JP2007252348A - Method for classifying expression form of skin color and method for selecting cosmetic based on the same - Google Patents
Method for classifying expression form of skin color and method for selecting cosmetic based on the same Download PDFInfo
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Abstract
Description
本発明は、ヒトメラノサイト刺激ホルモン１受容体のＤＮＡ配列のプロモーター部分の一塩基置換を利用した皮膚の色の発現形態の分類法及び該分類法を利用した化粧料の選択方法に関する。なお、本発明においては、コーディング部位の直前の部位をプロモーター１位と定義する。 The present invention relates to a method for classifying skin color expression using single base substitution of the promoter portion of the DNA sequence of human melanocyte-stimulating hormone 1 receptor, and a method for selecting a cosmetic using the classification method. In the present invention, the site immediately before the coding site is defined as promoter position 1.
メラノサイト刺激ホルモンは、メラニンの産生や炎症に関わるホルモンであり、そのうちメラノサイト刺激ホルモン１は主としてメラニンの産生に関わっていると言われている。メラノサイト刺激ホルモン１受容体（以後、単にＭＣ１Ｒと称する場合がある。）はこの意味で、生体のメラニン産生と深く関わっていると言える。又、かかるＭＣ１ＲをコードしたＤＮＡについても、その塩基配列は明らかにされている。かかるＭＣ１Ｒの954塩基対（合計317アミノ酸）からなるコーディング領域については、アミノ酸９２位のバリン、同１６３位のアルギニン等に一塩基置換（以後、単にＳＮＰと称することもある。）が存在することが知られている（例えば、非特許文献１、非特許文献２、非特許文献３、非特許文献４）し、この様なＳＮＰの中には、肌色や、毛髪の色などに影響を及ぼすものも存することが知られている。しかしながら、ヒトメラノサイト刺激ホルモン１受容体をコードした核酸のコーディング部位の直前の部位をプロモーター１位と定義した場合の、プロモーター部分の塩基配列の一塩基置換の存在の有無、又、その利用については全く知られていないのが現状であった。 Melanocyte-stimulating hormone is a hormone involved in melanin production and inflammation, and melanocyte-stimulating hormone 1 is said to be mainly involved in melanin production. In this sense, the melanocyte stimulating hormone 1 receptor (hereinafter sometimes simply referred to as MC1R) can be said to be deeply related to melanin production in the living body. The base sequence of the DNA encoding MC1R has also been clarified. With respect to the coding region consisting of 954 base pairs of MC1R (total of 317 amino acids), a single base substitution (hereinafter sometimes simply referred to as SNP) exists in valine at amino acid position 92, arginine at position 163, and the like. Are known (for example, Non-Patent Document 1, Non-Patent Document 2, Non-Patent Document 3, Non-Patent Document 4), and some of these SNPs affect skin color, hair color, etc. It is known that things exist. However, when the site immediately before the coding site of the nucleic acid encoding human melanocyte-stimulating hormone 1 receptor is defined as promoter position 1, the presence or absence of a single base substitution in the base sequence of the promoter portion, and its use The current situation is not known at all.
一方、皮膚についての個人差の大きさは、化粧品技術者の誰もが認識するところであり、この個人差を克服して、個人個人に適した化粧料を提供することは、化粧品技術者の誰もが見る夢であるが、どのように個人個人に適しているか否かを判別することについては、確固たる技術は存しておらず、経験によるところが少なくなかった。この意味で、客観的な化粧料の選択方法の開発が望まれていると言える。 On the other hand, the level of individual differences in skin is recognized by all cosmetic engineers, and it is nobody in cosmetic engineers to overcome this individual difference and provide cosmetics suitable for individuals. Although it is a dream that Momo sees, there is no firm technique for determining whether or not it is suitable for an individual, and there are not a few cases based on experience. In this sense, it can be said that development of an objective cosmetic material selection method is desired.
本発明は、この様な状況下為されたものであり、ＭＣ１ＲのＳＮＰを利用した、客観的に皮膚の色の発現形態を分類方法を提供し、以て、個々人に適した化粧料を選択する手段をも提供することを課題とする。 The present invention has been made under such circumstances, and provides a method for objectively classifying the appearance of skin color using MC1R SNPs, thereby selecting a cosmetic suitable for each individual. It is an object of the present invention to provide a means to do this.
