JP2006206611A - 微小管安定化剤を用いたアテローム性動脈硬化症または再狭窄症の治療方法 - Google Patents
微小管安定化剤を用いたアテローム性動脈硬化症または再狭窄症の治療方法 Download PDFInfo
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Abstract
【解決手段】特に、本発明は、タキソール(登録商標)または水溶性タキソール誘導体のような微小管安定化剤を低投与量用いて治療する動脈損傷後のアテローム性動脈硬化症または再狭窄症を予防または軽減する方法である。本発明で用いる低投与量は、医薬の好ましくない副作用を最小にしながら、動脈閉鎖を防止する。
【選択図】なし
Description
更に詳しくは、本発明はアテローム性動脈硬化症または再狭窄症の進行を防ぎ、または減ずるために低用量のタキソール(登録商標)溶液でこれらの患者を治療することに関する。
本発明のこれら及び他の目的に従って、タキソールたは水溶性タキソール誘導体のような微小管安定化剤の治療的に有効な量で治療することからなる、アテローム性動脈硬化症または再狭窄症を予防または軽減する方法を提供する。薬剤の治療的に有効な量とは、アテローム性動脈硬化症または再狭窄症の進行を予防または軽減するために十分な量である。
本発明の具体例の実施は、腹腔内または皮下注射、持続的な静脈内注入、経口的な摂取または局所的な(直接)投与、あるいはこれら2種以上の組み合せのような、数種の選択的な薬剤輸送経路を介して達成することができる。注射または持続注入のための溶液を処方する場合、最初にタキソール溶液を調製しなければならない。タキソールは、ポリオキシエチレンヒマシ油(CremophorEL)50%及び脱水アルコール、USP(50%)の担体中6mg/mlの濃縮溶液として、5mlのバイアル(30mg/バイアル)で、CTEP、DCT、NCI(IND#22850)を通じて供給されている。そのままのバイアルで冷凍保存し、使用に先立って希釈すべきである。5%のブドウ糖注射液あるいは0.9%の塩化ナトリウムで希釈する場合、0.3−1.2mg/mlのタキソール濃度のものが、室温で少なくとも12時間物理的及び化学的に安定である。(NCI研究薬;製薬学的データ(NCI Investigation Drugs;Pharmaceutical Data)(1990))。ポリオレフィン容器で調製した、D5WまたはNS希釈のタキソール濃度0.6mg/mlのもの、及びNS希釈の1.2mg/mlのものは、環境温度(20−23℃)で少なくとも25時間安定であることも報告された。(Waugh,et al.(1990)Am.J.Hosp.Pharm.48,1520)。これらの濃度で上記の時間の安定性が示されたが、これはタキソールのいずれの濃度を用いても良い本発明の実施を制限することを意味するものではない。
種々のタキソール濃度で前処理した培養VSMCの、再構成基底膜タンパク質で被覆されたフィルターに侵入するインビトロの能力を試験して、タキソール誘導された微小管束状化(bundling)がインビボでの新生動脈内膜形成に必要な細胞プロセスを損傷するかを評価した。
微小管の安定化と超重合がVSMC侵入のタキソール阻害に関与する決定的かつ十分な要因であることを確認するために、酸化デューテリウム(重水、2H2O)を用いて化学侵入(ボイデンチャンバー)アッセイを行った。酸化デューテリウムはタキソールとは異なるメカニズムで微小管/チューブリン重合を増強する。増強されたチューブリンの疎水性相互作用によってαβ−チューブリンヘテロダイマーの重合に決定的な濃度を減少すること(Itoh,T.J.,et al.,Biochim.Biophys.Acta.,800:21−27,1984)と、非重合チューブリン集団を重合形に変換すること(Takahashi,T.C.,et al.,Cell Struct.Funct.,9:45−52,1984)の両方によって、酸化デューテリウムのアイソトープと溶媒効果の組み合わせが、微小管の重合を可逆的に増加する。
細胞補充(recruitment)と移動に加えて、PDGFやbFGFなどの動脈損傷後に精巧に産生される各種の成長制御分子も分裂促進と細胞増殖に関係している。VSMC DNA合成に対するタキソールの効果を測定するために、[3H]チミジン取り込みを測定した。VSMCを24ウエルプレートに4.5x104でプレートした。10%FCS+DMEM中で5時間インキュベーとした後に、0.5mCi[3H]チミジンを添加し、さらに16時間インキュベーションを続けた。細胞をリン酸緩衝塩液で2回洗浄し、氷上で2時間10%TCAを用いて抽出し、次に2,000gで10分遠心した。上清を捨てて、ペレットを1N NaOH 0.5ml中に溶解した。1N HCl 0.5mlで中和した後、[3H]チミジンの取り込みをベックマン液体シンチレーションカウンターで測定した。チミジン添加18時間前とチミジン取り込み時の両方で、VSMCを種々の濃度のタキソールで処理した。これらの実験の各条件を3回実施した。
