JP5836666B2 - MITF-M production promoter, hair cosmetic composition and skin cosmetic composition containing the MITF-M production promoter - Google Patents
MITF-M production promoter, hair cosmetic composition and skin cosmetic composition containing the MITF-M production promoter Download PDFInfo
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- JP5836666B2 JP5836666B2 JP2011142472A JP2011142472A JP5836666B2 JP 5836666 B2 JP5836666 B2 JP 5836666B2 JP 2011142472 A JP2011142472 A JP 2011142472A JP 2011142472 A JP2011142472 A JP 2011142472A JP 5836666 B2 JP5836666 B2 JP 5836666B2
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- cosmetic composition
- hair
- production promoter
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Description
本発明は、メラノサイトにおける色素細胞の分化増殖因子である小眼球症関連転写因子Ｍ（ｍｉｃｒｏｐｈｔｈａｌｍｉａ−ａｓｓｏｃｉａｔｅｄ ｔｒａｎｓｃｒｉｐｔｉｏｎ ｆａｃｔｏｒ Ｍ：以下、ＭＩＴＦ−Ｍとする）の産生促進剤、及び該ＭＩＴＦ−Ｍ産生促進剤を含有する毛髪用化粧料組成物並びに皮膚用化粧料組成物に関する。 The present invention relates to a production promoter of microphthalmia-associated transcription factor M (hereinafter referred to as MITF-M), which is a differentiation growth factor of pigment cells in melanocytes, and the MITF-M production promoter. The present invention relates to a cosmetic composition for hair and a cosmetic composition for skin.
加齢による外観変化のひとつである白髪は周囲に老いの印象を与えるため、白髪染めなどで染毛されることが多い。しかし、白髪染め用酸化染毛剤の成分であるパラフェニレンジアミンは体質によってはアレルギーの原因となることがあり、含有されるアルカリ剤により接触性皮膚炎を起こすといった問題もある。そこで、毛髪自体の白髪化を防止する薬剤の開発が求められていた。また、褐色の肌に対する要望も強い。 Gray hair, which is one of the changes in appearance due to aging, gives the impression of aging in the surrounding area, and is often dyed with white hair dyeing. However, paraphenylenediamine, which is a component of an oxidative hair dye for hair dyeing, may cause allergies depending on the constitution, and there is also a problem that contact dermatitis is caused by the contained alkali agent. Therefore, there has been a demand for the development of a drug that prevents whitening of the hair itself. There is also a strong demand for brown skin.
毛髪の色や皮膚の色を決定するメラニン色素は、毛髪の毛母や皮膚の基底層に分布するメラノサイトで産生される色素成分であり、角質細胞に移送された後、細胞内で核の上方に集合し、紫外線から細胞のＤＮＡを保護するという重要な役割を担っている。 Melanin pigment, which determines the color of hair and skin, is a pigment component produced in melanocytes distributed in the hair matrix and the basal layer of the skin. After being transferred to the corneocytes, it is located above the nucleus in the cell. They gather and play an important role in protecting cellular DNA from ultraviolet light.
メラニン色素産生の場であるメラノサイトは、自己増殖した色素幹細胞が分化して生成したものであるが、メラニン色素産生能力の低下は、色素幹細胞の自己増殖機能が失われ、新たなメラノサイトを生み出せなくなったことに起因し、色素幹細胞の自己増殖機能の維持には、メラノサイト発生のマスター因子として知られるＭＩＴＦ−Ｍが必須の遺伝子であるとの報告がなされた（非特許文献１）。このことから、ＭＩＴＦ−Ｍの産生を促進し、色素幹細胞の自己増殖機能を高めることにより、メラノサイトが再生され、毛髪の白髪化予防及び肌の褐色化が達成できると考えられた。このＭＩＴＦ−Ｍは、メラノサイトの色素合成に関わる酵素であるチロシナーゼや、その関連蛋白質であるＴＲＰ−１（Ｔｙｒｏｓｉｎａｓｅ ｒｅｌａｔｅｄ ｐｒｏｔｅｉｎ−１）、ＴＲＰ−２（Ｔｙｒｏｓｉｎａｓｅ ｒｅｌａｔｅｄ ｐｒｏｔｅｉｎ−２）の転写制御を行うことも知られている。 Melanocytes, the place where melanin pigments are produced, are produced by differentiation of self-proliferating pigment stem cells, but a decrease in the ability to produce melanin pigments causes the loss of pigment stem cell self-proliferation functions and prevents the creation of new melanocytes. For this reason, it has been reported that MITF-M, which is known as a master factor for melanocyte generation, is an essential gene for maintaining the self-proliferating function of pigment stem cells (Non-patent Document 1). From this, it was thought that by promoting the production of MITF-M and enhancing the self-proliferating function of pigment stem cells, melanocytes are regenerated, and hair whitening prevention and skin browning can be achieved. MITF-M performs transcriptional control of tyrosinase, which is an enzyme involved in melanocyte pigment synthesis, and TRP-1 (Tyrosinase related protein-1) and TRP-2 (Tyrosinase related protein-2), which are related proteins. Is also known.
メラノサイトにおけるＭＩＴＦ−Ｍの発現量を高める試みとして、ホップ中の微量成分である特定のカルコン類、フラバノン類およびイソフムロン類などによる効果が報告されている（特許文献１）。しかし、これらの植物エキスで得られるＭＩＴＦ−Ｍ産生促進効果は、十分満足できるものではなかった。また、フォルスコリンのＭＩＴＦ−Ｍの発現量を高める効果についても報告されている（非特許文献２）が、フォルスコリンは水に極めて難溶性で、有機溶剤への溶解度も決して高くないため、優れたＭＩＴＦ−Ｍ産生促進効果を発揮する濃度でフォルスコリンを含有する化粧品組成物を得るのは困難であった。 As an attempt to increase the expression level of MITF-M in melanocytes, the effects of specific chalcones, flavanones and isohumulones, which are trace components in hops, have been reported (Patent Document 1). However, the MITF-M production promoting effect obtained with these plant extracts was not fully satisfactory. Moreover, although the effect of increasing the expression level of MITF-M of forskolin has been reported (Non-patent Document 2), forskolin is extremely poorly soluble in water and is not very high in solubility in organic solvents. In addition, it was difficult to obtain a cosmetic composition containing forskolin at a concentration that exerts the effect of promoting MITF-M production.
本発明は、メラノサイトにおけるＭＩＴＦ−Ｍ産生促進剤、及び該ＭＩＴＦ−Ｍ産生促進剤を用いた毛髪用化粧料組成物並びに皮膚用化粧料組成物の提供を目的とする。 An object of this invention is to provide the MITF-M production promoter in a melanocyte, the cosmetic composition for hair using the said MITF-M production promoter, and the cosmetic composition for skin.
本発明者らは、上記課題を解決するために鋭意研究を重ねた結果、フォルスコリンと、特定の植物エキスとを併用することにより、メラノサイトにおけるＭＩＴＦ−Ｍ産生量が飛躍的に増大することを見出し、この知見に基づき本発明を完成させるに至った。 As a result of intensive studies to solve the above problems, the present inventors have found that MITF-M production in melanocytes is dramatically increased by using forskolin in combination with a specific plant extract. Based on this finding, the present invention has been completed.
