JP4441083B2 - Melanocyte dendrite elongation inhibitor and cosmetics containing the same - Google Patents

Description

化学式（ＩＩ）
【０００８】
（２）デンドライトの伸長抑制が、マクロファージ由来のデンドライト伸長促進因子の抑制作用に起因することを特徴とする、（１）に記載のメラノサイトのデンドライト伸長抑制剤。
以下、本発明について、実施の形態を中心に詳細に説明を加える。
【０００９】
【発明の実施の形態】
（１）本発明のメラノサイトのデンドライトの伸長抑制剤
本発明のメラノサイトのデンドライト伸長抑制剤は、下記化学式（ＩＩ）に表されるマアキアイン（maackiain）及びその塩からなる。前記、本発明のメラノサイトのデンドライトの伸長抑制剤であるマアキアイン（maackiain）は、マメ科のSophora subprostrata(広豆根及び山豆根)の根茎から単離精製される。
【００１０】
【化３】

化学式（ＩＩ）
【００１１】
この化合物を得る方法として、常法に従えば良く、第一ステップとして各種有機溶媒により該化合物含有成分を抽出して、液液抽出し、有機溶媒画分を減圧乾固し、有機溶媒を除去する。場合によっては凍結乾燥をし粗分画を得る。第二ステップとして、粗分画を少量の有機溶媒に溶解し、シリカゲルクロマトグラフィー、ＯＤＳクロマトグラフィーや高速液体クロマトグラフィーにより、化学式（ＩＩ）に示されるマアキアイン（maackiain）を精製し単離する手段を取る。かかる植物の植物体それ自身、植物体を乾燥或いは細切、粉砕など加工した加工物、植物体乃至はその加工物を溶媒で抽出した抽出物、抽出物の溶媒を除去した、溶媒除去物、抽出物乃至はその溶媒除去物を液液抽出後、カラムクロマトグラフィー、ＯＤＳカラムクロマトグラフィーで精製した精製分画物である化学式（ＩＩ）に示されるマアキアイン（maackiain）を精製し単離することを意味する。粗分画を得るための溶媒としては、極性溶媒が特に好ましく例示できる。この様な極性溶媒としては、例えば、水、エタノール、メタノール、１，３−ブタンジオール、プロピレングリコールなどのアルコール類、酢酸エチルや蟻酸メチルなどのエステル類、アセトンやメチルエチルケトンなどのケトン類、クロロホルムや塩化メチレン等のハロゲン化炭化水素類、アセトニトリル等のニトリル類、ジエチルエーテルやテトラヒドロフランなどのエーテル類から選ばれる１種乃至は２種以上が好ましく例示できる。これらの内、特に好ましいものは、酢酸エチルや蟻酸メチルなどのエステル類である。この様な抽出物を作成するには、植物体乃至はその加工物に１〜１０倍量の溶媒を加え、室温であれば数日、沸点付近の温度であれば数時間浸漬すればよい。しかる後に、不溶物を濾過などで除去し、必要に応じて減圧濃縮や凍結乾燥により溶媒除去することが出来る。マメ科のSophora subprostrata(広豆根及び山豆根)の根茎の場合、根茎部分を酢酸エチルで抽出後、濾過し、減圧濃縮する、そして場合によって凍結乾燥し粗分画を得て、化学式（ＩＩ）に示されるマアキアイン（maackiain）を精製し単離することができる。それは、根茎の部分にこの様な物質が多く含まれており、特に好ましいからである。又この様な化合物はアシルハライドと塩基を用いたアシル化反応、アルキルハライドとアルカリを用いたアルキル化反応、アシル化糖と酸化銀触媒を用いた配糖化反応を行うことにより各種誘導体へと導くことができる。かくして得られた、本発明のメラノサイトのデンドライトの伸長抑制剤である、マメ科のSophora subprostrata(広豆根及び山豆根)の根茎から得られた化学式（ＩＩ）に示されるマアキアイン（maackiain）及び／又はその塩は、メラノサイトがデンドライトを伸長するのを抑制する作用に優れ、以て、メラノサイトより皮膚組織へメラニン顆粒が移動するのを抑制し、この様なメラニン顆粒の移動をメカニズムとする、光照射時に生じる、炎症を伴った黒化やソバカスなどの色素異常を予防或いは改善する作用を有する。この様な作用は、マクロファージが放出するメラノサイトのデンドライトの伸長因子がメラノサイトに働きかけるのを阻害することを機序としていると考えられる。勿論、色素異常が、メラニン顆粒の産生にあたってこの様なルートをとることから、本発明のメラノサイトのデンドライト伸長抑制剤は、光照射による炎症を伴った黒化やシミ・ソバカス以外の色素異常も抑制するが、この様な色素異常は他の手段でも予防や改善が可能であるため、本発明の効果の特徴は前記の光照射時に生じる、炎症を伴った黒化やシミ・ソバカスなどの色素異常を予防或いは改善する作用と言える。
