JP2022159521A - マウス衛星細胞の増殖のための小分子 - Google Patents
マウス衛星細胞の増殖のための小分子 Download PDFInfo
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Abstract
Description
この出願は、2016年2月1日に出願された米国特許出願第15/012,656号の一部継続であり、かつこれに対する優先権を主張する。米国特許出願第15/012,656号は、2012年6月18日に出願された国際出願第PCT/US2012/042964号の、米国特許法371の下の国内段階出願である、2014年6月13日に出願された米国特許出願第14/126,716号(これは、現在2016年2月2日に発行された米国特許第9,248,185号である)の一部継続であり、かつこれに対する優先権を主張する。国際出願第PCT/US2012/042964号は、米国特許法§119(e)の下、2011年6月16日に出願された米国仮出願第61/497,708号の利益を主張する。上記出願の内容は、それらの全体が参考として本明細書に援用される。
したがって、衛星細胞の増殖を誘導する組成物および方法が、当技術分野で必要とされている。
本明細書で使用される場合、用語「キナーゼ」は、ホスフェート基を転移する任意のホスホトランスフェラーゼ酵素を意味する。一部の実施形態では、キナーゼは、プロテインキナーゼである。プロテインキナーゼは、タンパク質中の特異的な残基のリン酸化を触媒する酵素のファミリーである。一般に、プロテインキナーゼは、いくつかの群;セリンおよび/またはトレオニン残基を優先的にリン酸化する群、チロシン残基を優先的にリン酸化する群、ならびにチロシンおよびSer/Thr残基の両方をリン酸化する群に分類される。
本明細書で使用される場合、ヘッジホッグシグナル伝達経路に関する用語「モジュレートする」は、ヘッジホッグシグナル伝達経路の構成成分の正常な機能を正に、または負に調節することを意味する。したがって、モジュレートする、という用語は、ヘッジホッグシグナル伝達経路の構成成分の正常な機能の増加、減少、マスキング、変更、無効化、または回復を指すために使用することができる。
本明細書で使用される場合、GPCRに関する用語「モジュレートする」は、GPCRシグナル伝達経路の正常な機能を正に、または負に調節することを意味する。したがって、モジュレートする、という用語は、GPCRの正常な機能の増加、減少、マスキング、変更、無効化、または回復を指すために使用することができる。GPCRモジュレーターは、GPCRアゴニストまたはGPCRアンタゴニストであってよい。
本明細書で使用される場合、用語「ドーパミン受容体モジュレーター」は、1種または複数のドーパミン受容体をモジュレートする化合物を指す。ドーパミン受容体モジュレーターは、ドーパミンアゴニストまたはドーパミンアンタゴニストであってよい。本明細書で使用される場合、用語「ドーパミンアゴニスト」は、1種または複数のドーパミン受容体を活性化する、および/もしくは刺激する、ならびに/またはドーパミンのレベルを上昇させる(L-ドーパまたはドーパミン代謝を阻害する薬物など)、ならびに/またはドーパミンシグナル伝達経路を刺激する、ならびに/またはノルエピネフリンのレベルを低下させる、ならびに/またはノルエピネフリンシグナル伝達経路を阻害する化合物を指す。用語「ドーパミンアゴニスト」にはまた、天然ヒトドーパミン受容体と共通する少なくとも一部の生物学的活性を示すドーパミン分子の類似体が含まれる。したがって、用語「ドーパミンアゴニスト」は、ドーパミン作動性薬剤を包含する。本明細書で使用される場合、用語「ドーパミン作動性薬剤」は、ドーパミンの作用を模倣する化合物を指す。したがって、ドーパミン作動性薬剤という用語は、ドーパミン、ドーパミンの誘導体、およびドーパミン受容体上でドーパミン様作用を有する化合物を包含することが意図されている。ドーパミンの例示的な類似体には、アポモルヒネ、ペルゴリド、ブロモクリプチンおよびリスリドなどのエルゴリンおよびアポルフィンが含まれる)。
