JP6921440B2 - アザミ抽出物を有効成分として含むメラニン生成促進用組成物 - Google Patents
アザミ抽出物を有効成分として含むメラニン生成促進用組成物 Download PDFInfo
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- JP6921440B2 JP6921440B2 JP2019552221A JP2019552221A JP6921440B2 JP 6921440 B2 JP6921440 B2 JP 6921440B2 JP 2019552221 A JP2019552221 A JP 2019552221A JP 2019552221 A JP2019552221 A JP 2019552221A JP 6921440 B2 JP6921440 B2 JP 6921440B2
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Description
タンの脂肪酸エステルが利用される。
て多様に処方される。本発明の薬学的組成物の経口投与量は、成人基準で0.001〜300mg/kg(体重)範囲内である。また、外用剤の場合、成人基準で1.0ないし3.0mlの量で1日1回ないし5回塗って1ヶ月以上継続するのが良い。ただし、上記投与量は、本発明の範囲を限定するものではない。
アザミの花を採取して異物および不純物が完全に除去されるようにきれいに洗浄した後、20ないし35℃で陰干しした後、粒子の大きさが1mm以下になるように粉砕した。その後、上記粉砕されたアザミ花粉末1kgを70%エタノール溶媒に浸した後、48時間超音波抽出して得た抽出液を濾過紙(Advantes, No.2)を用いて濾過した。上記濾過物を減圧濃縮してアザミ抽出物を製造した。
ヒトメラニン細胞をHMGS(Human Melanocytes Growth Supplement)が含まれたM254培地を用いて6−ウェルプレート(well plate)に1ウェル当たり1.5×105個で接種した後、細胞がウェルの底に約80%以上付着するまで5%CO2および37℃で培養した。培養後、培地を除去して実施例1の試料を適当濃度に希釈した培地に交換した後、5日間5%CO2および37℃で培養した。実施例1によるアザミ抽出物の濃度範囲は、細胞毒性がない1ppm、3ppm、5ppmに決定した。培地を除去した細胞をPBS(phosphated buffersalinee)で洗浄した後、細胞を回収した。回収された細胞は、血球計算盤を用いて細胞数を測定した後、10,000ないし13,000rpmで10分間遠心分離して上澄み液を除去し、ペレットを得た。得られた細胞ペレットを60℃で乾燥させた後、10%DMSOを含有した1M水酸化ナトリウム液100μlを入れて60℃の恒温槽で細胞内のメラニン溶液を得た。この溶液を用いてマイクロプレートリーダー(microplate reader)で450nmで吸光度を測定し、細胞の一定数当たりのメラニンの量を求めた。その結果を下表1と図1に示した。
B16メラノーマ細胞を10%のFBS(fetal bovine serum)を含有したDMEM培地で6−ウェルプレート(well plate)に1ウェル当たり1×105個で接種した後、細胞がウェルの底に約80%以上付着するまで5%CO2および37℃で培養した。培養後、培地を除去して実施例1の試料を適当濃度に希釈した培地に交換した後、3日間5%CO2および37℃で培養した。実施例1によるアザミ抽出物の濃度範囲は、細胞毒性がない10ppm、50ppm、100ppmに決定した。培地を除去した細胞をPBS(phosphated buffersalinee)で洗浄した後、細胞を回収した。回収された細胞は、血球計算盤を用いて細胞数を測定した後、10,000ないし13,000rpmで10分間遠心分離して上澄み液を除去し、ペレットを得た。得られた細胞ペレットを60℃で乾燥させた後、10%DMSOを含有した1M水酸化ナトリウム液100μlを入れて60℃の恒温槽で細胞内のメラニンを得た。この液を用いてマイクロプレートリーダー(microplate
reader)で450nmで吸光度を測定し、細胞の一定数当たりのメラニンの量を求めた。その結果を下表2と図2に示した。
3D skinを保存培地(maintenance medium)として12−ウェルプレート(well
plate)に5%CO2および37℃で培養した。培地を交換した後、アザミ抽出物を細胞毒性がない50ppm、100ppm濃度で処理した。10日間アザミ抽出物を濃度に合わせて処理した培地に3回交換した後、5%CO2および37℃で培養した。培地を除去した細胞をPBS(phosphated buffersalinee)で洗浄した後、細胞を回収した。10,000ないし13,000rpmで10分間遠心分離して上澄み液を除去し、ペレットを得た。得られた細胞ペレットを60℃で乾燥させた後、10%DMSOを含有した1M水酸化ナトリウム液100μlを入れて60℃の恒温槽で細胞内のメラニンを得た。この液を用いてマイクロプレートリーダー(microplate reader)で450nmで吸光度を測定し、細胞の一定数当たりのメラニンの量を求めた。その結果を下表3と図3に示した。