【００１２】
（２）本発明のメラノサイトとマクロファージが関与する皮膚現象対応用の皮膚外用剤
本発明のメラノサイトのデンドライト伸長抑制剤は、マクロファージが放出するメラノサイトのデンドライトの伸長因子がメラノサイトに働きかけるのを阻害することを機序としているので、メラノサイトとマクロファージとが協調的に働く生命現象を抑制することが出来、この様なメラノサイトのデンドライト伸長抑制剤を、皮膚外用剤に含有させることにより、メラノサイトとマクロファージが関与する皮膚現象へ対応する事が出来る。即ち、本発明の皮膚外用剤は、メラノサイトとマクロファージが関与する皮膚現象対応用であって、本発明のメラノサイトのデンドライト伸長抑制剤を含有することを特徴とする。ここで、本発明で言う皮膚外用剤とは、皮膚に外用で適用される組成物の総称であって、貼付剤を含む皮膚外用医薬や洗浄剤を含む化粧料が好ましく例示でき、これらの内では、化粧料であることが特に好ましい。これは、本発明のメラノサイトのデンドライト伸長抑制剤の安全性が高く、作用が穏やかであるためである。メラノサイトとマクロファージが関与する皮膚現象としては、特に好ましくは前述の光照射による炎症を伴った黒化やシミ・ソバカスなどの色素異常がまず一番重要な課題として挙げられるが、その他炎症反応なども含まれる。本発明のメラノサイトとマクロファージが関与する皮膚現象対応用の皮膚外用剤に於ける、メラノサイトのデンドライト伸長抑制剤の好ましい含有量は、皮膚外用剤全量に対して、０．００１重量％〜１０重量％であり、更に好ましくは０．０１重量％〜５重量％である。これは、少なすぎるとデンドライトの伸長抑制作用が発揮されない場合があり、多すぎても効果が頭打ちになり他の処方成分の自由度を損なうことがあるからである。
【００１３】
本発明のメラノサイトとマクロファージが関与する皮膚現象対応用の皮膚外用剤は、上記必須成分以外に、通常化粧料や皮膚外用医薬で使用される任意の成分を含有することが出来る。かかる任意成分としては、例えば、スクワラン、ワセリン、マイクロクリスタリンワックス等の炭化水素類、ホホバ油、カルナウバワックス，オレイン酸オクチルドデシル等のエステル類、オリーブ油、牛脂、椰子油等のトリグリセライド類、ステアリン酸、オレイン酸、リチノレイン酸等の脂肪酸、オレイルアルコール、ステアリルアルコール、オクチルドデカノール等の高級アルコール、スルホコハク酸エステルやポリオキシエチレンアルキル硫酸ナトリウム等のアニオン界面活性剤類、アルキルベタイン塩等の両性界面活性剤類、ジアルキルアンモニウム塩等のカチオン界面活性剤類、ソルビタン脂肪酸エステル、脂肪酸モノグリセライド、これらのポリオキシエチレン付加物、ポリオキシエチレンアルキルエーテル、ポリオキシエチレン脂肪酸エステル等の非イオン界面活性剤類、ポリエチレングリコール、グリセリン、１，３−ブタンジオール等の多価アルコール類、増粘・ゲル化剤、酸化防止剤、紫外線吸収剤、色剤、防腐剤、粉体等を含有することができる。本発明のメラノサイトとマクロファージが関与する皮膚現象対応用の皮膚外用剤は、抗炎症剤として知られるプレドニゾロン、ヒドロコルチゾン、インドメタシン、ジクロフェナックナトリウム等を配合させれば相乗効果により日光による炎症を伴う黒化症に有意義である。勿論、従来のメラニン産生抑制剤である、アスコルビン酸類やアルブチンなどのハイドロキノン類を含有することも相乗的な効果を発揮する場合があり、有利である。
【００１４】
【実施例】
以下に実施例を挙げて更に詳細に本発明について説明を加えるが、本発明がこれら実施例にのみ、限定を受けないことは言うまでもない。