VSMC DNA合成に及ぼす酸化デューテリウムの効果を決定するために、チミジン類似体であるブロモデオキシウリジン(BrDU)の取り込みを測定した。VSMCを24ウエルプレートに4.5x104でプレートした。種々の2H2O濃度で10%FCS+DMEM中、20時間インキュベートした後、10μM BrDUを加えてさらに4時間インキュベーションを続けた。細胞をリン酸緩衝塩液(PBS)で2回洗浄し、100%メタノール(−20℃)で10分固定した。細胞を1N HClで2時間インキュベートしてDNAを変性し、次いでPBSで4回洗浄した。2%BSA−PBS中のマウスモノクローナルBrDU抗体(Boehringer Mannheim)を細胞と一緒に1時間インキュベートした。PBSで洗浄後、アルカリホスファターゼと結合させたヤギ抗−マウス抗体を加えた。チミジンに代えたBrDUを含む細胞核はアルカリホスファターゼ基質で赤色に染色され、その他のすべての核は青色に染色される。BrDU−ポジティブな核の割合を、対照(100%として定義)と酸化デューテリウム処理群との間で比較した。
加齢動物保護および使用委員会に関する国立研究所(National Institute on Aging Animal Care and Use Committee)で認められたプロトコルに従い、GRCコロニーから得た6カ月齢のウイスターラットを20mg/kg体重ペンタバルビタール、2mg/kg体重ケタミン、および4mg/kg体重キシラジンで腹腔内麻酔した。左外側頸動脈に2−フレンチフォガーティ(2−French Fogarty)塞栓切除術カテーテルでカニューレ挿入し、食塩水で膨張させ、頸動脈を3回上下して通過させて膨張させた脱内皮化(deendothelializing)損傷を作出した。動物を2mg/kg体重のタキソール溶液で処理し、対照動物は賦形剤のみ(1日当たり13.4ml/kg体重の1:2:2:165のDMSO:クレモフォールEL:脱水エタノール:リン酸緩衝塩液)を損傷2時間後から始めて腹腔内注射した。タキソール溶液または賦形剤のみは次の4日間腹腔内注射として投与した。11日後に動物(タキソール処理8匹と賦形剤処理10匹)を上述のように麻酔し、頸動脈を単離して10%緩衝化ホルマリンで固定し、パラフィン中に保持した。頸動脈の断面切片を顕微鏡スライドに乗せてヘマトキシリンとエオシンで染色した。頸動脈の画像をディジタイジングボードに投影して動脈内膜(intima)と中膜(media)の断面席を測定した。結果を図2A〜2Cに示す。先行技術(Ferns,G.A.A.et al.,Science,253:1129−1132,1991)に示すように、再狭窄症のラット頸動脈損傷モデルはヒト再狭窄症研究に有用であり、ヒトの治療効果の可能性を示唆する。
は、ヒトの癌治療に元々用いられた投与量(3−6mg/kg)よりも有意に低く、前処理療法と最適の治療期間とを組み合わせて劇的に低い全身投与量で同等またはそれ以上の効果が得られる。さらに、治療の最終目的は「活性化」VSMCを抑制すること、あるいは好ましくは成長と移動のための刺激が分散するまでの間、(細胞死をもたらす細胞毒性を引き起こすことなく)まず活性化を防止することにあるのであるから、毒性の少ない短期間治療という最終目的がヒトについてもありうる。最終的には、局所的徐放性送達システムが血管形成術後の再狭窄症を防ぐ最良の解決策を与えるものであり、これによって送達医薬の局所的な高濃度が得られ全身的毒性の問題を本質的に排除できるであろう。医薬を含浸したポリマーで被覆した金属ステント、生分解性の医薬溶出性ポリマーステント、および金属ステントを被覆するための遺伝的にプライムされた内皮細胞などを含む医薬送達システムが有用であり、あるいは局所的に内皮細胞を被覆するものとして送達される(Muller,D.W.M.et al.,JACC 17:126b−131b,1991)。これらのシステムは全身性の副作用なしに化学治療剤を安全に使用することを可能にする。あるいは、治療には治療前(すなわち血管手術前)に一定期間持続静脈注入し、次いで別の治療時期(局所的、直接送達)または手術後(経口、注射)に行うことも含むことができる。
