JP7421695B2 - メラニン産生抑制剤及びこれを含有する美白剤又は皮膚外用剤 - Google Patents
メラニン産生抑制剤及びこれを含有する美白剤又は皮膚外用剤 Download PDFInfo
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- JP7421695B2 JP7421695B2 JP2019175742A JP2019175742A JP7421695B2 JP 7421695 B2 JP7421695 B2 JP 7421695B2 JP 2019175742 A JP2019175742 A JP 2019175742A JP 2019175742 A JP2019175742 A JP 2019175742A JP 7421695 B2 JP7421695 B2 JP 7421695B2
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- 239000011764 pyridoxine hydrochloride Substances 0.000 description 1
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- 239000011607 retinol Substances 0.000 description 1
- 150000004609 retinol derivatives Chemical class 0.000 description 1
- 235000020748 rosemary extract Nutrition 0.000 description 1
- 229940092258 rosemary extract Drugs 0.000 description 1
- 239000001233 rosmarinus officinalis l. extract Substances 0.000 description 1
- 229940109850 royal jelly Drugs 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 235000017709 saponins Nutrition 0.000 description 1
- 235000013580 sausages Nutrition 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 235000014102 seafood Nutrition 0.000 description 1
- 239000003352 sequestering agent Substances 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 229940045920 sodium pyrrolidone carboxylate Drugs 0.000 description 1
- HYRLWUFWDYFEES-UHFFFAOYSA-M sodium;2-oxopyrrolidine-1-carboxylate Chemical compound [Na+].[O-]C(=O)N1CCCC1=O HYRLWUFWDYFEES-UHFFFAOYSA-M 0.000 description 1
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Landscapes
- Cosmetics (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Description
1.スペルミジン、スペルミン、アセチルスペルミジン、ジアセチルスペルミジン、アセチルスペルミンから選択される1以上を有効成分として含有するメラニン産生抑制剤。
2.1.に記載のメラニン産生抑制剤を含有する美白剤。
3.1.に記載のメラニン産生抑制剤を含有する皮膚外用剤。
<メラニン産生抑制試験 >
B16メラノーマ細胞を用いて、メラニン産生抑制作用を確認した。
プトレシン二塩酸塩;(ナカライテスク社、商品コード29428-01)
スペルミジン三塩酸塩;(ナカライテスク社、商品コード32110-41)
スペルミン四塩酸塩;(ナカライテスク社、商品コード32113-11)
N8-アセチルスペルミジン二塩酸塩;(Sigma-Aldrich社、商品コードA3658)
N1,N8-ジアセチルスペルミジン;(Toronto Research Chemicals Inc、商品コードD367000)
N1-アセチルスペルミン三塩酸塩;(Sigma-Aldrich社、商品コード01467)
コウジ酸;(富士フイルム和光純薬社、商品コード118-00491)
合成メラニン;(Sigma-Aldrich社、商品コードM8631)
B16メラノーマ細胞を10%FBS(牛胎児血清)含有ダルベッコMEM培地を用いて培養した後、トリプシン処理により細胞を回収した。回収した細胞を10%FBSおよび1mmol/Lテオフィリン含有ダルベッコMEM培地で24.0×104cells/mLの濃度に希釈した後、48wellプレートに1well当たり300μLずつ播種し、6時間培養した。
