JP2018043999A - コラーゲン加水分解物及びその使用 - Google Patents
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Abstract
Description
本発明による使用のためのコラーゲン加水分解物を生産するために、(乾燥材料の)20重量%と40重量%の間の濃度での豚皮ゼラチン水溶液(タイプA、200g〜250gのブルーム(Bloom))を出発材料として使用する。ゼラチンは、120分〜180分間、50℃〜60℃にて、微生物起源の2つの異なるエンドプロテアーゼの連続作用によって酵素的に加水分解され、ここで、第1の酵素として、枯草菌又はバチルス・アミロリケファシエンス由来のエンドプロテアーゼを用い、第2の酵素として、バチルス・リケニホルミス由来のエンドプロテアーゼを用いる。続いて、酵素は熱的に非活性化され、溶液を噴霧乾燥する。
静的相:TSK 2000 SW XL(Tosoh Bioscience GmbH)
移動相:0.4mol/lのリン酸二水素ナトリウム(pH5.3)
流速:0.5ml/分
較正標準:画定されたI型コラーゲン断片(FILK,Freiberg)
検出:UV検出器Knauer K−2501(214nm)
セルライトの治療及び/又は予防のために、実施例1に従って生産した、低分子量のコラーゲン加水分解物の有効性を二重盲検無作為化プラセボ対照試験において調べた。試験被験者は、35.3〜55.4歳の69人の健康な女性であり、各23人の被験者の3つのグループに分けた。68人の被験者が首尾よく試験を完了した。
毎日25gのコラーゲン加水分解物、3番目はプラセボ投与を受けた。経口摂取のために、加水分解物を水又は冷たい飲み物(牛乳を除く)に溶かすことができた。
−Cutometer(登録商標)SEM575による皮膚弾力性
(3回の測定からの平均値)
−DermaLab(登録商標)デバイスによる経皮水分喪失(TEWL)
(3回の測定からの平均値)
−Corneometer(登録商標)CM825による皮膚水分含量
(10回の測定からの平均値)
コラーゲン(I型)の合成、及びプロテオグリカンのバイグリカンとバーシカンの合成の刺激は、ヒト真皮線維芽細胞(皮膚細胞)をインビトロで調べた。この目的のために、細胞は、0.5mg/mlの低分子量又は高分子量の加水分解物のいずれかとともに24時間インキュベートされ、次に、コラーゲンRNA、バイグリカンRNA及びバーシカンRNAの発現は、リアルタイムPCRによって決定され、半定量的に評価された(加水分解物を含まない対照と比較した)。
コラーゲン加水分解物を用いた皮膚水分量の影響を直接、無毛マウスを用いて調べた。無毛マウスは、多くの場合、皮膚科学的調査のために使用される確立されたモデル系であり、それから得られた知識は、原理的にはヒト皮膚に適用することができる(例えば、T.Fujimura et al.;J.Dermatol.Sci.2000(24)105−111、Y.Nishimori et al.;J.Invest.Dermatol.2001(117)1458−1463参照)。
いわゆる「角化膜」タンパク質は、病原微生物及び毒性物質の侵入に対する皮膚のバリア機能に重要な役割を果たす。CEタンパク質であるインボルクリン、ロリクリン及びフィラグリンの合成は、予め、毎日150μgコラーゲン加水分解物/kg体重(上記の通り)で5週間、食餌を与えられた無毛マウスにおいて決定された。(BSAを与えた)対照群を基準としたタンパク質の定量は、皮膚からのタンパク質の抽出後、SDSポリアクリルアミドゲル電気泳動、及び特異的抗体を用いたウェスタンブロットにより行われた。
約2,000Daの平均分子量を有する実施例1に従って生産された低分子量のコラーゲン加水分解物(以下、加水分解物Aと呼ぶ)は、約2,100Da(以下、加水分解物Bと呼ぶ)及び約2,900Da(以下、加水分解物Cと呼ぶ)の平均分子量を有する2つの市販のコラーゲン加水分解物と比較した。
コラーゲン(I型)及びプロテオグリカンのデコリン及びバーシカンの合成の刺激をヒト真皮線維芽細胞(皮膚細胞)においてインビトロで調べた。この目的のために、それぞれ加水分解物A、B及びCの0.5mg/mlとともに細胞を24時間インキュベートし、次に、コラーゲンRNA、デコリンRNA及びバーシカンRNAの発現をリアルタイムPCRによって決定し、半定量的に評価した。デコリンは、皮膚中のコラーゲン繊維の形成に重要な役割を果たしている。
コラーゲン加水分解物の本発明による使用のためのいくつかの例示的レシピを以下に示すが、これらは、通常、多くの方法において修正され得る。
グリシン 53.67重量%
コラーゲン加水分解物 21.95重量%
ゼラチン 10.08重量%
グアーガム 6.00重量%
レシチン 5.00重量%
クエン酸 2.00重量%
香味料(カシス) 0.50重量%
オレンジ油 0.50重量%
アセサルフェームK 0.30重量%
ココア塊 51.0重量%
ショ糖 22.4重量%
ココアバター 16.6重量%
コラーゲン加水分解物 10.0重量%
水 63.00重量%
アロエベラ濃縮物 31.00重量%
コラーゲン加水分解物 4.00重量%
ショ糖 1.50重量%
クエン酸 0.26重量%
香味料及び着色剤 0.24重量%