この様な状況に鑑みて、本発明者らは、個々人に適した化粧料を選択する手段を提供すべく、ＭＣ１ＲのＳＮＰを利用した、客観的に皮膚の色の発現形態を分類方法を求めて、鋭意研究努力を重ねた結果、ヒトメラノサイト刺激ホルモン１受容体のＤＮＡ塩基配列の内、前記プロモーター部分４９０位、４４５位及び２２６位の塩基の組み合わせを指標とすることにより、この様な分類ができることを見いだし、発明を完成させるに至った。即ち、本発明は以下に示す通りである。
（１）皮膚の色の発現形態を分類する場合において、刺激による皮膚の黒くなりやすさと、皮膚自体の色の黒さの２軸で特性を鑑別する方法であって、ヒトメラノサイト刺激ホルモン１受容体のＤＮＡ塩基配列の内、プロモーター部分４９０位、４４５位及び２２６位の塩基の組み合わせを指標とすることを特徴とする、皮膚の色の発現形態の分類法。
（２）ヒトメラノサイト刺激ホルモン１受容体のＤＮＡ塩基配列の内、プロモーター部分４９０位、４４５位及び２２６位の塩基の組み合わせによる分類が、１）４９０位Ｃ、４４５位Ｇ、２２６位Ａ／４９０位Ｃ、４４５位Ｇ、２２６位Ａと２）４９０位Ｃ、４４５位Ｇ、２２６位Ａ／４９０位Ｔ、４４５位Ａ、２２６位Ｔと３）４９０位Ｔ、４４５位Ａ、２２６位Ｔ／４９０位Ｔ、４４５位Ａ、２２６位Ｔの何れかに属するで分類することを特徴とする、（１）に記載の皮膚の色の発現形態の分類法。
（３）ヒトメラノサイト刺激ホルモン１受容体のＤＮＡ塩基配列の内、プロモーター部分に４９０位Ｃ、４４５位Ｇ、２２６位Ａの塩基配列を持つ蓋然性が高い場合、定常的なメラニン産生量が多い人と分類することを特徴とする、（１）又は（２）に記載の皮膚の色の発現形態の分類法。
（４）人より採取した毛髪より、ＤＮＡを抽出し、該ＤＮＡのメラノサイト刺激ホルモン１受容体に関するプロモーター領域とコーディング領域を含む８８２塩基対をttgacagctgagttgctgct（配列式１）のオリゴヌクレオチドとaggaagagcccgtcagagat（配列式２）のオリゴヌクレオチドをプライマーとして、これらの存在下増幅し、しかる後、acccctgtcctccctgag（配列式３）、caggaaggcaggagacagag（配列式４）、tcagagatggacacctccag（配列式５）、gacacctcctggcatctacc（配列式６）のオリゴヌクレオチドをプライマーして、これらの存在下シークエンス反応を行い、しかる後に配列を解読し、プロモーター部分４９０位、４４５位及び２２６位の塩基を同定し、該塩基のパターンより、１）４９０位Ｃ、４４５位Ｇ、２２６位Ａ／４９０位Ｃ、４４５位Ｇ、２２６位Ａと２）４９０位Ｃ、４４５位Ｇ、２２６位Ａ／４９０位Ｔ、４４５位Ａ、２２６位Ｔと３）４９０位Ｔ、４４５位Ａ、２２６位Ｔ／４９０位Ｔ、４４５位Ａ、２２６位Ｔの何れかに分類し、１）に分類される人には定常的にメラニンの生成を抑制する化粧料を勧め、２）に分類される人には、定常的にメラニンの生成を抑制する化粧料と、紫外線防護化粧料とを併用することを勧め、３）に分類される人には紫外線防護化粧料の定常的に使用することを勧めることを特徴とする、（１）〜（３）何れか１項に記載の皮膚の色の発現形態の分類法に基づいた化粧料の選択方法。
（５）ヒトメラノサイト刺激ホルモン１受容体をコードした核酸のプロモーター部分の塩基配列４９０位、４４５位及び２２６位の塩基の一塩基置換の化粧料の選択のための使用。
In view of such circumstances, the present inventors have sought a method for objectively classifying skin color expression using the MC1R SNP in order to provide means for selecting a cosmetic suitable for each individual. As a result of intensive research efforts, such a classification was made by using as an index the combination of the bases at positions 490, 445 and 226 in the promoter portion of the DNA sequence of the human melanocyte stimulating hormone 1 receptor. I found out that I could do it, and came to complete the invention. That is, the present invention is as follows.
(1) When classifying the expression form of the skin color, it is a method for distinguishing the characteristics by two axes, i.e., the ease of darkening of the skin by stimulation and the blackness of the color of the skin itself. A method for classifying the expression form of skin color, characterized in that a combination of bases at positions 490, 445 and 226 in the promoter portion of the body DNA base sequence is used as an index.
