JP2018511622A - 擬塑性ミクロゲルマトリクスの組成物およびキット - Google Patents
擬塑性ミクロゲルマトリクスの組成物およびキット Download PDFInfo
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Abstract
Description
実験セクションにおいて下記の材料と略語が使用される。
ADSC:脂肪由来幹細胞、初代ヒト腹部形成細胞
AHF:クリオプレシピテート抗血友病因子、南テキサス血液&組織センター、Lot W1409114 212930
BSA:ウシ血清アルブミン、bioWORLD,22070004−1,Lot V11121401
塩化カルシウム:シグマ社 C4901,Lot 110M0105V
カーボポール:ルブリゾール社、カーボポール(登録商標)Aqua SF−2,Lot 0101014502
I型コラーゲン:コーニング社(Corning Incorporated)、製品番号354236、ラット尾腱、Lot3298599
ECM:細胞外基質、Engelbreth-Holm-Swarmマウス肉腫、シグマ社E1270,Lot 093M4006V。
ジェランガム:CPケルコ社(CP Kelco),ケルコゲルCG−LA,Lot 1E0566A
グァーガム:メイキングコスメティック社(Making Cosmetics)、Lot 1092118
ヒトフィブリノゲン(FGN):シグマ社F3879,Lots 061M7010,071M7032V,SLBH0223V,SLBK3747V
ヒトトロンビン:シグマ社T7009,Lots 041M7007V,011M7009V,SLBB4394V
ヒドロキシエチルセルロース,カチオン性:ダウケミカル社(Dow Chemical Company),UCare(商標)ポリマーJR−30M,Lot XL2850GRXA
ヒドロキシプロピルメチルセルロース:Amerchol Methocel K15M,Lot WF15012N01
細片化羊膜:バイオディー社(BioD,LLC),BioDRestore(商標),組織ID R0925131
NaOH:水酸化ナトリウム,ピューリタンプロダクツ社(Puritan Products)7705,Lot 011043.
ニードル:18G,BD305195,Lot 2089215
PBS:リン酸緩衝生理食塩水:pH7.8−8.0(カルシウムおよびマグネシウム無し),INCELLZSOL:F,LotsA2014SEP10−01,Z2015JAN05−01;またはシグマアルドリッチ社D8537,Lots RNBB9451,RNBC1143,RNBC8400
PHMB:ポリ(ヘキサメチレンビグアニド)塩酸塩:アーチ・ケミカルズ社(Arch Chemicals),コスモシールCQ(商標),Lots 9PL211280,137261またはローションクラフタービグアニド20,Lot 14RC169159−6380
ポリ(エチレングリコール)ジアクリラート:MW 575,シグマアルドリッチ社437441,Lot MKBN7800V
ポリ(ビニルアルコール):デュポン社(DuPont),エルバノール(Elvanol)(登録商標),Lot 910113
PSG−(PEO)4:ペンタエリトリトールテトラ(スクシンイミジルオキシグルタリル)ポリオキシエチレン,4アーム(四官能基),NOFアメリカ社(NOF America Corporation),サンブライト(Sunbright)(登録商標)PTE−050GS (MW 5,000),Lot M8N526; PTE−100GS (MW 10,000),Lot M9D105;およびPTE200−GS (MW 20,000),Lot M119691
プルラン:林原,Lot 1G3021
SG−PEG−SG (PEG):NOFアメリカ社(NOF America Corporation),α−スクシンイミジルオキシグルタリル−ω−スクシンイミジルオキシグルタリルオキシポリオキシエチレン,(二官能性), NOFアメリカ社(NOF America Corporation); サンブライト(Sunbright)(登録商標)DE−034GS (MW 3400),Lot M83541; DE−100GS (MW 10,000),Lot M107543; DE−200GS (MW 20,000),Lot M115700
