Composition for alleviating skin pigmentation

Abstract

The present invention relates to a composition for alleviating skin pigmentation, and a method for providing information for selecting a personalized substance for alleviating skin pigmentation, comprising the step of identifying the nucleotide of a polymorphic site in a genetic polymorphism marker associated with the degree of skin pigmentation. In addition, the present invention relates to: a composition for diagnosing skin pigmentation-prone skin types, comprising a probe capable of detecting a genetic polymorphism marker associated with the degree of skin pigmentation, or an agent capable of amplifying the genetic polymorphism marker; a kit or microarray comprising the composition; and a method for providing information about skin pigmentation-prone skin types by using the genetic polymorphism marker or a combination of markers.

Description

Composition for improving skin pigmentation

[0001] The present invention relates to a composition for improving skin pigmentation and a method for providing information for selecting a customized substance for improving skin pigmentation, the method comprising a step of identifying the base of a polymorphic region of a genetic polymorphic marker having a significant correlation with the degree of skin pigmentation. In addition, the present invention relates to a composition for diagnosing whether a skin type is prone to skin pigmentation, the composition comprising a probe capable of detecting or an agent capable of amplifying a genetic polymorphic marker having a significant correlation with the degree of skin pigmentation, a kit or microarray comprising the composition, and a method for providing information on whether a skin type is prone to skin pigmentation using the genetic polymorphic marker or a combination of markers. The skin pigmentation relates to diffuse dark spots on the skin, dark skin tone, or spot-like pigmentation on the skin.

[0002]

[0003] The skin is located on the outermost part of our body and has the largest surface area (1.5 - 2 m 2 ) is an organ that is responsible for the defense function against the external environment (ultraviolet rays, blue light from various electronic devices) and foreign substances (air pollutants). Pigmentation is a type of this defense function. When the skin is exposed to ultraviolet rays, melanocytes produce melanin pigment and move around the nucleus of keratinocytes to create a melanin cap to protect the DNA in the nucleus and prevent skin aging and the occurrence of skin cancer.

[0004] However, excessive melanin production due to aging or exposure to ultraviolet rays can cause pigmentation such as dark spots (freckles), dark skin tone, and skin blotches.

[0005] Dark spots occur naturally due to UV exposure, pregnancy, and childbirth. However, these spots can cause skin unevenness and diminish radiance, leaving the skin dull and dull. Once dark spots appear, they are difficult to remove and have a high risk of recurrence, making it crucial to assess the risk of developing dark spots and take preventative measures. However, existing products focus solely on removing existing dark spots, offering standardized formulas and limited effectiveness, resulting in low consumer satisfaction.

[0006] Furthermore, dark skin tones are known to develop gradually with exposure to UV rays and with age. These external factors can alter the expression of specific genes and protein activity that regulate melanin production, and the resulting decrease in skin tone brightness is difficult and time-consuming to improve. Previous whitening products were standardized to improve not only skin tone but also freckles, blemishes, and sunburn, resulting in low satisfaction with their targeted improvement of skin tone. In particular, research on the genes and mechanisms that specifically darken skin tone is rare, and customized materials for this purpose have rarely been developed.

[0007] Naturally occurring spotty pigmentation on the skin due to factors such as aging or UV exposure can include senile lentigines, liver spots, and solar lentigines. Unlike diffuse pigmentation, which occurs when multiple pigments gather together, spotty pigmentation stands out from its surroundings. The darkened skin caused by spotty pigmentation can appear disharmonious with its surroundings and create a dull, unsightly appearance. Because spotty pigmentation is difficult to remove once it has developed and carries a high risk of recurrence, predicting the risk of spotty pigmentation and preemptively preventing it is necessary. However, existing products have focused on eliminating spotty pigmentation that has already occurred. Furthermore, the methods for removing pigmentation often focus on suppressing melanin production, resulting in low satisfaction with current treatments for spotty pigmentation.

[0008] While research has been conducted on genetic polymorphisms that determine skin color across races, very few studies have identified genetic polymorphisms that determine differences in skin pigmentation within the same race. Naturally, there is also a dearth of research on developing customized materials based on the analysis of genetic polymorphisms associated with individual skin pigmentation.

[0009]

[0010] The present inventors aimed to establish a skin characteristic classification system by building big data related to genetic information and skin type information. Furthermore, they identified genetic characteristics that determine individual skin characteristics, concretized scientific skin classification criteria, and based on this, discovered personalized active ingredients to develop customized cosmetics. As part of this effort, they identified specific single nucleotide polymorphism (SNP) markers significantly correlated with skin pigmentation and confirmed that these markers can be used to provide consumers with personalized prescriptions that can help improve skin pigmentation.

[0011]

[0012] One object of the present invention is to provide a composition for improving skin pigmentation.

[0013] One object of the present invention is to provide a composition for improving dark spots on the skin.

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

[0015] Another object of the present invention is to provide a method for providing information for selecting a customized substance for improving skin diffuse dark spots, which comprises a step of identifying a base at a polymorphic site of a single nucleotide polymorphism marker with a significant association with the degree of skin diffuse dark spots.

[0016] One object of the present invention is to provide a single nucleotide polymorphism (SNP) marker for diagnosing whether a skin type is prone to producing diffuse dark spots on the skin.

[0017] Another object of the present invention is to provide a composition for diagnosing whether a skin type is prone to the formation of diffuse dark spots on the skin, comprising a probe or an agent capable of detecting or amplifying a single nucleotide polymorphism (SNP) marker for diagnosing whether a skin type is prone to the formation of diffuse dark spots on the skin.

[0018] Another object of the present invention is to provide a kit or microarray for diagnosing whether a skin type is prone to producing skin-dispersed dark spots, including the composition.

[0019] Another object of the present invention is to provide a method for providing information on whether a skin type is prone to the creation of diffuse dark spots on the skin, including a step of identifying a polymorphic site of the single nucleotide polymorphism marker.

[0020] Another object of the present invention is to provide an information providing method for prescribing a customized substance to a high-risk group prone to developing skin-dispersed dark spots, including a step of identifying a polymorphic site of the single nucleotide polymorphism marker.

[0021] One object of the present invention is to provide a composition for improving dark skin tone.

[0022] Another object of the present invention is to provide a cosmetic composition or a quasi-drug composition containing the composition as an active ingredient.

[0023] Another object of the present invention is to provide an information-providing method for selecting a customized substance for improving dark skin tone, which comprises a step of identifying a base at a polymorphic site of a significant single nucleotide polymorphism marker associated with dark skin tone.

[0024] One object of the present invention is to provide a single nucleotide polymorphism (SNP) marker for diagnosing whether a person has a skin type that is prone to producing dark skin tones.

[0025] Another object of the present invention is to provide a composition for diagnosing whether a skin type is prone to producing dark skin tones, comprising a probe or an agent capable of detecting or amplifying a single nucleotide polymorphism (SNP) marker for diagnosing whether a skin type is prone to producing dark skin tones.

[0026] Another object of the present invention is to provide a kit or microarray for diagnosing whether a skin type is prone to producing a dark skin tone, including the composition.

[0027] Another object of the present invention is to provide a method for providing information on whether a skin type is prone to producing dark skin tones, including a step of identifying a polymorphic site of the single nucleotide polymorphism marker.

[0028] Another object of the present invention is to provide an information providing method for prescribing a customized substance targeting a high-risk group prone to developing dark skin tone, including a step of identifying a polymorphic site of the single nucleotide polymorphism marker.

[0029] One object of the present invention is to provide a composition for improving spot-like pigmentation.

[0030] Another object of the present invention is to provide a cosmetic composition or a quasi-drug composition containing the composition as an active ingredient.

[0031] Another object of the present invention is to provide an information-providing method for selecting a customized substance for improving spotty pigmentation, which comprises a step of identifying a base at a polymorphic site of a significant single nucleotide polymorphism marker associated with spotty pigmentation.

[0032] One object of the present invention is to provide a single nucleotide polymorphism (SNP) marker for diagnosing whether a skin type is prone to developing spot-like pigmentation.

[0033] Another object of the present invention is to provide a composition for diagnosing whether a skin type is prone to developing spotty pigmentation, comprising a probe or an agent capable of detecting or amplifying a single nucleotide polymorphism (SNP) marker for diagnosing whether a skin type is prone to developing spotty pigmentation.

[0034] Another object of the present invention is to provide a kit or microarray for diagnosing whether a skin type is prone to developing spot-like pigmentation, comprising the composition.

[0035] Another object of the present invention is to provide a method for providing information on whether a skin type is prone to producing spot-like pigmentation, including a step of identifying a polymorphic site of the single nucleotide polymorphism marker.

[0036] Another object of the present invention is to provide an information providing method for prescribing a customized substance targeting a high-risk group prone to developing spot-like pigmentation, including a step of identifying a polymorphic site of the single nucleotide polymorphism marker.

[0037]

[0038] The present invention can provide information on the degree of skin pigmentation of an individual through genetic polymorphism markers having a correlation with the degree of skin pigmentation, and furthermore, it will be possible to develop customized ingredients or products that can alleviate the degree of skin pigmentation based on the information on genetic polymorphism markers observed in an individual.

[0039]

[0040] Figure 1 illustrates a distributed dark spot gene expression material of the present invention.

[0041] Figure 2 illustrates a dark skin tone gene expression material of the present invention.

[0042] Figure 3 illustrates a spot-shaped pigmentation gene expression material of the present invention.

[0043] Figure 4 shows the results of analysis of the melanin production reduction efficacy and synergy of bisabolol, nutmeg extract, and DIPA of the present invention.

[0044] Figure 5 shows the results of analysis of the melanin production reduction efficacy and synergy of ferulic acid, soluble licorice extract, and / or bakuchiol of the present invention.

[0045] Figure 6 shows the results of analysis of the melanin production reduction efficacy and synergy of polydatin and / or quercetin of the present invention.

[0046] Figure 7 shows the results of analysis of the melanin production reduction efficacy and synergy of hexyldecanol, tocopherol, and / or hexylresorcinol of the present invention.

[0047] Figure 8 shows the pigmentation improvement rate of the distributed dark spot improvement synergy combination of the present invention.

[0048] Figure 9 confirms the protein carbonylation inhibition effect of gamma-oryzanol, Aspergillus ferment, or isostearic acid of the present invention.

[0049] Figure 10 shows a representative image of the distributed dark spot improvement of the present invention.

[0050] Figure 11 shows the pigmentation improvement rate of the distributed dark spot customized prescription of the present invention.

[0051] Figure 12 shows a representative image of the dark skin tone improvement of the present invention.

[0052] Figure 13 shows the rate of improvement in pigmentation of the dark skin tone customized prescription of the present invention.

[0053] Figure 14 shows a representative image of improvement in spot-like pigmentation in a high-risk group of the present invention.

[0054] Figure 15 shows the improvement rate of AI spot recognition by the customized prescription for spot-type pigmentation of the present invention.

[0055]

[0056] 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.

[0057] 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.

[0058]

[0059] As another aspect for achieving the purpose of the present invention, the present invention provides a composition for improving skin pigmentation. For example, Myristica Fragrans (Nutmeg) Extract, Polygonum Cuspidatum Extract, Bisabolol, Diisopropyl adipate (DIPA), Cysteine, cysteamine HCl, Sodium Hyaluronate, Ultra-low molecular weight hyaluronic acid, Niacinamide, Glycolic acid, Gluconolactone, Salicylic acid, Polydatin, Ferulic acid, Bakuchiol, Genistein, Oil soluble licorice extract, Aspergillus ferment, Quercetin, Tannic acid, Zinc PCA, Gamma Oryzanol, Chrysin, Tranexamic acid, Sunflower oil, Isostearic acid, Ascorbyl Glucoside, 3-O-Ethyl Ascorbic Acid, Ascorbic Acid, Glycyrrhiza Uralensis (Licorice) Extract, Tocopherol, Sodium Pyruvate, Ecklonia Cava Extract, Hexyldecanol,It may contain as an active ingredient one or more substances selected from the group consisting of asiaticoside and hexylresorcinol, but is not limited thereto.

[0060] For example, the skin pigmentation may be a diffuse dark spot, a dark skin tone, or a blotchy skin pigmentation.

[0061]

[0062] In another aspect for achieving the object of the present invention, the present invention provides a composition for improving dark spots by dispersing them on the skin. For example, the composition comprises, as an active ingredient, at least one substance selected from the group consisting of Myristica Fragrans (Nutmeg) Extract, Polygonum Cuspidatum Extract, Bisabolol, Diisopropyl adipate (DIPA), Cysteine, cysteamine HCl, Sodium Hyaluronate, Hyaluronic acid, Niacinamide, Glycolic Acid, Gluconolactone, and Salicylic Acid. As another example, the composition may include, but is not limited to, (i) cysteine, cysteamine HCL, nutmeg extract, high molecular weight hyaluronic acid, and ultra-low molecular weight hyaluronic acid; (ii) bisabolol, hollyhock root extract, and diisopropyl adipate; or (iii) niacinamide, glycolic acid, gluconolactone, and salicylic acid as active ingredients.

[0063]

[0064] The above term 'nutmeg extract (Myristica Fragrans (Nutmeg) Extract)' refers to the extraction of nutmeg. Nutmeg (Nutmeg, Myristica fragrans) is the dried mature fruit of the nutmeg tree (Myristica fragrans Houtt) belonging to the nutmeg family (Myristicaceae). Its main producing areas are Malaysia, Indonesia, and Sri Lanka, and it is usually harvested in April to June or November to December, and the skin is peeled before use. Nutmeg has been used to improve blood circulation and lung function and has an antibacterial effect, so it has been used to treat respiratory disorders and skin diseases, and it is also used as a fragrance because it has aromatic components.

[0065] Nutmeg extract contains lignan compounds such as nectandrin B, tetrahydrofurogualacin B, and fragransin C1, and flavonoid components such as myricetin and myricitrin. In addition, nutmeg extract is known to contain phenylpropanoid components such as myristicin, elemicin, and safrole, which are substances that are toxic to the human body. The nutmeg extract is not limited to an extraction method and may be extracted by a method known in the art.

[0066]

[0067] The above term 'Polygonum Cuspidatum Extract' refers to the extract of Polygonum cuspidatum. Polygonum cuspidatum is a perennial plant of the Polygonaceae family that grows in the mountains and fields of Korea and is distributed in Korea, Japan, Taiwan, China, etc. Polygonum cuspidatum has been used in the private sector as a diuretic, laxative, and sedative, and has been used in traditional medicine in Korea and other East Asian countries to treat various diseases such as sedation, diuresis, diarrhoea, purulent dermatitis, urethritis, cystitis, hypertension, cancer, and arteriosclerosis. The above Polygonum cuspidatum extract is not limited to the extraction method and may be extracted by a method known in the art.

[0068]

[0069] The above term 'bisabolol' refers to a type of natural monocyclic sesquiterpene alcohol, also known as α-(-)-bisabolol or levomenol. α-(-)-bisabolol has a structure represented by the following chemical formula 1, its chemical name is 6-methyl-2-(4-methylcyclohex-3-en-1-yl)hept-5-en-2-ol, and its molecular weight is 222.372 g·mol-1. Bisabolol is also known as a colorless, viscous oil that is the main component of the essential oil of the Brazilian candeia tree (Eremanthus erythropappus or Vanillosmopsis erythoropappa) or German chamomile (Matricaria recutita). Most commercially available bisabolol is produced by distillation of the bark of the Candeia tree. Bisabolol can be synthesized chemically and obtained using various extraction methods known in the art, with further purification if necessary.

