the degree of stimulation by vitamin C, comprising the composition for diagnosing the degree of stimulation by vitamin C.

[0154] The above microarray may comprise DNA or RNA polynucleotides. The above microarray is composed of a conventional microarray except that the probe polynucleotides comprise the polynucleotides of the present invention.

[0155] Methods for manufacturing microarrays by immobilizing probe polynucleotides on a substrate are well known in the art. The probe polynucleotide refers to a hybridizable polynucleotide, and refers to an oligonucleotide capable of sequence-specific binding to complementary strands of nucleic acids. The probe of the present invention is an allele-specific probe, which hybridizes to a DNA fragment derived from one member but not to a fragment derived from the other member due to the presence of a polymorphic site in the nucleic acid fragments derived from two members of the same species. In this case, the hybridization conditions must be sufficiently stringent to ensure that the probe hybridizes to only one of the alleles, as there is a significant difference in hybridization intensity between the alleles. This can induce a good hybridization difference between different allele forms. The probe of the present invention can be used for detecting alleles and for diagnosing skin irritation relief caused by vitamin C, etc. The above diagnostic methods include detection methods based on nucleic acid hybridization, such as Southern blot, and may be provided in a form pre-bound to the DNA chip substrate in methods using DNA chips. The above hybridization can usually be performed under stringent conditions, for example, a salt concentration of 1 M or less and a temperature of 25°C or higher. For example, conditions of 5x SSPE (750 mM NaCl, 50 mM Na Phosphate, 5 mM EDTA, pH 7.4) and 25-30°C may be suitable for allele-specific probe hybridization.

[0156] The process of immobilizing the probe polynucleotide associated with the degree of stimulation by vitamin C of the present invention onto a substrate can also be easily manufactured using such conventional techniques. Furthermore, hybridization of nucleic acids on microarrays and detection of hybridization results are well known in the art. For example, the detection can be accomplished by labeling a nucleic acid sample with a label capable of generating a detectable signal, such as a fluorescent substance such as Cy3 or Cy5, hybridizing the nucleic acid sample onto a microarray, and detecting the signal generated from the label.

[0157]

[0158] In another aspect, the present invention provides a method for providing information on the degree of stimulation by vitamin C, comprising the steps of (a) amplifying or hybridizing a polymorphic site of the single nucleotide polymorphism marker with a probe in DNA obtained from a sample isolated from an individual; and (b) identifying a base of the amplified or hybridized polymorphic site in step (a).

[0159] The above terms, 'object', (a) step, (b) step' are as described above.

[0160] The above method may additionally determine, but is not limited to, a high-risk group with a high risk of skin irritation caused by vitamin C when the risk score obtained by multiplying and adding the effect size (β) of the amplified or hybridized single nucleotide polymorphism marker (c) exceeds 1.

[0161]

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

[0163]

[0164] Example 1: Derivation of genetic polymorphism markers significantly associated with vitamin C skin irritation.

[0165]

[0166] We aimed to identify genomic regions (genetic variations) that exhibit differences in vitamin C stimulation (questionnaire responses) based on genetic information. To identify genomic regions (genetic variations) associated with vitamin C stimulation, we selected candidate genes suspected of being associated with vitamin C stimulation, particularly genes related to burning sensation, itching, stinging, keratin barrier function, vitamin C metabolism, redness, and rash, and utilized them for analysis. A microarray genotyping chip (Illumina) was used as a device for screening the genome.

[0167] In order to evaluate the presence and degree of vitamin C irritation in the present invention, subjects were asked to directly apply 20% of vitamin C to their facial skin (cheeks) after washing their face in the evening for two days, and a scale-type questionnaire response was collected at 1-point intervals from no irritation (0 points) to irritation (maximum 6 points). The collected vitamin C skin irritation scale was used as a phenotype.

[0168] To determine the correlation between genomic regions (genetic mutations) and skin irritation levels (scales), linear regression analysis was used to quantify correlation significance and genetic effects.