本明細書で使用される場合、セロトニン受容体に関する用語「モジュレートする」は、セロトニン受容体の正常な機能を正に、または負に調節することを意味する。したがって、モジュレートする、という用語は、セロトニン受容体の正常な機能の増加、減少、マスキング、変更、無効化、または回復を指すために使用することができる。セロトニン受容体モジュレーターは、セロトニン受容体のアゴニストまたはアンタゴニストであってよい。
本明細書で使用される場合、ヒスタミン受容体に関する用語「モジュレートする」は、ヒスタミン受容体の正常な機能を正に、または負に調節することを意味する。したがって、モジュレートする、という用語は、ヒスタミン受容体の正常な機能の増加、減少、マスキング、変更、無効化、または回復を指すために使用することができる。ヒスタミン受容体モジュレーターは、ヒスタミン受容体のアゴニストまたはアンタゴニストであってよい。
例示的なヒスタミンモジュレーターには、これらに限定されないが、A-349,821;ABT-239;アクリバスチン;アリメマジン(トリメプラジン);アンタゾリン;アステミゾール;アザタジン;アゼラスチン;ベポタスチン;ビラスチン;ビスフェンチジン(Bisfentidine);BL-6341A;BL-6548;BMY-25271;BMY-25405;BMY-52368;ブロムフェニラミン;カルビノキサミン;セチリジン;クロルシクリジン;クロルフェナミン(クロルフェニラミン);クロルプロマジン;シメチジン;シプロキシファン;クレマスチン;クロベンプロピット;クロザピン;シクリジン;シプロヘプタジン;D-16637;DA-4634;デスロラタジン;デクスクロルフェニラミン;ジメボン;ジメンヒドリネート;ジメチンデン;ジフェンヒドラミン;ドネチジン;エバスチン;エブロチジン;エンブラミン;エチンチジン;ファモチジン;ファモチジン;フェキソフェナジン;FRG-8701;FRG-8813;ハロペリドール;HB-408;HE-30-256;ヒドロキシジン;ICI-162846;ICIA-5165;インプロミジン;JNJ7777120;ケトチフェン;L-643728;ラフチジン;ランチジン(lamtidine);レボカバスチン;レボセチリジン;ロラタジン;ロクスチジン;ルピチジン;メクロジン;メピラミン;ミフェンチジン;ミゾラスチン;ニザチジン;ニザチジン;オロンザピン;オロパタジン;ORF-17578;フェニラミン;ピファチジン;プロメタジン;クエチアピン;キフェナジン(Quifenadine);ラミキソチジン(ramixotidine);ラニチジン;リタンセリン;ロキサチジン;SKF-94482;SR-58042;スホチジン;テルフェナジン;チオペラミド;チオチジン;VUF-6002;Wy-45727;ザルチジン;ならびにその類似体、誘導体、鏡像異性体、代謝産物、プロドラッグ、および薬学的に許容される塩が含まれる。
本明細書で使用される場合、HDACに関する用語「モジュレートする」は、HDACの正常な機能を正に、または負に調節することを意味する。したがって、モジュレートする、という用語は、HDACの正常な機能の増加、減少、マスキング、変更、無効化、または回復を指すために使用することができる。HDACモジュレーターは、セロトニン受容体のアゴニストまたはアンタゴニストであってよい。