スクラロース 0.0031重量%
Claims (17)
- セルライトを治療及び/又は予防するための有効成分としてのコラーゲン加水分解物を含む医薬組成物であって、該コラーゲン加水分解物の平均分子量は1,700Da〜2,300Daの範囲にあって、該コラーゲン加水分解物の分子量分布が、5重量%未満のコラーゲン加水分解物が7,500Da超であり、12〜18重量%が3,500〜7,500Daであり、25〜31重量%が1,500〜3,500Daであり、40〜46重量%が500〜1,500Daであり、及び5〜10重量%が500Da未満であり、該コラーゲン加水分解物が、ブタ皮膚のゼラチンの酵素的加水分解によって製造され、前記加水分解に使用される酵素が、微生物バチルス・サブチリス(Bacillus subtilis)、バチルス・リケニホルミス(Bacillus licheniformis)、バチルス・アミロリケファシエンス(Bacillus amyloliquefaciens)、アスペルギルス・オリザエ(Aspergillus oryzae)、及びアスペルギルス・メレウス(Aspergillus melleus)由来の酵素から選択されるメタロプロテアーゼ及び/又はセリンプロテアーゼである、上記医薬組成物。
- 少なくとも90重量%のコラーゲン加水分解物が3,500Da未満の分子量を有する、請求項1に記載の医薬組成物。
- 少なくとも45重量%のコラーゲン加水分解物が1,500Da未満の分子量を有する、請求項2に記載の医薬組成物。
- コラーゲン加水分解物が、600Da〜1,200Daの分子量を有する少なくとも4つの特徴的なペプチドを含む、請求項1〜3のいずれか1項に記載の医薬組成物。
- MALDI質量分析によって見出された分子量分布における少なくとも4つの特徴的なペプチドが、それらの周辺と比較して少なくとも2倍の強度を有する、請求項4に記載の医薬組成物。
- MALDI質量分析によって見出された分子量分布における少なくとも4つの特徴的なペプチドが、それらの周辺と比較して少なくとも4倍の強度を有する、請求項5に記載の医薬組成物。
- コラーゲン加水分解物が、620Da〜690Daのペプチド、790Da〜860Daのペプチド、980〜1,050Daのペプチド、及び1,175Da〜1,245Daのペプチドを含む、請求項1〜6のいずれか1項に記載の医薬組成物。
- コラーゲン加水分解物が、1,500Da〜3,500Daの分子量を有する特徴的なペプチドをさらに含む、請求項1〜7のいずれか1項に記載の医薬組成物。
- コラーゲン加水分解物が、12重量%以上のヒドロキシプロリン含量を有する、請求項1〜8のいずれか1項に記載の医薬組成物。
- コラーゲン加水分解物のN末端アミノ酸の少なくとも50%が、疎水性アミノ酸、特にアラニン、ロイシン及びイソロイシンである、請求項1〜9のいずれか1項に記載の医薬組成物。
- コラーゲン加水分解物が、腸内使用、特に経口投与のために与えられる、請求項1〜10のいずれか1項に記載の医薬組成物。
- コラーゲン加水分解物が、栄養補助食品であり、特に、錠剤、カプセル、糖衣丸薬、トローチ、サシェ、ゲル、又は溶液の形態で存在する、請求項11に記載の医薬組成物。
- 1日の摂取量として約1.5g〜5gのコラーゲン加水分解物が与えられる、請求項11又は12に記載の医薬組成物。
- 1日の摂取量として約2g〜3gコラーゲン加水分解物が与えられる、請求項13に記載の医薬組成物。
- 1日の摂取量として約2.3g〜2.7gのコラーゲン加水分解物が与えられる、請求項13に記載の医薬組成物。
- ビタミン、ミネラル、オメガ−3脂肪酸、オメガ−6脂肪酸、オメガ−9脂肪酸、ビオチン、ルテイン、リコピン、カフェイン、グルコサミン、コンドロイチン、ヒアルロナン、葉酸、アミノ酸、ユビキノン−10、スーパーオキシドジスムターゼ、及びローズヒップ、レモンバーベナ又は緑茶からの植物抽出物から選択される1つ以上のさらなる有効成分と組み合わせられる、請求項1〜15のいずれか1項に記載の医薬組成物。
- コラーゲン加水分解物が、50歳を超える女性、特に閉経後の女性に投与するために与えられる、請求項1〜16のいずれか1項に記載の医薬組成物。
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CN101518645A (zh) * | 2008-02-29 | 2009-09-02 | 李勇 | 海洋胶原肽在制备保护肾脏功能、延缓慢性肾衰进程药物、保健食品或食品中的用途 |
FR2930154B1 (fr) * | 2008-04-21 | 2012-04-20 | Oreal | Utilisation d'un extrait de baie, et plus particulierement de wolfberry, pour maintenir et/ou restaurer la tonicite et/ou la fermete de la peau |
CN102131492B (zh) * | 2008-08-27 | 2013-04-24 | 株式会社太平洋 | 含胶原肽的皮肤美容用组合物 |
DE102009030351A1 (de) * | 2009-06-22 | 2010-12-23 | Gelita Ag | Zusammensetzung zur Behandlung von degenerativen Gelenkerkrankungen |