[0169]

[0170] 1-1. Vitamin C stimulation level evaluation method

[0171] To derive genetic polymorphism markers that can explain the degree of vitamin C stimulation, a total of 84 healthy Koreans aged 20-70 were recruited. For the first evaluation, 51 subjects applied vitamin C preparations distributed as the first step of skin care after washing their faces in the evening. The degree of irritation was collected on a scale (0 to 6) and detailed irritation was collected by distinguishing between burning, tingling, and tickling.

[0172]

[0173] 1-2. Gene collection and analysis methods

[0174] Human genomic DNA was extracted from saliva for genetic analysis using the QIAmp mini prep kit (QIAGEN). The quality was confirmed by absorbance (OD 260 / 280) > 1.7, concentration 50 ng / ul, and band inspection using 1x TAE 1% agarose gel. Genetic analysis was performed only on samples that passed the quality test.

[0175] Genetic analysis was conducted using Illumina's microarray genotyping chip, and specifically, the genes of the subjects were analyzed using the same company's global screening array product.

[0176] Illumina's microarray genotyping chip genetic analysis experiment was conducted according to the provided manual, and the processes of genomic DNA amplification, DNA fragmentation, precipitation, hybridization, staining, washing, coating, and scanning were performed using the provided reagents.

[0177] The microarray genotyping chip for which the experiment was completed was scanned using iScan Control Software (Illumina). Upon completion of scanning, an idat file was automatically generated, and data quality control (sample call rate > 98%, marker call rate > 98%) and genetic information verification were performed using the GenomeStudio (Illumina) program.

[0178] In the present invention, only data that passed data quality control after genetic analysis was used.

[0179]

[0180] 1-3. Derivation of genetic polymorphism markers associated with vitamin C stimulation level

[0181] To identify genetic polymorphisms significantly associated with vitamin C stimulation, linear regression analysis was performed using the genetic polymorphisms of the subjects. Only genetic markers with a minor allele frequency of 2% or higher in the subject group were included in the linear regression analysis.

[0182] The significance of genetic polymorphism markers showing association with the level of vitamin C stimulation was evaluated using the T statistic of the results of linear regression analysis, and the criterion was set at p-value < 0.1. A number of genetic polymorphism markers showing significant association with the level of vitamin C stimulation were derived (Table 1).

[0183]

[0184] Vitamin C stimulation-associated gene polymorphism marker number SNP 1) rsID 2) GeneBeta 3) MAF 4) Majorallele 6) Minor allele 5)12:45981142rs940052PRKCE0.64250.402AG217:32250107rs902557ASIC20.71390 .2157CT317:31833531rs12602965ASIC2-0.49570.451CT49:73437824rs7038646T RPM30.49650.3039GA52:46040237rs79738529PRKCE0.45640.2059GA61:11246437 6rs17029069KCND3-0.45010.3922TC72:46078964rs7581914PRKCE-0.41950.3627G A817:32297419rs9902778ASIC20.4050.3824AG917:31677062rs452439ASIC20.42 490.2059CT102:45982030rs867286PRKCE0.42290.3333GA1119:51500192rs172256 2KLK80.43650.2549AC121:112356676rs641724KCND30.4260.27AG1317:32266753 rs1490895ASIC20.71880.2157AG1417:32014117rs10512451ASIC20.34870.4706GA

[0185] 1) SNP: National Institutes of Health (NIH) ID, sequence can be checked on the website

[0186] 2) rsID: Reference SNP

[0187] 3) Beta: Linear regression coefficient

[0188] 4) MAF (Minor allele frequency): Frequency of minor allele within the subject group

[0189] 5) Minor allele: lower allele

[0190] 6) Major allele: superior allele

[0191]

[0192] Example 2: Results of analysis of the effects of various substances on regulating the expression of genes associated with vitamin C skin irritation.

[0193]

[0194] Among various substances with skin-improving effects, a screening process was conducted to select substances that affect the expression of genes associated with vitamin C skin irritation.

[0195] Human keratinocyte cells (HaCaT) were purchased from ATCC. The purchased cells were cultured in a medium containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin in DMEM (Dulbecco's Modified Eagle Medium-GIBCO) at 37°C in a 5% CO2 incubator.