[0070] [Chemical Formula 1]

[0071]

[0072]

[0073] The term "diisopropyl adipate (DIPA)" refers to a compound having the following chemical formula (2), which acts as a lubricant on the skin surface, leaving it soft and smooth. It can be used as a carrier for other cosmetic ingredients and can also soften synthetic compounds by reducing brittleness and cracking. It is used in a variety of applications, such as the production of moisturizers, shaving lotions, and cleansers. The method of obtaining diisopropyl adipate is not limited, and it can be chemically synthesized by methods known in the art, or a commercially available material can be used.

[0074] [Chemical Formula 2]

[0075]

[0076]

[0077] The above term 'Cysteine' is a type of sulfur-containing α-amino acid with the structure HS-CH2CH(NH2)-COOH, and has a sulfhydryl group, so it has the characteristic of forming a disulfide bond with other cysteines. The method of obtaining the cysteine ​​is not limited, and it can be chemically synthesized by a method known in the art, or a commercially available material can be used.

[0078]

[0079] The above term 'cysteamine HCl' is a compound having the same structure as HSCH2CH2NH2HCl and is used as a reducing agent in cosmetic compositions. The method of obtaining the above cysteamine HCl is not limited, and it can be chemically synthesized by a method known in the art or a commercially available material can be used.

[0080]

[0081] The term "hyaluronic acid" refers to a natural, biosynthetic substance found in abundance in the skin of animals and other organisms. Because it contains many hydroxyl groups (-OH), it is a hydrophilic substance and acts as a moisturizer on animal skin. It is also found in human skin, and is particularly abundant in the skin of earthworms. It is primarily prescribed as a moisturizer in cosmetics. It reacts with the CD44 protein of epithelial cells to regulate various physiological functions. It is a linear, anionic natural polysaccharide composed of alternating β-DN-acetylglucosamine and β-D-glucuronic acid, and exists in salt form. The molecular structure of hyaluronic acid can be represented by the following chemical formula 3.

[0082] [Chemical Formula 3]

[0083]

[0084] In addition, the above hyaluronic acid has a molecular weight of 1.5 to 2.0 * 10 6 Da high molecular weight hyaluronic acid (sodium hyaluronate) and molecular weight 0.5 to 1 * 10 4 It can be divided into high molecular weight hyaluronic acid and ultra-low molecular weight hyaluronic acid. High molecular weight hyaluronic acid has a high viscosity, which forms a moisture film on the skin and prevents moisture loss. Ultra-low molecular weight hyaluronic acid has a low viscosity similar to water, so it is absorbed deep into the dermis layer, helping to supply moisture to the skin when used in a product. For the purpose of the present invention, hyaluronic acid may be high molecular weight hyaluronic acid and ultra-low molecular weight hyaluronic acid. The hyaluronic acid 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 material may be used.

[0085]

[0086] The above term 'niacinamide' is a water-soluble derivative of niacin, a water-soluble vitamin B3. It is a white powder that dissolves well in water and has a chemical formula of C6H6N2O and a molecular weight of 122. It is known to exhibit whitening efficacy by inhibiting the movement of melanin from melanocytes to keratinocytes, and is a raw material notified by the Ministry of Food and Drug Safety that is recognized for its whitening functionality when contained in cosmetics at 2-5%. The above niacinamide is not limited to the method of obtaining it, and can be chemically synthesized by a method known in the art or a commercially available substance can be used.

[0087] [Chemical Formula 4]

[0088]

[0089]

[0090] The term 'glycolic acid' above is a colorless, odorless, hygroscopic crystalline solid that is quite soluble in water. Its chemical formula is C2H4O3. It is used in various skin care products. Glycolate is a salt or ester form of glycolic acid. Glycolic acid is a type of AHA (alpha hydroxy acid). AHA mainly exists in fruits and refers to an organic acid derived from lactose. It is a substance that has an exfoliating effect and a skin moisturizing effect, and in addition to glycolic acid, there are lactic acid, citric acid, malic acid, etc. The glycolic acid is not limited to the method of obtaining it, and can be synthesized by a method known in the art or a commercially available substance can be used.

[0091]

[0092] The term "gluconolactone" above, also known as glucono-delta-lactone (GDL), is a food additive used as a sequestrant, acidifier, curing agent, acidifier, or leavening agent. Pure gluconolactone is a white, odorless, crystalline powder. When added to water, gluconolactone partially decomposes into gluconic acid, and the balance between the lactone form and the acid form is achieved through chemical equilibrium. Gluconolactone is a type of PHA (poly hydroxy acid). PHA has the advantage of having a large molecular weight, so that less is absorbed into the skin, resulting in less irritation. In addition to gluconolactone, there is lactobionic acid derived from corn starch. The gluconolactone is not limited by the method of obtaining it, and can be chemically synthesized by a method known in the art, or a commercially available substance can be used.

[0093]

[0094] The term "salicylic acid" above refers to an organic acid, also known as salicylic acid, and is widely used in the synthesis of organic acids. It is a plant hormone and is also frequently used as a pharmaceutical. It is very acidic with a pH of 2.4. Its chemical formula is C7H6O3. Salicylic acid is a type of BHA (beta hydroxy acid). BHA is fat-soluble and is used to remove sebum and waste products. Salicylic acid extracted from the bark of the western white willow is a representative example. The method of obtaining the salicylic acid is not limited, and it can be chemically synthesized by a method known in the art or a commercially available substance can be used.

[0095]

[0096] 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 skin-dispersible dark spot improvement material of the present invention may be contained in an amount of about 0.0001 to 10 wt%, about 0.0005 to 5 wt%, about 0.001 to 5 wt%, about 0.01 to 5 wt%, about 0.0001 to 3 wt%, about 0.0005 to 3 wt%, about 0.001 to 3 wt%, about 0.01 to 3 wt%, about 0.0001 to 1 wt%, about 0.0005 to 1 wt%, about 0.001 to 1 wt%, or about 0.01 to 1 wt% based on the total weight of the composition.

[0097]

[0098] For the purpose of the present invention, the above material has a skin-dispersing dark spot improvement effect.

[0099] For the purpose of the present invention, the term 'dark spot' means that some areas of an individual's skin become pigmented due to melanin overproduction, melanin accumulation, keratin accumulation, oxidized protein and lipid accumulation, etc., resulting in dark spots with lower skin saturation than other areas. When such spots are spread symmetrically across the entire face, such as on the cheeks, cheekbones, and under the eyes, they are called 'diffuse dark spots'. 'Diffuse dark spots' may include melasma as a representative example, but are not limited thereto, and may include, but are not limited to, spots formed after inflammation, black spots (alveoli, warts, lentigo), freckles, etc.

[0100] Typically, methods for assessing the severity of skin dark spots involve measuring skin tone brightness or color, or skin tone unevenness. However, it is virtually impossible to measure the brightness or color of individual, small, "distributed dark spots" spread over a wide area. Measuring skin tone unevenness can make it difficult to distinguish dark spots from wrinkles, raised dead skin cells, and pores. Therefore, defining "distributed dark spots" using skin tone brightness, color, or unevenness has limitations. Visual assessment is the most appropriate way to express skin concerns perceived by subjects, but it lacks objectivity and is difficult for a small number of evaluators to evaluate a large number of subjects. To address this, the present invention defines "distributed dark spots" by building a diagnostic AI that learns from visual evaluation results. Evaluation using diagnostic AI is better suited to show the subjective visual extent of "distributed dark spots" than objective quantitative assessments, and has the advantage of being able to evaluate a variety of subjects more consistently than visual evaluation.

[0101] For the purposes of the present invention, the above substance may be characterized by a reduction in melanin production. Specifically, treatment with bisabolol, nutmeg extract, or diisopropyl adipate among the above substances may promote a reduction in melanin.

[0102] For example, when a cream containing a mixture of the substance of the present invention, including bisabolol, nutmeg extract, and diisopropyl adipate, which are effective in reducing melanin production, was applied to the face, it was confirmed that a superior rate of improvement in pigmentation was observed compared to the control group, vitamin C.

[0103]

[0104] For the purpose of the present invention, the substance may be characterized by inhibiting the expression of AKAP1 or promoting the expression of RAB11FIP2 or PPARGC1B. Specifically, when cysteine, cysteamine HCL, nutmeg extract, high molecular weight hyaluronic acid, and / or ultra-low molecular weight hyaluronic acid are treated among the substances, the expression of AKAP1 can be inhibited, when bisabolol, Korean knotweed extract, and / or diisopropyl adipate are treated, the expression of RAB11FIP2 can be promoted, and when niacinamide, glycolic acid, gluconolactone, and / or salicylic acid are treated, the expression of PPARGC1B can be promoted, but is not limited thereto. It can be seen that the responsiveness to the material of the present invention varies depending on a specific genetic polymorphism type, and it is suggested that the efficiency of improving skin diffuse dark spots can be increased by providing a customized prescription to a high-risk group with a high risk level of developing skin diffuse dark spots.

[0105]

[0106] 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.

[0107]

[0108] The above composition is characterized by being personalized. Specifically, it can provide a personalized substance for an individual having a single nucleotide polymorphism (SNP) marker significantly associated with a specific degree of skin-dispersed dark spots.

[0109] For example, the single nucleotide polymorphism (SNP) markers significantly associated with the degree of skin diffuse dark spots may be, but are not limited to, one or more selected from Table 1.

[0110] 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.

[0111] 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.

[0112] Specifically, the genetic polymorphism markers of the present invention are significantly associated with skin-dispersed dark spots, 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.

[0113] In the case where an individual has an allele of a polymorphic marker with a significant association with diffuse dark spots on the skin, the effect of improving diffuse dark spots on the skin 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 degree of improvement in diffuse dark spots on the skin, can be measured to have a high or low skin characteristic, and thus, information can be provided for selecting a customized substance for increasing improvement in diffuse dark spots on the skin in an individual that includes one or more of the minor alleles.

[0114]

[0115] 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.

[0116] The above single nucleotide polymorphism marker may be one or more single nucleotide polymorphism markers selected from the single nucleotide polymorphism markers shown in Table 1. The single nucleotide polymorphism markers shown in Table 1 may be used to determine the degree of correlation with the degree of skin-dispersed dark spots.

[0117] The degree of skin-dispersed dark spots of the single nucleotide polymorphism markers of the present invention was determined by measuring the frequency of each marker. Such significance is characterized by a p-value such as, but not limited to, a p-value of less than 0.05, less than 0.01, less than 0.001, less than 0.0001, less than 0.00001, less than 0.0000001, 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.

[0118]

[0119] The single nucleotide polymorphism (SNP) marker of the present invention may be one or more of the markers shown in Table 1, but is not limited thereto. The single nucleotide polymorphism (SNP) marker may be one or more, and may be used in combinations of two or more, three or more, four or more, five or more, or six or more, as a number capable of determining skin-dispersed dark spots, but is not limited thereto.

[0120] 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.

[0121] For example, the single nucleotide polymorphism marker significantly associated with the degree of skin diffuse dark spots may be one or more single nucleotide polymorphism markers present in a gene encoding a protein of AKAP1, RAB11FIP2, PPARGC1B, BNC2, MC1R or MITF, and specifically, may be one or more selected from the group consisting of rs11079281, rs10444110, rs251466, rs16935073, rs2228479 and rs7430957, but is not limited thereto.

[0122] For example, if the SNP ID is rs11079281, Chr.Position (GRCh ver. 37) is described as "17:55231787" and the Allele is disclosed as T>C, this indicates that the 55231787th base on the 17th chromosome of humans is T or C, 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.

[0123] In one specific example, the marker selected from Table 1 is a polynucleotide comprising 5-100 contiguous DNA sequences including base 55231787 of chromosome 17 of a human, wherein base 55231787 is T or C (rs11079281); a polynucleotide comprising 5-100 contiguous DNA sequences including base 119563401 of chromosome 10 of a human, wherein base 119563401 of chromosome 10 of a human, wherein base 119563401 of chromosome 10 of a human, wherein base 119563401 of chromosome 10 of a human, wherein base 149195603 of chromosome 5 ... The present invention may be, but is not limited to, a polynucleotide comprising 5 to 100 contiguous DNA sequences including the 16795790th base of human chromosome 9, wherein the 16795790th base is A or C (rs16935073); a polynucleotide comprising 5 to 100 contiguous DNA sequences including the 89985940th base of human chromosome 16, wherein the 89985940th base is G or A (rs2228479); a polynucleotide comprising 5 to 100 contiguous DNA sequences including the 69978466th base of human chromosome 3, wherein the 69978466th base is A or G (rs7430957); and a polynucleotide selected from the group consisting of one or more polynucleotides and one or more polynucleotides selected from the group consisting of complementary polynucleotides thereof.

[0124]

[0125] The composition of the present invention can be used as a cosmetic composition for alleviating or improving skin-dispersible dark spots, 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 solutions, external ointments, creams, foams, nourishing toners, emollients, packs, emollients, emulsions, makeup bases, essences, soaps, liquid cleansers, bath products, sunscreen creams, sun oils, suspensions, emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansers, oils, powder foundations, emulsion foundations, wax foundations, patches, and sprays, but is not limited thereto.

[0126] 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.

[0127] The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation.

[0128] 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.

[0129] 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.

[0130] 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.

[0131] 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.

[0132] When the formulation of the present invention is soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolysates, isethionates, lanolin derivatives, fatty alcohols, vegetable oils, glycerol, sugars, etc. can be used as carrier components.

[0133]

[0134] In another aspect, the composition provides an over-the-counter drug composition for alleviating or improving skin-dispersing dark spots.

[0135] 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.

[0136] 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.

[0137]

[0138] In another aspect, the present invention provides a method for providing information for selecting a customized substance for improving skin-dispersed dark spots, comprising the steps of: (a) obtaining a biological sample from an individual; (b) amplifying or hybridizing with a probe a polymorphic region of at least one single nucleotide polymorphism marker selected from the group consisting of rs11079281, rs10444110, rs251466, rs16935073, rs2228479, and rs7430957 from the biological sample; and (c) identifying a base of the amplified or hybridized polymorphic region of the step (b).

[0139] The above-mentioned selected customized material can be applied to an individual having a specific polymorphic area to improve diffuse dark spots on the skin.

[0140]

[0141] In the present invention, the term "subject" refers to a subject for diagnosing the degree of diffuse dark spots on the skin. DNA may be obtained from samples such as, but not limited to, hair, urine, blood, various body fluids, isolated tissue, isolated cells, or saliva.

[0142] Any method known to those skilled in the art can be used to obtain genomic DNA in the above step (a).

[0143] 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.

[0144] 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.

[0145] The above substance may be at least one selected from the group consisting of phloretin, nutmeg extract, Korean knotweed extract, bisabolol, diisopropyl adipate, cysteine, cysteamine HCL, high molecular weight hyaluronic acid, ultra-low molecular weight hyaluronic acid, niacinamide, glycolic acid, gluconolactone, and salicylic acid, but is not limited thereto.

[0146] For the purpose of the present invention, the significant genes associated with the degree of the above-mentioned diffuse dark spots on the skin 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 AKAP1, RAB11FIP2, PPARGC1B, BNC2, MC1R or MITF protein, when one or more substances selected from the group consisting of nutmeg extract, Japanese knotweed extract, bisabolol, diisopropyl adipate, cysteine, cysteamine HCL, high molecular weight hyaluronic acid, ultra-low molecular weight hyaluronic acid, niacinamide, glycolic acid, gluconolactone and salicylic acid are treated, the improvement of diffuse dark spots on the skin can be increased, but the optimal combination of substances can be provided according to the genetic characteristics of each individual.

