JP4615064B2 - 不完全な組織修復を治療するための組成物および低侵襲的方法 - Google Patents
不完全な組織修復を治療するための組成物および低侵襲的方法 Download PDFInfo
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Description
本出願は、2002年4月13日に提出された米国仮特許出願第60/372,682号(これはその全体が参照として本明細書に組み入れられる)の優先権を請求する。
1つの態様において、本発明は、血小板リッチな血漿(platetet-rich plasma)を含む組成物を用いる、損傷組織の治療のための方法を対象とする。治療される組織は、結合組織、心筋、骨格筋、椎間板(disc material)、椎体、膵臓およびその他の内臓、脳組織もしくは脊髄組織、または血管組織のいずれでもよい。1つの好ましい態様において、組織は結合組織である。記載される組成物および方法は、創傷治癒および感染症にも有用である。
抗炎症薬療法、装具、安静および理学療法などの標準的な治療に反応しない結合組織損傷に対する治療プロトコールに対しては需要がある。柔軟で比較的血管に乏しい結合組織(以下では「結合組織」または「結合組織」)に対する損傷またはその他の障害は、治癒までに非常に長い時間(数カ月、場合によっては数年)がかかることが知られている。多くの症例では、結合組織に対する損傷が適切に治癒することはなく、外科的介入が必要になる。結合組織の損傷および障害は社会に大きな影響を及ぼす。これらの異常の全有病率は、米国立保健統計センター(National Center for Health Statistics)による1995年の調査によれば、米国では1000人当たり約140人である。この同じ調査では、直接的なコストは887億ドルであり、生産性の損失による間接的なコストは最大1119億ドルに上ると推計している。
ontrolled study)では有用なことは証明されていない。いくつかの研究からは、非ステロイド薬療法は実際には靱帯損傷の治癒過程に有害な影響を及ぼす可能性が示唆されている。また、外側上顆炎の症例の病理試料中に急性炎症性細胞は認められていない。コルチゾン注射は腱炎の治療に対しては実のところ意見が分かれており、急性靱帯損傷に対しては禁忌である。いくつかの研究で、コルチゾンを投与した患者の短期的な経過観察下における改善が言及されている。しかし、1年以上にわたる成績では、症状の再発率が高く、有効率も不確かなものに過ぎないことが示されている。また、これらの注射には腱断裂、感染、皮膚色素脱失、皮下萎縮、および糖尿病患者における高血糖のリスクもある。手術的手段には壊死組織切除および付随する病的な腱の修復が含まれる。しかし、開放手術または関節鏡下手術には、深部感染、神経血管構造への損傷、および瘢痕形成といった多くの合併症がみられる。さらに外科手術は費用もかかる上、局所麻酔または全身麻酔に伴うリスクも加わる。
本発明者は、驚くべきことに、本発明のある種の血小板組成物が、複数の種類の組織障害または組織損傷の治療に有用であることを見いだした。本明細書で用いる「損傷」という用語は、広義の用語であり、創傷、外傷もしくは病変を含む任意の組織障害または任意の組織変性を非制限的に指すことを目的に通常の意味で用いられる。特に、本発明の血小板組成物は、さまざまな結合組織の不完全修復を治療するために用いうる。
らに含む方法に関する。同じく有用なものには、脱顆粒化または血小板の保存を促進する物質、別の増殖因子または増殖因子阻害剤、NSAIDS、ステロイドおよび抗感染剤などの低分子調合薬(small molecule pharmaceutical)がある。1つの局面において、本発明は、この方法において患者の結合組織が、腱、靱帯、関節包および筋膜組織より選択される方法に関する。1つの局面において、本発明は、この方法において血小板組成物の入手が、ヒトから血液を採取すること;および血液を遠心処理して多血漿(plasma-rich)画分を得ることを含む方法に関する。1つの局面において、本発明は、この方法において血小板組成物が血小板リッチな血漿を含む方法に関する。1つの局面において、本発明は、この方法において血小板組成物が、患者の結合組織中の不完全修復領域の内部および周囲への導入前には外因性活性化物質を実質的に含まないことを条件とする方法に関する。1つの局面において、本発明は、この方法において血小板組成物が患者から入手した血小板を含む方法に関する。