SG−PEG:α−スクシンイミジルオキシグルタリル−ω−メトキシポリオキシエチレン,(単官能性), NOFアメリカ社(NOF America Corporation),サンブライト(Sunbright)(登録商標)ME−050GS (MW5,000),Lot M10N587
キサンタンガム:ボブズレッドミル,Lot 143454
ポリ(エチレングリコール)−ヒトフィブリノゲン(PEG−FGN)
種々のモル比のPEG化フィブリノゲン(PEG−フィブリノゲン,PEG−FGN)の調製について、20:1モルPEG−FGNの調製を用いて説明する。フィブリノゲン(FGN,MW 340kDa)を、pH7.8〜8.0の滅菌PBSに、80mg/mLで溶解させた(カルシウムおよびマグネシウム無し)。フィブリノゲンを常温または37℃で溶解させた。SG−PEG−SG(MW 3.4kDa)を常温で(〜22℃)pH7.8−8.0の滅菌PBS(カルシウムおよびマグネシウム無し)に16mg/mLで溶解させた。試験した全ロットのフィブリノゲンも同様に行った。SG−PEG−SGの反応性二官能PEG溶液は0.20−0.22μmフィルターを用いて滅菌濾過された。その後、SG−PEG−SGはFGNと1:1v/v(等容積)で混和された。常温または37oCで5〜30分のうちにゲル化が起こった。
PEGジアクリレート(二官能性);
SG−PEG−SG,Sunbright DE−034GS (MW 3400),DE−100GS(MW 10,000),DE−200GS (MW 20,000)(二官能性);
SG−PEG: Sunbright(登録商標)ME−050GS (MW 5,000)(単官能性);
PSG−(PEO)4,PTE−050GS(MW 5,000),Sunbright PTE−100GS(MW 10,000),PTE200−GS (MW 20,000)(四官能性);
ここで、
DE:NHS−OCO(CH2)3COO−PEG−CO(CH2)3COO−NHS;
ME:PEG−CO(CH2)3COO−NHS;
PTE:PEG−(CO(CH2)3COO−NHS)4;
NHS:N−ヒドロキシスクシンイミド。
PEG−FGNハイドロゲル(モル比20:1)を、PHMBの量が0から1000ppmまでの範囲に存在するように添加して調製した。PEG−FGN−PHMB複合材料の調製は、100ppmのPHMB(ミクロゲルが50mg/mLで再水和された濃度)を取り込むことにより説明され、他の濃度を有するPHMBも同様にして調製される。
下記水溶性ポリマーを取り込むことにより、PEG−FGN−水可溶性ポリマーハイドロゲル(PEGとFGNが20:1のモル比)が調製された:27.6mg/mLのカーボポール、27.6mg/mLのプルラン、20.7mg/mLのグァーガム、3.4mg/mLのヒドロキシエチルセルロース、1.4mg/mLのI型コラーゲン。カーボポールはアクリル酸をベースとした架橋合成ポリマーであり、プルランとグァーガムは天然多糖類であり、ヒドロキシエチルセルロースは修飾多糖類であり、I型コラーゲンはタンパクであって、人体に最も豊富に存在するコラーゲンである。PEG化の前に水溶性ポリマーが添加された。
PEG−フィブリンゲルを10:1から50:1までのモル比で調製した。20:1を超えてPEG化が高まると、トロンビンの活性とフィブリンゲルの形成が阻害され、より長い反応時間が必要とされる。フィブリノゲンを架橋させるため、2.5−12.5U/mLゲルのトロンビン濃度で、PEG−フィブリノゲンにトロンビンが添加された。
PEG化フィブリン(10:1)ハイドロゲルは、2mg/mLで取り込まれた下記の水性ポリマー:キサンタンガム、ジェランガム、グァーガム、カチオン性ヒドロキシエチルセルロース、ヒドロキシプロピルメチルセルロース、およびポリ(ビニルアルコール)により調製された。キサンタンガム、ジェランガムおよびグァーガムは天然由来の多糖類、カチオン性のヒドロキシエチルセルロースおよびヒドロキシプロピルメチルセルロースは変性多糖類、ポリ(ビニルアルコール)は合成ビニル系ポリマーである。
実施例1と同様の手順を用いて、下記のタンパクを20:1のモル比でPEG化した:フィブリノゲン、ウシ血清アルブミン、I型コラーゲン、クリオプレシピテート抗血友病因子(AHF)、および細胞外基質。1MのNaOHを用いてpHが>7.0まで上昇すると、PEG化によりゲル形成が誘発された。In vivoでの使用およびタンパク安定性を考えると、6.6から8.0までの範囲にあるpHを使用することができる。水和PEG化ゲルは、急速に機械攪拌することよりミクロゲル粒子に変換された。得られたミクロゲル粒子の擬塑性は、水和ミクロゲル粒子をシリンジに装填し、このPBSを含むミクロゲルを18ゲージニードルから注入させることにより実証され、せん断力がなくなるとミクロゲルのクラスターが形成された。