「エピジェネティック修飾因子」は、DNA配列の変化以外の機構に起因する細胞のエピジェネティック状態、すなわち、細胞における表現型または遺伝子発現を修飾する薬剤を指す。細胞のエピジェネティック状態には、例えば、DNAメチル化、ヒストン修飾(複数可)およびRNA関連サイレンシングが含まれる。
神経ペプチドは、ニューロンが相互に情報伝達するために使用する、より大きな神経伝達物質とは別個の小タンパク質様分子である。これらは、ニューロンシグナル伝達分子であり、特異的な方法で脳の活性に影響を及ぼし、したがって、特に、無痛、報酬、食物摂取、学習および記憶などの脳機能に関係する。神経ペプチドは、ニューロンにより発現および放出され、細胞表面受容体での作用により、ニューロン情報伝達を媒介またはモジュレートする。ヒトゲノムは、神経ペプチドの前駆体をコードする約90の遺伝子を含有する。現在約100種の異なるペプチドが、哺乳動物の脳においてニューロンの種々の集団により放出されることが公知である。
本明細書で使用される場合、用語「イオノフォア」は、一部の例では、他のものを実質的に排除して、特定のイオンと共に複合体を形成することができる分子を含む。一般に、イオノフォアは、イオンと結合することにより、またはそれに対するバリアの透過性を上昇させることにより、脂質バリアを横切ってのイオンの透過を助長する。
本明細書で使用される場合、用語「イオンチャネルモジュレーター」は、イオンチャネルの少なくとも1つの活性をモジュレートする化合物を指す。用語「イオンチャネルモジュレーター」は、本明細書で使用される場合、チャネル細孔自体と相互作用するか、またはチャネル複合体上の部位と相互作用することにより、チャネルのアロステリックモジュレーターとして作用し得る薬剤を含むことが意図されている。用語「イオンチャネルモジュレーター」は、本明細書で使用される場合、間接的にイオンチャネルの活性をモジュレートする薬剤を含むことも意図されている。「間接的に」とは、イオンチャネルとのモジュレーター相互作用に関して使用される場合、イオンチャネルモジュレーターが、イオンチャネル自体と直接的に相互作用しない、すなわち、イオンチャネルモジュレーターが、仲介物を介してイオンチャネルと相互作用することを意味する。したがって、用語「間接的に」はまた、イオンチャネルと結合または相互作用するために、イオンチャネルモジュレーターが、別の分子を必要とする状況を包含する。
ル)-5-ニトロ-3-ピリジンカルボン酸、{2-[4-(2ピリミジニル)1-ピペラジニル]エチル}エステル;5-(4-クロロフェニル)-N-(3,5-ジメトキシフェニル)-2-フランカルボキサミド(A-803467);ならびにその類似体、誘導体、薬学的に許容される塩、および/またはプロドラッグからなる群より選択される。
本明細書で使用される場合、用語「アデノシン受容体モジュレーター」は、アデノシン受容体の少なくとも1つの活性をモジュレートする化合物を指す。用語「アデノシン受容体モジュレーター」は、本明細書で使用される場合、アデノシン受容体自体と相互作用するか、またはチャネル複合体上の部位と相互作用することにより受容体のアロステリックモジュレーターとして作用し得る薬剤を含むことが意図されている。用語「アデノシン受容体モジュレーター」は、本明細書で使用される場合、間接的にアデノシン受容体の活性をモジュレートする薬剤を含むことも意図されている。アデノシン受容体とのモジュレーターが相互作用に関して使用される場合の「間接的に」とは、モジュレーターが受容体自体と直接的に相互作用しない、すなわち、モジュレーターが、仲介物を介してイオンチャネルと相互作用することを意味する。したがって、用語「間接的に」はまた、モジュレーターが受容体に結合または相互作用するために別の分子を必要とする状況を包含する。