[0196] To analyze the efficacy of vitamin C on skin irritation-related gene mRNA expression changes, human keratinocytes were seeded at 1X10^5 cells / well in 12 wells and cultured for one day in a 37℃, 5% CO2 incubator. Then, substances were added to the medium at appropriate concentrations and cultured for two days under the same conditions. Total RNA was obtained using the AccuPrep Universal RNA Extraction Kit (Bioner, K-3140), and cDNA was synthesized using AccuPower® RocketScript™ Cycle (Bioner, K-2202). The expression level of each gene was analyzed by performing qPCR using the synthesized cDNA and taqman probe (assay ID: human ASIC2- Hs00153756_m1, human PRKCE- Hs00942886_m1).

[0197] As a result, as shown in Fig. 1, it was confirmed that the materials that reduce human ASIC2 expression are quercetin, phloretin, genistein, tannic acid, zinc PCA, madecassoside or resveratrol, and the materials that reduce human PRKCE expression are madecassoside, tannic acid, tocopherol, genistein, asiaticoside or green citrus extract.

[0198]

[0199] Example 3: Confirmation of the synergistic effect of pure vitamin C stimulation improvement materials.

[0200]

[0201] In Example 2, we attempted to determine whether the combination of selected substances had a synergistic effect in suppressing human ASIC2 expression. Human keratinocytes were seeded at 1X10^5 cells / well in a 12-well plate and cultured for one day in a 37℃, 5% CO2 incubator. Then, a medium containing the substances in appropriate concentrations and combinations was added and cultured in the incubator for another day. RNA was extracted in the same manner as in Example 2, cDNA was synthesized, and qPCR was performed to determine whether the expression of the corresponding gene was synergistically suppressed.

[0202]

[0203] It was confirmed that when treated with 15 ppm of quercetin and 15 ppm of tannic acid, ASIC2 expression was reduced by approximately 10% (X) and 36% (Y), respectively. In addition, when quercetin and tannic acid at the corresponding concentrations were treated simultaneously, gene expression was reduced by 94% (T, effect measurement value). Since this is greater than the Colby-type prediction equation (E1) of 47% for the combination of the two substances, it was determined that there was a synergistic effect in reducing ASIC2 expression (Fig. 2). As a specific example, it is not limited to the single nucleotide polymorphism marker rs10512451 (17:32014117:A:G), which can affect vitamin C skin irritation depending on the type of suballele within the ASIC2 gene (Fig. 3).

[0204]

[0205] Colby formula E1=X+Y-(XY / 100)

[0206]

[0207] Furthermore, when treated with different combinations of 15 ppm quercetin, 15 ppm tannic acid, or 25 ppm resveratrol, ASIC2 expression was reduced by 10%, 36%, and 31%, respectively. However, when quercetin, tannic acid, and resveratrol were simultaneously treated at the corresponding concentrations, ASIC2 was reduced by 94% (T, effect measurement). This was greater than the Colby equation predicted by 77% (E2) for the combination of the three substances, and thus it was determined that there was a synergistic effect in reducing ASIC2 expression. (Fig. 2)

[0208]

[0209] Colby E2=(X+Y+Z)-(XY+XZ+YZ / 100)+XYZ / 10000

[0210]

[0211] Example 4: Human application trial for the development of a customized, irritation-relieving prescription for a high-risk group with pure vitamin C skin irritation perception.

[0212]

[0213] Through Example 2, a substance that regulates the expression of a significant polymorphism marker associated with pure vitamin C-specific skin irritation was selected. Through this, the aim was to provide a substance that can increase the irritation improvement efficiency according to the level of risk of vitamin C irritation.

[0214]

[0215] 4-1. Subject selection

[0216] After approval from the Institutional Review Board (IRB, Korea) (LGHH-20220324-AB-04-03), the study was conducted in accordance with the standard operating guidelines and clinical trial standards of the Institutional Review Board of LG Household & Health Care Co., Ltd., and 84 healthy subjects aged 20-50 years with no major skin abnormalities were selected.