PEG化されたフィブリノゲン系およびフィブリン系のハイドロゲルが、24−96時間乾燥凍結された。乾燥凍結されたゲルはその後、臼と杵、またはスペックサンプルプレップ(Spex SamplePrep)6870(メアチェン、ニュージャージー)を用いて粉砕された。乾燥ミクロゲル粉末をその後、タイラー製またはレッチェ製のステンレス鋼試験用篩を用いて(<250μm、<150μm、<106μm、または<75μm)の粒子サイズに篩い分けすることができる。乾燥ミクロゲル粉末は、PBS(カルシウムおよびマグネシウムは含有していてもしていなくてもよい)、生理食塩水(0.9%塩化ナトリウム)、脱イオン水、細胞培地(細胞は含有していてもしていなくてもよい)、または細片化羊膜、により再水和された。
水和ミクロゲル粒子に剪断減粘性があることが分かった。この予期しない性質により、所定部位への流体注入および移植が容易になり、ミクロゲル粒子組成物が注入後に直ちに、粘弾性固体として機能し、生体周囲組織に適合する形状が維持されるようになる。ミクロゲル粒子組成物の擬塑性および粘弾性を調べるため、レオメータ(アントンパール社製MCR101、アントンパール社製MCR302、アッシュランド、バージニア)が使用された。例として、凍結粉砕(または臼と杵)により製造された、20:1モル比の再水和PEG−フィブリノゲンミクロゲル粒子および10:1モル比の再水和PEG−フィブリンミクロゲル粒子を利用すると、図2に図示されている粘性に対するせん断速度は、これらのプロットの絶対的な傾きの値により説明されるように、架橋PEG化タンパクミクロゲル粒子組成物は急速に剪断減することを示していて、PEG−フィブリノゲンの傾きが−0.90であって、またPEG−フィブリンの傾きは−0.98であり、−1に近い傾きは、完全な剪断減粘性物質であることを示唆している。
臼と杵および凍結粉砕により粉砕された50:1のPEG−フィブリノゲンミクロゲルの剪断減粘性を、粘性に対するせん断速度のプロットとして図5に図示した。臼と杵により得られた乾燥ミクロゲル粒子は、より扁平状でフレーク状に近かったのに対して、凍結粉砕により得られた乾燥粒子はより球状に近かった。ミクロゲルをリン酸緩衝生理食塩水で300mg/mLで再水和させた。プロットの傾きは、臼と杵に対して−0.86、凍結粉砕に対して−0.84であった。凍結粉砕された粒子は多少粘性度が高いが、これは小さな粒子サイズ分布および小さな平均粒子サイズに起因するものだと思われる。
人間の腹部脂肪からヒト脂肪由来幹細胞(ADSC)を分離し、MesenPRO RS(商標)培地(ライフテクノロジー社)にて、成長補助剤および1%ペニシリンストレプトマイシンと共に、継代培養した。10:1、20:1、および50:1のPEG−FGNとPEG−フィブリンの粉末サンプル(15mg)を、12ウェルプレート(n=3)の細胞培養インサート(透明PET膜 サイズ=8.0μm、BDバイオサイエンス社)の中で、400μLの2.5×105細胞/mL懸濁液と混ぜた。このインサートに別の600μL培地を添加して、別に1mLを外部から添加し、全部で2mLの培地容量となった。培地を毎日交換し、CellTiter 96(登録商標)Aqueous One Solution Cell Proliferation Assay (MTS)(プロメガ社)を用いて細胞増殖をメーカーの手順に従って調べた。その後、カルセイン−AM(4mM)生体細胞染色試薬を用いて45分間細胞が染色され、10%ホルマリンを用いて固定された。各サンプルの巨視的蛍光画像を、デジタルカメラと共焦点顕微鏡をそれぞれ用いて取得した。
PEG化ミクロゲル粒子組成物に抗微生物特性を提供するために、フィブリノゲンとのPEG化反応にPHMBが取り込まれた。図9は、1,000ppmのPHMBを含む20:1のPEG−FGN−PHMB三重複合体の擬塑性を、20:1のPEG−FGNと比較して示している。PEG−FGNの傾きの値は−1.0であり、PEG−FGN−PHMBの値は−1.06であって、−1に近い傾きは完全な剪断減粘性材料であることを示している。
