JP7442969B2 - 架橋タンパク質泡を発生するための粉末組成物およびその使用方法 - Google Patents
架橋タンパク質泡を発生するための粉末組成物およびその使用方法 Download PDFInfo
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- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 238000011179 visual inspection Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
- 239000008096 xylene Substances 0.000 description 1
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Description
前記組成物は多孔質骨格であり、
前記骨格の細孔は2~500ミクロンであり、前記組成物は、
a)コラーゲンおよびゼラチンからなる群から選択される架橋可能タンパク質、
b)前記架橋可能タンパク質の架橋を誘導する架橋剤、および
c)液体、
から成る。
a)架橋可能ゼラチン、
b)前記架橋可能ゼラチンの架橋を誘導するトランスグルタミナーゼ、および
c)液体、
ここで、前記組成物は2~500ミクロンの粒径を有する多孔質骨格であり、
前記架橋可能ゼラチンは5~200ミクロンの粒径を有する微粉化ゼラチン粉末として前記組成物に導入され、
前記架橋可能ゼラチンは200~300ブルームであり、
前記架橋可能ゼラチンは0.5重量部~25重量部の範囲で前記組成物中に存在し、
前記トランスグルタミナーゼは0.0001重量部~2重量部の範囲で前記組成物中に存在する、
から成る組成物を提供する。
a)組織欠損または疾患を治療するのに十分な量で、組織欠損部位において患者に前期組成物を導入すること、
ここで、前期組成物は欠損部位において組織に付着する
ことから成る。
a)組織欠損または疾患を治療するのに十分な量で、組織欠損部位において患者の体内で前期組成物を形成すること、
ここで、前期組成物は欠損部位において組織に付着する、
ことから成る。
a)患者の肺の標的領域を崩壊させること、
b)崩壊される領域の第1の部分を崩壊される領域の第2の部分に付着させるのに十分な量で崩壊される領域の部位において、患者の体内に前記組成物を導入すること、
ここで、崩壊される領域の第1の部分を、崩壊される領域の第2の部分に付着させることで、患者の肺容量を減らし、
前記組成物は、線維芽細胞接着およびコラーゲン合成を促進するように構成され、
前記線維芽細胞接着およびコラーゲン合成は炎症を防ぐことから成る。
a)肺容量を減らすのに十分な量で、肺領域の部位において患者の体内に前期組成物を導入すること、
前記組成物は、線維芽細胞接着およびコラーゲン合成を促進するように構成され、
前記線維芽細胞接着およびコラーゲン合成は炎症を防ぐことから成る。
a)患者の肺の標的領域を崩壊させること、
b)崩壊される領域の第1の部分を崩壊される領域の第2の部分に付着させるのに十分な量で、崩壊される領域の部位において、患者の体内で前記組成物を形成すること、
ここで、崩壊される領域の第1の部分を崩壊される領域の第2の部分に付着させることで、患者の肺容量を減らし、
前記組成物は、線維芽細胞接着およびコラーゲン合成を促進するように構成され、
前記線維芽細胞接着およびコラーゲン合成は炎症を防ぐことから成る。
a)肺容量を減らすのに十分な量で、肺領域の部位において患者の体内で前期組成物を形成すること、
ここで、前記組成物は、線維芽細胞接着およびコラーゲン合成を促進するように構成され、
前記線維芽細胞接着およびコラーゲン合成は炎症を防ぐことから成る。
架橋可能タンパク質
架橋剤
本発明の一部の実施形態に従った組成物
前記組成物は多孔質骨格であり、
前記骨格の細孔は2~500ミクロンであり、前記組成物は、