[0147] This suggests that it may be possible to identify specific single nucleotide polymorphism (SNP) markers that are significantly correlated with diffuse dark spots on the skin and provide information to select personalized substances that can maximize the effect of improving diffuse dark spots on the skin based on genetic information.

[0148] Specifically, if the risk score obtained by multiplying and adding the genotypes of rs11079281, rs10444110, rs251466, rs16935073, rs2228479, and rs7430957 by the effect size (β) is less than 0.28, the risk of developing diffuse dark spots on the skin may be classified as a low-risk group, and if the risk score is 0.28 or more, the risk of developing diffuse dark spots on the skin may be classified as a high-risk group. As an example, when the risk of developing diffuse dark spots on the skin is a high-risk group, an information providing method may be provided to select bisabolol and nutmeg extract as customized substances, but is not limited thereto.

[0149]

[0150] 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.

[0151] 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.

[0152] 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.

[0153] 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.

[0154] 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.

[0155]

[0156] 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 skin-dispersed dark spots.

[0157] In another aspect, the present invention provides a composition for diagnosing the degree of a skin-dispersed dark spot, comprising a probe or an agent capable of detecting a single nucleotide polymorphism (SNP) marker for diagnosing the degree of a skin-dispersed dark spot.

[0158]

[0159] A diffuse dark spot on the skin refers to, but is not limited to, the result of measuring the ratio of all target areas to the entire measured area when a specific area of ​​the skin has an uneven skin color due to the influence of ultraviolet rays or hormones and is observed to be relatively darker than the surrounding skin. Specifically, it refers to determining whether the skin is prone to the creation of diffuse dark spots using one or more single nucleotide polymorphism (SNP) markers selected from Table 1. More specifically, a skin type prone to the creation of diffuse dark spots on the skin refers to a skin type that has a high rate of blemishes created per skin area.

[0160] 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.

[0161]

[0162] In the present invention, the term "probe capable of detecting a marker for diagnosing the degree of skin diffuse dark spots" refers to a composition capable of diagnosing the degree of skin diffuse dark spots by specifically hybridizing with a polymorphic region of the gene as described above, and the specific method of such gene analysis is not particularly limited and may be any gene detection method known in the technical field to which this invention pertains. In addition, the term may be used interchangeably with the terms "for diagnosing whether a skin type is prone to developing skin diffuse dark spots" or "for diagnosing whether a high-risk skin type is prone to developing skin diffuse dark spots."

[0163] In the present invention, the term "agent capable of amplifying a marker for diagnosing the degree of skin diffuse dark spots" refers to a composition capable of diagnosing the degree of skin diffuse dark spots by confirming the polymorphic region of the gene as described above through amplification, and specifically refers to a primer capable of specifically amplifying a polynucleotide of the marker for diagnosing the degree of skin diffuse dark spots. In addition, the term may be used interchangeably with the terms "for diagnosing whether a skin type is prone to developing skin diffuse dark spots" or "for diagnosing whether a high-risk skin type is prone to developing skin diffuse dark spots."

[0164] 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.

[0165] 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.

[0166] The probes or primers of the present invention can be chemically synthesized using the phosphoramidite solid support method or other well-known methods. These nucleic acid sequences can also be modified using many means known in the art. Non-limiting examples of such modifications include methylation, "capping," substitution with one or more homologs of a natural nucleotide, and modifications between nucleotides, such as modification with uncharged linkers (e.g., methyl phosphonate, phosphotriester, phosphoroamidate, carbamate, etc.) or charged linkers (e.g., phosphorothioate, phosphorodithioate, etc.).

[0167]

[0168] In another aspect, the present invention provides a kit for diagnosing the degree of a skin-dispersible dark spot, comprising the composition for diagnosing the degree of a skin-dispersible dark spot. The kit may be, but is not limited to, an RT-PCR kit or a DNA chip kit.

[0169] The kit of the present invention can diagnose the degree of skin diffuse dark spots by amplifying a single nucleotide polymorphism (SNP) marker, which is a marker for diagnosing the degree of skin diffuse dark spots, or by determining 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 skin diffuse dark spots 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 skin diffuse dark spots, 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 skin-dispersed dark spot diagnosis kit comprising the 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 on 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.

[0170]

[0171] In another aspect, the present invention provides a microarray for diagnosing the degree of a skin-dispersed dark spot, comprising the composition for diagnosing the degree of a skin-dispersed dark spot.

[0172] 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.

[0173] Methods for manufacturing microarrays by immobilizing probe polynucleotides on a substrate are well known in the art. The probe polynucleotide refers to a hybridizable polynucleotide, and refers to an oligonucleotide capable of sequence-specific binding to complementary strands of nucleic acids. The probe of the present invention is an allele-specific probe, which hybridizes to a DNA fragment derived from one member of the same species but not to a fragment derived from the other member due to the presence of a polymorphic site in the nucleic acid fragments derived from two members of the same species. In this case, hybridization conditions must be sufficiently stringent to ensure that only one of the alleles hybridizes, as there is a significant difference in hybridization intensity between alleles. This can induce a favorable hybridization difference between different allele forms. The probe of the present invention can be used in methods for detecting alleles and for diagnosing skin diffuse dark spots, etc. These diagnostic methods include detection methods based on nucleic acid hybridization, such as Southern blot, and in methods using DNA chips, the probe may be provided in a form pre-bound to the substrate of the DNA chip. The above hybridization can usually be performed under stringent conditions, for example, a salt concentration of 1 M or less and a temperature of 25°C or higher. For example, conditions of 5x SSPE (750 mM NaCl, 50 mM Na Phosphate, 5 mM EDTA, pH 7.4) and 25-30°C may be suitable for allele-specific probe hybridization.

[0174] The process of immobilizing the probe polynucleotide associated with the degree of skin-dispersible dark spots 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, followed by hybridization on a microarray and detection of the signal generated from the label.

[0175]

[0176] In another aspect, the present invention provides a method for providing information on the degree of skin-dispersed dark spots, 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).

[0177] The above terms, 'object', (a) step, (b) step' are as described above.

[0178] The above method may additionally determine, but is not limited to, a high-risk group with a high risk of developing skin-dispersed dark spots if the risk score obtained by multiplying and adding the effect size (β) of the (c) amplified or hybridized single nucleotide polymorphism marker is 0.28 or higher.

[0179]

[0180] In one embodiment to achieve the object of the present invention, the present invention provides a composition for improving dark skin tone. For example, the composition comprises, as an active ingredient, at least one substance selected from the group consisting of polydatin, ferulic acid, bakuchiol, genistein, oil-soluble licorice extract, Aspergillus ferment, quercetin, tannic acid, ZnPCA, gamma oryzanol, chrysin, tranexamic acid, sunflower oil, and isostearic acid. As another example, the composition may include, but is not limited to, (i) ferulic acid, polydatin, zinc PCA, tranexamic acid, quercetin, genistein, tannic acid, Aspergillus ferment, gamma-oryzanol, sunflower seed oil, and isostearic acid; (ii) ferulic acid, polydatin, bakuchiol, genistein, oil-soluble licorice extract, and Aspergillus ferment; (iii) ferulic acid, quercetin, genistein, tannic acid, polydatin, and zinc PCA, gamma-oryzanol; or (iv) chrysin, tannic acid, and zinc PCA as active ingredients.

[0181]

[0182] The above term 'polydatin' is also called piceid or 3,4',5-trihydroxystilbene-3-β mono-D glucoside, and is a stilbene polyphenol isolated from 'Polygonum cuspidatum', and a naturally derived compound, a chemically synthesized compound, or a mixture thereof can be used. In addition, the polydatin can be trans-polydatin, cis-polydatin, or a mixture thereof. The following chemical formula 5 represents polydatin in the trans form, and chemical formula 6 represents polydatin in the cis form. The polydatin 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 material may be used.

[0183] [Chemical Formula 5]

[0184]

[0185] [Chemical Formula 6]

[0186]

[0187] The term "ferulic acid" refers to a polyphenol compound found in many food crops, including whole grains, fruits, and seaweed. It is also called 4-hydroxy-3-methoxycinnamic acid. While research on the anti-diabetic and anti-hyperglycemic effects of ferulic acid has focused on peripheral tissues, its effects on the central nervous system have not yet been elucidated. The method of obtaining ferulic acid is not limited, and it can be chemically synthesized using methods known in the art, or a commercially available substance can be used. The INCI name for this substance is ferulic acid.

[0188] [Chemical Formula 7]

[0189]

[0190]

[0191] The term "Bakuchiol" above refers to a substance extracted from a wild plant used in Ayurveda and Chinese medicine in Asia. Bakuchiol is a plant-based retinol that does not cause skin irritation or side effects. Compared to conventional retinols that can cause skin irritation, sensitization, and dermatitis, it avoids stability issues, has high photostability, and can exhibit the same efficacy as retinol on aged skin. In addition, bakuchiol can exhibit a more excellent collagen synthesis effect and skin wrinkle improvement effect than retinol, and therefore, a cream-type cosmetic composition containing bakuchiol as an active ingredient can effectively improve problematic skin such as acne without skin irritation, and exhibit skin whitening and moisturizing effects. Bakuchiol can be chemically synthesized and obtained using various extraction methods known in the art, and, if necessary, can be obtained through an additional purification process.

[0192] [Chemical Formula 8]

[0193]

[0194]

[0195] 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.

[0196] [Chemical Formula 9]

[0197]

[0198]

[0199] The above term 'oil soluble licorice extract' is one of the raw materials for skin whitening notification, and is an oil soluble extract extracted from the roots of licorice, Glycyrrhiza glabra L. var. glandulifera Regal et Herder, Glycyrrhiza uralensis Fisher, or other closely related plants (leguminosae). It is mainly used in powdered form, but is not limited thereto.

[0200]

[0201] The above term 'Aspergillus ferment' refers to a substance obtained through the fermentation of the microorganism Aspergillus (Mold), and can be used as is or after filtering. Aspergillus ferment contains large amounts of ferulic acid and phenolic acid, which have DPPH radical scavenging properties, and thus have antioxidant effects. At the same time, they can inhibit tyrosinase and elastase activities, which can be effective in skin whitening and improving wrinkles.

[0202]

[0203] 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.

[0204] [Chemical Formula 10]

[0205]

[0206]

[0207] 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 utilized 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 methods known in the art, or a commercially available substance can be used. The INCI name of the substance is tannic acid.

[0208] [Chemical Formula 11]

[0209]

[0210]

[0211] 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.

[0212]

[0213] The above term 'gamma oryzanol' is a growth promoting substance contained in grains, isolated from rice bran. It has a strong antioxidant effect, which helps prevent the damaging effects of radiation exposure or chemotherapy. In addition, this component increases cerebral blood flow by regulating blood pressure and strengthening blood vessel walls, thereby improving brain cell metabolic function, improving autonomic nervous system dysfunction, preventing stroke and dementia, enhancing memory, and improving sleep quality, and has the effect of improving cerebrovascular diseases. In addition, it has the function of activating liver function and promoting alcohol metabolism, so it is used in hangover relief drinks. In addition, it is known to have an effect of relieving obesity and promoting kidney function. The above gamma oryzanol is not limited to the method of obtaining it, and can be chemically synthesized by a method known in the art, or a commercially available substance can be used. The INCI name of the above substance is Oryzanol.

[0214]

[0215] The above term 'Chrysin' is also called 5,7-dihydroxyflavone and is one of the flavonoids found in honey and propolis. Chrysin is C 15 H 10 It can be represented by the chemical formula O4 and the following chemical formula 12. The above chrysin 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.

[0216] [Chemical Formula 12]

[0217]

[0218]

[0219] The above term 'Tranexamic acid (TXA)' refers to trans-4-(aminomethyl)cyclohexanecarboxylic acid, C8H 15It has the molecular formula of NO2 and is known to have an anti-fibrinolytic effect, so it is used as a hemostatic or thrombolytic agent. Recently, its industrial value as a whitening ingredient has been proven as its inhibitory effect on the proliferation of skin pigment cells 'melanocytes' and melanin production induced by ultraviolet rays. The above tranexamic acid is not limited to the method of obtaining it, and it can be chemically synthesized by a method known in the art, or a commercially available substance can be used. The INCI name of the above substance is tranexamic acid.

[0220] [Chemical Formula 13]

[0221]

[0222]

[0223]

[0224] The term "sunflower oil" above refers to oil extracted from sunflower seeds. Sunflower seed oil is rich in nutrients that are good for the body, such as protein, unsaturated fatty acids such as linoleic acid and oleic acid, lecithin, carotenoids, tocopherols, vitamins A, C, and D, and dietary fiber. Sunflower seed oil is very high in vitamin E. Vitamin E is an antioxidant that prevents skin damage caused by harmful substances such as ultraviolet rays and helps alleviate scars and wrinkles, making the skin healthy and beautiful. It is also rich in beta-carotene, which is converted to vitamin A, an antioxidant, and is well known to delay aging and keep the skin young and firm. The sunflower seed oil is not limited by the method of acquisition, and may be extracted by a method known in the art or may use a commercially available substance.

[0225]

[0226] The above term 'Isostearic Acid' is manufactured through a multi-stage separation and purification process under high temperature and high pressure using fatty acids as raw materials. The mixed isostearic acid manufactured through this multi-stage process requires high purity and is generally manufactured into isostearyl alcohol rather than used as isostearic acid itself. Its main uses include cosmetics, lubricants, solvents, and metal treatment agents. The above isostearic 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 substance can be used. The INCI name of the above substance is isostearic acid.

[0227] 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 material for improving dark skin tone 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.

[0228]

[0229] For the purpose of the present invention, the above material has the effect of improving dark skin tone.

[0230] For the purposes of the present invention, the term "dark skin tone" means that an individual's skin becomes darker than the individual's original skin color due to pigmentation caused by excessive melanin production, melanin accumulation, keratin accumulation, oxidized protein and lipid accumulation, etc. The skin tone may be used interchangeably with skin color, and may refer to the overall skin tone including, but not limited to, skin color classified by brightness (skin brightness), redness, and yellowness. In addition, it may include skin texture including the degree of unevenness of skin color.

[0231] For the purposes of the present invention, the above-mentioned substance may be characterized by a reduction in melanin production. Specifically, when the above-mentioned substance is treated with i) ferulic acid, soluble licorice extract, and bakuchiol; or ii) polydatin and quercetin, a reduction in melanin can be promoted.

[0232] For example, when a cream containing a mixture of the substances of the present invention, including i) ferulic acid, a soluble licorice extract, and bakuchiol, which are effective in reducing melanin production; or ii) polydatin and quercetin, was applied to the face, it was confirmed that a synergistic effect in reducing melanin production was observed compared to when the substances were applied individually.

[0233]

[0234] For the purposes of the present invention, the above material may be characterized by a reduction in protein carbonylation. Specifically, when gamma-oryzanol, Aspergillus ferment, or isostearic acid is treated among the above materials, a reduction in carbonylation can be promoted.