【００１５】
＜実施例１＞
化学式（ＩＩ）に示されるマアキアイン（maackiain）の抽出・精製法を示す。マメ科のSophora subprostrata(広豆根及び山豆根)の根茎５００ｇに５Ｌの酢酸エチルを加え、攪拌しながら２時間、９０℃で加熱し、室温まで冷却した後、濾過して不溶物を取り除き、減圧濃縮後、その後、凍結乾燥し、粗分画を得た。そして、凍結乾燥物を水・クロロフォルムで液々抽出し、クロロホルム層を濃縮乾固した。この濃縮乾固物を少量のクロロホルムに溶解し、シリカゲルカラムクロマトグラフィーで溶出溶媒をクロロホルム：メタノール＝１００：０〜９：１の条件下で溶出させて、精製し、アセトンで再結晶し、マアキアイン（maackiain）を約５００ｍｇのを得た。
【００１６】
＜実施例２＞
上記実施例１のメラノサイトのデンドライト伸長抑制剤であるマアキアイン（maackiain）を用いて、デンドライト伸長抑制作用を調べた。即ち、予め常法に従い、マウス腹腔より、マクロファージを回収し、１０％ＦＢＳ添加ＭＥＳ培地で希釈し、２×１０ ⁶ セル／ｍｌの濃度のマクロファージ液を調製しておいた。このものを１３５μｌずつ３５ｍｍシャーレに分注し、これに０．０５ｍＷ／ｃｍ ² で２０分間の紫外線照射を行った。このメラノサイトのデンドライト伸長抑制剤であるマメ科のSophora subprostrata(広豆根及び山豆根)の根茎から得られたマアキアイン（maackiain）を乾燥重量％で０．００５％となるようＤＭＳＯに溶かして加え３７℃で１晩培養した。又、他方マウスの尾を切り、尾の表皮を細かく刻みシャーレに入れ０．５％トリプシンにて４℃で一晩処理し、ピンセットを用いて、表皮と真皮に分離し、表皮のみを回収し、０．５％トリプシンにて３７℃で２０分間処理し、フィルター濾過でメラノサイトのみを濾液として集めた。このメラノサイトを含む濾液を、ＭＥＳ培地に１０％ＦＢＳ、１０ ^-4 ＭのＩＢＭＸ及び１０ｎｇ／ｍｌのＴＰＡを加えた培地で、３７℃、７２時間培養した。これを同培地で懸濁させ、９６穴ウェルに１０００セル／ウェルずつ分注し、３７℃で一晩培養した。メラノサイトの培地を捨て、ＰＢＳで３回洗浄した後、１０％ＦＢＳ加イーグルの最少培地３５μｌに置換した。これに前記検体を含むマクロファージの培養上清３５μｌずつ添加し、３７℃で二晩培養し、光学顕微鏡下写真撮影を行い、この写真よりデンドライトの長さを測定した。結果を表１に示す。これより、本発明のメラノサイトのデンドライト伸長抑制剤であるマメ科のSophora subprostrata(広豆根及び山豆根)の根茎から得られたマアキアイン（maackiain）によるデンドライト伸長の抑制作用に優れることが分かる。
（検体）
１）ＵＶ照射のマクロファージにマアキアイン（maackiain）を０．０５％添加したもの２）ＵＶ照射のマクロファージ（ポジティブコントロール）
３）ＵＶ非照射のマクロファージ（ネガティブコントロール）
【００１７】
【表１】

【００１８】
＜実施例３＞
以下に示す処方で化粧水を作成した。即ち、処方成分を室温で攪拌可溶化して化粧水を得た。この化粧水について、シミ、そばかすに悩むパネラー1群３名を用いて、1ヶ月間、朝晩1日2回使用してもらいそのシミ、そばかすの予防及び改善効果を評価してもらった。評価基準は、評点2：著しい改善、評点1：明らかな改善、評点0.5：わずかな改善、評点0：改善なしの基準である。平均評点は１．２５であった。本発明のメラノサイトのデンドライドの伸長抑制効果のあるマアキアイン（maackiain）を含有する化粧水は、シミ、ソバカスの改善に効果のあることが認められた。
マアキアイン（maackiain） １ 重量部
１，３ブタンジオール ５ 重量部
グリセリン ３ 重量部
クエン酸ナトリウム ０．１重量部
メチルパラベン ０．２重量部
エタノール ８ 重量部
水 ８２．７重量部
【００１９】
＜実施例４＞
下記に示す処方に従って、皮膚外用医薬組成物を作成した。即ち、処方成分を攪拌分散して、皮膚外用剤を得た。このものは光による炎症を伴った、光による皮膚の黒化現象或いはシミ・ソバカスに対して著効を示した。