a)コラーゲンおよびゼラチンからなる群から選択される架橋可能タンパク質、
b)前記架橋可能タンパク質の架橋を誘導する架橋剤、および
c)液体
を含む。
骨格は、
1)細胞接着および移動を可能にする;
2)細胞および生化学的要因を送達および保持する;
3)必要不可欠な細胞栄養素および発現生成物の拡散を可能にする;または
4)細胞段階の挙動に特定の機械的および生物学的影響を働かせる
という目的のうちの少なくとも1つのために役割を果たすことができる。
a)架橋可能ゼラチン、
b)前記架橋可能ゼラチンの架橋を誘導するトランスグルタミナーゼ、および
c)液体、
ここで、前記組成物は2~500ミクロンの粒径を有する多孔質骨格であり、
前記架橋可能ゼラチンは5~200ミクロンの粒径を有する微粉化ゼラチン粉末として前記組成物に導入され、
前記架橋可能ゼラチンは200~300ブルームであり、
前記架橋可能ゼラチンは0.5重量部~25重量部の範囲で前記組成物中に存在し、
前記トランスグルタミナーゼは0.0001重量部~2重量部の範囲で前記組成物中に存在することから成る組成物を提供する。
治療を必要とする患者を治療する方法
a)患者の肺の標的領域を崩壊させること、
b)崩壊される領域の第1の部分を崩壊される領域の第2の部分に付着させるのに十分な量で崩壊される領域の部位において、患者の体内に前記組成物を導入すること。
ここで、崩壊される領域の第1の部分を崩壊される領域の第2の部分に付着させることで、患者の肺容量を減少させ、
前記組成物は、線維芽細胞接着およびコラーゲン合成を促進するように構成され、
前記線維芽細胞接着およびコラーゲン合成は炎症を防ぐことから成る。
a)肺容量を減少させるのに十分な量で、肺領域の部位において患者の体内に前期組成物を導入すること、
ここで、前記組成物は、線維芽細胞接着およびコラーゲン合成を促進するように構成され、
前記線維芽細胞接着およびコラーゲン合成は炎症を防ぐことから成る。
a)患者の肺の標的領域を崩壊させること、
b)崩壊される領域の第1の部分を崩壊される領域の第2の部分に付着させるのに十分な量で崩壊される領域の部位において、患者の体内で前記組成物を形成すること、
ここで、崩壊される領域の第1の部分を崩壊される領域の第2の部分に付着させることで、患者の肺容量を減らし、
前記組成物は、線維芽細胞接着およびコラーゲン合成を促進するように構成され、
前記線維芽細胞接着およびコラーゲン合成は炎症を防ぐことから成る。
a)肺容量を減らすのに十分な量で、肺領域の部位において患者の体内で前期組成物を形成すること。
ここで、前記組成物は、線維芽細胞接着およびコラーゲン合成を促進するように構成され、
前記線維芽細胞接着およびコラーゲン合成は炎症を防ぐことから成る。
実施例
実施例1:本発明の一部の実施形態に従った組成物の引き抜き試験
成分
方法
実施例3:追加の骨増生材料を用いた、および用いない骨欠損の治療における架橋ゼラチンの効果
ソケットタイプA:0.2グラムのmTG + 0.25gのゼラチン + 0.0125グラムのコンドロイチン硫酸A+C(GAG)に混合された4 BONE + 1.25mlの生理食塩水。
ソケットタイプB:4 BONE単独
ソケットタイプC:0.25グラムのゼラチンによる泡の中に混合された0.25グラムのmTG。
ソケットタイプD:陰性対照群としての機能を果たすように空のままにした
結果
組織学的分析
実施例4:本発明の一部の実施形態に従った組成物の性能に対するGAGの効果
材料
(i)0.034グラムのウシ気管由来コンドロイチン硫酸Aナトリウム塩(Sigma)
(ii)0.233グラムのゼラチン(Gelita AG)
(iii)0.233グラムのmTG(ACTIVA;1%mTG;味の素)
(iv)1mlの水
上記材料をシリンジ間の方法で混合し、架橋させた。
結果
実施例5:Surgifloを含む組成物
以下の実験では、ゼラチン顆粒に基づいたその他の市販製品の再溶解しmTGと架橋する能力を試験する
材料