[0235]

[0236] For the purpose of the present invention, the substance may be characterized by inhibiting the expression of OCA2, BNC2, RAB32 or KITLG. Specifically, when the substance is treated with ferulic acid, polydatin, zinc PCA, tranexamic acid, quercetin, genistein, tannic acid, Aspergillus ferment, gamma-oryzanol, sunflower seed oil and / or isostearic acid, the expression of OCA2 is inhibited, when polydatin, ferulic acid, bakuchiol, genistein, oil-soluble licorice extract and / or Aspergillus ferment, the expression of BNC2 is inhibited, when ferulic acid, quercetin, genistein, tannic acid, polydatin, zinc PCA and / or gamma-oryzanol are treated, the expression of RAB32 is inhibited, and when chrysin, tannic acid or zinc PCA is treated, the expression of KITLG may be inhibited, but is not limited thereto. It can be seen that the responsiveness to the material of the present invention varies depending on a specific genetic polymorphism type, suggesting that the efficiency of improving dark skin tone can be increased by providing a customized prescription to a high-risk group with a high risk level of developing dark skin tone.

[0237]

[0238] In the present invention, the term "about" is as described above.

[0239]

[0240] The composition is characterized by being personalized. Specifically, it can provide a personalized substance targeting an individual having a single nucleotide polymorphism (SNP) marker significantly associated with a specific degree of dark skin tone.

[0241] For example, the significant single nucleotide polymorphism (SNP) marker associated with dark skin tone may be, but is not limited to, one or more of those selected from Table 2.

[0242] In the present invention, the terms “polymorphism”, “allele” and “rs_id” are as described above.

[0243] Specifically, the genetic polymorphism markers of the present invention are significantly associated with dark skin tone, 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.

[0244] In the case where an individual has an allele of a significant polymorphism marker associated with dark skin tone, the effect of improving dark skin tone 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 degree of improvement in dark skin tone, can be measured to have a high or low skin characteristic, and thus, information can be provided for selecting a customized substance for increasing improvement in dark skin tone in an individual that includes one or more minor alleles.

[0245] 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 2. The single nucleotide polymorphism markers shown in Table 2 may be used to determine the degree of correlation with the degree of dark skin tone.

[0246] The degree of dark skin tone of the single nucleotide polymorphism marker of the present invention was determined by measuring the frequency of each marker. Such significance is characterized by a p-value such as, but not limited to, a p-value of less than 0.05, less than 0.01, less than 0.001, less than 0.0001, less than 0.00001, less than 0.0000001, 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.

[0247] The single nucleotide polymorphism (SNP) marker of the present invention may be one or more of the markers shown in Table 2, but is not limited thereto. The single nucleotide polymorphism (SNP) marker may be one or more, and may be used in a combination of two or more, three or more, four or more, or five or more, as a number capable of determining a dark skin tone, but is not limited thereto.

[0248] 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.

[0249] For example, the significant single nucleotide polymorphism marker associated with the degree of dark skin tone may be one or more single nucleotide polymorphism markers present in a gene encoding a protein of OCA2, BNC2, RAB32, KITLG or ASIP, and specifically, may be one or more selected from the group consisting of rs74653330, rs16935073, rs72620727, rs77310623 and rs6059711, but is not limited thereto.

[0250] For example, if the SNP ID is rs74653330, Chr.Position (GRCh ver. 37) is described as "15:28228553" and the Allele is disclosed as T>C, this indicates that the 28228553 base on the 15th chromosome of humans is T or C, 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.

[0251] In one specific example, the marker selected from Table 2 is a polynucleotide comprising 5-100 contiguous DNA sequences including base 28228553 of chromosome 15 of a human, wherein base 28228553 is T or C (rs74653330); a polynucleotide comprising 5-100 contiguous DNA sequences including base 16795790 of chromosome 9 of a human, wherein base 16795790 is C or A (rs16935073); a polynucleotide comprising 5-100 contiguous DNA sequences including base 89110410 of chromosome 12 of a human, wherein base 89110410 is C or A (rs72620727); The present invention may be, but is not limited to, a polynucleotide comprising 5 to 100 contiguous DNA sequences including the 146871819th base of human chromosome 6, wherein the 146871819th base is A or G (rs77310623); a polynucleotide comprising 5 to 100 contiguous DNA sequences including the 32780822nd base of human chromosome 20, wherein the 32780822nd base is T or C (rs6059711); and a polynucleotide selected from the group consisting of one or more polynucleotides and one or more polynucleotides selected from the group consisting of complementary polynucleotides thereof.

[0252]

[0253] The composition of the present invention can be used as a cosmetic composition for alleviating or improving dark skin tone, as described above.

[0254]

[0255] In another aspect, the composition provides an over-the-counter drug composition for alleviating or improving dark skin tone. The term "over-the-counter drug" in the present invention is as described above.

[0256]

[0257] In another aspect, the present invention provides a method for providing information for selecting a customized substance for improving dark skin tone, comprising the steps of: (a) obtaining a biological sample from an individual; (b) amplifying or hybridizing with a probe a polymorphic site of at least one single nucleotide polymorphism marker selected from the group consisting of rs74653330, rs16935073, rs72620727, rs77310623, and rs6059711 from the biological sample; and (c) identifying a base of the amplified or hybridized polymorphic site of step (b).

[0258] The above-mentioned selected customized material can be applied to an individual having a specific polymorphic region to improve dark skin tone.

[0259] In the present invention, the term "subject" refers to a subject for diagnosing skin tone darkness. DNA may be obtained from, but is not limited to, samples such as hair, urine, blood, various bodily fluids, isolated tissue, isolated cells, or saliva. Steps (a) and (b) above are also as described above.

[0260] The above substance may be at least one selected from the group consisting of polydatin, ferulic acid, bakuchiol, genistein, soluble licorice extract, aspergillus ferment, quercetin, tannic acid, zinc PCA, gamma-oryzanol, chrysin, tranexamic acid, sunflower seed oil, and isostearic acid, but is not limited thereto.

[0261] For the purpose of the present invention, the significant genes associated with the degree of dark skin tone can provide information for selecting specific substances for each individual. For example, in the case of an individual having one or more single nucleotide polymorphism markers present in a gene encoding OCA2, BNC2, RAB32, KITLG or ASIP protein, when treating one or more substances selected from the group consisting of polydatin, ferulic acid, bakuchiol, genistein, soluble licorice extract, aspergillus ferment, quercetin, tannic acid, zinc PCA, gamma-oryzanol, chrysin, tranexamic acid, sunflower seed oil and isostearic acid, improvement in dark skin tone can be increased, but an optimal combination of substances can be provided according to the genetic characteristics of each individual.

[0262] This suggests that it may be possible to identify specific single nucleotide polymorphism (SNP) markers that are significantly correlated with dark skin tone and provide information to select personalized substances that can maximize the effect of improving dark skin tone based on genetic information.

[0263] Specifically, if the risk score obtained by multiplying and adding the effect size (β) to the genotypes of rs74653330, rs16935073, rs72620727, rs77310623, and rs6059711 is less than -0.193, the risk of developing a dark skin tone may be classified as a low-risk group, and if the risk score is -0.193 or more, the risk of developing a dark skin tone may be classified as a high-risk group. As an example, if the risk of developing a dark skin tone is a high-risk group, an information providing method may be provided to select isostearic acid, sunflower seed oil, and polydatin as customized substances, but is not limited thereto.

[0264]

[0265] In the present invention, the term “SNP chip” is as described above.

[0266]

[0267] As one aspect for achieving the object of the present invention, the present invention provides a single nucleotide polymorphism (SNP) marker for diagnosing the degree of dark skin tone.

[0268] In another aspect, the present invention provides a composition for diagnosing the degree of dark skin tone, comprising a probe or an agent capable of detecting or amplifying a single nucleotide polymorphism (SNP) marker for diagnosing the degree of dark skin tone.

[0269]

[0270] Dark skin tone refers to, but is not limited to, a darkening of the overall skin tone compared to an individual's original skin color due to factors such as ultraviolet rays, hormones, and aging. Specifically, it refers to determining whether a person is prone to developing a dark skin tone using one or more single nucleotide polymorphism (SNP) markers selected from Table 2. More specifically, a skin type prone to developing a dark skin tone refers to a skin type whose overall skin tone is more likely to darken due to factors such as ultraviolet rays, hormones, and aging.

[0271] 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.

[0272]

[0273] In the present invention, the term "probe capable of detecting a marker for diagnosing the degree of dark skin tone" refers to a composition capable of diagnosing the degree of dark skin tone by specifically hybridizing with a polymorphic region of the gene as described above, and the specific method of such genetic analysis is not particularly limited and may be any genetic detection method known in the technical field to which this invention pertains. In addition, the term may be used interchangeably with the terms "for diagnosing whether a skin type is prone to developing a dark skin tone" or "for diagnosing whether a high-risk skin type is prone to developing a dark skin tone."

[0274] In the present invention, the term "agent capable of amplifying a marker for diagnosing the degree of dark skin tone" refers to a composition capable of diagnosing the degree of dark skin tone by confirming the polymorphic site of the gene as described above through amplification, and specifically refers to a primer capable of specifically amplifying the polynucleotide of the marker for diagnosing the degree of dark skin tone. In addition, the term may be used interchangeably with the terms "for diagnosing whether a skin type is prone to developing a dark skin tone" or "for diagnosing whether a high-risk skin type is prone to developing a dark skin tone."

[0275] 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.

[0276] In the present invention, the term “primer” is as described above.

[0277]

[0278] In another aspect, the present invention provides a kit for diagnosing dark skin tone, comprising the composition for diagnosing dark skin tone. The kit may be, but is not limited to, an RT-PCR kit or a DNA chip kit.

[0279]

[0280] In another aspect, the present invention provides a microarray for diagnosing dark skin tone, comprising the composition for diagnosing dark skin tone.

[0281] The process of immobilizing the probe polynucleotide associated with the degree of dark skin tone of the present invention onto a substrate can also be easily manufactured using these conventional techniques. Furthermore, hybridization of nucleic acids on microarrays and detection of hybridization results are well known in the art. For example, the detection can be accomplished by labeling a nucleic acid sample with a label capable of generating a detectable signal, such as a fluorescent substance such as Cy3 or Cy5, hybridizing the nucleic acid sample onto a microarray, and detecting the signal generated from the label.

[0282]

[0283] In another aspect, the present invention provides a method for providing information on the degree of dark skin tone, 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).

[0284] The above terms, 'object', (a) step, (b) step' are as described above.

[0285] The above method may additionally determine, but is not limited to, a high-risk group with a high risk of developing dark skin tone when the risk score obtained by multiplying and adding the effect size (β) of the amplified or hybridized single nucleotide polymorphism marker (c) is -0.193 or higher.

[0286]

[0287] In one embodiment to achieve the object of the present invention, the present invention provides a composition for improving spot-like pigmentation. For example, the composition comprises, as an active ingredient, at least one substance selected from the group consisting of ascorbyl glucoside, 3-O-ethyl ascorbic acid, ascorbic acid, Glycyrrhiza uralensis (Licorice) extract, tocopherol, sodium pyruvate, Ecklonia cava extract, hexyldecanol, asiaticoside, and hexylresorcinol. In another example, the composition comprises (i) tocopherol, licorice extract, and Ecklonia cava extract; (ii) sodium pyruvate, hexyldecanol, asiaticoside and hexylresorcinol; or (iii) ascorbyl glucoside, 3-O-ethyl ascorbic acid and ascorbic acid; but is not limited thereto.

[0288]

[0289] The above term 'Ascorbyl Glucoside' is one of the representative derivatives of ascorbic acid, and is mostly synthesized through an enzymatic reaction process. However, as the production cost increases due to the need for optimization of reaction conditions and high-purity purification processes, it is being used as a high value-added cosmetic raw material. Ascorbyl glucoside can be largely divided into alpha (α) and beta (β) forms, and beta ascorbyl glucoside (AA-β-2G) exhibits superior physiological activity due to the difference in the absorption mechanism in the human body. The above ascorbyl glucoside is not limited to the method of obtaining it, and can be chemically synthesized by a method known in the art, or a commercially available substance can be used.

[0290] [Chemical Formula 14]

[0291]

[0292]

[0293] The term "3-O-ethyl ascorbic acid" above is a whitening active ingredient with excellent antioxidant, wrinkle-improving, and anti-wrinkle effects. 3-O-ethyl ascorbic acid has excellent stability due to its unique chemical structure, as the ethyl group forms an ether with the 3-hydroxyl group of ascorbic acid. The method for obtaining the 3-O-ethyl ascorbic acid is not limited, and chemical synthesis using a method known in the art or a commercially available material can be used.

[0294] [Chemical Formula 15]

[0295]

[0296]

[0297] The above term 'ascorbic acid' is a type of water-soluble vitamin, vitamin C. It is also an important ingredient that plays a role in preventing skin aging by exhibiting skin whitening effects, antioxidant effects, and promoting collagen production. Ascorbic acid has a sugar-like structure that is easily oxidized to the form of dihydro-L-ascorbic acid by accepting two electrons, thereby preventing the oxidation of biological components such as proteins by highly reactive free radicals and peroxides in the body. The above ascorbic acid can be chemically synthesized and obtained using various extraction methods known in the art, and can be obtained through an additional purification process if necessary.

[0298] [Chemical Formula 16]

[0299]

[0300]

[0301] The above term 'Glycyrrhiza Uralensis (Licorice) Extract' includes all forms of extracts that can be formed using the extract itself and the extract, such as an extract obtained by extracting the root, fruit, stem or whole licorice of licorice, which has been used as a traditional herbal medicine in the East for a long time, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, a conditioned or purified product of the extract, or a mixture thereof. The method for obtaining the licorice extract is not particularly limited, and extraction can be performed according to a method commonly used in the art.

[0302]

[0303] The above term 'Ecklonia Cava Extract' is an extract of Ecklonia cava, a brown algae belonging to the Laminariales order and the Undaria family. It is known to mainly inhabit the waters off the coast of Jeju Island at a depth of about 10 m in Korea and also inhabits Japan, and is 1 to 2 m long, has a cylindrical stem, and a root-like base. The method for obtaining the above Ecklonia cava extract is not particularly limited, and can be extracted according to a method commonly used in the relevant technical field.

[0304] 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.

[0305] 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.

[0306]

[0307] 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.

[0308]

[0309] The term "sodium pyruvate" refers to sodium pyruvate, a salt of the conjugated anion of pyruvate, also known as pyruvate. Sodium pyruvate has antioxidant properties, energy-generating effects, and the ability to cross the blood-brain barrier. The method of obtaining sodium pyruvate is not limited, and may be chemically synthesized using methods known in the art, or a commercially available material may be used.

[0310]

[0311] The term "hexyldecanol" refers to a synthetic, non-drying fatty alcohol used as an emollient and skin conditioning agent. The method of obtaining hexyldecanol is not limited, and it can be chemically synthesized using methods known in the art, or a commercially available material can be used.

[0312]

[0313] 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.

[0314]

[0315] The term "Hexylresorcinol" above refers to an organic compound with local anesthetic, antiseptic, and antiparasitic properties. Hexylresorcinol is not limited by its method of acquisition, and may be chemically synthesized using methods known in the art, or a commercially available material may be used.

[0316] [Chemical Formula 17]

[0317]

[0318]

[0319] 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 material for improving spot-like pigmentation 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.

[0320]

[0321] For the purpose of the present invention, the above material has an effect of improving spot-like pigmentation.