PEG−FGN(20:1モル)は単独で、または、自己移植の幹細胞(ラットADSC)もしくはBioDRestore(商標)細片化羊膜と組み合わせて評価された。エチレンオキシドで滅菌化されたミクロゲルが、21ゲージのニードルを用いてラット背部皮下領域に注入された。5つの試験グループが複製により評価された:ラットADSCs、BioDRestore(商標)、PEG−FGN、ラットADSC含有PEG−FGN、およびBioDRestore(商標)含有PEG−FGN。ミクロゲルが容易に注入され、凝集性の固体を形成し、7日目と14日目に注入部位に維持されていたという結果が示されている。
第1の実施例は、水和すると擬塑性を示し、架橋PEG化タンパク、架橋PEG化タンパク系生体高分子を含有する水不溶性ミクロゲル粒子を含む組成物であって、
水和するとミクロゲル粒子のクラスターの粘性がせん断力の印加により減少し、せん断力がなくなるとミクロゲルクラスターを再構成し、
水和すると水和した前記ミクロゲル粒子が粘弾性固体の性質を持ち、貯蔵弾性率が損失弾性率よりも大きく、損失正接の値が1未満となることを特徴とする組成物に関する。
前記刻まれた組織には、刻まれた皮膚組織、筋組織、血管組織、神経組織、脂肪組織、軟骨組織、骨組織、腱組織、膀胱組織、腸組織、心臓組織、肺組織、腎組織、肝組織、膵組織、および声帯組織が含まれ、
前記微粉末化組織および微粉末脱細胞化組織には、皮膚組織、筋組織、血管組織、神経組織、脂肪組織、軟骨組織、骨組織、腱組織、膀胱組織、腸組織、心臓組織、肺組織、腎組織、肝組織、膵組織、声帯組織、が含まれ、
前記合成または天然由来の細胞外基質成分には、コラーゲン、グリコサミノグリカン、フィブリン、ラミニン、フィブロネクチンが含まれ、
生分解性ポリマーには、ポリグリコリド、ポリラクチド、ポリ(ラクチド−co−グリコリド)、ポリジオキサノン、ポリカプロラクトン、ポリ(トリメチレンカルボナート)、ポリ(プロピレンフマラート)、ポリウレタン、ポリ(エステルアミド)、ポリ(オルトエステル)、ポリ無水物、ポリ(アミノ酸)、ポリホスファゼン、細菌ポリエステル、が含まれる生物成分をさらに含有することを特徴とする実施例1に記載の組成物に関する。
前記生物開始材料をPEG化剤と反応させるステップと、
前記生物開始材料を前記PEG化剤と架橋させて二官能性または多官能性のPEG化を形成することで前記生物開始材料をPEG化するステップと、
PEG化された前記生物開始材料を水和または脱水した条件で粉砕またはせん断することにより複数のミクロゲル粒子を生成するステップと、
を備えた水不溶性ミクロゲル粒子を調製する方法であって、
前記ミクロゲル粒子は水溶液中で擬塑性を示し、
水和した前記ミクロゲル粒子の溶液はせん断力の印加により粘性が減少し、せん断力がなくなるとミクロゲルクラスターを形成し、
PEG化剤の生物開始材料に対するモル比は1:1から100:1にあり、水和した前記ミクロゲル粒子は粘弾性固体の性質があり、貯蔵弾性率が損失弾性率より大きく、損失正接の値が1未満となることを特徴とする水不溶性ミクロゲル粒子を調製する方法に関する。
Claims (29)
- 水和すると擬塑性を示し、架橋PEG化タンパク、架橋PEG化タンパク系生体高分子を含有する水不溶性ミクロゲル粒子を含む組成物であって、
水和するとミクロゲル粒子のクラスターの粘性がせん断力の印加により減少し、せん断力がなくなるとミクロゲルクラスターを再構成し、
水和すると水和した前記ミクロゲル粒子が粘弾性固体の性質を持ち、貯蔵弾性率が損失弾性率よりも大きく、損失正接の値が1未満となることを特徴とする組成物。 - PEG化剤のタンパク質およびタンパク系生体高分子に対するモル比が1:1から100:1までにあることを特徴とする請求項1に記載の組成物。
- 前記ミクロゲル粒子が水和していることを特徴とする請求項1に記載の組成物。
- 前記組成物はせん断力の印加により粘性が減少し、また、せん断力がなくなるとミクロゲルクラスターを形成し、前記組成物には粘弾性固体の性質があり、貯蔵弾性率が損失弾性率より大きく、損失正接の値が1未満となることを特徴とする請求項1に記載の組成物。
- 前記タンパクおよびタンパク系生体高分子が、細胞外基質、糖タンパク、構造タンパク、線維性タンパク、酵素、プロテオグリカン、天然ポリペプチド、合成ポリペプチド、球状タンパク、膜タンパク、血漿タンパク、ペプチド、オリゴペプチド、抗微生物ペプチド、ペプチド・ホルモン、シャペロン、金属タンパク質、ヘムタンパク質、凝固タンパク質、免疫系タンパク、イオンチャンネルタンパク、細胞接着タンパク、神経ペプチド、核タンパク、硬タンパク、色素タンパク、共役タンパク、タンパク−タンパク複合体、タンパク多糖類複合体、タンパク・脂質複合体、モーター・タンパク、ムコタンパク、リンタンパク、収縮性タンパク、輸送タンパク、シグナル伝達タンパク、調節タンパク、増殖因子タンパク、感知タンパク、防御タンパク、貯蔵タンパク、受容タンパク、抗体、組み換えタンパク、フィブリノゲン、フィブリン、トロンビン、コラーゲン、エラスチン、アルブミン、ゼラチン、ケラチン、ラミニン、およびこれらの組み合わせからなる群より選択されることを特徴とする請求項1に記載の組成物。