[0217]

[0218] 4-2. Subject selection criteria

[0219] Research subjects who do not meet the conditions based on the [Human Material Research Consent Form (IRB No. LGHH-20180727-AA-03, LGHH-20201210-AB-03)] held by the main research institution (LG Household & Health Care)

[0220] i) Those who did not agree to the item ‘1. Permanent preservation’ in the ‘Human body material preservation period’ section of the consent contents

[0221] ii) Those who agreed to item '3. I do not agree' in the 'Provision for secondary use within the retention period' section of the consent contents

[0222]

[0223] 4-3. Subject exclusion criteria

[0224] Among the subjects above, 1) those who were irritated or allergic to normal saline solution, or 2) those who were deemed unsuitable for the test by the research director were excluded from the study.

[0225]

[0226] 4-4. Vitamin C Skin Irritation Relief Material Irritation Evaluation Measurement / Analysis Method

[0227] Of the 84 subjects, 33 in the second group, excluding the first group, applied the control group (20% vitamin C serum) and the experimental group (vitamin C skin irritation relief material-added serum) to the right and left cheeks, respectively. The evaluation was conducted for a total of two days, on the first and third days, and rest was recommended on the second day, considering that repeated exposure to stimulation may dull the sensation. For the same reason, the cheeks to which the control and experimental serums were applied were reversed on the first and third days.

[0228] The evaluation scale was set to be from 0 to 6, with 1-2 being weak (slightly bothersome, questionable), 3-4 being moderate (definitely felt stimulation), and 5-6 being strong (strong stimulation that makes you want to wash it off). The types of stimulation were divided into burning, stinging, tickling, and others (erythema, hives, etc.). The stimulation values ​​evaluated on the 1st and 3rd days were averaged according to the type of stimulation to calculate the ‘degree of improvement in stimulation.’ As a result, it was confirmed that burning significantly decreased from 1.82 in the control group to 1.32 (p<0.05), stinging from 2.06 to 1.64, and tickling from 1.58 to 1.40 (Fig. 4).

[0229]

[0230] In terms of perceived improvement in stimulation, the percentage of people who felt improvement among those who felt stimulation for each type of stimulation was calculated. As a result, the perceived improvement rates were 64% for burning, 62% for stinging, and 55% for tickling. Lastly, the degree of perceived improvement in response to the stimulation-relieving ingredient according to the degree of existing vitamin C stimulation was represented in a color chart, and as can be seen in the figure, it was found that the stronger the existing stimulation, the more effectively the stimulation was improved (Fig. 4).

[0231]

[0232] 4-4. Differences in stimulation improvement according to stimulation gene score when prescribing a customized vitamin C stimulation relief composition

[0233] Based on the beta values ​​of the skin irritation-associated genetic polymorphism markers in Table 1, a vitamin C stimulation gene score (PRS) was calculated (see the formula below). Based on the calculated scores, 32 of the 33 secondary subjects were classified into low-risk and high-risk groups, as follows (one subject was disqualified from the assessment).

[0234]

[0235] i = subject

[0236] j = jth SNP

[0237] SNP j = Number of suballeles of the jth SNP

[0238] m = total number of SNPs (here 13)

[0239]

[0240] The sum of the effect sizes (β) of 13 genetic markers exceeds 1 → high-risk group (n=19)

[0241] The sum of the effect sizes (β) of 13 genetic markers is 1 or less → low-risk group (n=13)

[0242] (For reference, the 13 genetic markers above 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.)

[0243]

[0244] When the low-risk group and high-risk group were divided based on the vitamin C stimulation gene score of 1, the higher the risk of vitamin C stimulation, the more significantly the burning stimulation was reduced when the vitamin C prescription including the irritation relief agent was applied than the low-risk group (Fig. 5) (the reduction was measured in comparison to the burning stimulation felt by the subject when the control prescription was applied). This means that the vitamin C irritation relief agent prescription was more effective for the subject group predicted to have a high vitamin C stimulation gene score, and therefore, it can be expected that the prescription will be more effective for the subject group that is likely to feel irritation in response to vitamin C.