すなわち、本発明は、
（１）（Ａ）フォルスコリンと、（Ｂ）黒砂糖エキス、オレンジエキス、チンピエキス、ウメエキス、及びワカメエキスからなる群より選ばれる少なくとも１種の植物エキスとからなるＭＩＴＦ−Ｍ産生促進剤、
（２）（Ａ）フォルスコリンと、（Ｂ）黒砂糖エキス、オレンジエキス、チンピエキス、ウメエキス、及びワカメエキスからなる群より選ばれる少なくとも１種の植物エキスとの配合比が、乾燥質量換算で１：９９〜９５：５であることを特徴とする前項に記載のＭＩＴＦ−Ｍ産生促進剤、
（３）第（１）項又は第（２）項のいずれか１項に記載のＭＩＴＦ−Ｍ産生促進剤を含有する毛髪用化粧料組成物、
（４）第（１）項又は第（２）項のいずれか１項に記載のＭＩＴＦ−Ｍ産生促進剤を、フォルスコリンの質量で、０.００１〜５質量％含有することを特徴とする毛髪用化粧料組成物、
（５）第（１）項又は第（２）項のいずれか１項に記載のＭＩＴＦ−Ｍ産生促進剤を含有する皮膚用化粧料組成物、
（６）第（１）項又は第（２）項のいずれか１項に記載のＭＩＴＦ−Ｍ産生促進剤を、フォルスコリンの質量で、０.００１〜５質量％含有することを特徴とする皮膚用化粧料組成物、
を提供するものである。
That is, the present invention
(1) MITF-M production promoter comprising (A) forskolin and (B) at least one plant extract selected from the group consisting of brown sugar extract, orange extract, chimpi extract, ume extract, and wakame extract,
(2) The blending ratio of (A) forskolin to at least one plant extract selected from the group consisting of (B) brown sugar extract, orange extract, chimpi extract, ume extract and wakame extract is 1 in terms of dry mass. : The MITF-M production promoter according to the preceding item, which is 99 to 95: 5,
(3) A cosmetic composition for hair containing the MITF-M production promoter according to any one of (1) or (2),
(4) The MITF-M production promoter according to any one of (1) or (2) is contained in an amount of 0.001 to 5% by mass of forskolin. Cosmetic composition for hair,
(5) A cosmetic composition for skin containing the MITF-M production promoter according to any one of (1) or (2),
(6) The MITF-M production promoter according to any one of (1) or (2) is contained in 0.001 to 5% by mass of forskolin. Skin cosmetic composition,
Is to provide.
メラノサイトにおけるＭＩＴＦ−Ｍ産生能力を飛躍的に増大する効果を有する本発明のＭＩＴＦ−Ｍ産生促進剤により、メラノサイトのメラニン色素産生能力を高めることが可能となる。また、これを毛髪用化粧料に配合することで白髪化予防効果が、皮膚用化粧料に配合することで、皮膚の褐色化効果（タンニング効果）が期待できる。 With the MITF-M production promoter of the present invention having the effect of dramatically increasing the MITF-M production ability in melanocytes, it becomes possible to enhance the melanin pigment production ability of melanocytes. In addition, it can be expected to have the effect of preventing whitening by blending it into the cosmetic for hair, and the browning effect (tanning effect) of the skin by blending into the cosmetic for skin.
本発明のＭＩＴＦ−Ｍ産生促進剤は、（Ａ）フォルスコリンと、（Ｂ）黒砂糖エキス、オレンジエキス、チンピエキス、ウメエキス、及びワカメエキスからなる群より選ばれた少なくとも１種の植物エキスとの混合物である。この両成分を併用することにより、メラノサイトにおけるＭＩＴＦ−Ｍ産生能力を飛躍的に高めることができる（以下、（Ｂ）黒砂糖エキス、オレンジエキス、チンピエキス、ウメエキス、及びワカメエキスからなる群より選ばれた少なくとも１種の植物エキスを（Ｂ）成分という）。
フォルスコリンとは、コレウスフォルスコリ（Coleus forskohlii）およびその近縁種であるシソ科植物の塊根の抽出物から分離、精製され、構造決定された物質である。本発明において、フォルスコリンとして、上記植物の塊根から抽出した植物エキスをそのまま用いることができ、また、医薬、生化学試薬等として市販された精製品を用いることもできる。
The MITF-M production promoter of the present invention comprises (A) forskolin and (B) at least one plant extract selected from the group consisting of brown sugar extract, orange extract, chimpan extract, ume extract, and wakame extract. It is a mixture. By using both of these components in combination, the ability to produce MITF-M in melanocytes can be dramatically increased (hereinafter selected from the group consisting of (B) brown sugar extract, orange extract, chimpi extract, ume extract, and wakame extract. And at least one plant extract is referred to as component (B)).
Forskolin is a substance that has been separated, purified, and structure-determined from extracts of tuberous roots of Coleus forskohlii and its related species Lamiaceae. In the present invention, as the forskolin, a plant extract extracted from the tuberous root of the plant can be used as it is, and a purified product commercially available as a medicine, a biochemical reagent or the like can also be used.
黒砂糖エキスとは、イネ科サトウキビ属の植物の一種であるサトウキビSaccharum officinarumを原料とする黒砂糖から抽出された植物エキスである。
オレンジエキスとは、ミカン科ミカン属の一種であるオレンジCitrus sinensisの成熟した果皮および果実から抽出された植物エキスである。
チンピエキスとは、ミカン科ミカン属の一種であるウンシュウミカンCitrus unshiuの成熟した果皮から抽出された植物エキスである。
ウメエキスとは、バラ科の一種であるウメ Prunus mume Sieb. et Zucc. の果実から抽出された植物エキスである。
ワカメエキスとは、褐藻綱コンブ目チガイソ科の一種であるワカメUndaria pinnatifidaから抽出された植物エキスである。
The brown sugar extract is a plant extract extracted from brown sugar made from sugarcane Saccharum officinarum, which is a kind of plant belonging to the genus Gramineae.
The orange extract is a plant extract extracted from mature pericarp and fruits of orange Citrus sinensis, a kind of citrus citrus.
Chimpi extract is a plant extract extracted from the mature pericarp of Citrus unshiu, a kind of Citrus mandarin.
Ume extract is a plant extract extracted from the fruit of the plum, Prunus mume Sieb. Et Zucc.
Wakame extract is a plant extract extracted from Wakame Undaria pinnatifida, which is a kind of the family of the brown alga, Coleoptera.
本発明が用いるこれらの植物エキスは、いずれも、医薬又は民間薬、食品、化粧料の成分として一般に用いられているものを使用することができる。
フォルスコリン、オレンジエキス、チンピエキス、ウメエキス、ワカメエキスは前記各植物の各種部位から溶媒抽出して得ることができ、市販の各植物エキスを用いることもできる。
Any of these plant extracts used in the present invention may be those generally used as components of pharmaceuticals or folk medicines, foods, and cosmetics.
Forskolin, orange extract, chimpi extract, ume extract and wakame extract can be obtained by solvent extraction from various parts of each plant, and commercially available plant extracts can also be used.
黒砂糖エキスとしては、種々のものを使用することができる。例えば、黒砂糖を溶媒抽出して得たものをそのまま用いることができ、黒砂糖の着色成分を分離、濃縮して得られるエキスや、黒砂糖のエタノール抽出物を濃縮した後、水に溶かし、吸着剤で脱色するかあるいは脱色しないエキス、さらには、市販の黒砂糖エキスを用いることもできる。
本発明においては、このように黒砂糖エキスとして糖成分を用いることも、着色成分を用いることもできるが、糖成分を含有するものであることが好ましい。
Various types of brown sugar extract can be used. For example, what is obtained by solvent extraction of brown sugar can be used as it is, the extract obtained by separating and concentrating the colored components of brown sugar, the ethanol extract of brown sugar is concentrated, dissolved in water, An extract that is decolorized or not decolorized with an adsorbent, and a commercially available brown sugar extract can also be used.
In the present invention, a sugar component can be used as the brown sugar extract as described above, or a colored component can be used, but it is preferable to contain a sugar component.
前記植物エキスの抽出において、使用可能な抽出溶媒に特に制限はなく、水、メタノール、エタノール、プロパノール、ブタノールなどの低級１価アルコール、プロピレングリコール、ジプロピレングリコール、ブチレングリコール、グリセリンなどの多価アルコール類、酢酸メチル、酢酸エチルなどのエステル類、アセトン、メチルエチルケトンなどのケトン類、ジエチルエーテル、テトラヒドロフランなどのエーテル類、などの極性溶媒、クロロホルム、四塩化炭素などのハロゲン化炭化水素類、ヘキサン、ヘプタン、シクロヘキサンなどの炭化水素類を用いることができ、これらの溶媒の１種を用いることも、２種以上からなる混合溶媒を用いることもできる。特に極性溶媒を好ましく用いることができる。 In the extraction of the plant extracts, usable extraction solvents are not particularly limited, and lower monohydric alcohols such as water, methanol, ethanol, propanol and butanol, and polyhydric alcohols such as propylene glycol, dipropylene glycol, butylene glycol and glycerin. , Polar solvents such as esters such as methyl acetate and ethyl acetate, ketones such as acetone and methyl ethyl ketone, ethers such as diethyl ether and tetrahydrofuran, halogenated hydrocarbons such as chloroform and carbon tetrachloride, hexane and heptane Hydrocarbons such as cyclohexane can be used, and one kind of these solvents can be used, or a mixed solvent composed of two or more kinds can be used. In particular, a polar solvent can be preferably used.