(2) Among the DNA base sequences of human melanocyte stimulating hormone 1 receptor, the classification according to the combination of the bases at positions 490, 445 and 226 in the promoter portion is 1) 490, C, 445, G, 226, A / 490 Position C, 445th position G, 226th position A and 2) 490th position C, 445th position G, 226th position A / 490th position T, 445th position A, 226th position T and 3) 490th position T, 445th position A, 226th position T / Classification of expression form of skin color according to (1), characterized by belonging to any of / 490 position T, 445 position A, and 226 position T.
(3) A person with a large amount of steady melanin production when the promoter has a high probability of having a base sequence of positions 490, C, 445, and 226 in the DNA sequence of the human melanocyte stimulating hormone 1 receptor. (1) or (2), the method for classifying the expression form of the skin color according to (1) or (2).
(4) DNA is extracted from hair collected from a person, and 882 base pairs including a promoter region and a coding region for the melanocyte-stimulating hormone 1 receptor of the DNA are converted into oligonucleotides of ttgacagctgagttgctgct (Sequence Formula 1) and aggagagagcccgtcagagat (Sequence Formula) 2) Using the oligonucleotides of 2) as primers, amplification is performed in the presence of them, and then the oligonucleotides of acccctgtcctccctgag (Sequence Formula 3), caggaaggcaggagacagag (Sequence Formula 4), tcagagatggacacctccag (Sequence Formula 5), and gacacctcctggcatctacc (Sequence Formula 6) The primer is subjected to a sequencing reaction in the presence of these, and then the sequence is decoded to identify the bases at positions 490, 445, and 226 of the promoter portion. From the base pattern, 1) positions 490, C, and 445 G, 226th position A / 490th position C, 445th position G, 226th position A and 2) 4 90 C, 445 G, 226 A / 490 T, 445 A, 226 T and 3) 490 T, 445 A, 226 T / 490 T, 445 A, 226 T Cosmetics that constantly suppress the production of melanin are recommended for those classified into any one of the categories 1), and cosmetics that constantly suppress the production of melanin for those classified in 2) In addition, it is recommended to use UV protective cosmetics in combination with those classified in 3), and it is recommended to use UV protective cosmetics regularly. (1) to (3) A method for selecting a cosmetic based on the classification method for the appearance form of skin color according to claim 1.
(5) Use for the selection of cosmetics having a single base substitution at the base sequence of positions 490, 445 and 226 of the promoter part of the nucleic acid encoding human melanocyte stimulating hormone 1 receptor.
本発明によれば、ＭＣ１ＲのＳＮＰを利用した、客観的に皮膚の色の発現形態を分類方法を提供し、以て、個々人に適した化粧料を選択する手段をも提供することができる。 According to the present invention, it is possible to provide a method for objectively classifying the expression form of the color of the skin using the SNP of MC1R, and thus provide a means for selecting a cosmetic suitable for an individual.
本発明の中心部分は、ＭＣ１Ｒをコードする核酸のプロモーター部分にＳＮＰが存在し、該ＳＮＰの種類により、発現系である皮膚の色が影響を受けていること見いだしたこと、該ＳＮＰより、１）４９０位Ｃ、４４５位Ｇ、２２６位Ａ（以後、単にＣＧＡと記す場合もある）／４９０位Ｃ、４４５位Ｇ、２２６位Ａと２）４９０位Ｃ、４４５位Ｇ、２２６位Ａ／４９０位Ｔ、４４５位Ａ、２２６位Ｔ（以後、単にＴＡＴと記す場合もある）と３）４９０位Ｔ、４４５位Ａ、２２６位Ｔ／４９０位Ｔ、４４５位Ａ、２２６位Ｔの３分類とすることができ、この３分類の何れかに分類することができ、ＣＧＡの個数が多いほど定常的なメラニン産生能が高まり、ＴＡＴの個数が多いほど刺激に反応して産生されることを見いだしたことである。 The central part of the present invention is that the SNP is present in the promoter part of the nucleic acid encoding MC1R, and that the color of the skin, which is the expression system, is influenced by the type of the SNP. ) 490th position C, 445th position G, 226th position A (hereinafter sometimes referred to simply as CGA) / 490th position C, 445th position G, 226th position A and 2) 490th position C, 445th position G, 226th position A / 490th T, 445th A, 226th T (hereinafter sometimes simply referred to as TAT) and 3) 490th T, 445th A, 226th T / 490th T, 445th A, 226th T It can be classified into any one of these three classifications, and the higher the number of CGAs, the higher the ability to produce steady melanin, and the higher the number of TATs, the more they are produced in response to stimulation. I found that.