マアキアイン（maackiain） ０．５重量部
プレドニゾロン １ 重量部
ワセリン ８５ 重量部
【００２０】
＜実施例５＞
以下に示す処方に従ってクリームを作製した。即ち、イ、ロ、ハをそれぞれ８０℃に加熱溶解して、イにロを徐々に加え、更にハを加え乳化した後、ホモミキサーにより乳化粒子を均一化し、冷却してクリームを得た。このクリームは、炎症を伴うシミ・ソバカス等に優れた効果があった。
【００２１】
イ）
スクワラン １０ 重量部
セタノール ３ 重量部
ソルビタンセスキステアレート ２ 重量部
ポリオキシエチレン（２０）ベヘニルエーテル ２ 重量部
ビタミンＡ酸 １ 重量部
ロ）
１，３−ブタンジオール ５ 重量部
マアキアイン（maackiain） １ 重量部
カルボキシビニルポリマー ０．３重量部
水 ４０ 重量部
ハ）
水 ３７．３重量部
水酸化カリウム ０．２重量部
【００２２】
【発明の効果】
本発明によれば、炎症を伴った色素異常やシミ・ソバカスなどの色素異常に対して有効な予防或いは改善手段を提供することができる。[0001]
BACKGROUND OF THE INVENTION
The present invention, by dendrite elongation inhibitor of dendrite elongation inhibitor and main Ranosaito melanocytes, suitable for inhibiting the interaction of melanocytes and macrophages, to the skin external preparation of skin symptoms for the corresponding melanocytes and macrophages are involved.
[0002]
[Prior art]
It is already known that melanocytes are the leading role of life phenomena related to pigments in animals, but melanin, which is such a pigment, is produced in melanocytes and how it moves to epidermal cells. It is not yet known in detail, but it is only known that macrophages are often involved in the movement of such melanin granules. It has been pointed out that the involvement of such macrophages is caused by producing melanocyte dendrite elongation factor (DEF). No attempt has been made to suppress the growth of melanocytes and to suppress the movement of melanin granules to prevent the skin from becoming darkened by suppressing the extension of dendrites. Further, a compound represented in leguminous Sophora subprostrata (Hiromamene and Yamamamene) isolated formulas from rhizomes of (II) Maakiain (Maackiain)及 beauty / or of such melanocyte salt thereof It is not known at all that it has an elongation-inhibiting action.