(i)一方のシリンジの中で0.25グラムの乾燥Surgiflo止血基質(Ethicon Inc)を0.25グラムの粉末化mTG(ACTIVA WM;味の素)に混合させた
(ii)別のシリンジの中に1mlの生理食塩水を満たした
結果
実施例6:本発明の一部の実施形態に従った泡の長期機械的特性
材料
結果
実施例7:
結果
実施例8:顕微鏡検査
結果
実施例9:胃腸封止剤としての本発明の一部の実施形態に従った組成物の使用
方法
結果
実施例10:本発明の一部の実施形態に従ったさまざまな組成物の試験
実施例12:便失禁の治療としての本発明の一部の実施形態に従った組成物の使用
結果:前記泡は柔軟性のある体積中で生体位で安定し、組織に付着した。前記組織片は肛門の導管の直径を減少させ、管の人工的な接合(狭窄)を生じさせた。前記注入部位を100日後および120日後に採取した。前記ブタを安楽死させ、組織をホルマリン固定し、組織学的分析のために調製されたパラフィンブロックおよびスライドに埋め込んだ。どの動物も有害事象を示さなかった。前記生体材料は優れた耐性を示し、分解可能である。壊死も、空洞形成も、生体材料の移動も示さなかった。前記生体材料は線維芽細胞を移植部位に引き付けた。すなわち、増殖し、線維性組織を生成した繊維芽細胞。注入後、急性段階に前記材料はグレード2の炎症反応を生じ、その後の慢性段階で、線維芽細胞が生体材料の体積に置換するように働いている間、軽度(グレード1~2)の炎症が続く。
実施例14:尿失禁の治療としての本発明の一部の実施形態に従った組成物の使用
ブタによる慢性試験
結果
原発性括約筋機構不全症(PMSI)を罹患するイヌの治療
結果
実施例15:本発明の一部の実施形態に従ったさまざまな組成物の試験
(A):d(0.1)=4.24μm、d(0.5)=16.61μm、d(0.9)=31.51μm。
(B):d(0.1)=4.415μm、d(0.5)=13.064μm、d(0.9)=29.621μm。
(C):d(0.1)=13.451μm、d(0.5)=94.66μm、d(0.9)=423.785μm。
実施例18:ゼラチン成分との混合後の細胞の生存
群A:生体材料を調製するために10回、オスのルアーロックシリンジに入れられた120mgの無菌mTGに混合され、1mlの細胞原液培地(Dulbeccoの修飾イーグル培地、ロット番号1530279)に混合された200mgの無菌ゼラチンの組成を持ち、未処理プラスチック(Corning Inc.、Costar、参照番号3736、ロット番号30015036)の6ウェルプレート上に注入されたゼラチン生体材料。
群B:細胞を含まない培地に同じ組成の生体材料を混合し、40分間架橋させ、1x106個の細胞を3mlの培地(Dulbeccoの修飾イーグル培地、高グルコース、Biological Industries、ロット番号1530279、10%認定ウシ胎仔血清、カタログ番号04-001-1Aおよび1%ペニシリン-ストレプトマイシン溶液、Biological Industries、03-031-1Cを配合)に添加し、37℃で2時間シェーカーに入れた後、5%のCO2を含み、37℃設定のインキュベーターに移した。この方法によって、細胞はゼラチン骨格上に付着できるようになり、その体積内に埋め込まれるよりも表面上で成長する。
群C:対照として使用され、細胞を含まないが、群AおよびBの生物材料と同じ組成である。
5%のCO2を含み、37℃に設定されたインキュベーターで群A、B、Cを培養した。
実施例20:肺気腫治療のための手術用封止剤としての使用の実施形態
結果
実施例22:本発明の一部の実施形態に従った組成物の皮下注射
結果
Claims (25)
- 必要とする患者の体内で組織欠損または疾患を治療することに使用するための組成物であって、
a.前記組成物は、組織欠損または疾患を治療するのに十分な量で、必要とする患者の組織欠損または疾患の部位でイン・サイチュで形成されるか;または
b.前記組成物は、イン・ビトロ(in vitro)で形成され、組織欠損または疾患を治療するのに十分な量で、必要とする患者の組織欠損または疾患の部位に導入され;
