JP2020125254A - 化粧料、紫外線に対する細胞保護剤、光老化防止剤 - Google Patents
化粧料、紫外線に対する細胞保護剤、光老化防止剤 Download PDFInfo
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Abstract
Description
ヒトの身体は光老化に対する防御機能として損傷を元通りに修復するDNA修復機能のほか、IL-1αなどのインターロイキン類を産生しての炎症を促進する機能や、エンドセリンを放出しメラニン産生を亢進する機能、アポトーシスとよばれる細胞死を引き起こし、損傷した遺伝情報を次世代の細胞に引き継がれるのを防ぐ機構を有している。しかしながらそれでも十分ではなく、慢性的な紫外線の影響で光老化が進行し、皮膚のきめ細やかさが失われたり、しわやシミなどが生じてしまう。
[1] N(1)-アセチル-5-メトキシキヌラミンまたはその塩を含む、化粧料。
[2] 前記化粧料が水性化粧料であり、
そのpHが6.0以上である、[1]に記載の化粧料。
[3] N(1)-アセチル-5-メトキシキヌラミンまたはその塩を含む、紫外線に対する細胞保護剤。
[4] N(1)-アセチル-5-メトキシキヌラミンまたはその塩を含む、光老化防止剤。
本実施形態は化粧料に関し、N(1)-アセチル-5-メトキシキヌラミン(AMK)またはその塩を含有する。
AMKは以下の構造を有する化合物である。脳内においてはメラトニンが代謝されてN(1)-acetyl-N(2)-formyl-5-methoxykynuramine(AFMK)が生じるが、AMKはメラトニンの最終代謝産物である。
AMKまたはその塩としては、特に限定されず、天然由来のものであってもよく、また、合成品などの市販品を利用するようにしてもよい。例えばToronto Research Chemicals社製のものなどを挙げることができる。
例えば、本実施形態の化粧料は、医薬品、医薬部外品、化粧料等に使用されている任意の成分、例えば、脂肪酸セッケン、セチル硫酸ナトリウム等の陰イオン性界面活性剤、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、ポリオキシエチレン多価アルコール脂肪酸エステル、ポリオキシエチレン硬化ヒマシ油、多価アルコール脂肪酸エステル、ポリグリセリン脂肪酸エステル等の非イオン性界面活性剤、テトラアルキルアンモニウム塩等の陽イオン性界面活性剤、ベタイン型、スルホベタイン型、スルホアミノ酸型、N−ステアロイル−L−グルタミン酸ナトリウム等の両イオン性界面活性剤、レシチン、リゾフォスファチジルコリン等の天然系界面活性剤、フェノキシエタノール等の防腐剤、保湿剤、ポリマー類、油脂類、炭化水素類、高級脂肪酸、高級アルコール、紫外線吸収剤、アミノ酸誘導体、糖誘導体、香料、水、アルコール、増粘剤、タール系色素、酸化鉄等の着色顔料、金属イオン封鎖剤、酸化防止剤、薬剤等の成分を適宜配合することができる。
炭化水素類としては、流動パラフィン、スクワレン、スクワラン、マイクロクリスタリンワックス等が挙げられる。
高級アルコールとしては、例えばラウリルアルコール、ステアリルアルコール、セチルアルコール、セトステアリルアルコール等の直鎖アルコール、モノステアリルグリセリンエーテル、ラノリンアルコール、コレステロール、フィトステロール、オクチルドデカノール等の分枝鎖アルコール等が挙げられる。
ポリマー類としてはシリコーンなどが挙げられ、該シリコーンとしては、例えば、鎖状ポリシロキサンのジメチルポリシロキサン、メチルフェニルポリシロキサン等、環状ポリシロキサンのデカメチルシクロペンタシロキサン等が例示できる。
本実施形態の化粧料は適宜設定される含有量の原材料を混合するなど通常の方法に従って製造することができ、特に限定されない。例えば乳液等の場合、油相及び水相をそれぞれ加熱溶解したものを乳化分散して冷却する通常の方法により製造することができる。
水性化粧料である場合、他の形態の化粧料と比較して皮膚表面からの浸透性が高く、より効率的に紫外線による細胞の保護を高めることができる。また、このとき、pH6.0以上の水性化粧料とすることで、pH6.0未満である場合と比較してAMKまたはその塩の分解を抑えて紫外線に対する細胞保護効果をより高めることができる。