[0003]
On the other hand, resolving pigment abnormalities caused by abnormalities in melanin granules produced by melanocytes is a long-standing problem for humankind to embody beautiful white skin. Various mechanisms have been obtained, but none has focused on the suppression of melanocyte dendrite elongation. It is not known at all that it is useful for the prevention and improvement of pigmented dysfunctions associated with. In addition, melanin production inhibitors such as ascorbic acid, which are well-known in the past, are effective against pigment abnormalities associated with inflammation and pigment abnormalities such as spots and freckles. Prevention or improvement of such pigment abnormalities Development of means was desired.
[0004]
In addition, the rhizome essence of leguminous Sophora subprostrata (Hirzu and Yamazu) is known to have anti-inflammatory and antibacterial activity, but was isolated from it not at all known to have action to inhibit Maakiain (maackiain) and / or dendrite extension of melanocytes into a salt thereof is a compound represented by formula (II), therefore, it contains the ones It is well known that topical skin preparations such as cosmetics suppress dendrite elongation of melanocytes and are useful for the prevention and improvement of pigment abnormalities that involve pigment abnormalities, especially, light, and inflammation. It was not.
[0005]
[Problems to be solved by the invention]
The present invention has been made under such circumstances, and an object of the present invention is to provide an effective preventive or ameliorating means for pigment abnormalities associated with inflammation and pigment abnormalities such as stains and freckles.
[0006]
[Means for solving problems]
In view of such circumstances, the present inventors have sought for effective preventive or ameliorating means for pigment abnormalities associated with inflammation and pigment abnormalities such as spots and freckles, and as a result of earnest research, the chemical formula ( a compound represented in II) Maakiain (maackiain)及 beauty / or found outgrowth inhibition effect of dendrites excellent melanocytes into a salt thereof, a substance having such effects by containing the skin external preparation, with inflammation The present invention was found to be useful for the prevention and improvement of pigment abnormalities such as pigment abnormalities and spots and freckles. That is, the present invention is Ru der relates following technique.
[0007]
(1) made of Maakiain (Maackiain) and / or a salt thereof is a compound represented by the following chemical formula (II), Dendorai preparative Shin length inhibitors of melanocytes.
[Chemical formula 2]

Chemical formula (II)
[0008]
(2) The dendrite elongation inhibitor for melanocytes according to (1), wherein dendrite elongation inhibition is caused by the inhibitory action of a macrophage-derived dendrite elongation promoting factor .
Hereinafter, the present invention will be described in detail with a focus on embodiments.
[0009]
DETAILED DESCRIPTION OF THE INVENTION
(1) dendrite elongation inhibitor for melanocytes elongation inhibitor invention dendrites of melanocytes present invention, Maakiain (Maackiain)及 beauty salt thereof or Ranaru represented by the following chemical formula (II). Before reporting, Maakiain an elongation inhibitor dendrites of melanocytes present invention (Maackiain) is isolated and purified from the rhizome of leguminous Sophora subprostrata (Hiromamene and Yamamamene).