前記組成物は、
a.ゼラチンである架橋可能タンパク質、
b.前記架橋可能タンパク質の架橋を誘導する架橋剤、および
c.液体
を混合することにより形成され、
前記架橋可能タンパク質は5~200ミクロンの平均粒子サイズを有する、微粉化タンパク質粉末として混合され、
前記組成物は多孔質骨格であり、
前記骨格の細孔は2~500ミクロンであり、
前記組成物は、生分解性の架橋タンパク質泡であり、
前記組成物は、閉細胞泡である、
組成物。 - 前記液体は生理学的緩衝液である、請求項1に記載の組成物。
- 前記架橋可能タンパク質は200~300ブルームのゼラチンから成る、請求項1に記載の組成物。
- 前記架橋剤がトランスグルタミナーゼであり、トランスグルタミナーゼは0.0001重量%~2重量%の範囲で組成物中に存在する、請求項1に記載の組成物。
- ゼラチンが吸湿性の粒子状ゼラチン粉である、請求項1に記載の組成物。
- 治療を必要とする患者の組織欠損または疾患を治療することに使用するための請求項1に記載の組成物であって、
a.組織欠損または疾患を治療するのに十分な量で、組織欠損または疾患の部位において患者の体内に組成物を導入することを含み;
ここで、前記組成物は組織欠損または疾患の部位において組織に付着し、
組織欠損は創傷であり;または
組織は正常に機能せず、再生が必要であり;または
組織欠損は骨欠損であり、ここに組成物は患者の骨の再生を誘導し;または
疾患は便失禁であり;または
疾患は尿失禁であり;または
疾患は肺気腫である
、組成物。 - 患者における肺容量を減少することによって肺気腫の治療に使用するための、請求項1に記載の組成物。
- 組織再構築薬剤として使用するための、請求項1に記載の組成物。
- 架橋可能なゼラチンが0.5重量%~25重量%の範囲で組成物に存在する、請求項1に記載の組成物。
- 治療を必要とする患者の組織欠損または疾患の治療のための組成物であって、
前記組成物は、
a.ゼラチンである架橋可能タンパク質、
b.前記架橋可能タンパク質の架橋を誘導する架橋剤、および
c.液体
を混合することにより形成され、
前記架橋可能タンパク質は5~200ミクロンの平均粒子サイズを有する、微粉化タンパク質粉末として混合され、
前記組成物は多孔質骨格であり、
前記多孔質骨格の細孔は2~500ミクロンであり、
前記組成物は生分解性の架橋タンパク質泡であり、
前記組成物は閉細胞泡であり、
前記治療は、形成された組成物を、
組織欠損または疾患を治療するのに十分な量で、組織欠損または疾患の部位において患者に導入すること
を含む、組成物。 - 組織欠損は創傷である、請求項10に記載の組成物。
- 組織は正常に機能せず、再生が必要である、請求項10に記載の組成物。
- 組織欠損は骨欠損である、請求項10に記載の組成物。
- 組成物は患者の骨の再生を誘導する、請求項13に記載の組成物。
- 疾患は便失禁である、請求項10に記載の組成物。
- 疾患は尿失禁である、請求項10に記載の組成物。
- 疾患は肺気腫である、請求項10に記載の組成物。
- 治療を必要とする患者の組織欠損または疾患を治療するための請求項1に記載の組成物であって、
前記組成物は、組織欠損または疾患を治療するのに十分な量で、組織欠損または疾患の部位においてイン・サイチュで形成され;
ここに、前記組成物は欠損または疾患の部位において組織に付着する、組成物。 - 組織欠損は創傷である、請求項18に記載の組成物。
- 組織は正常に機能せず、再生が必要である、請求項18に記載の組成物。
- 組織欠損は骨欠損である、請求項18に記載の組成物。
- 組成物は患者の骨の再生を誘導する、請求項21に記載の組成物。
- 疾患は便失禁である、請求項18に記載の組成物。
- 疾患は尿失禁である、請求項18に記載の組成物。
- 疾患は肺気腫である、請求項18に記載の組成物。
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