前記ＳＮＰを検出するに当たり、人よりＤＮＡを抽出し、ＭＣ１Ｒコード部分を増幅する必要が存するが、ヒトＭＣ１Ｒの塩基配列に関しては、既に公知であり（例えば、GenBank Accession No.NM_002386参照）、この配列を元にプライマーを常法に従って設計し、ＰＣＲにかけて増幅することができる。この様なプライマーとしては、ttgacagctgagttgctgct（配列式１）のオリゴヌクレオチドとaggaagagcccgtcagagat（配列式２）のオリゴヌクレオチドとプライマーとして用いることが好適に例示できる。又、ＤＮＡを抽出するためのサンプルとしては、口腔内粘膜細胞、毛髪、眉毛などが例示でき、眉毛をサンプルとして、市販のＤＮＡ抽出キットを用いて抽出することが好ましく例示できる。この様な抽出キットとしては、例えば、ISO HAIR kit（ニッポンジーン社製）等が好適に例示できる。斯くして、増幅されたＭＣ１ＲのＤＮＡより、ＳＮＰを検出するわけであるが、かかる検出には、通常用いられている方法が例示でき、例えば、ＭＣ１Ｒ遺伝子の全塩基配列を決定しても良いし、１塩基のみを指標にして解析しても良い。１塩基だけを指標として解析する方法としては、例えば、ＲＰＡ（Rnase Protection assay）法、ＤＧＧＥ（denaturing gradient gel electrophoresis）法、ＳＳＣＰ（single strand conformation polymorphism）法、ＭＡＳＡ（mutant-allele-specific amplification）法、ＡＳＯ（allele-specific-oligonucleotide）ハイブリダイゼーション法、オリゴヌクレオチドアレイ法などが好ましく例示できる。この一例を以下に示す。 In detecting the SNP, it is necessary to extract DNA from humans and amplify the MC1R coding portion. However, the base sequence of human MC1R is already known (see, for example, GenBank Accession No. NM_002386). Based on the above, primers can be designed according to a conventional method and amplified by PCR. As such a primer, it can be preferably exemplified as an oligonucleotide of ttgacagctgagttgctgct (Sequence Formula 1), an aggagaagagcccgtcagagat (Sequence Formula 2) and a primer. Further, examples of the sample for extracting DNA include oral mucosal cells, hair, eyebrows and the like. Preferably, the eyebrow is used as a sample and extracted using a commercially available DNA extraction kit. As such an extraction kit, for example, ISO HAIR kit (manufactured by Nippon Gene Co., Ltd.) and the like can be suitably exemplified. Thus, SNP is detected from the amplified MC1R DNA, and for this detection, a commonly used method can be exemplified. For example, the entire base sequence of the MC1R gene may be determined. The analysis may be performed using only one base as an index. Examples of the analysis method using only one base as an index include, for example, RPA (Rnase Protection assay) method, DGGE (denaturing gradient gel electrophoresis) method, SSCP (single strand conformation polymorphism) method, MASA (mutant-allele-specific amplification) method. ASO (allele-specific-oligonucleotide) hybridization method, oligonucleotide array method and the like can be preferably exemplified. An example of this is shown below.
増幅したＭＣ１ＲのＤＮＡはシークエンス反応に付す。即ち、ABI BigDye Terminator Cycle Sequencing kit(Ver.3.1 アプライドバイオ社製)を用い、プライマーはacccctgtcctccctgag（配列式３）、caggaaggcaggagacagag（配列式４）、tcagagatggacacctccag（配列式５）及びgacacctcctggcatctacc（配列式６）の４種を使用した。反応条件は９８℃５分、９８℃１０秒―５０℃５分―６０℃４分を２５サイクルで行った。シークエンス反応物をＤｙｅＥＸＴＭ Ｓｐｉｎ Ｋｉｔ（キアゲン社製）を用いてカラムによる未反応物の除去による精製を行った。シークエンス反応物をＡＢＩ ＰＲＩＳＭ ３１００ Ｇｅｎｅｔｉｃ Ａｎａｌｙｚｅｒ（アプライドバイオ社製）を用いて配列の解読をし、精度よく解読されたプロモーター領域７１８ｂｐについて、ＳｅｑＳｃａｐｅ Ｖｅｒ．２．（アプライドバイオ社製）を用いてＳＮＰ判定を行った。 The amplified MC1R DNA is subjected to a sequencing reaction. That is, ABI BigDye Terminator Cycle Sequencing kit (Ver.3.1 Applied Bio) was used, and the primers were acccctgtcctccctgag (Sequence Formula 3), caggaaggcaggagacagag (Sequence Formula 4), tcagagatggacacctccag (Sequence Formula 5) and gacacctcctggcatctacc (Sequence Formula 6). Four types were used. The reaction conditions were 98 ° C for 5 minutes, 98 ° C for 10 seconds-50 ° C for 5 minutes-60 ° C for 4 minutes in 25 cycles. The sequence reaction product was purified by removing unreacted material through a column using DyeEX ™ Spin Kit (Qiagen). The sequence reaction was sequenced using ABI PRISM 3100 Genetic Analyzer (Applied Bio Inc.), and the accurately decoded promoter region 718 bp was analyzed using SeqScape Ver. 2. SNP determination was performed using (Applied Bio).
