JP2020132546A - 細胞間接着低下抑制剤 - Google Patents
細胞間接着低下抑制剤 Download PDFInfo
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- JP2020132546A JP2020132546A JP2019025569A JP2019025569A JP2020132546A JP 2020132546 A JP2020132546 A JP 2020132546A JP 2019025569 A JP2019025569 A JP 2019025569A JP 2019025569 A JP2019025569 A JP 2019025569A JP 2020132546 A JP2020132546 A JP 2020132546A
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- 229910052623 talc Inorganic materials 0.000 description 1
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- DZKXJUASMGQEMA-UHFFFAOYSA-N tetradecyl tetradecanoate Chemical compound CCCCCCCCCCCCCCOC(=O)CCCCCCCCCCCCC DZKXJUASMGQEMA-UHFFFAOYSA-N 0.000 description 1
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- OULAJFUGPPVRBK-UHFFFAOYSA-N tetratriacontyl alcohol Natural products CCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCO OULAJFUGPPVRBK-UHFFFAOYSA-N 0.000 description 1
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- 229960000401 tranexamic acid Drugs 0.000 description 1
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- 150000003626 triacylglycerols Chemical class 0.000 description 1
- 239000001069 triethyl citrate Substances 0.000 description 1
- VMYFZRTXGLUXMZ-UHFFFAOYSA-N triethyl citrate Natural products CCOC(=O)C(O)(C(=O)OCC)C(=O)OCC VMYFZRTXGLUXMZ-UHFFFAOYSA-N 0.000 description 1
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- VLPFTAMPNXLGLX-UHFFFAOYSA-N trioctanoin Chemical compound CCCCCCCC(=O)OCC(OC(=O)CCCCCCC)COC(=O)CCCCCCC VLPFTAMPNXLGLX-UHFFFAOYSA-N 0.000 description 1
- LZTRCELOJRDYMQ-UHFFFAOYSA-N triphenylmethanol Chemical compound C=1C=CC=CC=1C(C=1C=CC=CC=1)(O)C1=CC=CC=C1 LZTRCELOJRDYMQ-UHFFFAOYSA-N 0.000 description 1
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Abstract
Description
本発明は、このような状況下為されたものであり、肌機能の低下の抑制作用を有する成分として、リンパ管細胞の細胞間接着低下を抑制する成分を提供することを課題とする。
すなわち、本発明の要旨は以下に関する。
[2] 細胞がリンパ管細胞である、[1]に記載の細胞間接着低下抑制剤。
[3] [1]または[2]に記載の細胞間接着低下抑制剤を含有する、むくみ改善用の皮膚外用剤。
[4] 化粧料または医薬部外品である、[3]に記載の皮膚外用剤。
本発明の一態様は、トウニンの抽出物からなる、細胞間接着低下抑制剤(以下、「本発明の細胞間接着低下抑制剤」と称することがある)に関する。
上記のとおり、トウニンは漢方生薬として用いられてきたが、細胞間接着低下抑制作用については知られておらず、本発明者等によりその作用が知見され、本発明がなされたものである。
本発明の細胞間接着低下抑制剤が対象とする細胞としては、リンパ管細胞であり、例えば、皮膚リンパ管内皮細胞等が挙げられる。