本明細書で使用される場合、ガンマセクレターゼに関する用語「モジュレートする」は、ガンマセクレターゼの正常な機能を正に、または負に調節することを意味する。したがって、モジュレートする、という用語は、ガンマセクレターゼの正常な機能の増加、減少、マスキング、変更、無効化、または回復を指すために使用することができる。ガンマセクレターゼモジュレーターは、ガンマセクレターゼアゴニストまたはガンマセクレターゼアンタゴニストであってよい。
本明細書で使用される場合、用語「コルチコステロイド」は、副腎皮質で産生されるか、または合成で生産される一群のステロイドホルモンを指す。コルチコステロイドは、ストレス応答、炎症の免疫応答および調節、炭水化物代謝、タンパク質異化作用、血液電解質レベル、および行動などの広範な生理的システムに関係している。コルチコステロイドは一般に、化学構造に基づき、4つの群に分類される。A群コルチコステロイド(短時間から中時間作用型グルココルチコイド)には、ヒドロコルチゾン、酢酸ヒドロコルチゾン、酢酸コルチゾン、チキソコルトールピバル酸塩、プレドニゾロン、メチルプレドニゾロン、およびプレドニゾンが含まれる。B群コルチコステロイドには、トリアムシノロンアセトニド、トリアムシノロンアルコール、モメタゾン、アムシノニド、ブデソニド、デソニド、フルオシノニド、フルオシノロンアセトニド、およびハルシノニドが含まれる。C群コルチコステロイドには、ベタメタゾン、リン酸ベタメタゾンナトリウム、デキサメタゾン、リン酸デキサメタゾンナトリウム、およびフルオコルトロンが含まれる。D群コルチコステロイドには、ヒドロコルチゾン-17-ブチレート、ヒドロコルチゾン-17-バレレート、ジプロピオン酸アルクロメタゾン(aclometasone dipropionate)、吉草酸ベタメタゾン、ジプロピオン酸ベタメタゾン、プレドニカルベート、クロベタゾン-17-ブチレート、クロベタゾール-17-プロピオネート、カプロン酸フルオコルトロン、ピバル酸フルオコルトロン、および酢酸フルプレドニデンが含まれる。
本発明者らはまた、ヒット化合物の多くが、成長因子と一緒に与えられると、衛星細胞の増殖に対して相乗効果を有することを発見している。したがって、本明細書に記載の態様の一部の実施形態では、本明細書に記載の化合物を、成長因子と共に、衛星細胞と接触させる。例示的な成長因子には、これらだけに限定されないが、塩基性上皮成長因子(bEGF)、線維芽細胞成長因子(FGF)、FGF-1、FGF-2(bFGF)、FGF-4、チモシン、血小板由来成長因子(PDGF)、インスリン結合成長因子(IGF)、IGF-1、IGF-2、上皮成長因子(EGF)、トランスフォーミング成長因子(TGF)、TGF-アルファ、TGF-ベータ、軟骨誘導因子-Aおよび-B、類骨誘導因子、オステオゲニン、骨形成タンパク質、および他の骨成長因子、コラーゲン成長因子、ヘパリン結合成長因子-1または-2、およびそれらの生物学的に活性な誘導体が含まれる。
本明細書において考察する場合、本発明者らは、特に、増殖増強剤の一部が、成長因子と一緒に使用される場合に、衛星細胞の増殖に対して相乗効果を示すことを発見している。したがって、本開示は、増殖増強剤および成長因子を含む相乗作用組成物も提供する。成長因子を選択することができる。
衛星細胞集団を、細胞培養物中で、例えば、in vitroまたはex vivoで増殖増強剤と接触させることができるか、または増殖増強剤を、被験体に、例えば、in vivoで投与することができる。本発明の一部の実施形態では、本明細書に記載の増殖増強剤を、損傷筋組織を修復または再生するために被験体に投与することができる。