[0010]
[Chemical 3]

Chemical formula (II)
[0011]
As a method for obtaining this compound, a conventional method may be followed. As a first step, the compound-containing component is extracted with various organic solvents, liquid-liquid extraction is performed, and the organic solvent fraction is dried under reduced pressure, and the organic solvent is removed. To do. In some cases, the crude fraction is obtained by lyophilization. As a second step, the crude fraction was dissolved in a small amount of organic solvents, silica gel chromatography, ODS chromatography or high performance liquid Ri by the chromatography, a single purified the Maakiain (maackiain) represented by Structural Formula (II) Take a means to release. The plant body itself, a processed product obtained by drying, chopping, or crushing the plant body, an extract obtained by extracting the plant body or the processed product with a solvent, a solvent-removed product obtained by removing the solvent of the extract, after the extract to liquid-liquid extraction and the solvent removed substance by column chromatography, purified isolate Maakiain (maackiain) shown in ODS column purification fractions was purified by chromatography der Ru Structural formula (II) that it means to. As a solvent for obtaining a crude fraction, a polar solvent can be particularly preferably exemplified. Examples of such polar solvents include alcohols such as water, ethanol, methanol, 1,3-butanediol, and propylene glycol, esters such as ethyl acetate and methyl formate, ketones such as acetone and methyl ethyl ketone, chloroform, and the like. Preferred examples include one or more selected from halogenated hydrocarbons such as methylene chloride, nitriles such as acetonitrile, and ethers such as diethyl ether and tetrahydrofuran. Of these, particularly preferred are esters such as ethyl acetate and methyl formate. In order to prepare such an extract, a 1 to 10-fold amount of a solvent is added to a plant body or a processed product, and it is immersed for several days at room temperature or for several hours at a temperature near the boiling point. Thereafter, the insoluble matter is removed by filtration or the like, and the solvent can be removed by vacuum concentration or lyophilization as necessary. For rhizome leguminous Sophora subprostrata (Hiromamene and Yamamamene), after extracting the rhizome parts of ethyl acetate, filtered and vacuum concentrated, and to give a lyophilized crude fraction optionally, chemical The maackiain represented by formula (II) can be purified and isolated. This is because a lot of such substances are contained in the rhizome part, which is particularly preferable. Such compounds lead to various derivatives by performing acylation reaction using acyl halide and base, alkylation reaction using alkyl halide and alkali, and glycosylation reaction using acylated sugar and silver oxide catalyst. be able to. Thus obtained, a stretch inhibitor dendrites of melanocytes present invention, Maakiain shown in leguminous Sophora subprostrata (Hiromamene and Yamamamene) rhizomes from the resulting chemical formula (II) (maackiain ) And / or a salt thereof is excellent in the action of suppressing the extension of dendrites by melanocytes, and thus suppresses the movement of melanin granules from the melanocytes to the skin tissue. It has the effect of preventing or ameliorating pigment abnormalities such as blackening and buckwheat accompanied by inflammation that occur during light irradiation. Such an action is considered to be based on the mechanism by which the dendritic elongation factor of melanocytes released by macrophages inhibits melanocytes from acting. Of course, since pigment abnormalities take such a route in the production of melanin granules, the melanocyte dendrite elongation inhibitor of the present invention also suppresses pigment abnormalities other than blackening and spots and freckles associated with inflammation caused by light irradiation. However, since such pigment abnormalities can be prevented or improved by other means, the characteristics of the effect of the present invention are pigment abnormalities such as blackening accompanied by inflammation and spots and freckles that occur during the light irradiation. It can be said that this is an action to prevent or improve the above.
[0012]
(2) Skin external preparation for dealing with skin phenomenon involving melanocytes and macrophages of the present invention The dendritic elongation inhibitor of melanocytes of the present invention inhibits the melanocyte dendritic elongation factor released from macrophages from acting on melanocytes. Therefore, it is possible to suppress the life phenomenon in which melanocytes and macrophages work in a coordinated manner. By including such a dendritic elongation inhibitor of melanocytes in a topical skin preparation, melanocytes and macrophages are involved. Can cope with skin phenomenon. That is, the external preparation for skin of the present invention is for dealing with skin phenomena involving melanocytes and macrophages, and is characterized by containing the dendrite elongation inhibitor of melanocytes of the present invention. Here, the topical skin preparation as used in the present invention is a general term for compositions applied externally to the skin, and can be preferably exemplified by a skin external medicine containing a patch and a cosmetic containing a cleansing agent. Then, it is especially preferable that it is cosmetics. This is because the melanocyte dendrite elongation inhibitor of the present invention has high safety and mild action. As the skin phenomenon involving melanocytes and macrophages, particularly preferably, the above-mentioned blackening accompanied by inflammation caused by light irradiation and pigment abnormalities such as spots and freckles are listed as the most important issues. included. In the skin external preparation for dealing with skin phenomena involving melanocytes and macrophages of the present invention, the preferred content of the dendritic elongation inhibitor for melanocytes is 0.001% by weight to 10% by weight with respect to the total amount of the external preparation for skin. More preferably, it is 0.01% by weight to 5% by weight. This is because if the amount is too small, the dendrite elongation-inhibiting action may not be exhibited. If the amount is too large, the effect will reach its peak and the degree of freedom of other prescription ingredients may be impaired.