なお、pHの上限値については特に限定されないが、皮膚刺激の観点から例えばpH8.0以下が好ましい。
本実施形態において、化粧料のpHは公知の方法により調整することができ特に限定されず、例えば、公知のpH調整剤、緩衝剤等を用いてpHを調整することができる。pHの調整に用いられるpH調整剤、緩衝剤としては、化粧料等に通常使用されるpH調整剤、緩衝剤を挙げることができる。また、化粧料のpHは、pHメータを用いるなど周知の方法で測定することができる。化粧料のpHは、例えば室温(25℃)にて測定した値である。
また、本発明の一態様として、AMKまたはその塩を含む紫外線に対する細胞保護剤、AMKまたはその塩を含む光老化防止剤も提供することができる。該細胞保護剤および光老化防止剤は化粧料であってもよく、また、医薬部外品、医薬品等に分類されるものであってもよい。該細胞保護剤および光老化防止剤はその形態に応じて適宜設定できるが、例えば上記化粧料と同様の構成とすることができる。
以下のA液にB液を撹拌しながら加えて調製した。
<A液>
AMK・HCl 0.1重量%
グリセリン 3.00重量%
ソルビトール(70%水溶液) 2.00重量%
キサンタンガム 0.05重量%
ヒドロキシエチルセルロース 0.05重量%
アセチルヒドロキシプロリン(AHYP) 5.00重量%
アルギニン 5.00%
ヒアルロン酸Na(1%水溶液) 1.00重量%
クエン酸 0.05重量%
クエン酸Na 0.01重量%
PCA-Na(50%水溶液) 1.00重量%
水 100重量%とする量
<B液>
防腐剤 適量
BG 5.00重量%
水 10.00重量%
以下のA液にB液を撹拌しながら加えて調製した。
<A液>
AMK・HCl 0.3重量%
ヒドロキシプロリン 0.10重量%
BG 1.00重量%
DPG 1.00重量%
PEG-32 0.30重量%
PEG-150 0.20重量%
ペンチレングリコール 2.50重量%
クエン酸 0.03重量%
クエン酸Na 0.04重量%
EDTA-2Na 0.10重量%
防腐剤 適量
水 100.00重量%とする量
<B液>
脂溶性ビタミンC誘導体含有ナノエマルジョンベース 5.00重量%
水 10.00重量%
以下のC液に撹拌しながら組成物Bを添加し、さらにA液に撹拌しながら得られた混合物を加えて調製した。
<A液>
AMK・HCl 0.3重量%
キサンタンガム 0.40重量%
ヒドロキシエチルセルロース 0.40重量%
BG 3.00重量%
グリセリン 3.00重量%
ヒアルロン酸(1%水溶液) 5.00重量%
防腐剤 適量
水 100.00重量%とする量
<組成物B>
リン酸アスコルビルMg 3.00重量%
<C液>
クエン酸Na 0.50重量%
EDTA-4Na 0.10重量%
水 30.00重量%
以下の成分を攪拌しながら混合し、調製した。
AMK・HCl 0.1重量%
エタノール 1.0重量%
ソルビット(60%) 7.5重量%
グリセリン(85%) 7.5重量%
プロピレングリコール 1.5重量%
アルギン酸ナトリウム 0.4重量%
ポリソルベート80 1.5重量%
リン酸一水素ナトリウム 適量
リン酸二水素ナトリウム 適量
トラネキサム酸 0.05重量%
ステビアエキス 0.1重量%
ラウリルジメチルアミノ酪酸ベタイン 0.1重量%
香料 1.0重量%
水 100重量%とする量
以下のA液、B液を室温で溶解後、B液を攪拌しながらA液に加えて均一に調製した。
<A液>
AMK・HCl 0.3重量%
グリチルリチン酸ジカリウム 0.5重量%
PEG-60水添ヒマシ油 1.5重量%
エタノール 15.0重量%
防腐剤 適量
<B液>
PEG-8 1.0重量%
BG 5.0重量%
リン酸一水素ナトリウム 適量
リン酸二水素ナトリウム 適量
水 100重量%とする量
正常ヒト表皮角化細胞を用い、50%生存率となるUV-B照射条件(170 mJ)で照射試験を行い、細胞生存率測定、IL-1α量測定を行った。