[0322] For the purposes of the present invention, the term 'spotty pigmentation' means that some areas of an individual's skin become pigmented due to factors such as excessive melanin production, melanin accumulation, keratin accumulation, oxidized protein and lipid accumulation due to aging, UV exposure, etc., and the skin becomes darker and has lower saturation than other areas, resulting in spots. In addition, unlike diffuse pigmentation that occurs when multiple pigments gather together, such as freckles, spotty pigmentation individually causes a difference from the surrounding area. Skin that has become noticeably dark due to spotty pigmentation creates a disharmony with the surrounding area and gives a dull and dirty impression. Spotty pigmentation is not limited to spots caused by melanin, but may also include spots formed after inflammation, lentigos (black warts, age spots), etc.

[0323] Specifically, the above-mentioned spot-like pigmentation may be at least one selected from the group consisting of senile lentigines, liver spots, solar lentigines, black moles, and lentigos, but is not limited thereto.

[0324] Typically, to assess the degree of spot-like pigmentation, methods are used to measure the brightness or color of the skin tone of the spot-like area, or to measure the unevenness of the skin tone of the spot-like area. However, it is virtually impossible to measure the brightness or color of all areas of the face, and it is difficult to distinguish the unevenness of the skin tone of the spot-like area from dark spots such as wrinkles, raised dead skin cells, and pores. Therefore, there are limitations in defining 'skin tone of the spot-like area' using the brightness, color, or unevenness of the skin tone of the spot-like area. Although visual evaluation is most suitable for expressing skin concerns perceived by the subject, it has limitations in that it lacks objectivity and it is difficult for a small number of evaluators to evaluate a large number of subjects. To address this, the present invention defined 'spot-like pigmentation' by building a diagnostic AI that learned the results of visual evaluation. Evaluation through diagnostic AI is more suitable for showing the degree of visual 'spot-like pigmentation' that is subjectively felt than objective quantitative evaluation, and has the advantage of being able to consistently evaluate various subjects compared to visual evaluation.

[0325] For the purposes of the present invention, the above material may be characterized by a reduction in melanin production. Specifically, when treating with i) hexylresorcinol and hexyldecanol; or ii) hexyldecanol and tocopherol, the reduction in melanin can be promoted.

[0326] For example, when a cream containing a mixture of the substances of the present invention, i) hexylresorcinol and hexyldecanol; or ii) hexyldecanol and tocopherol, which are effective in reducing melanin production, was applied to the face, it was confirmed that there was a synergistic effect in reducing melanin production compared to when the substances were applied individually.

[0327]

[0328] For the purpose of the present invention, the substance may be characterized by inhibiting the expression of ASCL2 or TP63, or promoting the expression of PPARGC1B. Specifically, when treated with tocopherol, licorice extract, and / or perilla leaf extract, the expression of ASCL2 may be inhibited, when treated with sodium pyruvate, hexyldecanol, asiaticoside, and / or hexylresorcinol, the expression of TP63 may be inhibited, and when treated with ascorbyl glucoside, 3-O-ethyl ascorbic acid, and / or ascorbic acid, the expression of PPARGC1B may be promoted, 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 improving spotty pigmentation can be increased by providing a customized prescription to a high-risk group with a high risk level of developing spotty pigmentation.

[0329]

[0330] In the present invention, the term "about" is as described above.

[0331]

[0332] The above composition is characterized by being personalized. Specifically, it can provide a personalized substance targeting an individual having a single nucleotide polymorphism (SNP) marker significantly associated with a specific degree of spot-like pigmentation.

[0333] For example, the significant single nucleotide polymorphism (SNP) marker associated with the degree of spot-like pigmentation may be, but is not limited to, one or more selected from Table 3.

[0334] In the present invention, the terms “polymorphism”, “allele” and “rs_id” are as described above.

[0335] Specifically, the genetic polymorphism markers of the present invention are significantly associated with spot-like pigmentation, 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.

[0336] In the case where an individual has an allele of a polymorphic marker with a significant association with spot-like pigmentation, the effect of improving spot-like pigmentation 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 degree of improvement in spot-like pigmentation, can be measured to have skin characteristics with a high or low degree, and thus, information can be provided for selecting a customized substance for increasing improvement in spot-like pigmentation in an individual that includes one or more of the minor alleles.

[0337]

[0338] The above single nucleotide polymorphism marker may be one or more single nucleotide polymorphism markers selected from the single nucleotide polymorphism markers shown in Table 3. The single nucleotide polymorphism markers shown in Table 3 may be used to determine the degree of correlation with the degree of spot-like pigmentation.

[0339] The degree of spot-like pigmentation of the single nucleotide polymorphism marker of the present invention was determined by measuring the frequency of each marker. Such significance is characterized by a p-value such as, but not limited to, a p-value of less than 0.05, less than 0.01, less than 0.001, less than 0.0001, less than 0.00001, less than 0.000001, less than 0.00000001, 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.

[0340]

[0341] The single nucleotide polymorphism (SNP) marker of the present invention may be one or more of the markers shown in Table 3, 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 of markers capable of determining spot-like pigmentation, but is not limited thereto.

[0342] 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.

[0343] For example, the significant single nucleotide polymorphism marker associated with the degree of spot-like pigmentation may be one or more single nucleotide polymorphism markers present in a gene encoding a protein of ASCL2, TP63 or PPARGC1B, and specifically, may be one or more selected from the group consisting of rs251466, rs12222609 and rs9865771, but is not limited thereto.

[0344] For example, if the SNP ID is rs251466, Chr.Position (GRCh ver. 37) is described as "5:149195603" and the Allele is initiated as C>G, this indicates that the 149195603rd base of human chromosome 5 is C 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.

[0345] In one specific example, the marker selected from Table 3 is a polynucleotide comprising 5-100 contiguous DNA sequences including base 149195603 of chromosome 5 of a human, wherein base 149195603 is C or G (rs251466); a polynucleotide comprising 5-100 contiguous DNA sequences including base 2297093 of chromosome 11 of a human, wherein base 2297093 is C or T (rs12222609); a polynucleotide comprising 5-100 contiguous DNA sequences including base 189491453 of chromosome 3 of a human, wherein base 189491453 is A or G (rs9865771); and may be composed of one or more polynucleotides selected from the group consisting of their complementary polynucleotides and one or more polynucleotides selected from the group consisting of their complementary polynucleotides, but is not limited thereto.

[0346]

[0347] The composition of the present invention can be used as a cosmetic composition for alleviating or improving spot-like pigmentation, as described above.

[0348]

[0349] In another aspect, the composition provides an over-the-counter drug composition for alleviating or improving spot-like pigmentation. The term "over-the-counter drug" in the present invention is as described above.

[0350]

[0351] In another aspect, the present invention provides a method for providing information for selecting a customized substance for improving spot-like pigmentation, comprising the steps of: (a) obtaining a biological sample from an individual; (b) amplifying or hybridizing a polymorphic site of at least one single nucleotide polymorphism marker selected from the group consisting of rs251466, rs12222609, and rs9865771 from the biological sample with a probe; and (c) identifying a base of the amplified or hybridized polymorphic site of step (b).

[0352] The above-mentioned selected customized material can be applied to an individual having a specific polymorphic site to improve spot-like pigmentation.

[0353]

[0354] In the present invention, the term "subject" refers to a subject for diagnosis of skin spot-like pigmentation. DNA may be obtained from, but is not limited to, samples such as hair, urine, blood, various body fluids, isolated tissue, isolated cells, or saliva. Steps (a) and (b) above are also as described above.

[0355] The above substance may be one or more selected from the group consisting of ascorbyl glucoside, 3-O-ethyl ascorbic acid, ascorbic acid, licorice extract, tocopherol, sodium pyruvate, kelp extract, hexyldecanol, asiaticoside, and hexylresorcinol, but is not limited thereto.

[0356] For the purpose of the present invention, the significant genes associated with the degree of spot-like pigmentation 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 ASCL2, TP63 or PPARGC1B protein, when treating one or more substances selected from the group consisting of ascorbyl glucoside, 3-O-ethyl ascorbic acid, ascorbic acid, licorice extract, tocopherol, sodium pyruvate, Gamjatzu extract, hexyldecanol, asiaticoside and hexylresorcinol, improvement in spot-like pigmentation can be increased, but an optimal combination of substances can be provided depending on the genetic characteristics of each individual.

[0357] This suggests that it may be possible to identify specific single nucleotide polymorphism (SNP) markers that are significantly correlated with macular pigmentation and provide information to select personalized substances that can maximize the effect of improving macular pigmentation based on genetic information.

[0358] Specifically, if the risk score obtained by multiplying and adding the effect size (β) to the genotypes of rs251466, rs12222609, and rs9865771 is less than -0.009, the risk of developing spotty pigmentation may be classified as a low-risk group, and if the risk score is 0.000 or more, the risk of developing spotty pigmentation may be classified as a high-risk group. For example, in the case where the risk of developing spotty pigmentation is a high-risk group, an information providing method may be provided to select tocopherol, hexylresorcinol, and hexyldecanol as customized substances, but is not limited thereto.

[0359]

[0360] In the present invention, the term “SNP chip” is as described above.

[0361]

[0362] 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 spot-like pigmentation.

[0363] In another aspect, the present invention provides a composition for diagnosing the degree of spotty pigmentation, comprising a probe or an agent capable of detecting or amplifying a single nucleotide polymorphism (SNP) marker for diagnosing the degree of spotty pigmentation.

[0364]

[0365] Spot-like pigmentation refers to, but is not limited to, the skin tone of a specific area becoming darker than the individual's original skin tone due to factors such as ultraviolet rays, hormones, and aging. Specifically, it refers to determining whether or not a person is prone to spot-like pigmentation using one or more single nucleotide polymorphism (SNP) markers selected from Table 3. More specifically, a skin type prone to spot-like pigmentation refers to a skin type in which the overall skin tone of the spot-like area becomes more frequently darkened due to factors such as ultraviolet rays, hormones, and aging.

[0366] 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.

[0367]

[0368] In the present invention, the term "probe capable of detecting a marker for diagnosing the degree of spotty pigmentation" means a composition capable of diagnosing the degree of spotty pigmentation by specifically hybridizing with a polymorphic site of the gene as described above, and the specific method of such gene analysis is not particularly limited and may be any gene detection method known in the technical field to which this invention pertains. In addition, the term may be used interchangeably with the terms "for diagnosing whether a skin type is prone to developing spotty pigmentation" or "for diagnosing whether a high-risk skin type is prone to developing spotty pigmentation."

[0369] In the present invention, the term "agent capable of amplifying a marker for diagnosing the degree of spotty pigmentation" refers to a composition capable of diagnosing the degree of spotty pigmentation by confirming the polymorphic site of the gene as described above through amplification, and specifically refers to a primer capable of specifically amplifying a polynucleotide of the marker for diagnosing the degree of spotty pigmentation. In addition, the term may be used interchangeably with the terms "for diagnosing whether a skin type is prone to developing spotty pigmentation" or "for diagnosing whether a high-risk skin type is prone to developing spotty pigmentation."

[0370] 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.

[0371] In the present invention, the term “primer” is as described above.

[0372]

[0373] In another aspect, the present invention provides a kit for diagnosing the degree of spot-like pigmentation, comprising the composition for diagnosing the degree of spot-like pigmentation. The kit may be, but is not limited to, an RT-PCR kit or a DNA chip kit.

[0374]

[0375] In another aspect, the present invention provides a microarray for diagnosing the degree of spot-like pigmentation, comprising the composition for diagnosing the degree of spot-like pigmentation.

[0376] The process of immobilizing the probe polynucleotide associated with the degree of spot-like pigmentation 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, followed by hybridization on a microarray and detection of the signal generated from the label.

[0377]

[0378] In another aspect, the present invention provides a method for providing information on the degree of spot-like pigmentation, 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).

[0379] The above terms, 'object', (a) step, (b) step' are as described above.

[0380] The above method may additionally determine, but is not limited to, a high-risk group with a high risk of developing spot-like pigmentation when the risk score obtained by multiplying and adding the effect size (β) of the amplified or hybridized single nucleotide polymorphism marker (c) is 0.000 or more.

[0381]

[0382] 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.

[0383]

[0384] Example 1: Derivation of genetic polymorphism markers associated with skin pigmentation

[0385]

[0386] This study aimed to identify genomic regions (genetic variations) that are associated with differences in the presence and severity (numerical value) of facial skin pigmentation in Korean women, based on genetic information. The present invention utilizes a microarray genotyping chip (Illumina), which enables genome-wide screening without prior selection of candidate genes, to identify genomic regions (genetic variations) associated with facial skin pigmentation.

[0387] In order to evaluate the presence and degree of diffuse dark spots on the skin and / or the presence and degree of skin spot-like pigmentation in the present invention, an image-based diagnostic AI was developed to extract a novel phenotype, and a high correlation (0.876) was confirmed with visual evaluation. The novel AI-based phenotype was used, and in order to minimize external effects that may affect the presence and degree (numerical value) of diffuse dark spots on the skin, it was utilized after being adjusted for age, sex, BMI, and cluster principal component values.

[0388] To assess the presence and degree of dark skin tone in the present invention, skin tone brightness (L*) was extracted using a skin diagnostic device (Janus III, PIE). The extracted skin tone brightness value was used as a phenotype, and to minimize external factors that could affect dark skin tone and degree (numerical value), it was adjusted for age, sex, BMI, and cluster principal component values ​​before use.

[0389] To determine the association between genomic regions (genetic variants) and the presence and severity (numerical) of diffuse dark spots, dark skin tone and severity (numerical), and / or the presence and severity (numerical) of skin mottling, Bolt-LMM was used to quantify correlation significance and genetic effects. The Bolt-LMM method maximizes the analytical effectiveness while minimizing the influence of genetic relatedness among the study participants.

[0390]

[0391] 1-1. Evaluation method for diffuse dark spots, dark skin tone, and spot-like pigmentation on the skin

[0392] To derive genetic polymorphism markers that can explain the degree of diffuse dark spots on the cheeks and cheekbones, the degree of dark skin tone, and / or the degree of spot-like pigmentation, healthy Koreans aged 20 to 70 were recruited. In addition, for skin measurements, all analysis subjects washed their faces with cleanser or soap and waited in a constant temperature and humidity chamber for 20 minutes without applying any products to allow their skin to adapt to the measurement environment. Then, the degree of skin pigmentation was evaluated. For facial images, a high-resolution camera of a skin diagnosis device (PIE Janus3) was used. Measurements and analyses were performed according to the device manufacturer's manual. Specifically, the analysis subjects wore a headband and a black cloth over their clothes. They were instructed to fix their face to the forehead and chin rests built into the device and close their eyes. The camera built into the Janus3 device captured high-resolution images of the analysis subjects' faces. For high-resolution images between the under-eye area and the cheekbone, facial landmark detection technology was used to extract features in a way that makes personal identification impossible and minimizes interference with other areas.

[0393] To extract the degree of distributed dark spots on the skin, AI using the ResNet (Residual network) structure among CNN (Convolutional Neural Network, the basic structure of image deep learning) was used. Its performance was selected after comparing it with other well-known structures. The built AI was additionally evaluated using flipped images, and the average value was used as the final result, thereby improving accuracy and stability. To confirm the reliability of the AI ​​analysis value, it was compared with the results of visual evaluation, and since it showed a very high correlation coefficient of 0.876, it was applied to 50,000 data to extract a new phenotype, the degree of distributed dark spots.

[0394] To extract the degree of dark skin tone, the cheek area image was extracted from the captured image using a built-in algorithm. If the cheek area selection was not accurate, the evaluator manually adjusted it. The skin color average RGB and CIE-LAB color space conversion was performed using a built-in algorithm.