被験体に投与するため、増殖増強剤を、薬学的に許容される組成物で提供することができる。これらの薬学的に許容される組成物は、1種または複数の薬学的に許容される担体(賦形剤)および/または希釈剤と一緒に製剤化された治療有効量の1種または複数の増殖増強剤を含む。下記で詳細に記載するとおり、本発明の医薬組成物を、次のために適合するものを含めて、固体または液体形態で投与するために特別に製剤化することができる:(1)経口投与、例えば、水薬(水性または非水性の溶液または懸濁液)、強制経口投与剤(gavage)、ロゼンジ剤、糖剤、カプセル剤、丸剤、錠剤(例えば、頬側、舌下、および全身吸収を標的とするもの)、ボーラス剤、散剤、顆粒剤、舌に適用するためのペースト剤;(2)例えば、皮下、筋肉内、静脈内または硬膜外注射による非経口投与、例えば、滅菌溶液または懸濁液、または持続放出製剤;(3)局所塗布、例えば、皮膚に塗布されるクリーム剤、軟膏剤、または制御放出パッチ剤または噴霧剤として;(4)膣内または直腸内、例えば、膣坐剤、クリーム剤または泡沫として;(5)舌下;(6)眼;(7)経皮;(8)経粘膜;または(9)経鼻。加えて、化合物を、患者に埋植するか、または薬物送達系を使用して注射することができる。例えば、そのすべての内容が参照により本明細書に組み込まれるUrquhartら、Ann. Rev. Pharmacol. Toxicol.、24巻:199~236頁(1984年);Lewis編、「Controlled Release of Pesticides and Pharmaceuticals」(Plenum Press、New York、1981年);米国特許第3,773,919号;および米国特許第353,270,960号を参照されたい。
また別の態様では、本発明は、衛星細胞集団において増殖を刺激する、または増加させるために、候補化合物をスクリーニングする方法であって、
(a)衛星細胞の集団を試験化合物と接触させるステップと;
(b)衛星細胞の増殖を評価するステップと;
(c)衛星細胞の増殖を誘導する、増加させる、または増強する化合物を選択するステップと
を含む方法を提供する。
別の態様では、本発明は、筋修復または再生のためのキットを提供する。一部の実施形態では、キットは、本明細書に記載の化合物、例えば、キナーゼ阻害剤、Gタンパク質共役受容体(GPCR)モジュレーター、ヒストンデアセチラーゼ(HDAC)モジュレーター、ヘッジホッグシグナル伝達経路モジュレーター、神経ペプチド、ドーパミン受容体モジュレーター、セロトニン受容体モジュレーター、ヒスタミン受容体モジュレーター、イオノフォア、イオンチャネルモジュレーター、ガンマ-セクレターゼモジュレーター、およびその任意の組み合わせからなる群より選択される化合物を含む。化合物を、投与のための医薬製剤に予め製剤化することができるか、または医薬製剤に処方するための成分をキットに提供することができる。
1. 衛星細胞の増殖を増加させる方法であって、衛星細胞を、キナーゼ阻害剤、Gタンパク質共役受容体(GPCR)モジュレーター、ヒストンデアセチラーゼ(HDAC)モジュレーター、エピジェネティック修飾因子、ヘッジホッグシグナル伝達経路モジュレーター、神経ペプチド、ドーパミン受容体モジュレーター、セロトニン受容体モジュレーター、ヒスタミン受容体モジュレーター、アデノシン受容体(receprtor)アゴニスト、イオノフォア、イオンチャネルモジュレーター、ガンマ-セクレターゼモジュレーター、コルチコステロイド、およびその任意の組み合わせからなる群より選択される化合物と接触させるステップを含む、方法。
2. 化合物が、小さな有機または無機分子;サッカリン;オリゴ糖;多糖;ペプチド、タンパク質、ペプチド類似体および誘導体;抗体、抗体断片、ペプチド模倣物質;核酸;核酸類似体および誘導体;生物材料から作製される抽出物;天然に存在する組成物または合成組成物;ならびにその任意の組み合わせからなる群より選択される、パラグラフ1に記載の方法。
3. 化合物が、Flt3キナーゼ、PDGFR/EGFR、Bcr-abl、Jak3、またはSRCキナーゼ阻害剤である、パラグラフ1~2のいずれかに記載の方法。