[0245]

[0246] Example 5: Analysis of Vitamin C discoloration and antioxidant activity improvement due to high temperature of vitamin C irritation-relieving materials

[0247]

[0248] We confirmed the improvement of vitamin C discoloration and antioxidant activity for several substances that appear to have the effect of improving the stimulation of vitamin C.

[0249] First, when 10% and 15% vitamin C solutions dissolved in water were boiled at 80 degrees for 8 hours, the color turned yellow and it was confirmed that the antioxidant activity decreased (Fig. 6).

[0250] At this time, the discoloration was compared through the degree of yellowing when viewed with the naked eye and the absorbance at 390 nm, and the antioxidant activity was measured through DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging reaction. Similarly, solutions containing the candidate substances mixed in 10% and 15% vitamin C solutions were boiled at 80 degrees for 8 hours, and the changes in discoloration and antioxidant activity were measured (Figs. 7 and 8).

[0251] As a result, when vitamin C and various substances were treated together, it was confirmed that the discoloration of vitamin C was improved in some substances, and the decreased antioxidant activity was also improved to the level of the non-heated vitamin C solution. Specifically, in the presence of 500 ppm of resveratrol, discoloration was improved by 57% and 72% for 10% and 15% vitamin C, respectively, and in the case of antioxidant activity, it was 27% for 10% vitamin C alone, and 45% when 500 ppm of resveratrol was added to vitamin C, not only did the antioxidant activity improve, but the synergistic effect of the combination was confirmed compared to when resveratrol was treated alone (1%).

[0252] Likewise, it showed improvement in discoloration at 30% and 33% at 500 ppm of zinc PCA, and the antioxidant activity of vitamin C was 21%, zinc PCA 10%, and the combination was 55%, confirming the improvement in antioxidant activity and the synergistic effect of the combination.

[0253] At 200 ppm of madecassoside, discoloration was improved by 62% and 73%, and the antioxidant activity of vitamin C was 27%, madecassoside was 5%, and the combination was 43%, confirming the improvement in antioxidant activity and the synergistic effect of the combination.

[0254] At 200 ppm of quercetin, no improvement in discoloration was observed, and the antioxidant activity of 10% vitamin C, 3% quercetin, and 48% in combination were confirmed to have improved antioxidant activity and synergistic effects.

[0255] At 100 ppm of genistein, discoloration was improved by 50% and 60%, and the antioxidant activity of vitamin C was 37%, genistein 3%, and the combination was 54%, confirming the improvement in antioxidant activity and synergistic efficacy.

[0256] At 100 ppm of tannic acid, discoloration was improved by 19% and 25%, and the antioxidant activity of vitamin C was 21%, tannic acid was 5%, and the combination was 52%, confirming the improvement in antioxidant activity and synergistic efficacy.

[0257]

[0258] Example 6: Confirmation of the pigmentation / antioxidant boosting effects of a material that alleviates nerve stimulation and improves stability of pure vitamin C.

[0259]

[0260] In Example 2, we sought to confirm whether the two materials selected (tannic acid and tocopherol) could boost the whitening effect and antioxidant efficacy of vitamin C at the cellular level.

[0261] First, for whitening evaluation, mouse melanoma cells (B16F10) were seeded at 1X10^5 cells / well in a 12-well plate and cultured for one day in a 37℃, 5% CO2 incubator, and then treated with 10 nM a-MSH. Medium containing appropriate concentrations and combinations of substances was added, and cultured for another day in a 37℃, 5% CO2 incubator to confirm whether the substances inhibited melanin production increased by a-MSH. Cells attached to the plate were collected in PBS and cell pellets were obtained through centrifugation. The cell pellets were visually inspected and photographed. The cell pellets were dissolved in a 1N NaOH (sodium hydroxide) solution containing 10% DMSO (dimethyl sulfoxide) at 80℃ for 10 minutes, placed in a 96-well plate, and the absorbance at 405 nm was measured using an Epoch (BioteK) device. Melanin was quantified using the measured absorbance and normalized to the protein concentration of the sample to determine the melanin concentration. The melanin production inhibition rate of the sample was calculated by dividing the melanin concentration of the sample-treated group by the melanin concentration increased by a-MSH treatment.