培養終了後、10%FBSおよび1mmol/Lテオフィリン含有ダルベッコMEM培地で溶解した所定濃度の被験試料を各wellに300μLずつ添加し、4日間培養した。培養終了後、各wellから培地を取り除き、2mol/LのNaOH溶液200μLを添加して超音波破砕器により細胞を破壊し、波長475nmにおける吸光度を測定した。測定した吸光度の値から、合成メラニンを用いて作成した検量線をもとにメラニン量を算出した。また、細胞生存率の測定のため、同様に培養後、400μLのPBS(-)で洗浄し、終濃度0.05mg/mLで10%FBS含有ダルベッコMEMに溶解したニュートラルレッドを各wellに200μL添加した。2.5時間培養した後、ニュートラルレッド溶液を捨て、エタノール・酢酸溶液(エタノール:酢酸:水=50:1:49)を各wellに200μL添加し、色素を抽出した。抽出後、波長540nmにおける吸光度を測定した。空試験コントロールとして、10%FBSおよび1mmol/Lテオフィリン含有ダルベッコMEM培地のみで培養した細胞を同様の方法で試験した。試験は、各試料n=3で行った。
メラニン産生抑制率(%)=(1-(B/D)/(A/C))×100
A:被験試料無添加におけるメラニン量
B:被験試料添加におけるメラニン量
C:被験試料無添加での540nmにおける吸光度
D:被験試料添加での540nmにおける吸光度
なお、(D/C)×100(%)を細胞生存率とし、細胞生存率が80%未満の試料については、メラニン産生抑制率の算出をしていない。
スペルミジン、スペルミン、アセチルスペルミジン、ジアセチルスペルミジン、アセチルスペルミンはメラニン産生抑制作用が確認できた。プトレシンにはメラニン産生抑制作用は確認できなかった。
また、コウジ酸をポジティブコントロールとして同様に試験した結果、200μg/mLにおいて、44.7±1.6%のメラニン産生抑制作用を確認できた。
すなわち、スペルミジン、スペルミン、アセチルスペルミジン、アセチルスペルミンは、メラニン産生抑制作用が知られていたコウジ酸よりも、より低濃度で同等以上のメラニン産生抑制作用があることが確認できた。
(化粧水)
1.精製水 残余
2.グリセリン 5.0%
3.ジグリセリン 7.0%
4.1,3-ブチレングリコール 5.0%
5.ベタイン 1.0%
6.スペルミン 0.001%
7.フェノキシエタノール 0.3%
1~7を撹拌混合し、化粧水を得た。
1.精製水 残余
2.ジプロピレングリコール 6.0%
3.PEG-75 0.5%
4.ラフィノース 2.0%
5.フェノキシエタノール 0.2%
6.ペンチレングリコール 1.0%
7.キサンタンガム 0.1%
8.スクワラン 3.0%
9.SIMULGEL NS 2.5%
10.スペルミジン 0.0001%
11. エタノール 2.0%
6に7を分散させる(分散物)。8と9を混合する(混合物)。分散物を加熱溶解した1から5に徐々に添加し、さらに混合物を添加したのちホモミクサーで撹拌する。冷却後、10、11を加えた後、再度ホモミクサーで撹拌させる。
※SIMULGEL NS(SEPPIC社製;(アクリル酸ヒドロキシエチル/アクリロイルジメチルタウリンNa)コポリマー、スクワラン、ポリソルベート60、水、イソステアリン酸ソルビタンの混合物)
1.オリーブ油 5.0%
2.ホホバ油 1.0%
3.ジメチコン 0.5%
4.ステアリン酸ポリグリセリル-10 1.0%
5.グリセリン 7.0%
6.ジプロピレングリコール 6.0%
7.フェノキシエタノール 0.5%
8.1,3-ブチレングリコール 3.0%
9.キサンタンガム 1.0%
10.カルボキシビニルポリマー 0.1%
11.水酸化カリウム 0.02%
12.アセチルスペルミン 0.002%
13.精製水 残余
8に9を分散させる(分散物)。10を13に撹拌溶解させたものに、分散物を添加、さらに5から7を添加し加熱溶解させる(溶解物)。1から4を加熱溶解させ、溶解物を徐々に添加し、ホモミクサーで撹拌する。次いで、13の一部に11を溶解させたものを添加、冷却後、13の一部に溶解させた12を加えた後、再度ホモミクサーで撹拌させる。
1.精製水 残余
2.1,3-ブチレングリコール 5.0%
3.グリセリン 3.0%
4.ベタイン 1.5%
5.フェノキシエタノール 0.1%
6.カルボキシビニルポリマー 0.1%
7.水酸化カリウム 0.02%
8.ヒアルロン酸Na 0.001%
9.アセチルスペルミジン 0.0005%
6を1で攪拌溶解させたのち、2から5、8、9を添加する。溶解を確認した後、1の一部で溶解した7を添加し、再度ホモミクサーで撹拌させる。
Claims (2)
- アセチルスペルミジン、ジアセチルスペルミジン、アセチルスペルミンから選択される1以上を有効成分として含有するメラニン産生抑制剤(ただし、アセチルスペルミンとポリアミンオキシダーゼを含むものを除く)。
- 請求項1に記載のメラニン産生抑制剤を含有する美白剤。
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