この結果、前記のＣＧＡ／ＣＧＡ、ＣＧＡ／ＴＡＴ及びＴＡＴ／ＴＡＴの３種のＳＮＰの組み合わせが存することがわかった。２４５名の男女（年齢２０〜５５歳）のパネラーより採取した眉毛より求めた、ＳＮＰの組み合わせは表１に示すとおりであった。 As a result, it was found that there were three combinations of SNPs, CGA / CGA, CGA / TAT and TAT / TAT. The combinations of SNPs determined from the eyebrows collected from 245 male and female (aged 20-55) panelists were as shown in Table 1.
又、既に知られているＭＣ１Ｒ遺伝子のコーディング領域１６３位のＳＮＰの組み合わせとの相関性を見てみると以下の表２に示すようになる。即ち、コーディング領域アミノ酸Gln/Glnとプロモーター部TAT/TATが、アミノ酸Gln/ArgとCGA/TATが、アミノ酸Arg/ArgとCGA/CGAとが非常に相関していることがわかる。コーディング領域のＳＮＰの発現への影響を除去するために、ＣＧＡ／ＴＡＴについては、Ｇｌｎ／Ａｒｇの人だけを、ＣＧＡ／ＣＧＡについては、Ａｒｇ／Ａｒｇの人だけを解析対象とした。 Further, looking at the correlation with the already known combination of SNPs at the 163rd position of the coding region 163 of the MC1R gene, it is as shown in Table 2 below. That is, it can be seen that the coding region amino acids Gln / Gln and the promoter portion TAT / TAT are highly correlated with the amino acids Gln / Arg and CGA / TAT, and the amino acids Arg / Arg and CGA / CGA. In order to eliminate the influence on the SNP expression of the coding region, only Cln / Arg people were analyzed for CGA / TAT, and only Arg / Arg people were analyzed for CGA / CGA.
この様な前提のもとに、ＣＧＡ／ＣＧＡのタイプの人と、ＣＧＡ／ＴＡＴタイプの人の肌の特性の比較を行った。
（１） 顔色 (ITA 値）をコニカミノルタ社製の色彩色差計を用いて計測した。結果を表３に示す。TAT／OGAタイプの人の方がＣＧＡ／OGAタイプの人よりＩＴＡ値が低く、顔の色が定常的に黒いことがわかる。TATパターンのＳＮＰ組み合わせを持つことにより、定常的な紫外線刺激に対するメラニン産生能が高いことが示唆される。
Based on these assumptions, the skin characteristics of a CGA / CGA type person and a CGA / TAT type person were compared.
(1) The face color (ITA value) was measured using a color difference meter manufactured by Konica Minolta. The results are shown in Table 3. It can be seen that the TAT / OGA type person has a lower ITA value than the CGA / OGA type person, and the face color is constantly black. By having a SNP combination of TAT pattern, it is suggested that the ability to produce melanin with respect to steady UV stimulation is high.
（２）スキンタイプの鑑別を行った。スキンタイプは照射ドーズを振って、紫外線を照射し、最少紅斑量を求めることにより、最少紅斑量の多少と、照射後２４時間、４８時間、９６時間の皮膚反応より、下記の基準に従って、スキンタイプを決定した。ＳＮＰの組み合わせとスキンタイプの関係を表４に示す。これよりＴＡＴ／ＣＧＡの人はＣＧＡ／ＣＧＡタイプの人より、スキンタイプ３の性質を有する蓋然性が高いことがわかる。
スキンタイプ１：照射後２４時間以内に紅斑が出現し、９６時間では黒化を経ずに紅斑が消衰する。最少紅斑量は少ない。
スキンタイプ２：照射後２４時間以内に紅斑が出現し、４８時間で黒化に転じ、９６時間では赤みがなく、黒くなる。最少紅斑量は多くも少なくもない。
スキンタイプ３：紅斑が出現せずに、そのまま黒化する。最少紅斑量は多い。
(2) The skin type was identified. The skin type is irradiated with ultraviolet rays, and the minimum amount of erythema is obtained by irradiating ultraviolet rays, and the skin is evaluated according to the following criteria from the amount of minimum erythema and the skin reaction at 24 hours, 48 hours, and 96 hours after irradiation. Decided the type. Table 4 shows the relationship between SNP combinations and skin types. From this, it can be seen that TAT / CGA people are more likely to have skin type 3 properties than CGA / CGA type people.