本発明によるアザミ抽出物が人体の皮膚に安全であることを確認するために、皮膚安全性検証実験を行った。このために、皮膚累積刺激試験を実施した。
平均反応度=[{(反応指数×反応度)/(総被験者数×最高点数(4点))}×100]/検査回数(9回)
反応度において±は1点、+は2点および++は4点の点数を与え、平均反応度が3未
満の場合、安全な組成物と判定される。
を示したので、本発明によるアザミ抽出物は明確な累積刺激の様相を現わさない人体の皮膚に安全な物質と判定された。
1−1.柔軟化粧水
下表5のとおりアザミ抽出物を有効成分に含む柔軟化粧水を通常の方法によって製造した。
下表6のとおりアザミ抽出物を有効成分に含む栄養化粧水を通常の方法によって製造した。
下表7のとおりアザミ抽出物を有効成分に含む栄養クリームを通常の方法によって製造した。
下表8のとおりアザミ抽出物を有効成分として含む栄養クリームを通常の方法によって製造した。
下表9のとおりアザミ抽出物を有効成分に含むパックを通常の方法によって製造した。
2−1.散剤の製造
Claims (6)
- アザミ(Cirsium japonicum)の花抽出物を有効成分として含むメラニン生成促進用組成物。
- 前記組成物が白斑、白髪または低色素症の予防または改善のための化粧料組成物である、請求項1に記載のメラニン生成促進用組成物。
- 前記アザミの花抽出物が、前記化粧料組成物の総重量に対して0.0001ないし15重量%含まれる、請求項2に記載のメラニン生成促進用組成物。
- 前記組成物が、溶液、懸濁液、乳濁液、ペースト、ゲル、クリーム、ローション、パウダー、石鹸、界面活性剤含有クレンジング、オイル、粉末ファンデーション、乳濁液ファンデーション、ワックスファンデーションおよびスプレーよりなる群から選ばれる剤形である、請求項2に記載のメラニン生成促進用組成物。
- 前記組成物が白斑、白髪または低色素症の予防または治療のための薬学的組成物である、請求項1に記載のメラニン生成促進用組成物。
- 前記アザミの花抽出物が、前記薬学的組成物の総重量に対して0.0001ないし70重量%含まれる、請求項5に記載のメラニン生成促進用組成物。
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JP5951273B2 (ja) * | 2012-02-08 | 2016-07-13 | 三省製薬株式会社 | 白髪防止用外用剤 |
KR101669359B1 (ko) | 2015-03-11 | 2016-10-26 | 경상대학교산학협력단 | 멜라닌 합성 촉진용 조성물 |
KR20170025375A (ko) * | 2015-08-28 | 2017-03-08 | 주식회사 엘지생활건강 | 피부 개선용 조성물 |
CN105434764A (zh) * | 2015-12-18 | 2016-03-30 | 高松武 | 一种治疗白癜风的中药胶囊 |
CN106421150A (zh) * | 2016-10-19 | 2017-02-22 | 罗锡山 | 广西白癜风壮药 |
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2017
- 2017-03-21 KR KR1020170035571A patent/KR101989407B1/ko active IP Right Grant
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2018
- 2018-03-12 CN CN201880033218.3A patent/CN110650724A/zh active Pending
- 2018-03-12 WO PCT/KR2018/002898 patent/WO2018174453A1/ko unknown
- 2018-03-12 JP JP2019552221A patent/JP6921440B2/ja active Active
- 2018-03-12 US US16/496,565 patent/US20210361559A1/en not_active Abandoned
- 2018-03-12 EP EP18770560.3A patent/EP3643295A4/en active Pending
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EP3643295A1 (en) | 2020-04-29 |
WO2018174453A1 (ko) | 2018-09-27 |
KR101989407B1 (ko) | 2019-06-14 |
JP2020511514A (ja) | 2020-04-16 |
EP3643295A4 (en) | 2020-12-30 |
US20210361559A1 (en) | 2021-11-25 |
KR20180106755A (ko) | 2018-10-01 |
CN110650724A (zh) | 2020-01-03 |
US20220287956A1 (en) | 2022-09-15 |
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