植物エキスの抽出方法に特に制限はなく、例えば、フォルスコリン、黒砂糖エキス、オレンジエキス、チンピエキス、ウメエキス、ワカメエキスなどの植物エキスの前記原料を、２０〜４０℃の前記溶媒中に、１時間〜７日間浸漬し、濾過する方法を挙げることができる。 There is no particular limitation on the extraction method of the plant extract. For example, the raw materials of the plant extract such as forskolin, brown sugar extract, orange extract, chimpi extract, ume extract, wakame extract are placed in the solvent at 20 to 40 ° C. for 1 hour. The method of immersing for -7 days and filtering can be mentioned.
前記植物エキスの原料は、乾燥物を用いるのが好ましく、乾燥粉砕物を用いることがさらに好ましい。また、抽出溶媒に浸漬している間、撹拌していてもよく、静置していてもよい。本発明の植物エキスは、抽出操作を繰り返して高濃度の植物エキスとすることもできる。 As the raw material of the plant extract, it is preferable to use a dried product, and it is more preferable to use a dried pulverized product. Moreover, while being immersed in an extraction solvent, you may stir and may stand still. The plant extract of this invention can also be made into a high concentration plant extract by repeating extraction operation.
本発明のＭＩＴＦ−Ｍ産生促進剤は、前記（Ａ）フォルスコリンと（Ｂ）成分、すなわち黒砂糖エキス、オレンジエキス、チンピエキス、ウメエキス、ワカメエキスからなる群より選ばれた少なくとも１種とからなるが、これらの各成分は抽出溶媒に溶解したそのままの状態で用いることができ、抽出溶媒を揮発させて得た濃縮液の状態で、または、凍結乾燥して得た粉末等の固体状で用いることもできる。さらに、液状の、または固体状の成分を適当な溶媒に再溶解して用いることもできる。 The MITF-M production promoter of the present invention comprises (A) forskolin and (B) component, that is, at least one selected from the group consisting of brown sugar extract, orange extract, chimpi extract, plum extract and seaweed extract. However, each of these components can be used as it is dissolved in the extraction solvent, and is used in the form of a concentrate obtained by volatilizing the extraction solvent or in the form of a solid such as a powder obtained by freeze-drying. You can also. Furthermore, liquid or solid components can be used by re-dissolving in an appropriate solvent.
本発明のＭＩＴＦ−Ｍ産生促進剤を毛髪用化粧料または皮膚用化粧料として用いる場合、あるいは毛髪用化粧料または皮膚用化粧料に配合する場合、ＭＩＴＦ−Ｍ産生促進効果は頭皮または皮膚に浸透したときにより発揮されることから、促進剤中の各成分は、溶解した状態にあることが好ましい。 When the MITF-M production promoter of the present invention is used as hair cosmetics or skin cosmetics, or when blended in hair cosmetics or skin cosmetics, the MITF-M production promoting effect penetrates the scalp or skin. Therefore, it is preferable that each component in the accelerator is in a dissolved state.
本発明のＭＩＴＦ−Ｍ産生促進剤を構成する成分のうち、（Ｂ）成分は水及び種々の有機溶剤に容易に溶解するが、（Ａ）フォスコリンは水に極めて難溶性である。したがって、固体状態の成分を再溶解するのに適当な溶媒としては、（Ａ）フォルスコリンを溶解する有機溶剤を用いることが好ましく、皮膚刺激性の低い毛髪用化粧料組成物または皮膚用化粧料組成物を得るために、（Ａ）フォルスコリンを溶解する有機溶剤と、（Ｂ）成分を溶解する水との混合溶媒を用いることがより好ましい。（Ａ）フォルスコリンを溶解する有機溶剤としては皮膚刺激性の低いエタノール、１,３−ブチレングリコール、及びこれらの混合溶媒であることが好ましい。水及び各有機溶剤の混合比は特に制限されず、成分の溶解状態及び所望の皮膚刺激性等に応じて当業者が適宜に設定することができる。 Among the components constituting the MITF-M production promoter of the present invention, the component (B) is easily dissolved in water and various organic solvents, but (A) forskolin is extremely poorly soluble in water. Therefore, it is preferable to use (A) an organic solvent that dissolves forskolin as a suitable solvent for redissolving the solid-state components, and a cosmetic composition for hair or skin that has low skin irritation. In order to obtain a composition, it is more preferable to use a mixed solvent of (A) an organic solvent that dissolves forskolin and water that dissolves component (B). (A) The organic solvent for dissolving forskolin is preferably ethanol, 1,3-butylene glycol, and a mixed solvent thereof having low skin irritation. The mixing ratio of water and each organic solvent is not particularly limited, and can be appropriately set by those skilled in the art according to the dissolved state of the components and desired skin irritation.
本発明がＭＩＴＦ−Ｍ産生促進剤の構成成分とする（Ａ）フォルスコリンは、単独でもＭＩＴＦ−Ｍの発現量を高める効果を有するが、水に難溶の物質であり、有機溶剤への溶解度も決して高くないため、毛髪用・皮膚用化粧料に対する配合量が制限され、ＭＩＴＦ−Ｍ産生促進効果もそれにより制限される。しかし、ここに（Ｂ）成分を併用することによってＭＩＴＦ−Ｍ産生促進効果が増大するため、（Ａ）フォルスコリンの濃度が低くてもＭＩＴＦ−Ｍ産生促進効果に優れた毛髪用化粧料組成物・皮膚用化粧料組成物を得ることができる。 (A) Forskolin, which is a constituent component of the MITF-M production promoter of the present invention, has the effect of increasing the expression level of MITF-M alone, but is a substance that is hardly soluble in water and has solubility in organic solvents. However, the blending amount for hair and skin cosmetics is limited, and the effect of promoting the production of MITF-M is also limited thereby. However, since the MITF-M production promoting effect is increased by using the component (B) in combination here, (A) a cosmetic composition for hair excellent in the MITF-M production promoting effect even if the concentration of forskolin is low. -A skin cosmetic composition can be obtained.
本発明のＭＩＴＦ−Ｍ産生促進剤において、（Ａ）フォルスコリンと（Ｂ）成分との比率に特に制限はないが、（Ａ）フォルスコリンの質量と（Ｂ）成分の乾燥質量との比が、１：９９〜９５：５であることが好ましく、３：９７〜８５：１５にあることがより好ましい。（Ａ）フォルスコリンと（Ｂ）成分との比率が前記範囲にあるとき、特に顕著なＭＩＴＦ−Ｍ産生促進効果を得ることができる。（Ｂ）成分の比率が前記下限より少ないと、フォルスコリンとの相乗効果があまり期待できず、また、（Ｂ）成分を前記上限を超えて配合しても、配合量に見合う効果が得られないおそれがある。 In the MITF-M production promoter of the present invention, the ratio of (A) forskolin to (B) component is not particularly limited, but the ratio of (A) forskolin mass to (B) component dry mass is 1:99 to 95: 5 is preferable, and 3:97 to 85:15 is more preferable. When the ratio of (A) forskolin to (B) component is in the above range, a particularly remarkable MITF-M production promoting effect can be obtained. When the ratio of the component (B) is less than the lower limit, a synergistic effect with forskolin cannot be expected so much, and even if the component (B) exceeds the upper limit, an effect commensurate with the blending amount is obtained. There is a risk of not.
（毛髪用化粧料、皮膚用化粧料）
本発明のＭＩＴＦ−Ｍ産生促進剤は、頭皮・頭髪に適用する医薬品、医薬部外品、薬用又は化粧用の製剤類をはじめとする各種の外用剤に配合することができ、整髪料、頭皮料、洗髪料、ヘアリンス、養毛料などに配合して、毛髪のメラニン色素形成を促進する毛髪用化粧料として用いることができる。
(Hair cosmetics, skin cosmetics)
The MITF-M production promoter of the present invention can be blended in various external preparations including pharmaceuticals, quasi-drugs, medicinal or cosmetic preparations applied to the scalp / hair. It can be used as a cosmetic for hair that can be blended into hair, hair wash, hair rinse, hair nourishing and the like to promote melanin pigment formation in hair.