- タンパクPEG化架橋剤が、α−スクシンイミジルオキシグルタリル−ω−スクシンイミジルオキシグルタリルオキシポリオキシエチレン(SG−PEG−SG)、α−アミノプロピル−ω−アミノプロポキシポリオキシエチレン、α−アミノプロピル−ω−カルボキシペンチルオキシポリオキシエチレン、α,ω−ビス{2−[(3−カルボキシ−1−オキソプロピル)アミノ]エチル}ポリエチレングリコール、α−[3−(3−マレイミド−1−オキソプロピル)アミノ]プロピル−ω−[3−(3−マレイミド−1−オキソプロピル)アミノ]プロポキシポリオキシエチレン、ペンタエリトリトールテトラ(アミノプロピル)ポリオキシエチレン、α−[3−(3−マレイミド−1−オキソプロピル)アミノ]プロピル−ω−(スクシンイミジルオキシカルボキシ)ポリオキシエチレン、ペンタエリトリトールテトラ(スクシンイミジルオキシグルタリル)ポリオキシエチレン、ペンタエリトリトールテトラ(メルカプトエチル)ポリオキシエチレン、ヘキサグリセリンオクタ(スクシンイミジルオキシグルタリル)ポリオキシエチレン、ヘキサグリセリンオクタ(4−ニトロフェノキシカルボニル)ポリオキシエチレン、4−アームポリ(エチレングリコール)テトラアクリラート、4−アームスクシンイミジルオキシグルタリル」ポリオキシエチレン、ビス(ポリオキシエチレンビス[イミダゾイルカルボニル])、O−(3−カルボキシプロピル)−O′−[2−(3−メルカプトプロピオニルアミノ)エチル]ポリエチレングリコール、O,O′−ビス[2−(N−スクシンイミジルスクシニルアミノ)エチル]ポリエチレングリコール、O,O′−ビス(2−アジドエチル)ポリエチレングリコール、ポリ(エチレングリコール)ジアクリラート、ポリ(エチレングリコール)ジグリシジルエーテル、ポリ(エチレングリコール)二(炭酸p−ニトロフェニル)、ポリ(エチレングリコール)ジ(ビニルスルホン)、ポリ(エチレングリコール)ジ(プロプリオンアルデヒド)、ポリ(エチレングリコール)ジ(炭酸ベンゾトリアゾリル)、およびこれらの組み合わせ、からなる群から選択されることを特徴とする請求項1に記載の組成物。
- 前記組成物、水和した前記水不溶性ミクロゲル粒子、またはこれら両方の貯蔵弾性率の値が10Paから250,000Paまでの間にあり、損失弾性率の値が5Paから100,000Paまでの間にあることを特徴とする請求項1に記載の組成物。
- 抗菌剤、抗真菌薬、抗原虫薬、抗ウイルス薬、抗生物質、モノアシルグリセリン、モノアルキルグリコール、ビス(ビグアニド)およびその塩、ポリ(ヘキサメチレンビグアニド)およびその塩、グリセリンモノラウラート、クロルヘキシジン、クロルヘキシジンジグルコン酸塩、クロルヘキシジン二酢酸塩、アレキシジン、アレキシジン塩酸塩、銀塩、塩化ベンザルコニウム、塩化ベンゼトニウム、硫酸ゲンタマイシン、ヨード、ポビドンヨード、でんぷん・ヨード、硫酸ネオマイシン、ポリミキシンB、バシトラシン、テトラサイクリン、クリンダマイシン、ゲンタマイシン、ニトロフラゾン、酢酸マフェニド、スルファジアジン銀、テルビナフィン塩酸塩、硝酸ミコナゾール、ケトコナゾール、クロトリマゾール、イトラコナゾール、メトロニダゾール、抗微生物ペプチド、ポリクオタニウム−1、ポリクオタニウム−6、ポリクオタニウム−10、カチオン性グアー、水溶性キトサン誘導体、これらの塩、またはこれらの組み合わせをさらに含有することを特徴とする請求項1に記載の組成物。
- 水溶性ポリマーを0.01重量%から25重量%までの濃度で含有し、前記水溶性ポリマーは、ポリ(エチレングリコール)、ポリ(エチレンオキサイド)、ポリ(ビニルアルコール)およびその共重合体、ポリ(N−ビニルピロリドン)およびその共重合体、メチルセルロース、ヒドロキシエチルセルロース、ヒドロキシプロピルセルロース、ヒドロキシプロピルメチルセルロース、カルボキシメチルセルロース、グァーガム、ヒドロキシエチルグァー、ヒドロキシプロピルグァー、ゼラチン、アルブミン、ヒドロキシプロピルメチルグァー、カルボキシメチルグァー、カルボキシメチルキトサン、ローカストビーンガム、カラギナン、キサンタンガム、ジェランガム、プルラン、デキストラン、硫酸デキストラン、アロエベラゲル、スクレログルカン、シゾフィラン、アラビアゴム、タマリンドゴム、ポリ(メチルビニルエーテル)、エチレン酸化プロピレン酸化エチレンオキサイドブロック共重合、ヒアルロナン、コンドロイチン硫酸、ケラタン硫酸、デルマタン硫酸、ヘパラン硫酸、デキストラン、カルボマーおよびその塩、ポリ(アクリル酸)およびその塩、ポリ(メタクリル酸)およびその塩、ポリ(エチレン−co−アクリル酸)、ポリ(ビニルメチルエーテル)、ポリ(ビニルリン酸)塩、ポリ(ビニルスルホン酸)塩、ナトリウムポリ(2−アクリルアミド−2−メチルプロパンスルホナート)、ポリアクリルアミド、ポリ(N,N−ジメチルアクリルアミド)、ポリ(N−ビニルアセトアミド)、ポリ(N−ビニルホルムアミド)、ポリ(2−ヒドロキシメタクリル酸エチル)、ポリ(メタクリル酸グリセリル)、ポリ(N−イソプロピルアクリルアミド)およびポリ(N−ビニルカプロラクタム)であってこの2つは臨界共溶温度未満で水和、ポリクオタニウム−1、ポリクオタニウム6、ポリクオタニウム−10、イオネンポリマー、カチオン性グアー、ピリジニウム重合体、イミダゾリウム重合体、ジアリルジメチルアンモニウム重合体、ポリ(エルリジン)、アクリロイル/メタクリロイル/スチリルトリメチルアンモニウム重合体、アクリルアミド/メタクリルアミド−トリメチルアンモニウム重合体、および抗微生物ペプチド、からなる群より選択されることを特徴とする請求項1に記載の組成物。
- 前記組成物が乾燥または水和した膜状であることを特徴とする請求項9に記載の組成物。
- 細胞、幹細胞、細片化された羊膜組織、胎盤組織、刻まれた組織、微粉末化組織および微粉末脱細胞化組織、合成または天然由来の細胞外基質成分、ヒドロキシアパタイト、粒状架橋ウシ腱コラーゲンおよびグリコサミノグリカン、はちみつ、多糖類、生分解性ポリマー、およびこれらの組み合わせ、からなる群より選択され、
前記刻まれた組織には、刻まれた皮膚組織、筋組織、血管組織、神経組織、脂肪組織、軟骨組織、骨組織、腱組織、膀胱組織、腸組織、心臓組織、肺組織、腎組織、肝組織、膵組織、および声帯組織が含まれ、
前記微粉末化組織および微粉末脱細胞化組織には、皮膚組織、筋組織、血管組織、神経組織、脂肪組織、軟骨組織、骨組織、腱組織、膀胱組織、腸組織、心臓組織、肺組織、腎組織、肝組織、膵組織、声帯組織、が含まれ、
前記合成または天然由来の細胞外基質成分には、コラーゲン、グリコサミノグリカン、フィブリン、ラミニン、フィブロネクチンが含まれ、
生分解性ポリマーには、ポリグリコリド、ポリラクチド、ポリ(ラクチド−co−グリコリド)、ポリジオキサノン、ポリカプロラクトン、ポリ(トリメチレンカルボナート)、ポリ(プロピレンフマラート)、ポリウレタン、ポリ(エステルアミド)、ポリ(オルトエステル)、ポリ無水物、ポリ(アミノ酸)、ポリホスファゼン、細菌ポリエステル、
が含まれる生物成分をさらに含有することを特徴とする請求項1に記載の組成物。 - 前記幹細胞が、成体幹細胞、胚性幹細胞、羊膜幹細胞、誘導多能性幹細胞、胎児幹細胞、組織幹細胞、脂肪由来幹細胞、骨髄幹細胞、ヒト臍帯血幹細胞、血液前駆細胞、間充織幹細胞、造血幹細胞、表皮幹細胞、内皮前駆細胞、上皮幹細胞、胚盤葉上層幹細胞、心臓幹細胞、膵臓幹細胞、神経幹細胞、角膜輪幹細胞、周産期幹細胞、衛星細胞、サイドポピュレーション細胞、多能性幹細胞、全能性幹細胞、単能性幹細胞、およびこれらの混合物、からなる群から選択されることを特徴とする請求項11に記載の組成物。
- 前記細胞が、線維芽細胞、ケラチノサイト、ニューロン、グリア細胞、星状膠細胞、シュワン細胞、後根神経節、含脂肪細胞、内皮細胞、上皮細胞、軟骨細胞、繊維状軟骨細胞、筋細胞、心筋細胞、筋芽細胞、肝細胞、腱細胞、腸上皮細胞、平滑筋細胞、間質細胞、好中球、リンパ球、骨髄細胞、周皮細胞、血小板、およびこれらの混合物、からなる群から選択されることを特徴とする請求項11に記載の組成物。