4. 化合物が、プロテインキナーゼ阻害剤および受容体キナーゼ阻害剤からなる群より選択される、パラグラフ1~3のいずれかに記載の方法。一部の実施形態では、キナーゼ阻害剤は、レスタウルチニブ(CEP701、
5. 化合物を、衛星細胞と約0.01nM~約100μMの濃度で接触させる、パラグラフ1~4のいずれかに記載の方法。
6. 前記接触させるステップが少なくとも1時間にわたる、パラグラフ1~5のいずれかに記載の方法。
7. 前記接触させるステップが1~7日間にわたる、パラグラフ1~6のいずれかに記載の方法。
8. 接触がin vitroである、パラグラフ1~7のいずれかに記載の方法。
9. 接触がex vivoである、パラグラフ1~8のいずれかに記載の方法。
10. 接触がin vivoである、パラグラフ1~9のいずれかに記載の方法。
11. in vivoでの接触が哺乳動物における接触である、パラグラフ10に記載の方法。
12. in vivoでの接触がヒトにおける接触である、パラグラフ10または11に記載の方法。
13. in vivoでの接触が被験体における接触であり、被験体が損傷筋組織のための処置を必要としている、パラグラフ10~12のいずれかに記載の方法。
14. 損傷筋組織が、物理的傷害または事故、疾患、感染、濫用、血液循環の喪失、または筋萎縮もしくは消耗の結果である、パラグラフ13に記載の方法。
15. 損傷筋組織が、ジストロフィーの筋肉または老化している筋肉である、パラグラフ13または14に記載の方法。
16. 損傷筋組織が筋萎縮/消耗の結果である、パラグラフ13~15のいずれかに記載の方法。
17. 被験体において筋肉を修復または再生するための方法であって、治療有効量の化合物を被験体に投与するステップを含み、被験体が損傷筋組織を有し、化合物が、キナーゼ阻害剤、Gタンパク質共役受容体(GPCR)モジュレーター(modulaotrs)、ヒストンデアセチラーゼ(HDAC)モジュレーター、エピジェネティック修飾因子、ヘッジホッグシグナル伝達経路モジュレーター、神経ペプチド、ドーパミン受容体モジュレーター、セロトニン受容体モジュレーター、ヒスタミン受容体モジュレーター、イオノフォア、イオンチャネルモジュレーター、ガンマ-セクレターゼモジュレーター、およびその任意の組み合わせからなる群より選択される、方法。
18. 損傷筋組織が、物理的傷害または事故、疾患、感染、濫用、血液循環の喪失、または筋萎縮もしくは消耗の結果である、パラグラフ17に記載の方法。
19. 損傷筋組織が、ジストロフィーの筋肉または老化している筋肉である、パラグラフ17~18のいずれかに記載の方法。
20. 損傷筋組織が筋萎縮/消耗の結果である、パラグラフ17~19のいずれかに記載の方法。
21. 被験体が哺乳動物である、パラグラフ17~20のいずれかに記載の方法。
22. 被験体がヒトである、パラグラフ17~21のいずれかに記載の方法。
23. 化合物を治療剤と共投与する、パラグラフ17~22のいずれかに記載の方法。24. 化合物および治療剤を同じ製剤中で投与する、パラグラフ23に記載の方法。
25. 化合物を1μg/kg~150mg/kgの投薬量で投与する、パラグラフ17~24のいずれかに記載の方法。
26. 前記投与するステップが、注射、注入、点滴注入、吸入、または摂取による、パラグラフ17~25のいずれかに記載の方法。
27. 前記投与するステップが1日1回である、パラグラフ17~26のいずれかに記載の方法。
28. 被験体を筋修復または再生のために処置する前に、被験体を筋損傷または筋萎縮/消耗について診断するステップをさらに含む、パラグラフ17~27のいずれかに記載の方法。
29. 化合物が、
30. 衛星細胞の増殖を誘導する、刺激する、増強する、または増加させる化合物をスクリーニングするためのハイスループットアッセイであって、