[0013]
The skin external preparation for dealing with skin phenomena involving melanocytes and macrophages according to the present invention can contain any components that are usually used in cosmetics and external skin medicines in addition to the above essential components. Such optional components include, for example, hydrocarbons such as squalane, petrolatum, microcrystalline wax, esters such as jojoba oil, carnauba wax, octyldodecyl oleate, triglycerides such as olive oil, beef tallow, coconut oil, stearic acid, etc. , Fatty acids such as oleic acid, lithinoleic acid, higher alcohols such as oleyl alcohol, stearyl alcohol, octyldodecanol, anionic surfactants such as sulfosuccinic acid ester and sodium polyoxyethylene alkylsulfate, and amphoteric surfactants such as alkylbetaine salts Agents, cationic surfactants such as dialkylammonium salts, sorbitan fatty acid esters, fatty acid monoglycerides, polyoxyethylene adducts thereof, polyoxyethylene alkyl ethers, polyoxyethylenes Nonionic surfactants such as fatty acid esters, polyhydric alcohols such as polyethylene glycol, glycerin, 1,3-butanediol, thickening / gelling agents, antioxidants, ultraviolet absorbers, colorants, preservatives , Powder and the like can be contained. The skin external preparation for dealing with skin phenomena involving melanocytes and macrophages of the present invention is melanosis with inflammation caused by sunlight by synergistic effects when prednisolone, hydrocortisone, indomethacin, diclofenac sodium, etc., known as anti-inflammatory agents are combined. Is meaningful. Of course, the inclusion of hydroquinones such as ascorbic acids and arbutin, which are conventional melanin production inhibitors, may also exhibit a synergistic effect and is advantageous.
[0014]
【Example】
Hereinafter, the present invention will be described in more detail with reference to examples, but it is needless to say that the present invention is not limited to these examples.
[0015]
<Example 1>
Shown in the formula (II) shows the extraction and purification of luma Akiain (maackiain). Add 5 L of ethyl acetate to 500 g of rhizome of leguminous Sophora subprostrata (Hirzu and Yam), heat at 90 ° C. with stirring for 2 hours, cool to room temperature, and filter to remove insolubles. The solution was concentrated under reduced pressure, and then freeze-dried to obtain a crude fraction. The freeze-dried product was extracted liquid-liquid with water / chloroform, and the chloroform layer was concentrated to dryness. This concentrated dried product is dissolved in a small amount of chloroform, and the silica gel column chromatography is used to elute the elution solvent under the conditions of chloroform: methanol = 100: 0-9: 1, purify, recrystallize with acetone, and maakiain. About 500 mg of (maackiain) was obtained.