[0395] In order to extract the degree of skin spot pigmentation, we created a tool that can generate one data with one click to build an AI for spot-type OX diagnosis, and created numerous spotty and non-spotty images. This was used to train the first AI, and after applying it to actual images, images similar to false positive and false negative images were included in the training data again to enable maximum performance improvement with minimal data addition. The AI ​​was implemented to operate by scanning images of the same size as the training data across the entire face, and since spot pigmentation accounts for almost all cases where there is one or none on the face, the maximum value was presented as the final diagnosis result.

[0396] The AI ​​for partial images used a ResNet (Residual network) structure among CNNs (Convolutional Neural Networks, the basic structure of image deep learning). This structure was selected through performance evaluations with existing well-known structures. By additionally evaluating left-right flipped images and using the average as the final result, the AI ​​improved accuracy and evaluation stability. The final result showed an extremely high accuracy of over 99%, and was applied to 50,000 data sets to extract a new phenotype, the degree of spotty pigmentation.

[0397]

[0398] 1-2. Gene collection and analysis method

[0399] To conduct genetic analysis, genomic DNA was extracted from the subject's saliva using the QIAmp mini prep kit (QIAGEN). The quality was confirmed by DNA gel band test using 1X TAE 1% agarose gel with absorbance (OD 260 / 280) > 1.7, concentration 50 ng / ul, and genetic analysis was performed only on those that passed the quality test.

[0400] Genetic analysis was performed using Illumina's microarray genotyping chip, and specifically, the genes of the subjects were analyzed using the same company's global screening array product.

[0401] 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.

[0402] The microarray genotyping chip for which the above experimental process was completed was scanned using iScan Control Software (Illumina), and after scanning was completed, the automatically generated idat file was used to check the data quality control level (sample call rate > 98%, marker call rate > 98%) and genetic information using the GenomeStudio (Illumina) program.

[0403] In the present invention, only data that passed data quality control after genetic analysis was used.

[0404]

[0405] 1-3. Derivation of genetic polymorphism markers associated with the degree of diffuse dark spots, dark skin tone, and spotty pigmentation on the skin.

[0406] In order to derive genetic polymorphism markers that have significant associations with the degree of diffuse dark spots on the skin, the degree of dark skin tone, and the degree of skin spot-type pigmentation, a linear-mixed model (linear regression + genetic association penalty model between samples) was analyzed using the genetic polymorphism markers of the analysis subjects, and the BOLT-LMM v2.3.5 program was used for the analysis. Age, sex, BMI, and 10 PCs (correcting for genetic differences due to racial differences) were used as covariates during the analysis.

[0407] To secure additional genetic polymorphism markers, we conducted an imputation analysis (using the Beagle v 5.1 program) using the preceding genetic polymorphism markers, and the reference data used was the information registered in the international public genome database 1000 Genomes project.

[0408] Imputation is a statistical technique for inferring new genetic polymorphism marker information based on genetic polymorphism marker information obtained through experiments.

[0409] The significance of genetic polymorphism markers showing association with the degree of skin diffuse dark spots was evaluated using the chi-square statistic of the BOLT-LMM results, and the criterion was set as p-value < 1.0 * e-4.

[0410] As a result, a number of genetic polymorphism markers with significant association with the degree of skin-dispersed dark spots, as shown in Table 1 below, were derived.

[0411]

[0412]

[0413]

[0414] 1) National Institutes of Health (NIH) ID, sequence can be checked on the website

[0415] 2) (major allele) > (minor allele) meaning

[0416] 3) Minor allele frequency = (2 mm + Mm) / 2(MM + Mm + mm)

[0417] 4) Statistical significance of phenotypic differences for three genotypes (M / M, M / m, m / m) (M: major allele, m: minor allele)

[0418] 5) A number indicating how many times higher the risk of being a case (having a distributed dark spot sensitivity) is when carrying a minor allele compared to when not carrying a minor allele.

[0419]

[0420] The significance of genetic polymorphism markers showing association with skin tone brightness was evaluated using the chi-square statistic of the BOLT-LMM results, and the criterion was set at p-value < 5e-08.

[0421] As a result, a number of genetic polymorphism markers significantly associated with dark skin tone were identified, as shown in Table 2 below.

[0422]

[0423]

[0424]

[0425] 1) National Institutes of Health (NIH) ID, sequence can be checked on the website

[0426] 2) (major allele) > (minor allele) meaning

[0427] 3) Minor allele frequency = (2 mm + Mm) / 2(MM + Mm + mm)

[0428] 4) Statistical significance of phenotypic differences for three genotypes (M / M, M / m, m / m) (M: major allele, m: minor allele)

[0429] 5) A number indicating how many times higher the risk of being a case (having dark skin tone) is when carrying the minor allele compared to when not carrying the minor allele.

[0430]

[0431] The significance of genetic polymorphism markers showing association with the degree of skin spot-like pigmentation was evaluated using the chi-square statistic of the BOLT-LMM results, and the criterion was set at p-value < 5e-8.

[0432] As a result, a number of genetic polymorphism markers with significant association with the degree of spot-like pigmentation, as shown in Table 3 below, were derived.

[0433]

[0434]

[0435]

[0436] 1) National Institutes of Health (NIH) ID, sequence can be checked on the website

[0437] 2) (major allele) > (minor allele) meaning

[0438] 3) Minor allele frequency = (2 mm + Mm) / 2(MM + Mm + mm)

[0439] 4) Statistical significance of phenotypic differences for three genotypes (M / M, M / m, m / m) (M: major allele, m: minor allele)

[0440] 5) As the minor allele increases by one, the degree of change in phenotypic risk (-: decreased risk of spotty pigmentation, +: increased risk of spotty pigmentation)

[0441] Presence of Spot-like Pigmentation = AI-Based Skin Facial Image Analysis

[0442]

[0443] Example 2: Results of analysis of the effects of various substances on regulating the expression of genes associated with skin pigmentation.

[0444] 2-1. Results of analysis of the effects of various substances on regulating the expression of genes associated with skin-dispersed dark spots.

[0445] Among various substances with skin-improving properties, we conducted a screening process to identify those that influence the expression of genes associated with diffuse dark spots. We identified changes in the expression of genes associated with diffuse dark spots in cells where UVB exposure increased melanin synthesis, and selected substances that restored expression to pre-UVB levels.

[0446] Mouse melanoma cells (B16F10; mouse melanoma cells, CRL-6475) were purchased from ATCC and used. The purchased cells were cultured in a medium made with 10% FBS (fetal bovine serum) and 1% Penicillin / Streptomycin in DMEM (Dulbecco's Modified Eagle Medium-GIBCO) in a 5% CO2 incubator at 37°C.

[0447] To analyze the efficacy of changes in the mRNA expression of skin diffuse dark spot-related genes, mouse melanoma cells were seeded in 6-well plates at 2 x 10 5 Cells were seeded per well and cultured in a 37°C, 5% CO2 incubator for one day. After replacing the medium with PBS, the cells were exposed to UVB (312 nm) 30 mJ / cm using an RMX 3W (Vilber Lourmat) device. 2 The substances were added to the medium at appropriate concentrations and cultured for one day under the same conditions. Afterwards, total RNA was obtained using the AccuPrep Universal RNA Extraction Kit (Bioneer, K-3140) and analyzed by AccuPower® RocketScript. TM cDNA was synthesized using Cycle (Bioneer, K-2202). The expression levels of each gene were analyzed by qPCR using the synthesized cDNA and taqman probes (assay ID: AKAP1- Mm00501539_m1, RAB11FIP2- Mm01261268_m1, PPARGC1B- Mm00504730_m1).

[0448] As a result, as shown in Fig. 1, UVB increased AKAP1 expression, while RAB11FIP2 and PPARGC1B decreased. Substances that decrease AKAP1 expression include cysteine, cysteamine HCL, nutmeg extract, high molecular weight hyaluronic acid, and ultra-low molecular weight hyaluronic acid, and substances that increase RAB11FIP2 expression include bisabolol, Korean knotweed extract, and diisopropyl alcohol (DIPA). In addition, it was confirmed that PPARGC1B expression was increased by niacinamide, glycolic acid (AHA), gluconolactone (PHA), and salicylic acid (BHA).

[0449]

[0450] 2-2. Results of analysis of the effect of various substances on regulating the expression of genes associated with dark skin tone.

[0451] Among various substances with skin-improving properties, we conducted a screening process to identify those that influence the expression of genes associated with dark skin tone. We identified changes in gene expression associated with dark skin tone in cells where melanin synthesis was increased with a-MSH, and selected substances that restored expression to pre-a-MSH levels.

[0452] Mouse melanoma cells (B16F10; mouse melanoma cells, CRL-6475) were purchased from ATCC and used. The purchased cells were cultured in a medium prepared by adding 10% FBS (fetal bovine serum) and 1% penicillin / streptomycin to DMEM (Dulbecco's Modified Eagle Medium-GIBCO) in a 5% CO2 incubator at 37°C.

[0453] RT-qPCR experiments were conducted to analyze the efficacy of changes in the mRNA expression of genes associated with dark skin tone. The cells were cultured in a 5% CO2 incubator at 37℃ for one day. To evaluate the expression change of KITLG by UVB, the medium of human keratinocytes (HaCaT) was replaced with PBS, and then irradiated with UVB (312 nm) 30 mJ / cm2 using an RMX 3W (Vilber Lourmat) device. The medium was replaced with fresh medium and cultured for one day. To evaluate the expression change of KITLG by substance treatment, the cells were treated with appropriate concentrations of substances along with fresh medium and cultured for one day. In addition, to evaluate the expression of OCA2, BNC2, and RAB32, melanocytes were treated with appropriate concentrations of substances and α-MSH (10 nM) in fresh medium and cultured for one day. Afterwards, 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 qPCR using the synthesized cDNA and taqman probe (assay ID: OCA2- Mm00498969_m1, BNC2- Mm00556284_m1, RAB32- Mm00458960_m1, KITLG- Hs00241497_m1).

[0454] As a result, as shown in Fig. 2, OCA2, BNC2, and RAB32 were increased by a-MSH, and the expression of KITLG was also increased by UVB.

[0455] Specifically, it was confirmed that substances that reduce OCA2 expression include ferulic acid, polydatin, zinc PCA, tranexamic acid, quercetin, genistein, Aspergillus ferment, gamma-oryzanol, sunflower seed oil, and isostearic acid, and substances that reduce BNC2 expression include polydatin, ferulic acid, bakuchiol, genistein, soluble licorice extract, and a combination of soluble licorice extract and Aspergillus ferment.

[0456] In addition, it was confirmed that substances that reduce RAB32 expression include ferulic acid, quercetin, genistein, tannic acid, polydatin, zinc PCA, and gamma-oryzanol, and substances that reduce KITLG expression include chrysin, zinc PCA, and tannic acid.

[0457]

[0458] 2-3. Results of analysis of the effect of various substances on the expression of genes associated with skin spot-like pigmentation.

[0459] Among various substances with skin-improving properties, a screening process was conducted to identify those that influence the expression of genes associated with spotty pigmentation. We identified changes in the expression of these genes in cells where melanin synthesis was increased with α-MSH, and selected substances that restored expression to pre-α-MSH levels.

[0460] Mouse melanoma cells (B16F10; mouse melanoma cells, CRL-6475) were purchased from ATCC and used. The purchased cells were cultured in a medium prepared by adding 10% FBS (fetal bovine serum) and 1% penicillin / streptomycin to DMEM (Dulbecco's Modified Eagle Medium-GIBCO) in a 5% CO2 incubator at 37°C.

[0461] To analyze the efficacy of changes in the mRNA expression of genes associated with skin spot-like pigmentation, mouse melanoma cells were seeded in 6-well plates at 2X10 5Cells were seeded per well and cultured in a 37℃, 5% CO2 incubator for one day. The medium was replaced with 10 nM α-MSH, and the substances were added to the medium at appropriate concentrations and cultured for one day under the same conditions. After that, total RNA was obtained using the AccuPrep Universal RNA Extraction Kit (Bioneer, K-3140) and cDNA was synthesized using AccuPower® RocketScript™ Cycle (Bioneer, K-2202). The expression level of each gene was analyzed by qPCR using the synthesized cDNA and taqman porbe (assay ID: ASCL2- Mm01268891_m1, PPARGC1B- Mm00504730_m1, TP63- Hs00978349_m1).

[0462] As a result, as shown in Fig. 3, ASCL2 expression was increased and PPARGC1B was decreased by α-MSH in B16F10 cells. In human keratinocytes, HaCaT, UVB 10 mJ / cm 2 The expression of TP63 was increased by . Substances that again decreased ASCL2 expression included tocopherol, licorice extract, and perilla leaf extract, and PPARGC1B expression was again increased by AA2G, ascorbic acid, and SINOWHITE EA (3-O-Ethyl Ascorbic Acid). It was confirmed that sodium pyruvate, hexyldecanol, asiaticoside, and hexylresorcinol treated to HaCaT cells decreased the expression of TP63 (Fig. 3).

[0463]

[0464] Example 3: Analysis of the melanin reduction efficacy and synergy of a customized material for improving skin pigmentation.

[0465]

[0466] 3-1. Analysis of the Melanin Reduction Efficacy and Synergy of a Customized Skin-Dispersing Dark Spot Improvement Material

[0467] In Example 2, it was confirmed whether the combination of selected substances for improving skin-dispersed dark spots had a synergistic effect in suppressing melanin production increased by UVB.

[0468] Specifically, 2*10 mouse melanoma cells were seeded in a 6-well plate. 5 After dispensing cells / well and culturing them in a 37℃, 5% CO2 incubator for one day, UVB (312 nm) 30 mJ / cm 2 was investigated. The medium containing the appropriate concentration and combination of substances was added and cultured again in a 37℃, 5% CO2 incubator for one day to confirm whether the substances inhibited the melanin production increased by UVB. The cells attached to the plate were collected in PBS and centrifuged to obtain a cell pellet. The cell pellet was visually assessed and photographed. The cell pellet was 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 treatment group by the melanin concentration increased by UVB.

[0469] As a result, it was confirmed that melanin production was reduced by approximately 5.2% (X) when treated with 100 ppm nutmeg extract and 10 ppm bisabolol, and that melanin production was reduced by approximately 1.5% (Y) when treated with 25 ppm DIPA alone. In addition, it was confirmed that melanin production was reduced by approximately 32.6% (T, effect measurement value) when treated with a combination of 100 ppm nutmeg extract, 10 ppm bisabolol, and 25 ppm DIPA. This is greater than the Colby equation predicted value (E) of 6.6%, suggesting a synergistic effect in reducing melanin production (Fig. 4).

[0470] Colby formula E=(X+Y)-(X*Y / 100)

[0471]

[0472] 3-2. Analysis of the Melanin Reduction Efficacy and Synergy of Customized Materials for Dark Skin Tone Improvement

[0473] In Example 2, it was confirmed whether the combination of substances selected for improving dark skin tone had a synergistic effect in suppressing melanin production increased by a-MSH treatment.

[0474] Specifically, mouse melanoma cells and human keratinocytes were seeded at 2Х10^5 cells / well in 6-well plates 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 the cells were cultured for another two days 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. 1 N NaOH (sodium hydroxide) solution containing 10% DMSO (dimethyl sulfoxide) was added to the cell pellet, and the pellets were dissolved at 80℃ for 10 minutes. The pellets were placed in 96-well plates and the absorbance at 405 nm was measured using an Epoch (BioteK) instrument. 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.