Skin type 1: Erythema appears within 24 hours after irradiation, and in 96 hours, erythema disappears without blackening. The minimum amount of erythema is small.
Skin type 2: Erythema appears within 24 hours after irradiation, turns black after 48 hours, has no redness and becomes black after 96 hours. The minimum amount of erythema is not too much.
Skin type 3: Black spots appear without appearance of erythema. The minimum amount of erythema is large.
（３）これらのパネラーについて、黒子の存在状況を第三者によって観察した。観察の基準は、距離１００ｃｍで顔を観察して、目立つ黒子の数を計数し、１０個未満の場合には「黒子が少ない」に、１０以上２０未満の場合に「黒子が中程度」に、２０個以上の場合には「黒子が多い」に分類した。客観性を維持するために、観察者は３名とし、観察結果が割れた場合には多数の方に分類した。結果を表５に示す。これより、黒子のたくさん存在する蓋然性は、ＴＡＴ／ＣＧＡの人の方がＣＧＡ／ＣＧＡタイプの人より高いことがわかる。 (3) About these panelists, the existence situation of Kuroko was observed by the third party. The standard of observation is to observe the face at a distance of 100 cm, count the number of outstanding moles, and if it is less than 10, it is “small moles”, and if it is 10 or more and less than 20, it is “medium moles”. In the case of 20 or more, it was classified as “many moles”. In order to maintain objectivity, the number of observers was three, and when the observation result was broken, it was classified into a large number of people. The results are shown in Table 5. From this, it can be seen that the probability of having a lot of moles is higher in TAT / CGA people than in CGA / CGA types.
これらのデータより、ＴＡＴのＳＮＰの組み合わせを有することにより、ＵＶ刺激を受けて、メラニン産生を亢進させやすい基質を有するようになることが推察された。このこと検証する為に、更に、コーディング１６３位アミノ酸Gln/Argの人について、ＣＧＡ／ＣＧＡ、ＣＧＡ／ＴＡＴ及びＴＡＴ／ＴＡＴについての解析を同様に行った。 From these data, it was inferred that having a combination of SATs of TAT has a substrate that is likely to enhance melanin production under UV stimulation. In order to verify this, the analysis of CGA / CGA, CGA / TAT, and TAT / TAT was similarly performed on the person with coding amino acid Gln / Arg at position 163.
（４）腕の色についての解析
紫外線が当たりにくく、光の刺激のないところでの肌の色に代替する、上腕内側部の色を顔の色と同様に測定し、比較した。結果を表６に示す。ＣＧＡ／ＣＧＡとＴＡＴ／ＴＡＴの間では、Ｐ＝０．０３６８で有意差が認められた。これより、ＣＧＡのＳＮＰの組み合わせを有することにより、刺激を受けない基底状態でのメラニン産生が亢進する基質を有しやすいことがわかる。
(4) Analysis of the color of the arm The color of the inner side of the upper arm, which substitutes for the skin color where it is difficult to be exposed to ultraviolet light and where there is no light stimulation, was measured and compared in the same manner as the face color. The results are shown in Table 6. There was a significant difference between CGA / CGA and TAT / TAT at P = 0.0368. From this, it can be seen that having a combination of CGA SNPs tends to have a substrate that enhances melanin production in the basal state without stimulation.
（５）スキンタイプの解析
（２）と同様にスキンタイプの解析を行った。結果を表７に示す。これより、（２）と同様にＴＡＴのＳＮＰの組み合わせが紫外線に反応してメラニンを作りやすい形質を受け継ぐことがわかる。
(5) Analysis of skin type Skin type was analyzed in the same manner as in (2). The results are shown in Table 7. From this, it can be seen that the combination of SATs of TAT inherits the trait that makes it easy to produce melanin in response to ultraviolet rays, as in (2).
（６）シミの解析
（３）黒子の黒子の解析の手技と同様の手技で観察対象をシミにして、同様にシミの判定を行った。判定基準は次に示すものを用いた。即ち、観察の基準は、距離１００ｃｍで顔を観察して、目立つシミの数を計数し、シミが見あたらない場合は「シミスコア１」に、１個以上１０個未満の場合には「シミスコア２」に、１０以上１５未満の場合に「シミスコア３」に、１５個以上２０個未満の場合には「シミスコア４」に、２０個以上の場合には「シミスコア５」に分類した。結果を表８に示す。これより、ＣＧＡ／ＣＧＡ＞ＣＧＡ／ＴＡＴ＞ＴＡＴ／ＴＡＴの順でシミが重篤になることがわかる。ＣＧＡのＳＮＰ組み合わせを有することは、シミを重篤化させやすい形質を引き継ぐ傾向にあることがわかる。
(6) Spot analysis (3) Spots were similarly determined using the same technique as that of Kuroko's mole analysis. The following criteria were used. That is, the standard of observation is to observe the face at a distance of 100 cm, count the number of noticeable spots, “spot score 1” when no spots are found, “spot score 2” when 1 or more and less than 10 spots. In addition, when it was 10 or more and less than 15, it was classified as “spot score 3”, when it was 15 or more but less than 20, it was classified as “stain score 4”, and when it was 20 or more, it was classified as “stain score 5”. The results are shown in Table 8. From this, it can be seen that the spots become serious in the order of CGA / CGA> CGA / TAT> TAT / TAT. It can be seen that having a CGA SNP combination tends to inherit a trait that is likely to cause severe spots.