[0016]
<Example 2>
The dendrite elongation-inhibiting action was examined using maackiain, which is a dendrite elongation inhibitor of the melanocytes of Example 1 above. That is, according to a conventional method, macrophages were collected from the mouse abdominal cavity and diluted with MES medium supplemented with 10% FBS to prepare a macrophage solution having a concentration of 2 × 10 ⁶ cells / ml. 135 μl of this was dispensed into 35 mm petri dishes and irradiated with ultraviolet rays at 0.05 mW / cm ² for 20 minutes. The maackiain obtained from the rhizome of leguminous Sophora subprostrata, which is a dendrite elongation inhibitor of melanocytes, is added to DMSO so that the dry weight% is 0.005%. Cultured overnight at 37 ° C. Also, cut the tail of the other mouse, finely cut the tail epidermis into a petri dish, treat with 0.5% trypsin at 4 ° C overnight, separate it into epidermis and dermis using tweezers, and collect only the epidermis. The solution was treated with 0.5% trypsin at 37 ° C. for 20 minutes, and only melanocytes were collected as a filtrate by filtration. The filtrate containing melanocytes was cultured at 37 ° C. for 72 hours in a medium in which 10% FBS, 10 ⁻⁴ M IBMX and 10 ng / ml TPA were added to MES medium. This was suspended in the same medium, dispensed at 1000 cells / well into 96-wells, and cultured overnight at 37 ° C. The melanocyte medium was discarded, washed 3 times with PBS, and replaced with 35 μl of minimal medium with 10% FBS-added eagle. To this was added 35 μl each of the culture supernatant of macrophages containing the specimen, cultured at 37 ° C. overnight, photographed under an optical microscope, and the length of dendrites was measured from this photograph. The results are shown in Table 1. This shows that the dendrite elongation-inhibiting action by maackiain obtained from the rhizome of leguminous Sophora subprostrata (Hamazu-root and Yamazu-root), which is the melanocyte dendrite elongation inhibitor of the present invention, is excellent.
(Sample)
1) UV irradiated macrophages with 0.05% maackiain added 2) UV irradiated macrophages (positive control)
3) UV non-irradiated macrophages (negative control)
[0017]
[Table 1]

[0018]
<Example 3>
A lotion was prepared according to the following formulation. That is, the prescription ingredients were stirred and solubilized at room temperature to obtain a lotion. This skin lotion was used twice a day in the morning and evening for 1 month, using 3 people from a group of panelists suffering from stains and freckles, and evaluated the effect of preventing and improving the stains and freckles. The evaluation criteria are score 2: marked improvement, score 1: obvious improvement, score 0.5: slight improvement, score 0: no improvement. The average score was 1.25. It was confirmed that the skin lotion containing maackiain, which has the effect of suppressing the elongation of the melanocyte dendriide of the present invention, is effective in improving spots and freckles.
Maackiain 1 part by weight 1,3 butanediol 5 parts by weight glycerin 3 parts by weight sodium citrate 0.1 part by weight methylparaben 0.2 part by weight ethanol 8 parts by weight water 82.7 parts by weight
<Example 4>
A pharmaceutical composition for external application to the skin was prepared according to the formulation shown below. That is, the prescription components were stirred and dispersed to obtain a skin external preparation. This product showed a remarkable effect on light-induced skin darkening or spots or freckles accompanied by light-induced inflammation.
Maackiain 0.5 parts by weight prednisolone 1 part by weight petrolatum 85 parts by weight [0020]
<Example 5>
A cream was prepared according to the formulation shown below. That is, a, b, and c were heated and dissolved at 80 ° C. respectively, and b was gradually added to emulsified and further emulsified by adding c, and then the emulsion particles were homogenized with a homomixer and cooled to obtain a cream. This cream had an excellent effect on blemishes and freckles with inflammation.
[0021]
I)
Squalane 10 parts by weight Cetanol 3 parts by weight Sorbitan sesquistearate 2 parts by weight Polyoxyethylene (20) behenyl ether 2 parts by weight Vitamin A acid 1 part by weight b)
1,3-butanediol 5 parts by weight maackiain 1 part by weight carboxyvinyl polymer 0.3 part by weight water 40 parts by weight c)
Water 37.3 parts by weight Potassium hydroxide 0.2 parts by weight
【The invention's effect】
ADVANTAGE OF THE INVENTION According to this invention, the effective prevention or improvement means can be provided with respect to pigment abnormalities accompanying inflammation, and pigment abnormalities, such as a spot and a buckwheat.

Claims (2)

A melanocyte dendrite elongation inhibitor comprising maackiain represented by the following chemical formula (II) and / or a salt thereof.

Chemical formula (II)   The dendritic elongation inhibitor of melanocytes according to claim 1, wherein the dendrite elongation inhibition is caused by the inhibitory action of a macrophage-derived dendrite elongation promoting factor.