(a)衛星細胞を試験化合物と接触させるステップと;
(b)衛星細胞の増殖を評価するステップと;
(c)衛星細胞の複製または成長を誘導する、刺激する、増強する、または増加させる化合物を選択するステップと
を含む、ハイスループットアッセイ。
31. 衛星細胞の増殖を評価するステップが、細胞マーカーを検出するステップを含む、パラグラフ30に記載のアッセイ。
32. 細胞マーカーが、CXCR4、β1-インテグリン、Sca-1、Mac-1、CD45、PAX7、PAX3、Myf5、MyoD、デスミン、およびその任意の組み合わせからなる群より選択される、パラグラフ31に記載のアッセイ。
33. 試験化合物が0.1nM~1000mMの範囲の濃度を有する、パラグラフ30~32のいずれかに記載のアッセイ。
34. アッセイを約15℃~約55℃の範囲の温度で行う、パラグラフ30~33のいずれかに記載のアッセイ。
35. 試験化合物を、膵臓細胞と1時間から7日間にわたって接触させる、パラグラフ30から34のいずれかに記載のアッセイ。
36. 試験化合物が、衛星細胞の増殖を、非処置対照に対して少なくとも5%、10%、20%、30%、40%、50%、50%、70%、80%、90%、1倍、1.1倍、1.5倍、2倍、3倍、4倍、5倍、10倍、50倍、100倍またはそれより多く増加させる、パラグラフ30から35のいずれかに記載のアッセイ。
37. 衛星細胞を哺乳動物から単離する、パラグラフ30~36のいずれかに記載のアッセイ。
38. 衛星細胞を被験体から単離し、被験体が損傷筋組織のための処置を必要としている、パラグラフ30から37のいずれかに記載のアッセイ。
39. 損傷筋組織が、物理的傷害または事故、疾患、感染、濫用、血液循環の喪失、または筋萎縮もしくは消耗の結果である、パラグラフ38に記載の方法。
40. 損傷筋組織が、ジストロフィーの筋肉または老化している筋肉である、パラグラフ38または39に記載の方法。
41. 損傷筋組織が筋萎縮/消耗の結果である、パラグラフ38~40のいずれかに記載の方法。
42. 上述のパラグラフ1~41のいずれかに記載の一部の実施形態では、キナーゼ阻害剤は、
44. 衛星細胞の増殖を増加させる方法であって、衛星細胞を化合物と接触させるステップを含み、化合物がキナーゼ阻害剤である、方法。
45. キナーゼがプロテインキナーゼである、パラグラフ44に記載の方法。
46. プロテインキナーゼがタンパク質チロシンキナーゼである、パラグラフ44に記載の方法。
47. タンパク質チロシンキナーゼが受容体タンパク質チロシンキナーゼである、パラグラフ46に記載の方法。
48. 受容体タンパク質チロシンキナーゼがRETファミリーのメンバーである、パラグラフ47に記載の方法。
49. 受容体タンパク質チロシンキナーゼがRETである、パラグラフ47に記載の方法。
50. 化合物が受容体タンパク質チロシンキナーゼへのリガンドの結合に干渉する、パラグラフ47に記載の方法。
51. 化合物が、小さな有機または無機分子、サッカリン、オリゴ糖、多糖、ペプチド、タンパク質、ペプチド類似体および誘導体、抗体、抗体断片、ペプチド模倣物質、核酸、核酸類似体および誘導体、生物材料から作製された抽出物、天然に存在する組成物または合成組成物ならびにその任意の組み合わせからなる群より選択される、パラグラフ44に記載の方法。
52. 化合物が抗体である、パラグラフ51に記載の方法。
53. 化合物を、衛星細胞と約0.01nM~約100μMの濃度で接触させる、パラグラフ44に記載の方法。
54. 接触がin vivoである、パラグラフ44に記載の方法。
55. in vivo接触が哺乳動物における接触である、パラグラフ54に記載の方法。