- 塩形態または中性の形態にある、ナノ粒子、ミクロ粒子、消毒剤、抗感染症薬、抗微生物剤、殺胞子剤、駆虫剤、末梢神経障害治療薬、神経障害治療薬、走化性剤、鎮痛剤、抗炎症剤、抗アレルギー剤、抗高血圧薬、マイトマイシン型抗生物質、ポリエン系抗真菌薬、発汗抑制剤、充血除去剤、酔い止め薬、中枢神経系薬、創傷治療薬、抗VEGF薬、抗腫瘍薬、腐食剤、抗乾癬薬、抗糖尿病薬、抗関節炎薬、かゆみ止め、かゆみ止め薬、麻酔剤、抗マラリア剤、皮膚病薬、抗不整脈薬、抗けいれん薬、抗嘔吐薬、抗リウマチ薬、抗アンドロゲン剤、アントラサイクリン、禁煙剤、抗にきび薬、抗コリン剤、老化防止剤、抗ヒスタミン剤、駆虫剤、止血剤、血管収縮剤、血管拡張剤、血栓症薬、抗凝固薬、心・血管作動薬、抗狭心症薬、勃起不全薬、性ホルモン、成長ホルモン、イソフラボン、インテグリン結合シークエンス、生物活性リガンド、細胞付着メディエータ、免疫調節剤、腫瘍壊死因子アルファ、抗がん剤、抗腫瘍薬、抗鬱剤、鎮咳薬、抗−腫瘍薬、麻薬拮抗薬、抗高コレステロール薬、アポトーシス誘導剤、避妊薬、サンレス・タンニング剤、軟化剤、アルファヒドロキシ酸、マヌカ蜂蜜、局所レチノイド、ホルモン、腫瘍特異的抗体、アンチセンス・オリゴヌクレオチド、低分子干渉RNA(siRNA)、抗VEGFRNAアプタマー、核酸、DNA、DNA断片、DNAプラスミド、Si−RNA、トランスフェクション剤、ビタミン、精油、リポソーム、銀ナノ粒子、金ナノ粒子、薬物含有ナノ粒子、アルブミン系ナノ粒子、キトサン含有ナノ粒子、多糖系ナノ粒子、デンドリマーナノ粒子、リン脂質ナノ粒子、酸化鉄ナノ粒子、ビスマスナノ粒子、ガドリニウムナノ粒子、金属ナノ粒子、セラミックナノ粒子、石英系ナノ粒子、ウイルス由来ナノ粒子、ウイルス様ナノ粒子、抗生物質含有ナノ粒子、酸化窒素含有ナノ粒子、ナノシェル、ナノロッド、高分子ミセル、銀塩、亜鉛塩、量子ドットナノ粒子、高分子系微粒子、高分子系ミクロスフェア、薬物含有微粒子、薬物含有ミクロスフェア、抗生物質含有、抗生物質含有ミクロスフェア、抗菌性微粒子、抗菌性ミクロスフェア、サリチル酸、過酸化ベンゾイル、5−フルオロウウラシル、ニコチン酸、ニトログリセリン、クロニジン、エストラジオール、テストステロン、ニコチン、乗り物酔い止め薬、スコポラミン、フェンタニル、ジクロフェナク、ブプレノルフィン、ブピバカイン、ケトプロフェン、オピオイド、カンナビノイド、酵素、酵素阻害薬、オリゴペプチド、シクロペプチド、ポリペプチド、タンパク、プロドラッグ、プロテアーゼ阻害薬、サイトカイン、ヒアルロン酸、コンドロイチン硫酸、デルマタン硫酸、副交感神経遮断薬、キレート剤、育毛促進剤、脂質、糖脂質、糖タンパク、内分泌性ホルモン、成長ホルモン、増殖因子、分化因子、熱ショックタンパク、免疫反応修飾物質、単糖類、多糖類、インスリンおよびインスリン誘導体、ステロイド、コルチコステロイド、非ステロイド性抗炎症薬、およびこれらの組み合わせ、からなる群から選択される生物活性剤をさらに含有することを特徴とする請求項1に記載の組成物。
- 前記水不溶性ミクロゲル粒子が乾燥粉末状であることを特徴とする請求項1に記載の組成物。
- 水、等張食塩水、平衡塩類溶液、緩衝溶液、リンゲル液、細胞培地、幹細胞培地、血清、血漿、羊水、ワルトンゼリー、栄養培地、消毒液、またはこれらの組み合わせから選択される水性媒体をさらに含有することを特徴とする請求項1に記載の組成物。
- 溶液、懸濁液、クリーム、ローション、ジェル、ペースト、乳液、香料、スプレー、フォーム、エアロゾール、またはその他の形状をさらに含むことを特徴とする請求項16に記載の組成物。
- 前記タンパクが、フィブリノゲン、フィブリン、細胞外基質、血漿タンパク、およびコラーゲンからなる群から選択され、前記PEG化剤がα−スクシンイミジルオキシグルタリル−ω−スクシンイミジルオキシグルタリルオキシポリオキシエチレン(SG−PEG−SG)であることを特徴とする請求項1に記載の組成物。
- 前記タンパクはフィブリノゲンとフィブリンから選択され、前記ミクロゲル粒子が哺乳類の体内に血管形成を引き起こすことを特徴とする請求項1に記載の組成物。
- 前記タンパクがフィブリノゲンとフィブリンから選択され、前記PEG化剤がα−スクシンイミジルオキシグルタリル−ω−スクシンイミジルオキシグルタリルオキシポリオキシエチレン(SG−PEG−SG)であって、活性生物剤がポリ(ヘキサメチレンビグアニド)およびその塩であることを特徴とする請求項1に記載の組成物。
- タンパク質、タンパク系生体高分子、またはこれら両方の組み合わせを含有する生物開始材料を提供するステップと、
前記生物開始材料をPEG化剤と反応させるステップと、
前記生物開始材料を前記PEG化剤と架橋させて二官能性または多官能性のPEG化を形成することで前記生物開始材料をPEG化するステップと、
PEG化された前記生物開始材料を水和または脱水した条件で粉砕またはせん断することにより複数のミクロゲル粒子を生成するステップと、
を備えた水不溶性ミクロゲル粒子を調製する方法であって、
前記ミクロゲル粒子は水溶液中で擬塑性を示し、
水和した前記ミクロゲル粒子の溶液はせん断力の印加により粘性が減少し、せん断力がなくなるとミクロゲルクラスターを形成し、