[0262] As a result, it was confirmed that when tannic acid or tocopherol was treated simultaneously with vitamin C, melanin production was suppressed more effectively than when vitamin C was treated alone (Fig. 9).

[0263]

[0264] Next, to evaluate the antioxidant efficacy, mouse melanoma cells (B16F10) were seeded at 1X10^4 cells / well in a black clear bottom 96 well plate and cultured in a CO2 incubator for one day. The next day, the medium containing the substance was added and cultured again, and the DCF-DA assay was performed on the third day of culture. Specifically, first, the cell culture medium was removed, washed with PBS, and then the DCF-DA dye was mixed with serum-free medium and incubated with the cells for 30 minutes. The medium was removed again, washed with PBS, and then changed to HBSS (Hank's Balanced Salt Solution) and measured under the conditions of Plate reader Excitation 529 nm, Emission 529 nm.

[0265] As a result, it was confirmed that when tannic acid or tocopherol was treated simultaneously with vitamin C, the antioxidant activity was superior to that of the vitamin C treatment group alone (Fig. 9).

[0266]

[0267] From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without altering its technical spirit or essential characteristics. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as encompassing all changes or modifications derived from the meaning and scope of the following claims and their equivalent concepts, rather than the detailed description above.

Claims

1. A composition for alleviating skin irritation by vitamin C, comprising as an active ingredient at least one substance selected from the group consisting of quercetin, phloretin, genistein, tannic acid, zinc PCA, madecassoside, resveratrol, tocopherol, asiaticoside, and green citrus extract.

2. In the first paragraph, the composition (i) quercetin, phloretin, genistein, tannic acid, zinc PCA, madecassoside and resveratrol; or (ii) A composition comprising madecassoside, tannic acid, tocopherol, genistein, asiaticoside and citrus extract as active ingredients.

3. In paragraph 1, A composition characterized in that the above material is contained in an amount of 0.0001 to 10 wt% based on the total weight of the composition.

4. In paragraph 1, The composition is characterized by a decrease in melanin production.

5. In paragraph 4, A composition wherein the substance for reducing melanin production is tannic acid, tocopherol, and a combination thereof.

6. In paragraph 1, The composition is characterized by improved antioxidant capacity.

7. In paragraph 6, A composition 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 zinc PCA.

8. In paragraph 1, The composition is characterized by improved stability of vitamin C.

9. In paragraph 8, A composition wherein the stability of the above vitamin C is improved by at least one selected from the group consisting of resveratrol, tannic acid, madecassoside, genistein, and zinc PCA.

10. In paragraph 1, A composition for alleviating skin irritation by the above vitamin C, wherein the composition targets an individual that includes one or more single nucleotide polymorphism (SNP) markers for diagnosis associated with vitamin C irritation selected from Table 1.

11. In paragraph 10, A composition, wherein the single nucleotide polymorphism marker for the above vitamin C stimulation-related diagnosis is one or more single nucleotide polymorphism markers present in a gene encoding one or more proteins selected from the group consisting of ASIC2, PRKCE, TRPM3, KCND3, and KLK8.

12. In paragraph 10, A composition, wherein the single nucleotide polymorphism marker for diagnosis of vitamin C stimulation association is at least one selected from the group consisting of rs940052, rs902557, rs12602965, rs7038646, rs79738529, rs17029069, rs7581914, rs9902778, rs452439, rs867286, rs1722562, rs641724, rs1490895, and rs10512451.

