JP4836961B2 - 核内転写因子ap−1の発現抑制剤、それを用いた医薬品および製品 - Google Patents
核内転写因子ap−1の発現抑制剤、それを用いた医薬品および製品 Download PDFInfo
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
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- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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Description
ローマカミツレ(Anthemis nobilis Linn.)の乾燥ハーブ頭状花1kgを、50%(v/v)エタノール20kgに浸漬し、1週間抽出処理を行った。抽出処理後、吸引ろ過を行い、12.63kgのローマカミツレ抽出液を得た。
カマメロサイドは以下の手順で精製した。前記ローマカミツレ抽出液を凍結乾燥した。この乾燥物(エキス固形分)10.31gを、pH4に調整した水(H2O)750mLに懸濁した後、クロロホルムを用いて分配を行った。そして、クロロホルム層を除去した後、さらに、水層に酢酸エチルを添加して分配を行い、2.80gの酢酸エチル層(酢酸エチル分配エキス)を得た。pH4の酢酸エチル分配エキスを、メタノール50mLで溶解し、その中にシリカゲルを添加した。これによって、前記シリカゲルに、酢酸エチル層に含まれる前記ローマカミツレエキスを吸着させた。そして、これを、加温槽上において、スパーテルにより撹拌した後、減圧濃縮することにより、完全に有機溶媒を除去した。得られた濃縮物を、シリカゲル含有濃縮画分という。
カラム:商品名COSMOSIL 5C18−MS
250×10mm I.D.
溶離液:アセトニトリル/水/TFA(体積比20/80/0.1)
流速:4.7mL/min
検出:UV at 220nm
カラム温度:40℃
試料注入量:50μL
分取時間:14〜16min
HeLa細胞をトリプシン処理して、4℃、1000rpm、3分間遠心分離を行った。回収したHela細胞を、2×105cells/wellの密度となるように30mmシャーレにまき、検定プレートとした。前記検定プレートを、12時間、37℃、5%CO2存在下におき、細胞を培養した後、形質転換を行った。前記形質転換は、形質転換用試薬(商品名Effectene Transfection Reagent、QIAGEN社)を用いて、2種類のベクターを前記細胞に導入した。前記ベクターは、商品名pAP−1(PMA)−SEAP(CLONTECH社)(0.4μg)、および、商品名pLuc−control(CLONTECH社)(0.02μg)を使用した。そして、前記両ベクターを導入した細胞を、16時間、37℃、5%CO2存在下で培養した後、培養細胞を前記検定プレートからトリプシンを用いて剥離した。剥離した細胞を、96well plateに、0.2×104cellずつ分注した。分注した細胞を2時間培養した後、終濃度(エキス固形分重量)が10μg/mL、5.0μg/mL、2.5μg/mL、1.0μg/mLとなるように、前記カマメロサイド溶液を添加し、さらに、TPA(12−O−tetradecanoylphorbol acetate)を、最終濃度4μg/mLとなるよう添加した。前記添加から24時間後、培地を25μLずつ回収した。そして、回収した培地について、含まれる核内転写因子AP−1活性に基づくsecreated alkaline phosphatase(SEAP)活性を、キット(商品名Great EscAPe SEAP kit、CLONTECK社)を用いて評価した。また。遺伝子発現の補正は、Luciferase活性測定キット(Promega社)により測定したLuciferase活性測定値を用いて行った。この結果を、図2のグラフに示す。なお、核内転写因子AP−1発現の抑制評価は、カマメロサイド添加濃度ごとのLuciferase活性を100%とした場合のsecreated alkaline phosphatase(SEAP)活性の相対値%(阻害率%)で表した。
ジャーマンカモミールの抽出液を調製し、この抽出液について、前述の条件でHPLC分析を行い、カマメロサイドの存在を確認した。
カラム:商品名COSMOSIL 5C18−AR−II
250×4.6mm I.D.
溶離液:アセトニトリル/水/TFA(体積比20/70/0.1)
流速:1.0mL/min
検出:UV at 340nm
カラム温度:40℃
試料注入量:20μL
本参考例は、実施例1で得られたローマカミツレ抽出液およびカマメロサイドを用いて、NF−κBの活性抑制を評価した例である。
Claims (5)
- カマメロサイドからなることを特徴とする、核内転写因子AP−1の発現抑制用医薬。
- 前記AP−1の発現抑制が、遺伝子レベルでの発現抑制およびタンパク質レベルでの発現抑制の少なくとも一方である、請求項1記載のAP−1の発現抑制用医薬。
- 前記カマメロサイドが、ローマカミツレ由来カマメロサイドおよびジャーマンカモミール由来カマメロサイドの少なくとも一方である、請求項1または2記載の核内転写因子AP−1の発現抑制用医薬。
- 核内転写因子AP−1が関与する疾患の予防もしくは治療のための医薬品であって、請求項1から3のいずれか一項に記載の核内転写因子AP−1の発現抑制用医薬を含み、前記疾患が、再狭窄症、非悪性過剰増殖性疾患、肺繊維症、ドライアイ症候群またはパーキンソン病である、医薬品。
- 前記非悪性過剰増殖性疾患が、子宮内膜過形成、良性前立腺肥大または増殖性硝子体網膜症である、請求項4記載の医薬品。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2007548640A JP4836961B2 (ja) | 2006-06-22 | 2007-06-19 | 核内転写因子ap−1の発現抑制剤、それを用いた医薬品および製品 |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
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JP2006172997 | 2006-06-22 | ||
JP2006172997 | 2006-06-22 | ||
PCT/JP2007/062340 WO2007148697A1 (ja) | 2006-06-22 | 2007-06-19 | 核内転写因子ap-1の発現抑制剤、それを用いた医薬品および製品 |
JP2007548640A JP4836961B2 (ja) | 2006-06-22 | 2007-06-19 | 核内転写因子ap−1の発現抑制剤、それを用いた医薬品および製品 |
Publications (2)
Publication Number | Publication Date |
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JPWO2007148697A1 JPWO2007148697A1 (ja) | 2009-11-19 |
JP4836961B2 true JP4836961B2 (ja) | 2011-12-14 |
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JP2007548640A Active JP4836961B2 (ja) | 2006-06-22 | 2007-06-19 | 核内転写因子ap−1の発現抑制剤、それを用いた医薬品および製品 |
Country Status (5)
Country | Link |
---|---|
US (1) | US20100240603A1 (ja) |
EP (1) | EP2033650A4 (ja) |
JP (1) | JP4836961B2 (ja) |
CN (1) | CN101472597B (ja) |
WO (1) | WO2007148697A1 (ja) |
Families Citing this family (6)
Publication number | Priority date | Publication date | Assignee | Title |
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JP2008231031A (ja) * | 2007-03-20 | 2008-10-02 | Arkray Inc | デヒドロエピアンドロステロン産生促進剤およびその用途 |
WO2009141541A2 (fr) * | 2008-04-22 | 2009-11-26 | Centre National De La Recherche Scientifique | Nouvelles compositions inhibant la melanogenese et leurs utilisations |
GB0808974D0 (en) * | 2008-05-16 | 2008-06-25 | Veritron Ltd | Plant extract and its therapeutic use |
ITCS20080019A1 (it) * | 2008-10-09 | 2010-04-10 | Univ Calabria | Molecola naturale estratta da un agrume, processo di estrazione e uso farmaceutico |
RU2641014C2 (ru) | 2013-03-14 | 2018-01-18 | Марс, Инкорпорейтед | Ароматизирующая композиция, содержащая глюкозиды hmg |
CN111773215A (zh) * | 2020-07-30 | 2020-10-16 | 曾辉 | 一种治疗aml的药物及应用 |
Citations (6)
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JP2003212770A (ja) * | 2002-01-22 | 2003-07-30 | Ichimaru Pharcos Co Ltd | メイラード反応阻害剤 |
US6890569B1 (en) * | 2002-10-18 | 2005-05-10 | Ann M. Tanke | Restorative tea system and associated method |
JP2005194246A (ja) * | 2004-01-09 | 2005-07-21 | Ichimaru Pharcos Co Ltd | NF−κB活性化抑制剤 |
WO2005070440A1 (en) * | 2004-01-24 | 2005-08-04 | Eman Gaber Haggag | Herbal formula for the treatment of allergic asthma ( chronic bronchial asthma) |
JP2005343873A (ja) * | 2004-06-07 | 2005-12-15 | Kao Corp | アロマターゼ活性化剤 |
JP2006016312A (ja) * | 2004-06-30 | 2006-01-19 | Morishita Jintan Kk | 脂肪代謝改善組成物 |
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US6683047B1 (en) * | 1990-10-10 | 2004-01-27 | The Burnham Institute | Method of inhibiting transcription utilizing nuclear receptors |
US6025388A (en) * | 1995-04-26 | 2000-02-15 | Allergan Sales, Inc. | Method for inhibiting gene expression promoted by AP1 protein with RARβ selective retinoids and method for treatment of diseases and conditions with such retinoids |
CA2343101A1 (en) * | 1998-09-11 | 2000-03-23 | Ajinomoto Co., Inc. | Benzene derivatives and medicinal use thereof |
US6287567B1 (en) * | 2000-01-07 | 2001-09-11 | Dorothy J. Blount | Method of making an herbal drink |
JP4060554B2 (ja) | 2001-08-17 | 2008-03-12 | 株式会社 伊藤園 | 遺伝子転写制御因子のdna結合能抑制剤 |
US7115563B2 (en) * | 2002-05-29 | 2006-10-03 | Insignion Holding Limited | Composition and its therapeutic use |
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2007
- 2007-06-19 JP JP2007548640A patent/JP4836961B2/ja active Active
- 2007-06-19 CN CN2007800230904A patent/CN101472597B/zh active Active
- 2007-06-19 EP EP07767198A patent/EP2033650A4/en not_active Withdrawn
- 2007-06-19 US US12/161,229 patent/US20100240603A1/en not_active Abandoned
- 2007-06-19 WO PCT/JP2007/062340 patent/WO2007148697A1/ja active Application Filing
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2003212770A (ja) * | 2002-01-22 | 2003-07-30 | Ichimaru Pharcos Co Ltd | メイラード反応阻害剤 |
US6890569B1 (en) * | 2002-10-18 | 2005-05-10 | Ann M. Tanke | Restorative tea system and associated method |
JP2005194246A (ja) * | 2004-01-09 | 2005-07-21 | Ichimaru Pharcos Co Ltd | NF−κB活性化抑制剤 |
WO2005070440A1 (en) * | 2004-01-24 | 2005-08-04 | Eman Gaber Haggag | Herbal formula for the treatment of allergic asthma ( chronic bronchial asthma) |
JP2005343873A (ja) * | 2004-06-07 | 2005-12-15 | Kao Corp | アロマターゼ活性化剤 |
JP2006016312A (ja) * | 2004-06-30 | 2006-01-19 | Morishita Jintan Kk | 脂肪代謝改善組成物 |
Also Published As
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---|---|
US20100240603A1 (en) | 2010-09-23 |
CN101472597A (zh) | 2009-07-01 |
JPWO2007148697A1 (ja) | 2009-11-19 |
CN101472597B (zh) | 2012-10-03 |
EP2033650A4 (en) | 2009-10-21 |
EP2033650A1 (en) | 2009-03-11 |
WO2007148697A1 (ja) | 2007-12-27 |
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