また、本発明のＭＩＴＦ−Ｍ産生促進剤は、皮膚に適用する医薬品、医薬部外品、薬用又は化粧用の製剤類をはじめとする各種の外用剤に配合することができ、化粧水、乳液、ボディローション、ボディクリームなどに配合して、皮膚のメラニン色素形成を促進する皮膚用化粧料として用いることができる。 In addition, the MITF-M production promoter of the present invention can be blended in various external preparations including pharmaceuticals applied to the skin, quasi drugs, medicinal or cosmetic preparations, lotions, emulsions It can be blended into body lotions, body creams and the like and used as a skin cosmetic that promotes melanin pigment formation on the skin.
本発明のＭＩＴＦ−Ｍ産生促進剤を毛髪用化粧料、皮膚用化粧料に配合する場合の配合量に特に制限はないが、フォルスコリンの有機溶剤への溶解度から、毛髪用化粧料組成物及び皮膚用化粧料組成物中のフォルスコリンの濃度は、０.００１〜５質量％であることが好ましく、０.００１〜４質量部であることがより好ましい。フォルスコリンの濃度が前記上限を超えると、フォルスコリンが析出して、配合量に見合う効果が得られないおそれがある。一方、フォルスコリンの濃度が前記下限未満では、（Ｂ）成分を併用しても、メラノサイトでの十分なＭＩＴＦ−Ｍ産生促進効果が得られないおそれがある。 There are no particular restrictions on the amount of the MITF-M production promoter of the present invention incorporated into hair cosmetics and skin cosmetics. From the solubility of forskolin in organic solvents, hair cosmetic compositions and The concentration of forskolin in the cosmetic composition for skin is preferably 0.001 to 5% by mass, and more preferably 0.001 to 4 parts by mass. If the concentration of forskolin exceeds the above upper limit, forskolin may precipitate and an effect commensurate with the blending amount may not be obtained. On the other hand, if the concentration of forskolin is less than the lower limit, even if the component (B) is used in combination, a sufficient effect of promoting MITF-M production in melanocytes may not be obtained.
ＭＩＴＦ−Ｍ産生促進効果はフォルスコリンの濃度が高いほど、強くなる傾向にあるが、フォルスコリンの濃度を低く抑えることによって、有機溶剤の使用量を下げることができ、それによって皮膚刺激性の低い毛髪用化粧料組成物、皮膚用化粧料組成物を得ることができる。ＭＩＴＦ−Ｍ産生促進効果及び皮膚刺激性の観点からフォルスコリンのより好ましい配合量は、０.００１〜１質量％である。 The MITF-M production promoting effect tends to become stronger as the concentration of forskolin increases, but by reducing the concentration of forskolin, the amount of organic solvent used can be reduced, thereby reducing skin irritation. A cosmetic composition for hair and a cosmetic composition for skin can be obtained. From the viewpoint of the MITF-M production promoting effect and skin irritation, the more preferable amount of forskolin is 0.001 to 1% by mass.
本発明の毛髪用化粧料組成物、皮膚用化粧料組成物には、本発明の効果を損なわない範囲で、前記ＭＩＴＦ−Ｍ産生促進剤に加え、皮膚外用剤用として公知の機能成分、例えば、保湿剤、エモリエント剤、血行促進剤、細胞賦活化剤、抗酸化剤、抗炎症剤、抗菌剤、過酸化物抑制剤など、を配合することができる。 In the cosmetic composition for hair and the cosmetic composition for skin of the present invention, in addition to the MITF-M production promoter, functional components known for skin external preparations, for example, within a range not impairing the effects of the present invention. , Moisturizers, emollients, blood circulation promoters, cell activators, antioxidants, anti-inflammatory agents, antibacterial agents, peroxide inhibitors, and the like can be blended.
公知の機能成分として、より具体的には、グリセリン、尿素、アミノ酸、ヒアルロン酸類などの保湿剤；スクワラン、マカデミアナッツ油、オリーブ油、ホホバ油、シリコン油などのエモリエント剤；ビタミンＥ類、トウガラシチンキなどの血行促進剤；核酸などの細胞賦活化剤、ジブチルヒドロキシトルエン（ＢＨＴ）、ジブチルヒドロキシアニソール（ＢＨＡ）、酢酸トコフェロールなどの抗酸化剤；グリチルリチン、アラントインなどの抗炎症剤；ヒノキチオール、塩化ベンザルコニウム、クロルヘキシジン塩、パラヒドロキシ安息香酸エステルなどの抗菌剤；スーパーオキシドジスムターゼ（ＳＯＤ）などの過酸化物抑制剤など、種々の機能成分を配合することができ、オウゴンエキス、イチョウエキス、胎盤抽出物、乳酸菌培養抽出物などの植物・動物・微生物由来の各種抽出物なども制限なく添加して使用することができる。 More specifically, known functional ingredients include humectants such as glycerin, urea, amino acids, and hyaluronic acids; emollients such as squalane, macadamia nut oil, olive oil, jojoba oil, and silicon oil; vitamin Es, pepper tincture, and the like. Blood circulation promoters; cell activators such as nucleic acids, antioxidants such as dibutylhydroxytoluene (BHT), dibutylhydroxyanisole (BHA), tocopherol acetate; anti-inflammatory agents such as glycyrrhizin and allantoin; hinokitiol, benzalkonium chloride, Various functional ingredients such as antibacterial agents such as chlorhexidine salt and parahydroxybenzoate ester; peroxide inhibitors such as superoxide dismutase (SOD) can be blended. Orgon extract, Ginkgo biloba extract, placenta extract, lactic acid bacteria Culture extract Various extracts of plants, animals, and from microorganisms such objects can also be used by adding without limitation.
本発明のＭＩＴＦ−Ｍ産生促進剤を含有する毛髪用化粧料組成物、皮膚用化粧料組成物の剤型に特に制限はなく、例えば、ペースト剤、クリーム、ジェル、軟膏、ローション、乳液、パック、パウダー、ハップ剤などとして用いることができる。 There is no particular limitation on the dosage form of the cosmetic composition for hair and the cosmetic composition for skin containing the MITF-M production promoter of the present invention. For example, paste, cream, gel, ointment, lotion, emulsion, pack It can be used as a powder or a haptic agent.
また、本発明のＭＩＴＦ−Ｍ産生促進剤を含有する毛髪用化粧料組成物、皮膚用化粧料組成物には、本発明の効果を損なわない範囲で、その剤型化のために界面活性剤、油脂類などの基材成分や、必要に応じて増粘剤、防腐剤、等張化剤、酸化防止剤、紫外線吸収剤、キレート剤、香料、着色料などの種々の添加物を併用することができる。 Further, in the cosmetic composition for hair and the cosmetic composition for skin containing the MITF-M production promoter of the present invention, a surfactant is used for its formulation as long as the effects of the present invention are not impaired. , Base ingredients such as fats and oils, and various additives such as thickeners, preservatives, tonicity agents, antioxidants, ultraviolet absorbers, chelating agents, fragrances, and coloring agents, if necessary be able to.