[0475] As a result, it was confirmed that melanin production was reduced by approximately 5.04% (X), 6.47% (Y), and 2.07% (Z) when treated with 10 ppm of ferulic acid, 5 ppm of oil-soluble licorice extract, and 5 ppm of bakuchiol, respectively. In addition, when the ferulic acid, oil-soluble licorice extract, and bakuchiol at the corresponding concentrations were combined and treated together, melanin production was reduced by approximately 35.07% (T, effect measurement value). This was greater than the Colby prediction value (E1) of 13.03% for the combination of the three substances, so it was determined that there was a synergistic effect in reducing melanin production (Fig. 5).

[0476] Colby formula E1 = (X+Y)-(XY+XZ+YZ / 100)+XYZ / 10000

[0477]

[0478] In addition, when treated with another combination, 10 ppm polydatin and 20 ppm quercetin, melanin production was reduced by approximately 23.8% and 33.9%, respectively, but when polydatin and quercetin at the corresponding concentrations were treated simultaneously, melanin was reduced by 57.9% (T, effect measurement value). This was greater than the Colby equation prediction equation (E2) of 49.5% for the combination of the two substances, so it was determined that there was a synergistic effect in reducing melanin production (Fig. 6).

[0479] Colby formula E2 = X+Y-(XY / 100)

[0480]

[0481] 3-3. Analysis of the Melanin Reduction Efficacy and Synergy of Customized Materials for Spot-Type Pigmentation Improvement

[0482] In Example 2, it was confirmed whether the combination of substances selected for improving spot-like pigmentation had a synergistic effect in suppressing melanin production increased by UVB.

[0483] 2Х10 mouse melanoma cells B16F10 in 6-well plates 5Cells were seeded per well and cultured in a 37℃, 5% CO2 incubator for one day, and then treated with 10 nM α-MSH. Medium containing appropriate concentrations and combinations of substances was added, and cultured again in a 37℃, 5% CO2 incubator for one day to confirm whether the substances inhibited melanin production increased by α-MSH. Cells attached to the plate were collected with PBS and centrifuged to obtain a cell pellet. The cell pellet was 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) instrument. Melanin was quantified using the measured absorbance and normalized to the protein concentration of the sample to measure melanin. The inhibition rate of melanin production in the sample was calculated as the ratio of melanin produced by α-MSH to melanin produced by the sample-treated group. In addition, the inhibition rate of increased melanin production was calculated as the degree of inhibition of increased melanin production compared to the control group without α-MSH treatment.

[0484] As a result, when 1 ppm hexyldecanol and 10 ppm hexylresorcinol were simultaneously treated, the melanin production inhibition rate was 53.7%, which was higher than the Colby-type predicted value of 48.5% for both conditions, and when 5 ppm hexyldecanol and 1 ppm tocopherol were simultaneously treated, the melanin production increase inhibition rate was 45.1%, which was higher than the Colby-type predicted value of 27.1% for both conditions, and a synergy was observed (Fig. 7).

[0485] Colby formula E=(X+Y)-(XХY / 100)

[0486]

[0487] Example 4: Human application test to determine the degree of improvement in skin-dispersed dark spots compared to vitamin C.

[0488] In order to improve the skin-dispersed dark spots obtained through Example 3, we sought to confirm whether the combination showing synergistic efficacy in reducing melanin also showed excellent efficacy in humans.

[0489]

[0490] 4-1. Subject selection

[0491] After approval from the Institutional Review Board (IRB, Korea) (LG-SKEV-2022-09-A), 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. Ten healthy subjects in their 20s to 50s with no major skin abnormalities were selected.

[0492]

[0493] 4-2. Subject exclusion criteria

[0494] Among the subjects above, the following were excluded: 1) those with eczematous / infectious skin diseases, 2) those with allergic constitutions or hypersensitivity, 3) those who were pregnant or breastfeeding, 4) those with significant nutritional disorders, 5) those addicted to drugs or alcohol, 6) those with moles, acne, tattoos, erythema, burn marks, etc. on the test area, 7) those who had participated in the same test less than 1 month ago, and 8) those with hypertension or diabetes.

[0495]

[0496] 4-3. Skin Dispersive Dark Spot Measurement Device and Measurement / Analysis Method

[0497] The control group applied 8% vitamin C essence to the right cheek and under the eye twice a day, morning and evening, for four weeks. The experimental group applied 0.5% bisabolol (ingredient name: Alpha-Melight), 2% nutmeg extract (ingredient name: nutmeg extract L1), and 1% DIPA (CRODAMOL DA-LQ) to the left cheek and under the eye twice a day, morning and evening.

[0498] Using PIE's Janus3, the front and side of the face were photographed under standard, polarized, and UV light. Areas under both eyes and on both cheeks were designated to obtain the pigmentation values ​​under polarized light. The percentage improvement in pigmentation was calculated by calculating the percentage reduction in pigmentation after four weeks of use compared to the baseline polarized pigmentation values.

[0499]

[0500] 4-4. Check the degree of improvement in dark spots on the skin compared to vitamin C.

[0501] In order to improve the skin-dispersed dark spots obtained through Example 3, three substances of the present invention (bisabolol, nutmeg extract, DIPA) effective in inhibiting melanin production were mixed and applied to the left face, and the control essence (8% vitamin C) was applied to the right face.

[0502] As a result, it was confirmed that the rate of improvement in pigmentation after 4 weeks of application was significantly increased when applying the essence containing bisabolol, nutmeg extract, and DIPA compared to vitamin C. (Improvement rate increased by approximately 14%, p-value = 0.03, Student's T-test) (Fig. 8)

[0503]

[0504] Example 5: Experiment to confirm the inhibition of protein carbonylation of dark skin tone material to determine the degree of improvement in dark skin tone compared to vitamin C.

[0505] Previous studies have shown that skin dullness and yellowing are promoted by protein carbonylation in keratinocytes. Therefore, through a protein carbonylation assay, we examined whether materials tailored to dark skin tones could also improve dullness and yellowing.

[0506] Human keratinocytes (HaCaT) were seeded at 1X10^4 in a black clear bottom 96-well plate, and after 24 hours, customized materials were treated at appropriate concentrations. After another 24 hours, UVA was irradiated at 10 J / cm, washed with PBS, and incubated with fluolescein-5-thiosemicabarzide in MES buffer for 1 hour at RT. After washing once more with PBS, fluorescence was measured using a plate reader at Excitation 492 nm and Emission 562 nm. And, the reduction in protein carbonylation of other effective substances was measured based on the UVA only treatment group.

[0507] As a result, when the above materials, gamma-oryzanol, aspergillus ferment, and isostearic acid, were treated at 2.5 ppm, 1%, and 50 ppm, respectively, protein carbonylation was significantly reduced by 39%, 43%, and 31%, respectively (Fig. 9).

[0508]

[0509] Example 6: Human application trial for developing a customized pigmentation improvement prescription for high-risk groups with diffuse dark spots on the skin.

[0510] In Example 2, a substance that modulates the expression of a polymorphic marker significantly associated with diffuse dark spots, dark skin tone, and / or spotty pigmentation was selected. Through this, the aim was to provide a substance that can increase the efficacy of improving diffuse dark spots according to the risk level of developing diffuse dark spots, dark skin tone, and / or spotty pigmentation.

[0511] 6-1. Subject selection

[0512] After approval from the Institutional Review Board (IRB, Korea) (LG-SKEV-2022-09-A), 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. Ten healthy subjects in their 20s to 50s with no major skin abnormalities were selected.

[0513] Based on the genetic polymorphism markers associated with the presence or absence of distributed dark spots in Table 1, subjects were classified into low-risk and high-risk groups according to the risk level of developing distributed dark spots, as follows.

[0514] The risk score of the six genetic markers (the value obtained by multiplying the genotype by the effect size (β) and adding them together) is less than 0.28 → low-risk group (n=5).

[0515] The risk score of 6 genetic markers (the value obtained by multiplying the genotype by the effect size (β) and adding them together) is 0.28 or higher → high-risk group (n=5)

[0516]

[0517] 6-2. Subject exclusion criteria

[0518] Same as Example 4-2

[0519]

[0520] 6-3. Skin Dispersive Dark Spot Measurement Device and Measurement / Analysis Method

[0521] The control group applied 8% vitamin C essence to the right cheek and under the eye twice a day, morning and evening, for four weeks. The experimental group applied 0.5% bisabolol (ingredient name: Alpha-Melight), 5% nutmeg extract (ingredient name: nutmeg extract L1), and 2% niacinamide (ingredient name: niacinamide) to the left cheek and under the eye twice a day, morning and evening.

[0522] Using PIE's Janus3, the front and side of the face were photographed under standard, polarized, and UV light. Areas under both eyes and on both cheeks were designated to obtain the pigmentation values ​​under polarized light. The percentage improvement in pigmentation was calculated by calculating the percentage reduction in pigmentation after four weeks of use compared to the baseline polarized pigmentation values.

[0523]

[0524] 6-4. Confirmation of the improvement rate of pigmentation according to the material treatment of the present invention.

[0525] When the essence (customized prescription) containing the three substances of the present invention (bisabolol, nutmeg extract, and niacinamide) was applied, the rate of improvement in pigmentation was significantly better in the high-risk group than in the low-risk group (approximately 13.9%, p value 0.03, Student's T-test). Unlike the customized prescription, vitamin C alone did not improve pigmentation in either the low-risk or high-risk group. The customized prescription did not significantly increase the rate of improvement in pigmentation in the low-risk group, but it was confirmed that it significantly increased the rate of improvement in pigmentation in the high-risk group (approximately 24% increase, p value 0.03, Student's T-test).

[0526] This shows that the responsiveness to a substance varies depending on whether or not the subject possesses an allele of a specific genetic SNP, and that the efficiency of improving distributed dark spots can be increased by providing customized prescriptions to a high-risk group with a high level of risk for distributed dark spots (Figs. 10 and 11).

[0527]

[0528] Example 7: Human application trial for developing a customized pigmentation improvement prescription for high-risk dark skin tone groups.

[0529] In Example 2, a substance that regulates the expression of a polymorphic marker significantly associated with dark skin tone was selected. Through this, the aim was to provide a substance that can increase the efficacy of improving dark skin tone according to the level of risk of developing dark skin tone.

[0530]

[0531] 7-1. Subject selection

[0532] After approval from the Institutional Review Board (IRB, Korea) (LG-SKEV-2022-09-A), 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. Nine healthy subjects in their 20s to 50s with no major skin abnormalities were selected.

[0533] Based on the skin tone brightness-related genetic polymorphism markers in Table 2, subjects were classified into low-risk and high-risk groups according to the risk level of developing dark skin tone, as follows.

[0534] Risk score of 5 genetic markers (value calculated by multiplying the genotype by the effect size (β) and adding it) less than -0.193 → low-risk group (n=4)

[0535] Risk score of 5 genetic markers (the value obtained by multiplying the genotype by the effect size (β) and adding them together) -0.193 or higher → high-risk group (n=5)

[0536]

[0537] 7-2. Subject exclusion criteria

[0538] Same as Example 4-2

[0539]

[0540] 7-3. Skin tone brightness measurement device and measurement / analysis method

[0541] The control group applied 8% vitamin C essence to the right cheek and under the eye twice daily, morning and evening, for four weeks. The experimental group applied 1% isostearic acid, 1% sunflower seed oil, and 0.2% polydatin to the left cheek and under the eye twice daily, morning and evening.

[0542] Using PIE's Janus3, the front and side of the face were photographed under standard, polarized, and ultraviolet light. The areas under both eyes and on both cheeks were designated to obtain skin tone brightness values ​​under standard light. The difference (△L) between the baseline skin tone brightness value and the 4-week skin tone brightness value was calculated to determine the degree of improvement in pigmentation.

[0543]

[0544] 7-4. Confirmation of the rate of improvement in pigmentation according to the material treatment of the present invention.

[0545] When the essence (customized formulation) containing the three substances of the present invention (isostearic acid, sunflower seed oil, and polydatin) was applied, the rate of improvement in pigmentation was significantly better in the high-risk group compared to the low-risk group (△L approximately 1.3 increase, p value 0.01, Student's T-test). Unlike the customized formulation, vitamin C alone showed a slight improvement in the low-risk group, but did not improve pigmentation in the high-risk group. The customized formulation did not significantly increase the rate of improvement in pigmentation in the low-risk group, but did significantly increase the rate of improvement in pigmentation in the high-risk group (△L approximately 2.1 increase, p value 0.03, Student's T-test).

[0546] This demonstrates that responsiveness to a substance varies depending on the presence of a specific genetic SNP allele, suggesting that personalized treatment options for high-risk groups with a high risk of developing dark skin tone could enhance the effectiveness of improving dark skin tone (Figures 12 and 13).

[0547]

[0548] Example 8: Human application trial for developing a customized pigmentation improvement prescription for a high-risk group with skin spot-like pigmentation.

[0549] In Example 2, a substance that regulates the expression of a polymorphic marker associated with spot-like pigmentation was selected. Through this, the aim was to provide a substance that can increase the efficacy of improving spot-like pigmentation according to the level of risk of spot-like pigmentation development.

[0550]

[0551] 4-1. Subject selection

[0552] After approval from the Institutional Review Board (IRB, Korea) (LG-SKEV-2022-09-A), 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. Nine healthy subjects in their 20s to 50s with no major skin abnormalities were selected.

[0553] Based on the genetic polymorphism markers associated with the presence or absence of spot-like pigmentation in Table 3, subjects were classified into low-risk and high-risk groups according to the risk level of spot-like pigmentation development, as follows.

[0554] The sum of all minor allele effect sizes (β) of the three genetic markers is -0.009 or less → low-risk group (n=5)

[0555] The sum of all minor allele effect sizes (β) of the three genetic markers is 0.000 or higher → high-risk group (n=4)

[0556]

[0557] 4-2. Subject exclusion criteria

[0558] Same as Example 4-2

[0559]

[0560] 4-3. Skin spot-type pigmentation measurement device and measurement / analysis method

[0561] The experimental group essence contained 0.2% tocopherol (ingredient name: COVIOX-70), 0.9% hexylresorcinol (ingredient name: hexylresorcinol), and 5% hexyldecanol (ingredient name: Hexyldecanol) and applied it to the affected area twice a day, in the morning and evening.

[0562] Using PIE's Janus3, the front and side of the face were photographed under normal, polarized, and ultraviolet light. In the side images showing spots, the area near the spots was designated and AI analysis was used to determine the degree of spot recognition. Furthermore, a decrease in spot recognition was observed in the experimental group following the use of essence.

[0563]

[0564] 4-4. Confirmation of improvement in AI pigmentation recognition through material treatment of the present invention.

[0565] Three substances of the present invention (tocopherol, hexylresorcinol, and hexyldecanol) effective in inhibiting melanin production obtained through Example 3 were mixed and applied to the spot area.

[0566]

[0567] As shown in Figures 14 and 15, it was confirmed that the degree to which AI recognized spot improvement in the high-risk group significantly increased compared to the low-risk group after 8 weeks of application of the experimental group essence (AI's spot recognition improvement rate increased by approximately 53% compared to the low-risk group, p value 0.01 or less, Student's T-test).