剤などの低分子調合薬がある。1つの局面において、本発明は、血小板組成物の入手が、ヒトから血液を採取すること;および血液を遠心処理して多血漿画分を得ることを含む本方法に関する。1つの局面において、本発明は、血小板組成物が生理的pHまたはそれを上回るpHにある本方法に関する。1つの局面において、本発明は、血小板組成物が患者から入手した血小板を含む本方法に関する。
られており、このためウシトロンビンは手術で避けられている。ヒトのプールトロンビン(pooled thrombin)も同じようにウイルス、プリオン、細菌などの種々の物質による汚染の恐れがあるために避けられている。組換えヒトトロンビンを用いることもありうるが、これは非常に高価である。
リウムなど)、グルコン酸カルシウム、塩化コリン、デキストロース(d-グルコース)、エチレンビス(オキシエチレンニトリロ)四酢酸(EGTA)、4-(2-ヒドロキシエチル)-1-ピペラジンエタンスルホン酸(HEPES)、マレイン酸、4-モルホリンプロパンスルホン酸(MOPS)、1,4-ピペラジンビス(エタンスルホン酸)(PIPES)、スクロース、N-トリス(ヒドロキシメチル)メチル-2-アミノエタンスルホン酸(TES)、トリス(ヒドロキシメチル)アミノメタン(TRIS塩基)、トリス(ヒドロキシメチル)アミノメタン塩酸(TRIS.HCl)および尿素が含まれる。1つの好ましい態様において、pH調整剤は重炭酸緩衝液であり、より好ましくは重炭酸ナトリウムである。
PRPを、Harvest社(Plymouth, MA)により製造された遠心装置を用いて調製した(同様の装置はBiomet GPSシステム、Depuy Symphony器械およびMedtronic Magellan器械としても入手可能である)。血液約55ccを標準的な滅菌シリンジを用いて患者から採取し、凝固阻止のために5ccのクエン酸デキストロース溶液を配合した後に、製造者のプロトコールに従って、血小板を単離するため遠心により沈降させた。続いて、これらの血小板を約3ccの血漿中に再懸濁させた。その結果得られた血小板リッチな血漿溶液(PRP)は酸性度が高かったため、PRP 1cc当たり約0.05ccの8.4%重炭酸ナトリウム緩衝液を用いて滅菌条件下で中和し、ほぼ生理的なpH 7.4とした。PRPを外因性活性化物質の添加によって活性化することはしなかった。このPRP組成物を本明細書では自己血小板抽出物(APEX)と称する。
全血50ccを患者から採取した後、Knightonの方法(米国特許第5,165,938号、コラム3)
に従って調製する。PRPを、Knightonに従って組換えヒトトロンビンを用いて活性化する。脱顆粒化した血小板を遠心により沈降させ、放出物を含む上清を採取する。選択的には、重炭酸ナトリウム緩衝液を用いて放出物のpHをpH 7.4に調整してもよい。
全血30mlを患者から採取した。血小板組成物は、アルギン酸塩を血小板組成物に添加しない点を除き、Cochrumに対する米国特許第5,510,102号(これはその全体が参照として本明細書に組み入れられる)の実施例1に従って調製した。
本明細書に記載のPRP組成物の効果に関する試験を、外側上顆炎(テニス肘)を呈しており、非手術的治療(抗炎症薬療法、装具を付けた状態での安静および理学療法)が奏功しなかった患者を対象として行った。これらの患者の症状の平均持続期間は16.6カ月であった。インフォームドコンセントを得た後に、患者を形式に則って試験に組み入れ、ランダムにAPEX治療群または対照群のいずれかに割り当てた。
実施例4の患者標本を6カ月時点で再び評価した。本明細書に記載した本発明の方法を用いたAPEX組成物による治療から少なくとも6カ月を経過し、評価を受けた患者は現在までに5例に上る。その結果の概要を以下の表3および4に示した。これまでに検討した5例の患者に関する平均値を示している。
アキレス腱炎を呈する患者に対して、本発明の腱処置を行う30分前にバリウム5mgを経口投与する。次に患者の準備およびドレープかけを無菌的な様式で行う。エピネフリンを含む0.5%ブピビカインによる局所浸潤麻酔を皮膚、皮下構造およびアキレス腱に対して行う。局所麻酔薬が足関節の外側で維持されるように注意する。
肘の内側側副靱帯断裂を呈する患者に対して、本発明の腱処置を行う30分前にバリアム5mgを経口投与する。次に患者の準備およびドレープかけを無菌的な様式で行う。エピネフリンを含む0.5%ブピビカインによる局所浸潤麻酔を皮膚、皮下構造および肘の内側側副靱帯に対して行う。局所麻酔薬が足関節の外側で維持されるように注意する。
患者は急性の心筋機能不全(すなわち心発作)または慢性の心筋機能不全(すなわち、