本発明のＭＩＴＦ−Ｍ産生促進剤を含有する毛髪用化粧料組成物、皮膚用化粧料組成物の剤型化に使用可能な界面活性剤に特に制限はなく、例えば、高級アルコールのアルキレンオキシド付加物、高級アルキルアミンのアルキレンオキシド付加物、高級脂肪酸のアルキレンオキシド付加物、高級脂肪酸アミドのアルキレンオキシド付加物、多価アルコールの脂肪酸エステル、硬化ひまし油のアルキレンオキシド付加物、ポリエチレングリコールソルビタンアルキルエステル、ステロール等のアルキレンオキシド付加物などの非イオン界面活性剤；アルキル硫酸ナトリウム、アルキロイルメチルタウリンナトリウム、α−オレフィンスルホン酸ナトリウム、ポリオキシアルキレンアルキルエーテル硫酸ナトリウムなどの陰イオン界面活性剤；塩化アルキルピリジニウム、塩化ジステアリルジメリルアンモニウムなどの陽イオン界面活性剤；アミノプロピオン酸ナトリウム、アルキルポリアミノエチルグリシンなどの両性界面活性剤などを用いることができる。これらの界面活性剤は１種を用いることができ、２種以上を用いることもできる。 There is no particular limitation on the surfactant that can be used to formulate the cosmetic composition for hair and the cosmetic composition for skin containing the MITF-M production promoter of the present invention. For example, addition of an alkylene oxide to a higher alcohol , Alkylene oxide adduct of higher alkylamine, alkylene oxide adduct of higher fatty acid, alkylene oxide adduct of higher fatty acid amide, fatty acid ester of polyhydric alcohol, alkylene oxide adduct of hydrogenated castor oil, polyethylene glycol sorbitan alkyl ester, sterol Nonionic surfactants such as alkylene oxide adducts such as sodium alkylsulfate, sodium alkyloxyltaurate, sodium α-olefin sulfonate, sodium polyoxyalkylene alkyl ether sulfate, salts It can be used sodium aminopropionic acid, and amphoteric surfactants such as alkylpolyaminoethylglycine; alkyl pyridinium, cationic surfactants such as chloride distearyl di Merrill ammonium. These surfactants can be used alone or in combination of two or more.
本発明のＭＩＴＦ−Ｍ産生促進剤を含有する毛髪用化粧料組成物、皮膚用化粧料組成物の剤型化に使用可能な基材成分に特に制限はなく、例えば、オリーブ油、ツバキ油、アボカド油、マカデミアナッツ油、杏仁油、ホホバ油、スクワラン、スクワレン、馬油など、公知の油脂類を用いることができる。 There are no particular limitations on the base ingredient that can be used for formulating the cosmetic composition for hair and the cosmetic composition for skin containing the MITF-M production promoter of the present invention. For example, olive oil, camellia oil, avocado Known fats and oils such as oil, macadamia nut oil, apricot kernel oil, jojoba oil, squalane, squalene, and horse oil can be used.
本発明のＭＩＴＦ−Ｍ産生促進剤を含有する毛髪用化粧料組成物、皮膚用化粧料組成物の剤型化に使用可能な増粘剤にも特に制限はなく、例えば、ポリビニルアルコール、ポリビニルアクリルアミド、ポリエチレングリコール、及びこれらの各種誘導体；ヒドロキシアルキルセルロースなどのセルロース類及びその誘導体；デキストラン、ゼラチン、アラビアガム、トラガントガムなどのガム類；カルボキシビニルポリマーなどの水溶性高分子を用いることができる。 There is no particular limitation on the thickener that can be used for formulating the cosmetic composition for hair and the cosmetic composition for skin containing the MITF-M production promoter of the present invention. For example, polyvinyl alcohol, polyvinyl acrylamide , Polyethylene glycol, and various derivatives thereof; celluloses such as hydroxyalkyl cellulose and derivatives thereof; gums such as dextran, gelatin, gum arabic, and gum tragacanth; and water-soluble polymers such as carboxyvinyl polymer.
本発明のＭＩＴＦ−Ｍ産生促進剤を含有する毛髪用化粧料組成物、皮膚用化粧料組成物の剤型化に使用可能な防腐剤に特に制限はなく、例えば、パラヒドロキシ安息香酸エステル、パラオキシ安息香酸塩とその誘導体、フェノキシエタノール、ヒノキチオール、塩化ベンザルコニウム、クロルヘキシジン塩などを用いることができる。 There are no particular restrictions on the preservatives that can be used for formulating hair cosmetic compositions and skin cosmetic compositions containing the MITF-M production promoter of the present invention. For example, parahydroxybenzoic acid esters, paraoxy Benzoate and its derivatives, phenoxyethanol, hinokitiol, benzalkonium chloride, chlorhexidine salt and the like can be used.
本発明のＭＩＴＦ−Ｍ産生促進剤を含有する毛髪用化粧料組成物、皮膚用化粧料組成物の剤型化に使用可能な等張化剤に特に制限はなく、例えば、塩化ナトリウム、塩化カリウム、塩化カルシウムなどの無機塩類を用いることができる。 There are no particular limitations on the tonicity agent that can be used to formulate the cosmetic composition for hair and the cosmetic composition for skin containing the MITF-M production promoter of the present invention. For example, sodium chloride, potassium chloride Inorganic salts such as calcium chloride can be used.
本発明のＭＩＴＦ−Ｍ産生促進剤を含有する毛髪用化粧料組成物、皮膚用化粧料組成物に使用可能な紫外線吸収剤に特に制限はなく、例えば、パラアミノ安息香酸、ベンゾフェノン系紫外線吸収剤、ベンゾトリアゾール系紫外線吸収剤などを用いることができる。 There is no particular limitation on the ultraviolet absorber that can be used in the cosmetic composition for hair and the cosmetic composition for skin containing the MITF-M production promoter of the present invention. For example, paraaminobenzoic acid, benzophenone ultraviolet absorber, A benzotriazole ultraviolet absorber or the like can be used.
本発明のＭＩＴＦ−Ｍ産生促進剤を含有する毛髪用化粧料組成物、皮膚用化粧料組成物の剤型化に使用可能なキレート剤に特に制限はなく、例えば、エチレンジアミン四酢酸、フィチン酸、クエン酸及びこれらの水溶性塩などを用いることができる。 There is no particular limitation on the chelating agent that can be used to formulate the cosmetic composition for hair and the cosmetic composition for skin containing the MITF-M production promoter of the present invention. For example, ethylenediaminetetraacetic acid, phytic acid, Citric acid and water-soluble salts thereof can be used.
以下、実施例を挙げて本発明をさらに詳細に説明するが、本発明はこれらの実施例によりなんら限定されるものではない。 EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated further in detail, this invention is not limited at all by these Examples.
・フォルスコリン
以下に示す実施例においては、生化学試薬として市販のフォルスコリン（Ｓｉｇｍａ Ａｌｄｒｉｃｈ社製）または、コレウスホルスコリ根エキス（サビンサジャパンコーポレーション製、フォルスコリン含有量９５％）を用いた。
Forskolin In the examples shown below, commercially available forskolin (manufactured by Sigma Aldrich) or Coleus forskolin root extract (manufactured by Savinsa Japan Corporation, forskolin content 95%) was used as a biochemical reagent.
・黒砂糖エキス
黒砂糖を粉砕し、５０ｇを３７５ｍｌの５０質量％エタノール水溶液に室温（約２５℃）下で浸漬した。２４時間後これをろ過し、得られた濾液から水、エタノールを減圧留去し、さらに凍結乾燥し、粉末状の黒砂糖エキス（４３.３４ｇ）を得、以下に示す実施例に使用した。
-Brown sugar extract Brown sugar was pulverized and 50 g was immersed in 375 ml of 50% by mass ethanol aqueous solution at room temperature (about 25 ° C.). After 24 hours, this was filtered, water and ethanol were distilled off from the obtained filtrate under reduced pressure, and freeze-dried to obtain a powdered brown sugar extract (43.34 g), which was used in the examples shown below.
・オレンジエキス
成熟したオレンジを果皮ごと１ｃｍ角に刻み、これを乾燥後粉砕し、５０ｇを３７５ｍｌの５０質量％エタノール水溶液に室温（約２５℃）下で浸漬した。２４時間後これをろ過し、得られた濾液から水、エタノールを減圧留去し、さらに凍結乾燥し、粉末状のオレンジエキス（１６.０１ｇ）を得、以下に示す実施例に使用した。
-Orange extract The mature orange was cut into 1 cm squares together with the pericarp, dried and pulverized, and 50 g was immersed in 375 ml of a 50 mass% aqueous ethanol solution at room temperature (about 25 ° C). After 24 hours, this was filtered, water and ethanol were distilled off from the obtained filtrate under reduced pressure, and freeze-dried to obtain a powdery orange extract (16.01 g), which was used in the examples shown below.