[0568]

[0569] 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. Myristica Fragrans (Nutmeg) Extract, Polygonum Cuspidatum Extract, Bisabolol, Diisopropyl adipate (DIPA), Cysteine, Cysteine ​​HCl, Sodium Hyaluronate, Ultra-low molecular weight hyaluronic acid, Niacinamide, Glycolic acid, Gluconolactone, Salicylic acid, Polydatin, Ferulic acid, Bakuchiol, Genistein, Oil soluble licorice extract, Aspergillus ferment, Consists of Quercetin, Tannic acid, ZnPCA, Gamma Oryzanol, Chrysin, Tranexamic acid, Sunflower oil, Isostearic acid, Ascorbyl Glucoside, 3-O-Ethyl Ascorbic Acid, Ascorbic Acid, Glycyrrhiza Uralensis (Licorice) Extract, Tocopherol, Sodium Pyruvate, Ecklonia Cava Extract, Hexyldecanol, Asiaticoside and Hexylresorcinol. A composition for improving skin pigmentation, comprising one or more substances selected from the group as an active ingredient.

2. A composition according to claim 1, wherein the skin pigmentation is a skin diffuse dark spot, a dark skin tone, or a skin spot-like pigmentation.

3. A composition for improving dark spots by dispersing the skin, comprising as an active ingredient at least one substance selected from the group consisting of Myristica Fragrans (Nutmeg) Extract, Polygonum Cuspidatum Extract, Bisabolol, Diisopropyl adipate (DIPA), Cysteine, cysteamine HCl, Sodium Hyaluronate, Hyaluronic acid, Niacinamide, Glycolic acid, Gluconolactone, and Salicylic acid.

4. In the third paragraph, the composition (i) cysteine, cysteamine HCL, nutmeg extract, high molecular weight hyaluronic acid, and ultra-low molecular weight hyaluronic acid; (ii) bisabolol, hollyhock root extract, and diisopropyl adipate; or (iii) A composition comprising niacinamide, glycolic acid, gluconolactone, and salicylic acid as active ingredients.

5. In paragraph 3, 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.

6. In paragraph 3, The composition is characterized by a decrease in melanin production.

7. In paragraph 6, A composition wherein the substance for reducing melanin production is bisabolol, nutmeg extract, or diisopropyl adipate.

8. In paragraph 3, A composition characterized in that the above substance inhibits the expression of AKAP1 or promotes the expression of RAB11FIP2 or PPARGC1B.

9. In paragraph 8, A composition wherein the substance for inhibiting the expression of AKAP1 is cysteine, cysteamine HCL, nutmeg extract, high molecular weight hyaluronic acid, or ultra-low molecular weight hyaluronic acid.

10. In paragraph 8, A composition wherein the substance for promoting the expression of RAB11FIP2 is bisabolol, a root extract of Hojanggeun, or diisopropyl adipate.

11. In paragraph 8, A composition wherein the substance for promoting the expression of PPARGC1B is niacinamide, glycolic acid, gluconolactone, or salicylic acid.

12. In paragraph 3, The composition is customized for an individual having one or more single nucleotide polymorphism markers present in a gene encoding a protein of AKAP1, RAB11FIP2, PPARGC1B, BNC2, MC1R or MITF.

13. In paragraph 12, A composition wherein the single nucleotide polymorphism marker is at least one selected from the group consisting of rs11079281, rs10444110, rs251466, rs16935073, rs2228479, and rs7430957.

14. In paragraph 13, The single nucleotide polymorphism marker for the diagnosis of the above-mentioned skin-dispersed dark spot is a polynucleotide consisting of 5-100 consecutive DNA sequences including the 55231787th base of the 17th chromosome of a human being, wherein the 55231787th base is T or C (rs11079281); A polynucleotide consisting of 5 to 100 contiguous DNA sequences including the 119563401st base of human chromosome 10, wherein the 119563401st base is T or C (rs10444110); A polynucleotide consisting of 5-100 contiguous DNA sequences including the 149195603rd base of human chromosome 5, wherein the 149195603rd base is C or G (rs251466); A polynucleotide consisting of 5-100 contiguous DNA sequences including the 16795790th base of human chromosome 9, wherein the 16795790th base is A or C (rs16935073); A polynucleotide consisting of 5-100 contiguous DNA sequences including the 89985940th base of human chromosome 16, wherein the 89985940th base is G or A (rs2228479); A composition comprising a polynucleotide comprising 5 to 100 contiguous DNA sequences including base 69978466 of human chromosome 3, wherein base 69978466 is A or G (rs7430957); and one or more polynucleotides selected from the group consisting of complementary polynucleotides thereof. 15.(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 at least one single nucleotide polymorphism marker selected from the group consisting of rs11079281, rs10444110, rs251466, rs16935073, rs2228479, and rs7430957 from the biological sample; and (c) A method for providing information for selecting a customized substance for improving skin-dispersed dark spots, comprising a step of identifying the base of the amplified or hybridized polymorphic region of step (b).

16. In paragraph 15, A method for providing information, wherein the above substance is at least one selected from the group consisting of nutmeg extract, Korean knotweed extract, bisabolol, diisopropyl adipate, cysteine, cysteamine HCL, high molecular weight hyaluronic acid, ultra-low molecular weight hyaluronic acid, niacinamide, glycolic acid, gluconolactone, and salicylic acid.

17. In paragraph 15, An information provision method wherein the risk score obtained by multiplying and adding the effect size (β) to the genotypes of rs11079281, rs10444110, rs251466, rs16935073, rs2228479, and rs7430957 is less than 0.28, and the risk of developing diffuse dark spots on the skin is classified as a low-risk group, and the risk score is 0.28 or higher, and the risk of developing diffuse dark spots on the skin is classified as a high-risk group.

18. In paragraph 17, A method of providing information, wherein a customized substance is prescribed in a case where the risk of occurrence of the above-mentioned skin-dispersed dark spots is high-risk.

19. In paragraph 18, A method for providing information, wherein the above substances are bisabolol, nutmeg extract and niacinamide.

20. In paragraph 15, A method for providing information, wherein the sample is hair, urine, blood, various body fluids, isolated tissue, isolated cells or saliva.

21. In paragraph 15, A method for providing information, wherein amplification and confirmation of the above polymorphic site are performed using a SNP chip.

22. A composition for improving dark skin tone, comprising as an active ingredient at least one substance selected from the group consisting of polydatin, ferulic acid, bakuchiol, genistein, oil-soluble licorice extract, aspergillus ferment, quercetin, tannic acid, ZnPCA, gamma oryzanol, chrysin, tranexamic acid, sunflower oil, and isostearic acid.

23. In the 22nd paragraph, the composition (i) ferulic acid, polydatin, zinc PCA, tranexamic acid, quercetin, genistein, tannic acid, Aspergillus ferment, gamma-oryzanol, sunflower seed oil and isostearic acid; (ii) ferulic acid, polydatin, bakuchiol, genistein, soluble licorice extract and Aspergillus ferment; (iii) ferulic acid, quercetin, genistein, tannic acid, polydatin, and zinc PCA, gamma-oryzanol; or (iv) A composition comprising chrysin, tannic acid, and zinc PCA as active ingredients.

24. In paragraph 22, 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.

25. In paragraph 22, The composition is characterized by a decrease in melanin production.

26. In paragraph 25, A composition wherein the substances for reducing melanin production are i) ferulic acid, soluble licorice extract, and bakuchiol; or ii) polydatin and quercetin.

27. In paragraph 22, The composition is characterized by a decrease in protein carbonylation.

28. In paragraph 27, A composition wherein the substance for reducing the above protein carbonylation is gamma-oryzanol, aspergillus ferment, or isostearic acid.

29. In paragraph 22, A composition characterized in that the above substance inhibits the expression of OCA2, BNC2, RAB32 or KITLG.

30. In paragraph 29, A composition wherein the substance for inhibiting the expression of OCA2 is ferulic acid, polydatin, zinc PCA, tranexamic acid, quercetin, genistein, tannic acid, Aspergillus ferment gamma-oryzanol, sunflower seed oil, or isostearic acid.

31. In paragraph 29, A composition wherein the substance for inhibiting the expression of BNC2 is polydatin, ferulic acid, bakuchiol, genistein, a soluble licorice extract, or an Aspergillus ferment.

32. In paragraph 29, A composition wherein the substance for inhibiting the expression of RAB32 is ferulic acid, quercetin, genistein, tannic acid, polydatin, zinc PCA, or gamma-oryzanol.

33. In paragraph 29, A composition wherein the substance for inhibiting the expression of KITLG is chrysin, tannic acid, or zinc PCA.

34. In paragraph 22, The composition is tailored to target an individual having one or more single nucleotide polymorphism markers present in a gene encoding a protein of OCA2, BNC2, RAB32, KITLG or ASIP.

35. In paragraph 34, A composition wherein the single nucleotide polymorphism marker is at least one selected from the group consisting of rs74653330, rs16935073, rs72620727, rs77310623, and rs6059711.

36. In paragraph 35, The above single nucleotide polymorphism marker for diagnosing dark skin tone is a polynucleotide consisting of 5-100 consecutive DNA sequences including the 28228553rd base of human chromosome 15, which is T or C (rs74653330); A polynucleotide consisting of 5-100 contiguous DNA sequences including the 16795790th base of human chromosome 9, wherein the 16795790th base is C or A (rs16935073); A polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 89110410th base of human chromosome 12, wherein the 89110410th base is C or A (rs72620727); A polynucleotide consisting of 5 to 100 consecutive DNA sequences including the 146871819th base of human chromosome 6, wherein the 146871819th base is A or G (rs77310623); A composition comprising a polynucleotide comprising 5 to 100 contiguous DNA sequences including the 32780822nd base of human chromosome 20, wherein the 32780822nd base is T or C (rs6059711); and one or more polynucleotides selected from the group consisting of complementary polynucleotides thereof. 37.(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 at least one single nucleotide polymorphism marker selected from the group consisting of rs74653330, rs16935073, rs72620727, rs77310623, and rs6059711 from the biological sample; and (c) A method for providing information for selecting a customized substance for improving dark skin tone, comprising a step of identifying the base of the amplified or hybridized polymorphic site of step (b).

38. In paragraph 37, A method for providing information, wherein the above substance is at least one selected from the group consisting of polydatin, ferulic acid, bakuchiol, genistein, soluble licorice extract, aspergillus ferment, quercetin, tannic acid, zinc PCA, gamma-oryzanol, chrysin, tranexamic acid, sunflower seed oil, and isostearic acid.

39. In paragraph 37, An information provision method wherein the risk score obtained by multiplying and adding the effect size (β) to the genotypes of rs74653330, rs16935073, rs72620727, rs77310623, and rs6059711 is less than -0.193, and the risk of developing dark skin tone is classified as a low-risk group, and the risk score is more than -0.193, and the risk of developing dark skin tone is classified as a high-risk group.

40. In paragraph 39, A method of providing information, wherein a customized substance is prescribed in cases where the risk of developing the above dark skin tone is high.

41. In paragraph 40, A method for providing information, wherein the above substances are isostearic acid, sunflower seed oil and polydatin.

42. In paragraph 37, A method for providing information, wherein the sample is hair, urine, blood, various body fluids, isolated tissue, isolated cells or saliva.

43. In paragraph 37, A method for providing information, wherein amplification and confirmation of the above polymorphic site are performed using a SNP chip.

44. A composition for improving spot-like pigmentation, comprising as an active ingredient at least one substance selected from the group consisting of ascorbyl glucoside, 3-O-ethyl ascorbic acid, ascorbic acid, Glycyrrhiza uralensis (Licorice) extract, tocopherol, sodium pyruvate, Ecklonia cava extract, hexyldecanol, asiaticoside, and hexylresorcinol.

45. In paragraph 44, the composition (i) Tocopherol, licorice extract and perilla leaf extract; (ii) sodium pyruvate, hexyldecanol, asiaticoside and hexylresorcinol; or (iii) A composition comprising ascorbyl glucoside, 3-O-ethyl ascorbic acid and ascorbic acid as active ingredients.

46. ​​In paragraph 44, 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.

47. A composition according to claim 44, wherein the spot-like pigmentation is at least one selected from the group consisting of senile lentigines, melasma, solar lentigines, black warts, and lentigos.

48. In paragraph 44, The composition is characterized by a decrease in melanin production.

49. In paragraph 48, A composition wherein the substance for reducing melanin production is i) hexylresorcinol and hexyldecanol; or ii) hexyldecanol and tocopherol.

50. In paragraph 44, A composition characterized in that the above substance inhibits the expression of ASCL2 or TP63 or promotes the expression of PPARGC1B.

51. In paragraph 50, A composition wherein the substance for inhibiting the expression of ASCL2 is tocopherol, licorice extract or perilla leaf extract.

52. In paragraph 50, A composition wherein the substance for inhibiting the expression of TP63 is sodium pyruvate, hexyldecanol, asiaticoside or hexylresorcinol.

53. In paragraph 50, A composition wherein the substance for promoting the expression of PPARGC1B is ascorbyl glucoside, 3-O-ethyl ascorbic acid or ascorbic acid.

54. In paragraph 44, The composition is a customized composition targeting an individual having one or more single nucleotide polymorphism markers present in a gene encoding a protein of ASCL2, TP63 or PPARGC1B.

55. In paragraph 54, A composition wherein the single nucleotide polymorphism marker is at least one selected from the group consisting of rs251466, rs12222609, and rs9865771.

56. In paragraph 55, The single nucleotide polymorphism marker for the diagnosis of the above-mentioned spot-like pigmentation is a polynucleotide consisting of 5-100 consecutive DNA sequences including the 149195603rd base of the 5th chromosome of a human being, wherein the 149195603rd base is C or G (rs251466); A polynucleotide consisting of 5-100 contiguous DNA sequences including the 2297093rd base of human chromosome 11, wherein the 2297093rd base is C or T (rs12222609); A composition comprising a polynucleotide comprising 5 to 100 contiguous DNA sequences including the 189491453rd base of human chromosome 3, wherein the 189491453rd base is A or G (rs9865771); and one or more polynucleotides selected from the group consisting of complementary polynucleotides thereof. 57.(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 at least one single nucleotide polymorphism marker selected from the group consisting of rs251466, rs12222609, and rs9865771 from the biological sample; and (c) A method for providing information for selecting a customized substance for improving spot-like pigmentation, comprising a step of identifying the base of the amplified or hybridized polymorphic site of step (b).

58. In paragraph 57, A method for providing information, wherein the above substance is at least one selected from the group consisting of ascorbyl glucoside, 3-O-ethyl ascorbic acid, ascorbic acid, licorice extract, tocopherol, sodium pyruvate, kelp extract, hexyldecanol, asiaticoside, and hexylresorcinol.

59. In paragraph 57, An information provision method wherein the risk score obtained by multiplying and adding the effect size (β) to the genotypes of rs251466, rs12222609, and rs9865771 is -0.009 or less, and the risk of developing spotty pigmentation is classified as a low-risk group, and the risk score is 0.000 or more, and the risk of developing spotty pigmentation is classified as a high-risk group.

60. In paragraph 59, A method of providing information, wherein a customized substance is prescribed in cases where the risk of developing the above-mentioned spot-like pigmentation is high-risk.

61. In paragraph 60, A method for providing information, wherein the above substances are tocopherol, hexylresorcinol and hexyldecanol.

62. In paragraph 57, A method for providing information, wherein the sample is hair, urine, blood, various body fluids, isolated tissue, isolated cells or saliva.

63. In paragraph 57, A method for providing information, wherein amplification and confirmation of the above polymorphic site are performed using a SNP chip.

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