PEG化剤の生物開始材料に対するモル比は1:1から100:1にあり、水和した前記ミクロゲル粒子は粘弾性固体の性質があり、貯蔵弾性率が損失弾性率より大きく、損失正接の値が1未満となることを特徴とする水不溶性ミクロゲル粒子を調製する方法。 - 請求項1に記載の組成物を提供するステップと、前記組成物を軟組織に注入するステップとを備え、前記注入は、経皮、皮下、経口、筋内、粘膜下、経鼻、膣内、口腔、髄腔内、硬膜外、脳実質内、眼部、網膜下、歯科的、腫瘍内、心腔内、関節内、静脈内、陰茎海綿体内、骨内、腹腔内、腹腔内、筋膜内、臓器内、および硝子体内であることを特徴とすることを特徴とする軟組織の損傷または外傷を治療する方法。
- 請求項1に記載の水不溶性ミクロゲル粒子が乾燥粉末状であって、前記注入ステップの前にこの乾燥粉末を水和させるステップをさらに備えることを特徴とする請求項22に記載の方法。
- 注入後に前記組成物が血管形成を促進させることを特徴とする請求項22に記載の方法。
- 水和すると擬塑性を示す水不溶性ミクロゲル粒子と、前記水不溶性ミクロゲル粒子を哺乳類の体の内部または表面に留置するための指示とを含むキットであって、前記ミクロゲル粒子には架橋PEG化タンパク、架橋PEG化タンパク系生体高分子が含まれ、水和すると前記ミクロゲル粒子のベッドはせん断力の印加により粘性が減少し、せん断力がなくなるとミクロゲルクラスターを形成し、PEG化剤のタンパク系生体高分子に対するモル比が1:1から100:1までにあり、水和すると、水和した前記ミクロゲル粒子が粘弾性固体の性質を持ち、貯蔵弾性率が損失弾性率よりも大きく、損失正接の値が1未満となることを特徴とするキット。
- 前記水不溶性ミクロゲル粒子が乾燥粉末状であって、乾燥ミクロゲル粒子は1μmから250μmまでのサイズを有し、前記指示に前記水不溶性ミクロゲル粒子を乾燥粉末状に水和させることが含まれていることを特徴とする請求項25に記載のキット。
- 水和ミクロゲルが細胞と混合され、水和した細胞/ミクロゲル組成物が、使用の指示に従って、哺乳類の体の内部または表面に留置されることを特徴とする請求項25に記載のキット。
- 抗微生物剤をさらに含有し、前記指示には、前記水不溶性ミクロゲル粒子と前記抗微生物剤を含有する治療組成物を、使用の指示に従って、哺乳類の体の内部または表面に塗布することが含まれることを特徴とする請求項25に記載のキット。
- 前記ミクロゲルが哺乳類の体の内部または表面に留置されると血管形成を促進させることを特徴とする請求項25に記載のキット。
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EP3283057A1 (en) | 2018-02-21 |
CN107708675B (zh) | 2022-07-22 |
BR112017022045B1 (pt) | 2023-11-28 |
AU2016250012A1 (en) | 2017-11-02 |
US20160303281A1 (en) | 2016-10-20 |
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US11590259B2 (en) | 2023-02-28 |
JP6930733B2 (ja) | 2021-09-01 |
AU2016250012B2 (en) | 2020-12-03 |
PT3283057T (pt) | 2021-11-04 |
KR20170140274A (ko) | 2017-12-20 |
HK1245663A1 (zh) | 2018-08-31 |
EP3283057B1 (en) | 2021-07-28 |
CA2982352A1 (en) | 2016-10-20 |
WO2016168196A1 (en) | 2016-10-20 |
BR112017022045A2 (pt) | 2018-07-03 |
DK3283057T3 (da) | 2021-11-01 |
CN107708675A (zh) | 2018-02-16 |
KR102592242B1 (ko) | 2023-10-25 |
MX2017013002A (es) | 2018-02-01 |
ES2895947T3 (es) | 2022-02-23 |
EP3283057A4 (en) | 2018-12-12 |
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