・チンピエキス
乾燥したウンシュウミカンの果皮を粉砕し、５０ｇを３７５ｍｌの５０質量％エタノール水溶液に室温（約２５℃）下で浸漬した。２４時間後これをろ過し、得られた濾液から水、エタノールを減圧留去し、さらに凍結乾燥し、粉末状のチンピエキス（１４.３４ｇ）を得、以下に示す実施例に使用した。
-Chimpi extract The dried persimmon peel was pulverized, and 50 g was immersed in 375 ml of a 50% by mass ethanol aqueous solution at room temperature (about 25 ° C.). After 24 hours, this was filtered, and water and ethanol were distilled off from the obtained filtrate under reduced pressure, and further freeze-dried to obtain a powdery chimpi extract (14.34 g), which was used in the examples shown below.
・ウメエキス
乾燥したウメの果肉の粉砕物の５０ｇを、３７５ｍｌの５０質量％エタノール水溶液に室温（約２５℃）下で浸漬した。２４時間後これをろ過し、得られた濾液から水、エタノールを減圧留去し、さらに凍結乾燥し、粉末状のウメエキス（１０.４３ｇ）を得、以下に示す実施例に使用した。
-Ume Extract 50 g of the dried ume pulp was immersed in 375 ml of a 50% by mass ethanol aqueous solution at room temperature (about 25 ° C.). After 24 hours, this was filtered, and water and ethanol were distilled off from the obtained filtrate under reduced pressure, and further freeze-dried to obtain a powdery ume extract (10.43 g), which was used in the examples shown below.
・ワカメエキス
乾燥したワカメの粉砕物の５０ｇを、３７５ｍｌの５０質量％エタノール水溶液に室温（約２５℃）下で浸漬した。２４時間後これをろ過し、得られた濾液から水、エタノールを減圧留去し、さらに凍結乾燥して、粉末状のワカメエキス（１.９８ｇ）を得、以下に示す実施例に使用した。
-Wakame extract 50 g of the dried ground seaweed was immersed in 375 ml of a 50 mass% ethanol aqueous solution at room temperature (about 25 ° C). After 24 hours, this was filtered, water and ethanol were distilled off from the obtained filtrate under reduced pressure, and freeze-dried to obtain a powdered seaweed extract (1.98 g), which was used in the examples shown below.
・ホップエキス
乾燥したホップの雌花穂の粉砕物の５０ｇを、３７５ｍｌの５０質量％エタノール水溶液に室温（約２５℃）下で浸漬した。２４時間後これをろ過し、得られた濾液から水、エタノールを減圧留去し、さらに凍結乾燥して、粉末状のホップエキス（６.０１ｇ）を得、以下に示す比較例に使用した。
-Hop extract 50 g of the pulverized material of dried hop female flower spikes was immersed in 375 ml of a 50 mass% ethanol aqueous solution at room temperature (about 25 ° C). After 24 hours, this was filtered, and water and ethanol were distilled off from the obtained filtrate under reduced pressure, and further freeze-dried to obtain a powdery hop extract (6.01 g), which was used in Comparative Examples shown below.
（１）ＭＩＴＦ−Ｍ産生促進剤
フォルスコリン（Ｓｉｇｍａ Ａｌｄｒｉｃｈ社製）は、ジメチルスルホキシドに種々の濃度で溶解し、ジメチルスルホキシドの最終濃度が０.１質量％となるように培地中に添加した。
(1) MITF-M production promoter Forskolin (manufactured by Sigma Aldrich) was dissolved in dimethyl sulfoxide at various concentrations, and was added to the medium so that the final concentration of dimethyl sulfoxide was 0.1% by mass.
黒砂糖エキス、オレンジエキス、チンピエキス、ウメエキス、ワカメエキス、ホップエキスは、５０質量％エタノール水溶液に種々の濃度で溶解し、エタノールの最終濃度が０.５質量％となるように培地中に添加した。 Brown sugar extract, orange extract, chimpi extract, plum extract, wakame extract and hop extract were dissolved in 50% by mass ethanol aqueous solution at various concentrations, and added to the medium so that the final concentration of ethanol was 0.5% by mass. .
フォルスコリン及び／又は植物エキスを添加した培地は、予め０.２２μｍメンブランフィルターを用いて滅菌濾過したものを使用した。
フォルスコリン及び植物エキスの培地中の最終濃度を表２に示す。
As the medium to which forskolin and / or plant extract was added, a medium which had been sterilized and filtered using a 0.22 μm membrane filter in advance was used.
Table 2 shows the final concentrations of forskolin and plant extract in the medium.
１）細胞の培養とＲＮＡの精製
２４ウェルプレートの各ウェルにヒト正常メラノサイト専用培地（ＨＭＧＳ添加Ｍｅｄｉｕｍ２５４培地、クラボウ製）に懸濁したヒト正常メラノサイト（ＨＥＭｎ−ＭＰ、クラボウ製）を１０,０００ ｃｅｌｌｓ／ウェルずつ播種した。３７℃で７日間培養した後、表２に記載の濃度でフォルスコリン及び／又は各植物エキスを添加した同培地に交換し、３時間培養した。培養後、培地を除去し、ヒト正常メラノサイトをリン酸緩衝液で洗浄した後、Ｓｉｇｍａ社のプロトコルに従ってＴＲＩｚｏｌ試薬（ｓｉｇｍａ社製）を混合し、トータルＲＮＡを精製した。
1) Cell culture and RNA purification 10,000 cells of normal human melanocytes (HEMn-MP, Kurabo Industries) suspended in a medium dedicated to normal human melanocytes (Medium 254 medium supplemented with HMGS, Kurabo Industries) in each well of a 24-well plate Seeded per well. After culturing at 37 ° C. for 7 days, the medium was replaced with the same medium supplemented with forskolin and / or each plant extract at the concentrations shown in Table 2, and cultured for 3 hours. After the culture, the medium was removed and human normal melanocytes were washed with a phosphate buffer, and then TRIzol reagent (manufactured by Sigma) was mixed according to the protocol of Sigma to purify the total RNA.
２）ｃＤＮＡの合成
トータルＲＮＡ精製品を元に、Ｈｉｇｈ−Ｃａｐａｃｉｔｙ ｃＤＮＡ ｓｙｎｔｈｅｓｉｓ Ｋｉｔ（アプライドバイオシステムズ社製）を用い、アプライドバイオシステムズ社のプロトコルに従ってｃＤＮＡを合成した。
2) Synthesis of cDNA Based on the total RNA purified product, cDNA was synthesized according to the protocol of Applied Biosystems using High-Capacity cDNA synthesis Kit (Applied Biosystems).
ＰＣＲ用９６ウェルプレート（バイオ・ラッド ラボラトリーズ（株）製）の各ウェルへ、Ｈｉｇｈ−Ｃａｐａｃｉｔｙ ｃＤＮＡ ｓｙｎｔｈｅｓｉｓ Ｋｉｔの１０×ＲＴ
バッファー溶液を２.０μＬ、２５×ｄＮＴＰ Ｍｉｘ（１００ｍＭ）を０.８μＬ、１０×ＲＴランダムプライマーを２.０μＬ、ＭｕｌｔｉＳｃｒｉｂｅTM Ｒｅｖｅｒｓｅ
Ｔｒａｎｓｃｒｉｐｔａｓｅ （逆転写酵素）を１.０μＬ、ＲＮＡ分解酵素阻害剤を１.０μＬ、ＵＬＴＲＡＳＰＥＣ ＷＡＴＥＲ（バイオ・ラッド ラボラトリーズ（株）製）を３.２μＬ添加し、各植物エキスを添加したサンプルから精製したＲＮＡの１μｇを添加し混合し、ｉＱ サーマルサイクラー（５８２ＢＲ、バイオ・ラッド ラボラトリーズ（株）製）を用いて、２５℃で１０分間、３７℃で１２０分間保持の後、８５℃で５分間インキュベートした。これをＴＥバッファーにて２０倍に希釈し、ｃＤＮＡのＴＥバッファー溶液を得た。
10 × RT of High-Capacity cDNA synthesis Kit to each well of 96-well plate for PCR (manufactured by Bio-Rad Laboratories)
Buffer solution 2.0 μL, 25 × dNTP Mix (100 mM) 0.8 μL, 10 × RT random primer 2.0 μL, MultiScribe ™ Reverse
1.0 μL of transscriptase (reverse transcriptase), 1.0 μL of RNase inhibitor, and 3.2 μL of ULTRASPEC WATER (manufactured by Bio-Rad Laboratories) were added and purified from the sample to which each plant extract was added. 1 μg of RNA was added, mixed, and incubated at 25 ° C. for 10 minutes and at 37 ° C. for 120 minutes using an iQ thermal cycler (582BR, manufactured by Bio-Rad Laboratories), and then incubated at 85 ° C. for 5 minutes. . This was diluted 20 times with TE buffer to obtain a TE buffer solution of cDNA.
３）リアルタイムＰＣＲによるＭＩＴＦ−Ｍの定量
ＰＣＲ用９６ウェルプレートの各ウェルへ、ｉＱ ＳＹＢＲ Ｇｒｅｅｎ Ｓｕｐｅｒｍｉｘ（バイオ・ラッド ラボラトリーズ（株）製）を１２.５μＬ、目的遺伝子であるＭＩＴＦ−Ｍを増幅するためのフォワードプライマー（５ｐｍｏｌ／μＬ）を１.５μＬ、リバースプライマー（５ｐｍｏｌ／μＬ）を１.５μＬ、ｃＤＮＡのＴＥバッファー溶液を２μＬ、ＲＮａｓｅフリー水を７.５μＬ加えた。増幅の１サイクルの温度条件は、熱変性を９５℃で１５秒、アニーリングを５６℃で３０秒、伸長反応を７２℃で３０秒とした。
3) Quantification of MITF-M by real-time PCR To amplify 12.5 μL of iQ SYBR Green Supermix (manufactured by Bio-Rad Laboratories) to each well of a 96-well PCR plate, the target gene MITF-M 1.5 μL of the forward primer (5 pmol / μL), 1.5 μL of the reverse primer (5 pmol / μL), 2 μL of the TE buffer solution of cDNA, and 7.5 μL of RNase-free water were added. The temperature conditions for one cycle of amplification were heat denaturation at 95 ° C. for 15 seconds, annealing at 56 ° C. for 30 seconds, and extension reaction at 72 ° C. for 30 seconds.
ＭｙｉＱ ｓｉｎｇｌｅ−ｃｏｌｏｒリアルタイムＰＣＲ解析システム（バイオ・ラッド ラボラトリーズ（株）製）で、増幅過程の蛍光強度をモニタリングし、一定の蛍光強度が得られるサイクル数からＭＩＴＦ−ＭのｍＲＮＡ量を求め、これをＧＡＰＤＨのｍＲＮＡ量で補正した。 The MyQ single-color real-time PCR analysis system (manufactured by Bio-Rad Laboratories Co., Ltd.) monitors the fluorescence intensity during the amplification process, determines the amount of mRNA of MITF-M from the number of cycles that gives a certain fluorescence intensity, The amount of GAPDH mRNA was corrected.
表１に、ＭＩＴＦ−Ｍ増幅用プライマーおよび内部標準遺伝子として使用したグリセルアルデヒド３リン酸脱水素酵素（ＧＡＰＤＨ）の増幅用プライマー（インビトロジェン社製）を示す。また、表２には、本発明のＭＩＴＦ−Ｍ産生促進成分である（Ａ）フォルスコリン、（Ｂ）黒砂糖エキス、オレンジエキス、チンピエキス、ウメエキス、ワカメエキス、および比較例として用いたホップエキスの培地中の配合濃度（乾燥質量換算）とＭＩＴＦ−Ｍ産生促進剤無添加時のＭＩＴＦ−ＭのｍＲＮＡ量を１.００としたときの、相対発現量比を示す。 Table 1 shows primers for amplification of MITF-M and primers for amplification of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) used as an internal standard gene (manufactured by Invitrogen). Table 2 also shows (A) forskolin, (B) brown sugar extract, orange extract, chimpi extract, ume extract, wakame extract, and hop extract used as a comparative example, which are MITF-M production promoting components of the present invention. The relative expression level ratio when the compounding concentration in the culture medium (in terms of dry mass) and the amount of MITF-M mRNA without addition of the MITF-M production promoter is 1.00 is shown.
表２に示すように、本発明の（Ａ）フォルスコリンと（Ｂ）成分とからなるＭＩＴＦ−Ｍ産生促進剤は、いずれの組み合わせにおいても優れたＭＩＴＦ−Ｍ産生促進効果を示し、（Ａ）フォルスコリンと（Ｂ）成分の合計量が多いほど、ＭＩＴＦ−Ｍ産生促進効果が高まることが確認された。
一方、フォルスコリンとホップエキスとの併用に、ＭＩＴＦ−Ｍ産生量に対する相乗効果は認められなかった（比較例２１）。
As shown in Table 2, the MITF-M production promoter composed of (A) forskolin and (B) component of the present invention exhibits an excellent effect of promoting MITF-M production in any combination, (A) It was confirmed that the greater the total amount of forskolin and component (B), the higher the effect of promoting MITF-M production.
On the other hand, the combined use of forskolin and hop extract did not show a synergistic effect on the amount of MITF-M production (Comparative Example 21).
（２）毛髪用化粧料組成物の抗白髪効果
本発明のＭＩＴＦ−Ｍ産生促進剤として、（Ａ）フォルスコリン含有量９５％のコレウスホルスコリ根エキスと、（Ｂ）成分とを毛髪用化粧料（シャンプー、コンディショナー、ヘアトニック）に配合して、抗白髪効果を評価した。
(2) Anti-white hair effect of hair cosmetic composition As MITF-M production promoter of the present invention, (A) Coleus forskori root extract with 95% forskolin content and (B) component are used for hair makeup. The anti-white hair effect was evaluated by blending with ingredients (shampoo, conditioner, hair tonic).
毛髪用化粧料組成物（シャンプー、コンディショナー、ヘアトニック）の組成を以下に示し、（Ａ）コレウスホルスコリ根エキス、（Ｂ）黒砂糖エキス、オレンジエキス、チンピエキス、ウメエキス、ワカメエキス、およびホップエキスの添加量を表３に示す。コレウスホルスコリ根エキスは、サビンサジャパンコーポレーション製を、黒砂糖エキス、オレンジエキス、チンピエキス、ウメエキス、ワカメエキスおよびホップエキスは５０質量％エタノール水溶液で抽出した各植物エキスの凍結乾燥物を用いた。なお、配合量の単位は質量部で、精製水の「残量」とは全量を１００質量部とする量である。また、シャンプーにおけるクエン酸２５％水溶液の適量とは、ｐＨを約５.５に調整するに要する量、塩化ナトリウム１０％水溶液の適量とは、粘度を約１５００ｍＰａ・sに調整するに要する量である。 The composition of the cosmetic composition for hair (shampoo, conditioner, hair tonic) is shown below. (A) Coleus forskori root extract, (B) Brown sugar extract, orange extract, chimpi extract, plum extract, wakame extract, and hop extract Table 3 shows the amount of added. Coleus forskori root extract was manufactured by Savinsa Japan Corporation, and brown sugar extract, orange extract, chimpi extract, ume extract, wakame extract and hop extract were lyophilized products of plant extracts extracted with 50% by mass aqueous ethanol. The unit of the blending amount is parts by mass, and the “remaining amount” of purified water is an amount that makes the total amount 100 parts by mass. The appropriate amount of 25% aqueous citric acid solution in shampoo is the amount required to adjust the pH to about 5.5, and the appropriate amount of 10% aqueous sodium chloride solution is the amount required to adjust the viscosity to about 1500 mPa · s. is there.