本発明の細胞間接着低下抑制剤として用いられるトウニンの抽出物の原料としては、基源植物であるモモ(Prunus persica Batsch)自体、これを乾燥、粉砕、細切等加工した加工物等を用いることができる。植物の使用できる部位は特段の限定はないが、漢方生薬の使用部位に準じた部位を使用するのが特に好ましく、種子を特に好ましく用いることができる。起源植物としては、自生若しくは生育された植物、漢方生薬原料等として販売されるもの等を限定されず、用いることができる。
抽出溶媒としては、水、メタノール、エタノール、イソプロピルアルコール、ブタノール等のアルコール類、1,3-ブタンジオール、ポリプロピレングリコール等の多価アルコール類、アセトン、メチルエチルケトン等のケトン類、ジエチルエーテル、テトラヒドロフラン等のエーテル類等の極性溶媒から選択される1種または2種以上が好適なものとして例示することができる。好ましくは、水およびエタノールが挙げられる。
植物由来の極性溶媒による抽出物をろ過後、カラムクロマトグラフィー等にて分画し、有効成分濃度を高め、より効果の高い画分を選択してもよい。
さらなる本発明の一態様は、本発明の細胞間接着低下抑制剤を含有する、むくみ改善用の皮膚外用剤(以下、「本発明の皮膚外用剤」と称することがある)に関する。
さらなる本発明の一態様は、化粧料または医薬部外品である、本発明の皮膚外用剤(以下、「本発明の化粧料または医薬部外品」と称することがある)に関する。
本発明の皮膚外用剤、化粧料または医薬部外品は、有効成分として、本発明の細胞間接着低下抑制剤を含有する。
なお、後記実施例に示されるとおり、リンパ管内皮細胞はコルチゾールにより、細胞間接着が弱くなることが明らかとなった。リンパ管内皮細胞の細胞間接着が弱まることで、肌機能の低下につながると考えられている。
ここで、本発明の皮膚外用剤は、リンパ管細胞の細胞間接着低下を抑制する機能を有しており、限定されないが、例えばストレス等によるコルチゾールの亢進による細胞間接着低下に起因する肌機能低下を改善する用途に用いることができる。コルチゾールの亢進による肌機能低下による事象としては、限定されないが、例えば、むくみ等が挙げられる。
本発明の皮膚外用剤の使用対象としては、肌機能の低下の抑制が望まれる対象であれば限定されず、例えば哺乳類、具体的にはヒトが挙げられる。対象の年齢、性別等も限定されない。
投与量としては、特に限定されないが、所望の効果と安全性とを考慮して、例えば経皮投与する場合は、有効成分である細胞間接着低下抑制剤の固形物換算で0.0002〜0.05 mg/日が好ましい。
また、本発明の細胞間接着低下抑制剤の効果を妨げない限り、本発明の細胞間接着低下抑制剤以外の有効成分を含有してもよい。有効成分としては、限定されないが、美白成分、シワ改善成分、抗炎症成分、動植物由来の抽出物等が挙げられる。
コルビン酸、トラネキサム酸、アルブチン、1−トリフェニルメチルピペリジン、1−トリフェニルメチルピロリジン、2−(トリフェニルメチルオキシ)エタノール、2−(トリフェニルメチルアミノ)エタノール、2−(トリフェニルメチルオキシ)エチルアミン、トリフェニルメチルアミン、トリフェニルメタノール、トリフェニルメタンおよびアミノジフェニルメタン、N−(p−トルイル)システイン酸、N−(p−メトキシベンゾイル)システイン酸等が挙げられる。更にその他の美白成分として、N−ベンゾイル−セリン、N−(p−メチルベンゾイル)セリン、N−(p−エチルベンゾイル)セリン、N−(p−メトキシベンゾイル)セリン、N−(p−フルオロベンゾイル)セリン、N−(p−トリフルオロメチルベンゾイル)セリン、N−(2−ナフトイル)セリン、N−(4−フェニルベンゾイル)セリン、N−(p−メチルベンゾイル)セリン メチルエステル、N−(p−メチルベンゾイル)セリン エチルエステル、N−(2−ナフトイル)セリン メチルエステル、N−ベンゾイル−O−メチルセリン、N−(p−メチルベンゾイル)−O−メチルセリン、N−(p−メチルベンゾイル)−O−アセチルセリン、N−(2−ナフトイル)−O−メチルセリン等があげられる。
、アボガドエキス、アマチャエキス、アーモンドエキス、アルニカエキス、アロエエキス、アロニアエキス、アンズエキス、イチョウエキス、インドキノエキス、ウイキョウエキス、ウドエキス、エイジツエキス、エゾウコギエキス、エンメイソウエキス、オウゴンエキス、オウレンエキス、オタネニンジンエキス、オトギリソウエキス、オドリコソウエキス、オレンジエキス、カキョクエキス、カッコンエキス、カモミラエキス、カロットエキス、カワラヨモギエキス、カンゾウエキス、キウイエキス、キューカンバーエキス、グアバエキス、クジンエキス、クマザサエキス、クララエキス、クルミエキス、グレープフルーツエキス、黒米エキス、クロレラエキス、クワエキス、ケイケットウエキス、ゲットウヨウエキス、ゲンチアナエキス、ゲンノショウコエキス、紅茶エキス、ゴボウエキス、コメエキス、コメ発酵エキス、コメヌカ発酵エキス、コメ胚芽油、コケモモエキス、サルビアエキス、サボンソウエキス、ササエキス、サンザシエキス、サンシャエキス、サンショウエキス、シイタケエキス、ジオウエキス、シソエキス、シナノキエキス、シモツケソウエキス、シャクヤクエキス、ショウキョウエキス、ショウブ根エキス、シラカバエキス、スギナエキス、ステビアエキス、ステビア発酵物、セイヨウキズタエキス、セイヨウサンザシエキス、セイヨウニワトコエキス、セイヨウノコギリソウエキス、セイヨウハッカエキス、セージエキス、ゼニアオイエキス、センキュウエキス、センブリエキス、ソウハクヒエキス、ダイオウエキス、ダイズエキス、タイソウエキス、タイムエキス、タンポポエキス、チョウジエキス、チンピエキス、甜茶エキス、トウガラシエキス、トウキエキス、トウキンセンカエキス、トウヒエキス、ドクダミエキス、トマトエキス、納豆エキス、ニンジンエキス、ニンニクエキス、ノバラエキス、ハイビスカスエキス、バクモンドウエキス、ハスエキス、ハス胚芽エキス、パセリエキス、バーチエキス、ハマメリスエキス、ヒキオコシエキス、ヒノキエキス、フキタンポポエキス、フキノトウエキス、ブクリョウエキス、ブッチャーブルームエキス、ブドウエキス、ブドウ種子エキス、ヘチマエキス、ベニバナエキス、ペパーミントエキス、ボダイジュエキス、ボタンエキス、ホップエキス、マツエキス、マロニエエキス、ミズバショウエキス、ムクロジエキス、メリッサエキス、モズクエキス、モモエキス、ヤグルマギクエキス、ユーカリエキス、ユキノシタエキス、ユズエキス、ユリエキス、ヨクイニンエキス、ヨモギエキス、ラベンダーエキス、リンゴエキス、ルイボス茶エキス、レイシエキス、レタスエキス、レモンエキス、レンギョウエキス、レンゲソウエキス、ローズエキス、ローズマリーエキス、ローマカミツレエキス、ローヤルゼリーエキス、ワレモコウエキス等のエキスが好ましいものとして挙げられる。化粧料中における動植物由来抽出物の含有量は、通常0.0001〜30質量%であり、0.001〜10質量%が好ましく、0.01〜5質量%がより好ましい。
化粧料中における抗炎症成分の含有量は、通常0.01〜30質量%であり、0.1〜10質量%が好ましく、1〜5質量%がより好ましい。
極性油としては、合成エステル油として、ミリスチン酸イソプロピル、オクタン酸セチル、ミリスチン酸オクチルドデシル、パルミチン酸イソプロピル、ステアリン酸ブチル、ラウリン酸ヘキシル、ミリスチン酸ミリスチル、オレイン酸デシル、オレイン酸オクチルドデシル、ジメチルオクタン酸ヘキシルデシル、乳酸セチル、乳酸ミリスチル、酢酸ラノリン、ステアリン酸イソセチル、イソステアリン酸イソセチル、12−ヒドロキシステアリル酸コレステリル、ジ−2−エチルヘキシル酸エチレングリコール、ジペンタエリスリトール脂肪酸エステル、モノイソステアリン酸N−アルキルグリコール、ジカプリン酸ネオペンチルグリコール、リンゴ酸ジイソステアリル、ジ−2−ヘプチルウンデカン酸グリセリン、トリ−2−エチルヘキシル酸トリメチロールプロパン、トリイソステアリン酸ト
リメチロールプロパン、テトラ−2−エチルヘキシル酸ペンタンエリスリトール、トリ−2−エチルヘキシル酸グリセリン、トリイソステアリン酸トリメチロールプロパンを挙げることができる。
OP2−デシルテトラデシルエーテル等)、テトロニック類、POEヒマシ油・硬化ヒマシ油誘導体(POEヒマシ油、POE硬化ヒマシ油等)、ショ糖脂肪酸エステル、アルキルグルコシド等の非イオン界面活性剤類、等が挙げられる。
<評価サンプル>
トウニンエキス:トウニン抽出液‐J(丸善製薬株式会社)
<細胞>
細胞名:皮膚リンパ管微小血管内皮細胞(HMVEC-dLyAd、ロンザ)
<培地>
培地名:EGM(登録商標)-2MV(ロンザ)
<試薬類>
・PBS
・0.05 % トリプシン‐EDTA
・4 % パラホルムアルデヒド
・0.5 % TritonX-100/PBS
・10 % ブロックエース
・DAPI Fluoromount-G
<抗体>
タイトジャンクション関連タンパク質であるZO-1に対する以下の抗体を用いて免疫組織染色を行った。
一次抗体:Rabbit anti ZO-1(Invitrogen, 61-7300)(10 % ブロックエースで200倍希釈)
二次抗体:Goat anti-Rabbit IgG Cross-Adsorbed Secondary Antibody, Alexa 488(Life tec., A11008)(10 % ブロックエースで250倍希釈)
試験は、以下の方法で行った。
[方法]
・凍結HMVECを75 cm2フラスコ3本に播種し(2.5×105 cells/本)、翌日培地交換を実施
・3日後、細胞を回収し4 wellスライドチャンバーへ播種(7.0×104 cells/well)
・2日後、Cortisol含有培地(0, 0.5, 1, 5 μg/ml)を添加し (1 ml/well)、5日あるいは7日培養(37℃)
・PBS(-) で洗浄、0.5 % TritonX-100/PBSを添加し、静置(10min)
・10%ブロックエースを添加し、静置(30 min)、一次抗体を添加し4 ℃で一晩静置