13. In paragraph 1, A composition characterized in that the above substance inhibits the expression of ASIC2 or PRKCE.

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

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

16. In paragraph 12, The single nucleotide polymorphism marker for diagnosing stimulation by the above vitamin C is a polynucleotide consisting of 5-100 consecutive DNA sequences including the 45981142nd base of chromosome 2 of a human, wherein the 45981142nd base is A or G (rs940052); a polynucleotide consisting of 5-100 consecutive DNA sequences including the 32250107th base of chromosome 17 of a human, wherein the 32250107th base is C or T (rs902557); a polynucleotide consisting of 5-100 consecutive DNA sequences including the 31833531st base of chromosome 17 of a human, wherein the 31833531st base is C or T (rs12602965); A polynucleotide comprising 5-100 contiguous DNA sequences including base 73437824 of human chromosome 9, wherein base 73437824 is G or A (rs7038646); A polynucleotide comprising 5-100 contiguous DNA sequences including base 46040237 of human chromosome 2, wherein base 46040237 is G or A (rs79738529); A polynucleotide comprising 5-100 contiguous DNA sequences including base 112464376 of human chromosome 1, wherein base 112464376 is T or C (rs17029069); A polynucleotide comprising 5-100 contiguous DNA sequences including base 46078964 of human chromosome 2, wherein base 46078964 is G or A (rs7581914); A polynucleotide comprising 5-100 contiguous DNA sequences including base 32297419 of human chromosome 17, wherein base 32297419 is A or G (rs9902778); A polynucleotide comprising 5-100 contiguous DNA sequences including base 31677062 of human chromosome 17, wherein base 31677062 is C or T (rs452439);A polynucleotide comprising 5-100 contiguous DNA sequences including base 45982030 of human chromosome 2, wherein base 45982030 is G or A (rs867286); A polynucleotide comprising 5-100 contiguous DNA sequences including base 51500192 of human chromosome 19, wherein base 51500192 is A or C (rs1722562); A polynucleotide comprising 5-100 contiguous DNA sequences including base 112356676 of human chromosome 1, wherein base 112356676 is A or G (rs641724); A composition comprising at least one polynucleotide selected from the group consisting of a polynucleotide comprising 5 to 100 contiguous DNA sequences including the 32266753rd base of human chromosome 17, wherein the 32266753rd base is A or G (rs1490895); a polynucleotide comprising 5 to 100 contiguous DNA sequences including the 32014117th base of human chromosome 17, wherein the 32014117th base is A or G (rs10512451); and complementary polynucleotides thereof. 17.(a) A step of obtaining a biological sample from an individual; (b) a step of amplifying or hybridizing with a probe a polymorphic region of a single nucleotide polymorphism marker consisting of rs940052, rs902557, rs12602965, rs7038646, rs79738529, rs17029069, rs7581914, rs9902778, rs452439, rs867286, rs1722562, rs641724, rs1490895 and rs10512451 from the biological sample; and (c) A method for providing information for selecting a customized substance for alleviating irritation by vitamin C, comprising a step of identifying the base of the amplified or hybridized polymorphic site of step (b).

18. In paragraph 17, A method for providing information, wherein the above substance is at least one selected from the group consisting of quercetin, phloretin, genistein, tannic acid, zinc PCA, madecassoside, resveratrol, tocopherol, asiaticoside, and green citrus extract.

19. In paragraph 17, An information provision method wherein the risk score obtained by multiplying and adding the effect size (β) to the genotypes of rs940052, rs902557, rs12602965, rs7038646, rs79738529, rs17029069, rs7581914, rs9902778, rs452439, rs867286, rs1722562, rs641724, and rs1490895 is classified as a low-risk group for the occurrence of irritation by vitamin C if the risk score is 1 or less, and the risk score is classified as a high-risk group for the occurrence of irritation by vitamin C if the risk score is greater than 1.

20. In paragraph 19, A method of providing information, wherein a customized substance is prescribed in cases where the risk of skin irritation caused by the above vitamin C is high.

21. In paragraph 17, A method for providing information, wherein the sample is hair, urine, blood, various body fluids, isolated tissue, isolated cells or saliva.

22. In paragraph 17, A method for providing information, wherein amplification and confirmation of the above polymorphic site are performed using a SNP chip.

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