評価
実施例３６〜４０、比較例２２〜３０の頭髪用化粧料組成物（シャンプーとコンディショナー、ヘアトニック）について、それぞれ３０〜６０歳代の白髪のある男女１０名を被験者とした。
シャンプー及びコンディショナーについては毎日の入浴時に、ヘアトニックについては毎朝および毎日の入浴後に適量を使用した。このモニター試験を６ヶ月間継続し、使用前後の頭頂部の毛髪２００本あたりの白髪の本数を比較して、下記の基準にて評価した。
５点（著効）：使用前の白髪の本数に対し、使用後の白髪の本数が８０％未満となった場合
４点（有効）：使用前の白髪の本数に対し、使用後の白髪の本数が８０％以上８５％未満となった場合
３点（有効）：使用前の白髪の本数に対し、使用後の白髪の本数が８５％以上９０％未満となった場合
２点（有効）：使用前の白髪の本数に対し、使用後の白髪の本数が９０％以上９５％未満となった場合
１点（やや有効）：使用前の白髪の本数に対し、使用後の白髪の本数が９５％以上１００％未満の場合
０点（無効）：使用前の白髪の本数に対し、使用後の白髪の本数が１００％以上となった場合
Evaluation About the cosmetic compositions for hair of Examples 36 to 40 and Comparative Examples 22 to 30 (shampoo and conditioner, hair tonic), 10 men and women with white hair of 30 to 60 years old were subjects.
Appropriate amounts were used at the time of daily bathing for shampoos and conditioners, and every morning and after daily bathing for hair tonics. This monitoring test was continued for 6 months, and the number of white hairs per 200 hairs at the top of the head before and after use was compared and evaluated according to the following criteria.
5 points (significant): When the number of white hair after use is less than 80% of the number of gray hair before use 4 points (valid): The number of white hair before use against the number of white hair before use When the number is 80% or more and less than 85% 3 points (effective): When the number of white hair after use is 85% or more and less than 90% with respect to the number of white hair before use 2 points (effective): When the number of gray hair after use is 90% or more and less than 95% with respect to the number of gray hair before use 1 point (somewhat effective): The number of gray hair after use is 95 against the number of white hair before use % Or more and less than 100% 0 point (invalid): When the number of gray hair after use is 100% or more compared to the number of gray hair before use
次に、各毛髪用化粧料に対する被験者１０名の評価点を合計し、以下の基準により効果を判定した。
Ａ： 評価点の合計が ４１〜５０点
Ｂ： 評価点の合計が ３１〜４０点
Ｃ： 評価点の合計が ２１〜３０点
Ｄ： 評価点の合計が １１〜２０点
Ｅ： 評価点の合計が ６〜１０点
Ｆ： 評価点の合計が ５点以下
評価結果を表４−１、表４−２に示す。尚、この期間中、前記以外の頭髪用化粧料については、通常使用しているものをそのまま使用した。使用期間中に皮膚の異常を訴えたものはいなかった。
Next, the evaluation scores of 10 test subjects for each hair cosmetic were added together, and the effect was determined according to the following criteria.
A: Total score is 41-50 points B: Total score is 31-40 points C: Total score is 21-30 points D: Total score is 11-20 points E: Total score 6 to 10 points F: The total of the evaluation points is 5 or less. The evaluation results are shown in Tables 4-1 and 4-2. During this period, the hair cosmetics other than those described above were used as they were. No one complained of skin abnormalities during use.
本発明のＭＩＴＦ産生促進剤を配合した実施例３６〜４０の毛髪用化粧料に、白髪の改善効果が認められた。 In the hair cosmetics of Examples 36 to 40, in which the MITF production promoter of the present invention was blended, an effect of improving gray hair was observed.
（３）皮膚用化粧料組成物のタンニング効果
本発明のＭＩＴＦ−Ｍ産生促進剤として、（Ａ）コレウスホルスコリ根エキスと、（Ｂ）成分とを皮膚用化粧料（ボディローション、ボディクリーム）に配合して、タンニング効果を評価した。
(3) Tanning effect of skin cosmetic composition As a MITF-M production promoter of the present invention, (A) Coleus forskori root extract and (B) component are used as a skin cosmetic (body lotion, body cream). And tanning effect was evaluated.
皮膚用化粧料組成物（ボディローション、ボディクリーム）の組成を以下に示し、（Ａ）コレウスホルスコリ根エキス、（Ｂ）黒砂糖エキス、オレンジエキス、チンピエキス、ウメエキス、ワカメエキス、およびホップエキスの添加量を表３に示す。コレウスホルスコリ根エキスは、サビンサジャパンコーポレーション製を、黒砂糖エキス、オレンジエキス、チンピエキス、ウメエキス、ワカメエキスおよびホップエキスは５０質量％エタノール水溶液で抽出した各植物エキスの凍結乾燥物を用いた。なお、配合量の単位は質量部で、表中の精製水の「残量」とは全量を１００質量部とする量である。 The composition of the cosmetic composition for skin (body lotion, body cream) is shown below. (A) Coleus forskori root extract, (B) Brown sugar extract, orange extract, chimpi extract, ume extract, wakame extract, and hop extract The amount added is shown in Table 3. Coleus forskori root extract was manufactured by Savinsa Japan Corporation, and brown sugar extract, orange extract, chimpi extract, ume extract, wakame extract and hop extract were lyophilized products of plant extracts extracted with 50% by mass aqueous ethanol. In addition, the unit of compounding quantity is a mass part, and the "remaining amount" of the purified water in a table | surface is the quantity which makes the whole quantity 100 mass parts.
評価
実施例４１〜４５、比較例３１〜３９の皮膚用化粧料組成物（ボディローション、ボディクリーム）について、それぞれ男女５名を被験者とした。
実施例および比較例の皮膚用化粧料を、１日２回上腕内側部に塗布し、１ヶ月間連続使用した後の試験部位とその周りの皮膚の明暗度を比較して下記の基準にて評価した。
２点：無塗布部と比較して褐色に変化
１点：無塗布部と比較してやや褐色に変化
０点：無塗布部と比較して変化なし
Evaluation About the cosmetic compositions for skin (body lotion, body cream) of Examples 41 to 45 and Comparative Examples 31 to 39, 5 men and women were subjects.
The skin cosmetics of Examples and Comparative Examples were applied to the inner side of the upper arm twice a day, and the brightness of the test site and the surrounding skin were compared after the continuous use for one month according to the following criteria. evaluated.
2 points: Change to brown compared to uncoated area 1 point: Change to slightly brown compared to uncoated area 0 point: No change compared to uncoated area
次に、各皮膚用化粧料に対する被験者５名の評価点を合計し、以下の基準により効果を判定した。
Ａ： 評価点の合計が ７〜１０点
Ｂ： 評価点の合計が ３〜６点
Ｃ： 評価点の合計が ２点以下
評価結果を表５−１、表５−２に示す。尚、この期間中に皮膚の異常を訴えたものはいなかった。
Next, the evaluation points of five subjects for each skin cosmetic were totaled, and the effect was determined according to the following criteria.
A: Total score of 7 to 10 points B: Total score of 3 to 6 points C: Total score of 2 or less Evaluation results are shown in Tables 5-1 and 5-2. No one complained of skin abnormalities during this period.
本発明のＭＩＴＦ産生促進剤を配合した実施例４１〜４５の皮膚用化粧料に、良好なタンニング効果が認められた。 A good tanning effect was observed in the skin cosmetics of Examples 41 to 45 containing the MITF production promoter of the present invention.
本発明のＭＩＴＦ−Ｍ産生促進剤は、メラノサイトにおけるＭＩＴＦ−Ｍ産生量の増大に優れた効果を発揮するため、これを頭髪用化粧料組成物に配合することにより、白髪の予防、緩和、改善の効果が、皮膚用化粧料組成物に配合することにより、タンニング効果が期待できる。 Since the MITF-M production promoter of the present invention exhibits an excellent effect in increasing the amount of MITF-M production in melanocytes, it is prevented, alleviated and improved by blending it into a cosmetic composition for hair. The effect of the above can be expected to be a tanning effect by blending into the cosmetic composition for skin.
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| JP6655299B2 (en) * | 2015-04-03 | 2020-02-26 | クラシエホームプロダクツ株式会社 | Hair cosmetic composition |
| JP2020527604A (en) * | 2017-07-14 | 2020-09-10 | ライフ・サイエンス・インベストメンツ・リミテッドＬＩＦＥ ＳＣＩＥＮＣＥ ＩＮＶＥＳＴＭＥＮＴＳ Ｌｔｄ | Cosmetic compositions containing Coleus barbatus and Cassia occidentalis and / or Cassia Arata, and their use in the treatment of vitiligo |
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