・PBS(-) で洗浄し(5 min×3)、二次抗体を添加し、静置(rt, 30 min)
・PBS(-) で洗浄後(5 min×4)、Milli-Q(登録商標)水で洗浄(2回)
・DAPI Fluoromount(登録商標)-Gで封入して顕微鏡観察
観察結果を図1に示す。5 μg/mlのCortisol添加群で、緑色の蛍光が減少したことから、リンパ管細胞にCortisolを添加すると、細胞間接着が弱くなることが明らかとなった。
皮膚リンパ管微小血管内皮細胞にトウニンエキスを添加した際の細胞間接着関連タンパク質ZO-1の発現量変化により、トウニンエキスのリンパ管細胞の細胞間接着への影響を試験した。
試験は、以下の方法で行った。
<細胞起眠〜回収>
・凍結HMVECを75cm2フラスコ3本に播種し(1.9×105 cells/本)、翌日、培地交換を実施した。
・5日後、細胞を回収し4 wellスライドチャンバー3枚へ播種した(6.0×104 cells/well)。
・2日後、トウニンエキス添加培地(トウニンエキス0.05 %、0.1 %、0.125 %の3濃度)への交換を実施した。
・トウニンエキス添加後、37℃で4日培養後回収した(n=3)。
<固定>
・PBSで洗浄
・4 %パラホルムアルデヒドで固定(10分)
・PBSで洗浄
<免疫染色>
・0.5 % TritonX-100/PBSを添加し、静置(10分)
・10 %ブロックエースを添加し、静置(30分)
・一次抗体を添加し4 ℃一晩静置
・PBSで洗浄(5分×3回)
・二次抗体を添加し、静置(30分)
・PBSで洗浄(5分×4回)
・脱イオン蒸留水で洗浄(2回)
・DAPI Fluoromount(登録商標)-Gで封入
・乾燥
・顕微鏡で観察
トウニンエキス添加4日後の結果を図2に示す。トウニンエキスを添加した細胞の方が、無添加(0%)よりも細胞間の染色強度が強く、トウニンエキスは、リンパ管細胞の接着を強化することが明らかとなった。
Claims (4)
- トウニン(Prunus persica Batsch)の抽出物からなる、細胞間接着低下抑制剤。
- 細胞がリンパ管細胞である、請求項1に記載の細胞間接着低下抑制剤。
- 請求項1または2に記載の細胞間接着低下抑制剤を含有する、むくみ改善用の皮膚外用剤。
- 化粧料または医薬部外品である、請求項3に記載の皮膚外用剤。
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH11335235A (ja) * | 1998-05-22 | 1999-12-07 | Shiseido Co Ltd | 抗老化剤 |
JP2006056845A (ja) * | 2004-08-23 | 2006-03-02 | Teikoku Seiyaku Co Ltd | バラ科植物抽出物を含有する貼布剤 |
JP2009256244A (ja) * | 2008-04-17 | 2009-11-05 | Maruzen Pharmaceut Co Ltd | クローディン産生促進剤、及びオクルディン産生促進剤、並びに、皮膚バリア機能改善剤 |
CN105816406A (zh) * | 2016-04-07 | 2016-08-03 | 景东旭 | 营养美容液 |
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- 2019-02-15 JP JP2019025569A patent/JP7391517B2/ja active Active
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JPH11335235A (ja) * | 1998-05-22 | 1999-12-07 | Shiseido Co Ltd | 抗老化剤 |
JP2006056845A (ja) * | 2004-08-23 | 2006-03-02 | Teikoku Seiyaku Co Ltd | バラ科植物抽出物を含有する貼布剤 |
JP2009256244A (ja) * | 2008-04-17 | 2009-11-05 | Maruzen Pharmaceut Co Ltd | クローディン産生促進剤、及びオクルディン産生促進剤、並びに、皮膚バリア機能改善剤 |
CN105816406A (zh) * | 2016-04-07 | 2016-08-03 | 景东旭 | 营养美容液 |
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Title |
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粧技誌, 2012, VOL.46 NO.3, P.188-196, JPN6022050140, ISSN: 0005021289 * |
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