うっ血性心不全)を呈している。APEX組成物を実施例1の通りに調製する。抽出物1cc当たり約0.05ccの8.4%重炭酸ナトリウム緩衝液を用いて、pHを7.4またはそれよりも幾分高いpHにする。抽出物を外因性活性化物質の添加によって活性化することはしない。
患者は骨格筋の筋力低下または萎縮を呈している。これは損傷または手術処置による結果である可能性がある。実施例1の技法を用いて、自己由来の血小板抽出物(APEX)を入手し、緩衝剤で処理して生理的pHにする。
患者は腰痛を呈しており、MRIスキャンでは***性または低輝度の椎間板が認められる。実施例1の技法を用いて、自己由来の血小板抽出物(APEX)を入手し、緩衝剤で処理して生理的pHにする。
患者は糖尿病またはインスリン産生能の低下を呈している。実施例1の技法を用いて、自己由来の血小板抽出物(APEX)を入手し、緩衝剤で処理して生理的pHにする。
患者は脊髄損傷または脳卒中などの急性神経障害を呈している。実施例1の技法を用いて、自己由来の血小板抽出物(APEX)を入手し、緩衝剤で処理して生理的pHにする。
患者は下肢に血管過少の領域を呈している。この患者は末梢血管閉塞症と診断されている。実施例1の技法を用いて、自己由来の血小板抽出物(APEX)を入手し、緩衝剤で処理して生理的pHにする。
患者は適切に治癒していない慢性創傷を呈している。これは糖尿病性足部潰瘍である可能性がある。実施例1の技法を用いて、自己由来の血小板抽出物(APEX)を入手し、緩衝剤で処理して生理的pHにする。
患者は良性または悪性の腫瘍または***(process)を呈している。実施例1の技法を用いて、自己由来の血小板抽出物(APEX)を入手し、緩衝剤で処理して生理的pHにする。APEXをインビボまたはインビトロで用いて、腫瘍細胞死を惹起または誘導することができる。
患者は表在性感染または深部感染を呈している。実施例1の技法を用いて、自己由来の血小板抽出物(APEX)を入手し、緩衝剤で処理して生理的pHにする。
研究者または臨床医は、線維芽細胞または骨関節炎軟骨細胞の細胞培養物を増殖させることを望んでいる。実施例1の技法を用いて、自己由来の血小板抽出物(APEX)を入手し、緩衝剤で処理して生理的pHにする。
Claims (6)
- 血小板リッチな血漿組成物を含む、個体における心筋機能不全部位の内部および周囲にカテーテルによって導入して用いられるための心筋梗塞治療剤であって、
該血漿組成物は、滴定によりpHが7.3〜7.5に調節され、かつ
前記導入に先だって前記血小板が活性化されていないものである、心筋梗塞治療剤。 - 血漿組成物が、前記個体から入手した血小板を含む、請求項1に記載の心筋梗塞治療剤。
- 前記個体がヒトである、請求項1又は2に記載の心筋梗塞治療剤。
- 滴定が重炭酸緩衝液を用いて行われる、請求項1〜3の何れか一項に記載の心筋梗塞治療剤。
- 前記導入は移植用器具と組み合わせて行われる、請求項1〜4の何れか一項に記載の心筋梗塞治療剤。
- 移植用器具がステントである、請求項5に記載の心筋梗塞治療剤。
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JP2010100648A (ja) | 2010-05-06 |
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EP1494535A4 (en) | 2005-10-26 |
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US20160206657A1 (en) | 2016-07-21 |
US20060263407A1 (en) | 2006-11-23 |
AU2003221946A1 (en) | 2003-11-03 |
EP2258379B1 (en) | 2014-12-17 |
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US20120183520A1 (en) | 2012-07-19 |
EP1494535B1 (en) | 2017-01-18 |
US6811777B2 (en) | 2004-11-02 |
WO2003088905A2 (en) | 2003-10-30 |
EP2258379A1 (en) | 2010-12-08 |
KR20050029118A (ko) | 2005-03-24 |
US20070264245A1 (en) | 2007-11-15 |
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