以上の解析結果より、ＭＣ１Ｒのプロモーター領域にＣＧＡのＳＮＰ組み合わせによって発現される形質は、定常的にメラニンの産生が亢進している形質で光などの刺激の非存在下でもメラニン産生が亢進され、肌の色が黒くなりやすい形質となる。この様な形質に対しては普段の肌の手入れにおいて、常に美白作用を有する化粧料を使用することが好ましい。取り分け、美白作用に於いては、メラニン産生抑制作用のみならず、例えば、メラノサイトのデンドライトの伸長を抑制する成分や、メラニン顆粒を分解する成分を併せて含有させ、メラニンの産生を多面的に抑制することが好ましい。メラニン産生抑制成分としては、アルブチン、トラネキサム酸、アスコルビン酸リン酸エステル、アスコルビン酸グルコシド或いはこれらの塩等が好ましく例示でき、デンドライトの伸長を抑制する成分としては、センタウレイジンやメチルオフィオポゴナノンＢ等が好ましく例示でき、メラニン産生抑制成分としてはイソフラバノン類が好ましく例示できる。これらの成分の含有量は、それぞれ０．１〜５質量％が好ましい。ＴＡＴのＳＮＰ組み合わせによって発現される形質は光に敏感に反応して、メラニンの産生を亢進させる形質であり、この様な形質に於いては、光の刺激を生体が受けないことが肝要であり、二酸化チタンや酸化亜鉛を配合した紫外線防護化粧料で紫外線からの防護に注力するとともに、光刺激によってメラニンの生じるのを速やかに抑制するための美白化粧料を使用することが好ましい。この場合、メラニンの生成抑制については、定常的にはメラニンの産生は高いわけではないので、多面的な抑制は要しない。この様な基準で化粧料を選択し、勧めることで、個々人に適合して、白く美しい肌を実現し、これを維持することができる。総括すれば、ＣＧＡ／ＣＧＡに分類される人には定常的にメラニンの生成を抑制する化粧料を勧め、ＣＧＡ／ＴＡＴに分類される人には、定常的にメラニンの生成を抑制する化粧料と、紫外線防護化粧料とを併用することを勧め、ＴＡＴ／ＴＡＴに分類される人には紫外線防護化粧料の定常的に使用することを勧めることが好ましいと言える。 From the above analysis results, the trait expressed by the CGA SNP combination in the promoter region of MC1R is a trait in which the production of melanin is steadily enhanced, and melanin production is enhanced even in the absence of stimuli such as light, The skin color tends to be black. For such traits, it is preferable to always use a cosmetic material that has a whitening effect in normal skin care. In particular, in the whitening effect, not only the melanin production inhibitory action, but also contains, for example, a component that suppresses melanocyte dendrite elongation and a component that decomposes melanin granules, and multifacetedly suppresses melanin production. It is preferable to do. Preferred examples of the melanin production-suppressing component include arbutin, tranexamic acid, ascorbic acid phosphate ester, ascorbic acid glucoside, and salts thereof. Examples of the component that suppresses dendrite elongation include centaureidine and methylophiopogononone. B and the like can be preferably exemplified, and isoflavanones can be preferably exemplified as the melanin production inhibiting component. The content of these components is preferably 0.1 to 5% by mass. The traits expressed by the SAT combination of TAT are those that respond sensitively to light and enhance the production of melanin. In such traits, it is important that the living body is not stimulated by light. In addition, it is preferable to use a whitening cosmetic for emphasizing protection from ultraviolet rays with an ultraviolet protective cosmetic blended with titanium dioxide or zinc oxide and for quickly suppressing the formation of melanin by light stimulation. In this case, since melanin production is not constantly high in terms of melanin production inhibition, multifaceted inhibition is not required. By selecting and recommending cosmetics based on such criteria, it is possible to achieve and maintain white and beautiful skin that suits the individual. In summary, cosmetics that steadily suppress the production of melanin are recommended for people classified as CGA / CGA, and cosmetics that steadily suppress the production of melanin for those classified as CGA / TAT. It is recommended to use UV protective cosmetics in combination, and it is preferable to recommend the regular use of UV protective cosmetics to those classified as TAT / TAT.