WO2011011808A1 (en) * | 2009-07-30 | 2011-02-03 | Roman Buga | A cosmetic composition comprising sodium chloride in combination with one or more of protein, collagen, gelatin or amino acid |
DE102010060564A1 (de) * | 2010-11-15 | 2012-05-16 | Gelita Ag | Verwendung von Kollagenhydrolysat zur Verbesserung der Gesundheit der menschlichen Haut, Haare und/oder Nägel |
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2012
- 2012-11-21 RU RU2014126216A patent/RU2635995C2/ru active
- 2012-11-21 ES ES12794246.4T patent/ES2669318T3/es active Active
- 2012-11-21 PL PL12794246T patent/PL2785420T3/pl unknown
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- 2012-11-21 JP JP2014543839A patent/JP2015504440A/ja active Pending
- 2012-11-21 WO PCT/EP2012/073180 patent/WO2013079373A1/de active Application Filing
- 2012-11-21 CN CN201280058817.3A patent/CN104010697B/zh active Active
- 2012-11-21 BR BR112014011481-1A patent/BR112014011481B1/pt active IP Right Grant
- 2012-11-21 MX MX2014006420A patent/MX359181B/es active IP Right Grant
- 2012-11-21 EP EP12794246.4A patent/EP2785420B1/de active Active
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2014
- 2014-05-20 US US14/282,239 patent/US20140255485A1/en not_active Abandoned
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Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
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JP2005053798A (ja) * | 2003-08-04 | 2005-03-03 | Nonogawa Shoji Kk | フィブロネクチン生成促進剤 |
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US20140255485A1 (en) | 2014-09-11 |
CA2854856A1 (en) | 2013-06-06 |
MX2014006420A (es) | 2014-07-30 |
MX359181B (es) | 2018-09-17 |
JP2015504440A (ja) | 2015-02-12 |
ES2669318T3 (es) | 2018-05-24 |
WO2013079373A1 (de) | 2013-06-06 |
CA2854856C (en) | 2020-03-10 |
RU2014126216A (ru) | 2016-01-27 |
BR112014011481A2 (pt) | 2017-05-09 |
AU2012344135A1 (en) | 2014-06-26 |
EP2785420A1 (de) | 2014-10-08 |
RU2635995C2 (ru) | 2017-11-17 |
AU2012344135B2 (en) | 2017-01-05 |
BR112014011481B1 (pt) | 2019-02-05 |
CN104010697A (zh) | 2014-08-27 |
EP2785420B1 (de) | 2018-03-14 |
JP6462101B2 (ja) | 2019-01-30 |
CN104010697B (zh) | 2017-09-08 |
PL2785420T3 (pl) | 2018-08-31 |
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