細胞を、2×104 cells/0.1 mL/ウェルとなるよう96ウェルプレートに播種し、CO2インキュベーター(5%CO2、37℃、湿潤)で培養した。翌日、被験物質添加/非添加培地に置換し、6時間CO2インキュベーター(5%CO2、37℃、湿潤)で培養した後、UV-B照照射装置を用いてUV-B 約1500 μW/cm2(紫外線強度計を用いて強度測定)を、予備試験で決定した50%致死率を与える紫外線強度となるよう、114秒照射した(照射量170 mJ)。なお、別プレートにUV-B非照射条件を設定した。照射後18時間培養した後、wst-8法で生細胞数測定を行った。IL-1αELISA用に培養上清を回収した後、生細胞数測定試薬SFを10%添加した培地に交換し、CO2インキュベーター内(5%CO2、37℃)でインキュベートし、30分後および90分後に吸光度(450 nm、参照波長630 nm)をプレートリーダーで測定し、60分間の吸光度差から相対生細胞数を算出した。
培養上清中のIL-1α濃度をHuman IL-1 alpha/IL-1F1 Quantikine ELISA Kitを用いて測定した。培養上清はKit付属のCalibrator Diluent RD5-5で4倍希釈して測定に用いた。方法はkit付属のプロトコルに従い、スタンダードの吸光度から検量線を作成し、希釈液中IL-1α濃度を算出し、希釈倍率(4倍)を乗じて培養上清中IL-1α濃度を算出した。
UV-B照射により、細胞生存率がおよそ50%に低下した。被験物質処理群に関しては、MELは10 μMで効果が見られなかったのに対し、AMKは10 μMで効果が見られた(Dunnett’s testおよびStudent’s t test)。
なお、UV-B非照射条件では、被験物質による細胞生存率変化は見られなかった。
次にUV-B照射条件、培養上清中IL-1α濃度測定結果を図2に示す。
UV-B照射により、培養上清中IL-1α濃度がおよそ5倍に増加した。被験物質処理群に関しては、MELは10 μM処理条件において、vehicleコントロールと比較して、培養上清中IL-1α濃度の有意な減少がみられなかったがAMKは10 μMで有意差が見られた(Student’s t test)。
<溶液の調製>
10mgのAMK・HClを正確に秤量し、濃度が0.1%となるようにリン酸緩衝溶液(NaHPO4+NaH2PO4から調整)を加え全体で10.000gになるように調整した。
室温(21℃)、遮光インキュベーター内で各サンプルを3か月整置保存した。
カラム:waters XBridge C18 3.5um 4.6x150mm
ガードカラム:住化分析センター(SCAS)SUMIPAX Filter PG-ODS
温度:室温(21℃)
展開溶液A:0.1vol%HCOOH H2O 0.1%ギ酸水溶液
展開溶液B:0.1vol%HCOOH CH3CN 0.1%アセトニトリル
Flow rate :1.0ml/min
グラジェントプログラム(A:B):
0〜1.0min hold 95:5
1.0〜21.0min linear gradient 5:95
21.0〜30.0min hold 5:95
検出:254nm
装置:waters 600S HPLC system
検出器:日本分光 UV-970
溶液をpH6.0以上とすることで、AMK・HClの残存率を低下が大きく抑制されていることが理解できる。
Claims (4)
- N(1)-アセチル-5-メトキシキヌラミンまたはその塩を含む、化粧料。
- 前記化粧料が水性化粧料であり、
そのpHが6.0以上である、請求項1に記載の化粧料。 - N(1)-アセチル-5-メトキシキヌラミンまたはその塩を含む、紫外線に対する細胞保護剤。
- N(1)-アセチル-5-メトキシキヌラミンまたはその塩を含む、光老化防止剤。
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JPH0995409A (ja) * | 1995-10-02 | 1997-04-08 | Yamahatsu Sangyo Kk | 抗光酸化外用剤 |
JP2001335457A (ja) * | 2000-05-26 | 2001-12-04 | Yakult Honsha Co Ltd | 抗酸化剤およびこれを含有する皮膚外用剤 |
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