以下に、実施例を挙げて、本発明について更に詳細に説明を加えるが、本発明が、かかる実施例にのみ限定されないことは言うまでもない。 Hereinafter, the present invention will be described in more detail with reference to examples, but it goes without saying that the present invention is not limited to such examples.
本発明の皮膚の色の発現形態の分類法と、それを利用した化粧料の選択方法の効果を確かめるために、パネラーを用いて使用テストを行った。パネラーは１群２０名とし、本発明の皮膚の色の発現形態の分類法と、それを利用した化粧料の選択方法群（本発明群）と、対照としての自由選択群（対照群）の２群の計４０名（男性、２５〜５５歳）を用意した。各パネラーは試験開始前に眉毛を採取され、この眉毛より、前述の方法でＭＣ１Ｒのプロモーター領域のＳＮＰの組み合わせを判定しておいた。４０人のパネラーはパネラー属性に差が生じないように又、化粧料としては、各人２種を選択してもらう試験設計とし、１種として紫外線防護化粧料を「紫外線防護化粧料１」と「紫外線防護化粧料２」（プラシーボ）の何れかを、残る１種として、美白化粧料を「美白化粧料１」（多面的メラニン産生抑制効果を有する）と「美白化粧料２」（メラニン産生抑制剤としてアルブチンのみを含有）の何れかを選択してもらった。選択に際しては、本発明群は、ＣＧＡ／ＣＧＡの人は紫外線防護化粧料２と美白化粧料１を、ＣＧＡ／ＴＡＴの人は、紫外線防護化粧料１と美白化粧料１を、ＴＡＴ／ＴＡＴの人は紫外線防護化粧料１と美白化粧料２を渡した。対照群は、個人の選択に任せて紫外線防護化粧料と美白化粧料とを選択してもらった。提供した化粧品以外の化粧品の使用は控えてもらった。選択した化粧料は２８日間連日使用してもらった。試験開始前と２８日間の連用試験の終了後に肌の色を前述の方法と同じ方法で測定した。試験前のＩＴＡ値から試験後のＩＴＡ値を減じたΔＩＴＡを算出し、各群ごとに平均と偏差を求めた。結果を表１３に示す。これより、本発明の皮膚の色の発現形態の分類法を利用した化粧料の選択方法によって、化粧料を選択した群の方がΔＩＴＡの値が大きく、肌色改善効果に優れることがわかる。 In order to confirm the effect of the classification method of the skin color expression form of the present invention and the selection method of cosmetics using the same, a use test was conducted using a panel. There are 20 panelists per group. The classification method of the skin color expression mode of the present invention, the selection method group of cosmetics using the same (the present invention group), and the free selection group (control group) as a control. A total of 40 people (male, 25-55 years old) in two groups were prepared. Each panelist collected eyebrows before the start of the test, and the combination of SNPs in the promoter region of MC1R was determined from the eyebrows by the method described above. The panelists of 40 people do not have a difference in the paneler attributes, and the cosmetics are designed so that each person can select two types of UV protection cosmetics as “UV protection cosmetics 1”. One of “UV-protective cosmetic 2” (placebo) is the remaining one, and whitening cosmetic is “whitening cosmetic 1” (which has a multifaceted melanin production inhibitory effect) and “whitening cosmetic 2” (melanin production) Any one of arbutin contained as an inhibitor) was selected. In selecting the present invention group, the CGA / CGA person applies UV protection cosmetic 2 and whitening cosmetic 1, the CGA / TAT person applies UV protection cosmetic 1 and whitening cosmetic 1, and TAT / TAT. The person handed UV protection cosmetic 1 and whitening cosmetic 2. The control group was allowed to choose between UV protection cosmetics and whitening cosmetics depending on individual selection. We refrained from using cosmetics other than the provided cosmetics. The selected cosmetics were used every day for 28 days. The skin color was measured by the same method as described above before the start of the test and after the 28-day continuous test. ΔITA was calculated by subtracting the ITA value after the test from the ITA value before the test, and the average and deviation were obtained for each group. The results are shown in Table 13. From this, it can be seen that according to the method for selecting cosmetics using the method for classifying the appearance form of skin color of the present invention, the group in which cosmetics are selected has a larger ΔITA value and is superior in skin color improvement effect.
本発明は、個人個人に適した化粧料を選択するカウンセリング販売に応用できる。 The present invention can be applied to counseling sales for selecting cosmetics suitable for individuals.
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