Claims (23)
- 酸化デューテリウム、パクリタキセルまたは水溶性パクリタキセル誘導体を含む、患者の再狭窄症を予防しまたは軽減するための医薬組成物。
- 前記水溶性パクリタキセル誘導体が、2’−サクシニル−パクリタキセル;2’−サクシニル−パクリタキセルトリエタノールアミン;2’−グルタリル−パクリタキセル;2’−グルタリル−パクリタキセルトリエタノールアミン塩;N−(ジメチルアミノエチル)グルタミドとの2’−O−エステル;そしてN−(ジメチルアミノエチル)グルタミド塩酸塩との2’−O−エステルからなる群から選択される、請求項1の医薬組成物。
- 前記予防しまたは軽減することが前記医薬組成物の全身性送達により達成される、請求項1の医薬組成物。
- 前記予防しまたは軽減することが前記医薬組成物の局所送達により達成される、請求項1の医薬組成物。
- 前記局所送達が局所的持続性放出送達システムを含む、請求項4の医薬組成物。
- 前記局所的持続性放出送達がステントを含む、請求項5の医薬組成物。
- 前記ステントが、ポリマーで被覆した金属ステント、生分解性の薬物溶出ポリマーステント、および内皮細胞で被覆した金属ステントからなる群から選択される、請求項6の医薬組成物。
- 前記予防しまたは軽減することが再狭窄症の進行を予防しまたは軽減することを含む、請求項1の医薬組成物。
- 治療的に有効な量の酸化デューテリウム、パクリタキセルまたは水溶性パクリタキセル誘導体を局所的に送達して、患者における再狭窄症を予防しまたは軽減するための手段を含む、薬物送達システム。
- 前記薬物送達システムがステントを含む、請求項9の薬物送達システム。
- 前記ステントが、ポリマーで被覆した金属ステント、生分解性の薬物溶出ポリマーステント、および内皮細胞で被覆した金属ステントからなる群から選択される、請求項10の薬物送達システム。
- 前記水溶性パクリタキセル誘導体が、2’−サクシニル−パクリタキセル;2’−サクシニル−パクリタキセルトリエタノールアミン;2’−グルタリル−パクリタキセル2’−グルタリル−パクリタキセルトリエタノールアミン塩;2’−(ジメチルアミノエチル)グルタミドとの2’−O−エステル;そしてN−(ジメチルアミノエチル)グルタミド塩酸塩との2’−O−エステルからなる群から選択される、請求項9の薬物送達システム。
- 前記予防しまたは軽減することが再狭窄症の進行を予防しまたは軽減することを含む、請求項9の薬物送達システム。
- 前記パクリタキセルの治療的に有効な量が2mg/kgに等しい、またはそれよりも少ない、請求項9の薬物送達システム。
- 患者の再狭窄症の予防または軽減のための、活性成分として酸化デューテリウム、パクリタキセルまたは水溶性パクリタキセル誘導体を含む医薬組成物を製造する際の、酸化デューテリウム、パクリタキセルまたは水溶性パクリタキセル誘導体の使用方法。
- 前記水溶性パクリタキセル誘導体が、2’−サクシニル−パクリタキセル;2’−サクシニル−パクリタキセルトリエタノールアミン;2’−グルタリル−パクリタキセル;2’−グルタリル−パクリタキセルトリエタノールアミン塩;N−(ジメチルアミノエチル)グルタミドとの2’−O−エステル;そしてN−(ジメチルアミノエチル)グルタミド塩酸塩との2’−O−エステルからなる群から選択される、請求項15の方法。
- 前記予防しまたは軽減することが前記医薬組成物の全身性送達によって達成される、請求項15の方法。
- 前記予防しまたは軽減することが前記医薬組成物の局所送達によって達成される、請求項15の方法。
- 前記局所送達が局所的持続性放出送達システムを含む、請求項18の方法。
- 前記局所的持続性放出送達システムがステントを含む、請求項19の方法。
- 前記ステントが、ポリマーで被覆した金属ステント、生分解性の薬物溶出ポリマーステント、および内皮細胞で被覆した金属ステントからなる群から選択される、請求項20の方法。
- 前記予防しまたは軽減することが再狭窄症の進行を予防しまたは軽減することを含む、請求項15の方法。
- 前記パクリタキセルの治療的に有効な量が2mg/kgに等しい、またはそれよりも少ない、請求項15の方法。
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JP2006128856A Expired - Lifetime JP4615478B2 (ja) | 1993-07-29 | 2006-05-08 | 微小管安定化剤を用いたアテローム性動脈硬化症または再狭窄症の治療方法 |
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