56. in vivo接触が哺乳動物における接触であり、哺乳動物が損傷筋組織の処置を必要としている、パラグラフ55に記載の方法。
57. 損傷筋組織が、物理的傷害または事故、疾患、感染、濫用、血液循環の喪失、筋萎縮、筋消耗、ジストロフィーの筋肉または老化している筋肉の結果である、パラグラフ56に記載の方法。
58. 化合物が、キザルチニブ(AC220)、またはその任意の塩、エステルまたはキレートを含む、パラグラフ44~57のいずれかに記載の方法。
59. 損傷筋組織を有する被験体において筋肉を修復または筋肉を再生するための方法であって、治療有効量の化合物を被験体に投与するステップを含み、化合物がキナーゼ阻害剤である、方法。
60. キナーゼがプロテインキナーゼである、パラグラフ59に記載の方法。
61. プロテインキナーゼがタンパク質チロシンキナーゼである、パラグラフ60に記載の方法。
62. タンパク質チロシンキナーゼが受容体タンパク質チロシンキナーゼである、パラグラフ61に記載の方法。
63. 受容体タンパク質チロシンキナーゼがRETファミリーのメンバーである、パラグラフ62に記載の方法。
64. 受容体タンパク質チロシンキナーゼがRETである、パラグラフ61に記載の方法。
65. 化合物が受容体タンパク質チロシンキナーゼへのリガンドの結合に干渉する、パラグラフ61に記載の方法。
66. 化合物が、小さな有機または無機分子、サッカリン、オリゴ糖、多糖、ペプチド、タンパク質、ペプチド類似体および誘導体、抗体、抗体断片、ペプチド模倣物質、核酸、核酸類似体および誘導体、生物材料から作製された抽出物、天然に存在する組成物または合成組成物ならびにその任意の組み合わせからなる群より選択される、パラグラフ59に記載の方法。
67. 損傷筋組織が、物理的傷害または事故、疾患、感染、濫用、血液循環の喪失、筋萎縮、筋消耗、ジストロフィーの筋肉または老化している筋肉の結果である、パラグラフ59に記載の方法。
68. 被験体が哺乳動物である、パラグラフ59に記載の方法。
69. 化合物が、キザルチニブ(AC220)、またはその任意の塩、エステルまたはキレートを含む、パラグラフ59~68のいずれかに記載の方法。
70. 化合物が、
71. 上述のパラグラフ1~41、44、51~57、59~60、66、および67~68のいずれかに記載の一部の実施形態では、キナーゼ阻害剤は、1種または複数のB-Raf阻害剤、JAK3阻害剤、p38MAPK阻害剤、C-Raf1阻害剤、Akt阻害剤、ERK阻害剤、BMK1/ERK5阻害剤、p38MAPK阻害剤、RTK阻害剤、ERK5阻害剤、Bcr-Abl阻害剤、RhoK阻害剤、p38阻害剤、p110阻害剤、FAK阻害剤、ATP競合JNK阻害剤、MELK阻害剤もしくは表5に特定されている経路の阻害剤またはその塩、エステルもしくはキレートを含む。
72. 上述のパラグラフ1~41、44、51~57、59~60、66、および67~68のいずれかに記載の一部の実施形態では、キナーゼ阻害剤は、BAY-439006(すなわち、ソラフェニブ;HMSL10008-101-1);HG-6-64-01(すなわち、HMSL10017-101-1);HKI-272(すなわち、ネラチニブ;HMSL10018-101-1);KIN001-055(すなわち、HY-11067;HMSL10033-101-1);SB239063(すなわち、HMSL10036-101-1);KIN001-242(すなわち、HMSL10044-104-1);SB590885(すなわち、GSK2118436;HMSL10046-101-1);AZ-628(すなわち、HMSL10050-101-1);MK2206(すなわち、HMSL10057-102-1);XMD11-50(すなわち、LRRK2-in-1;HMSL10086-101-1);XMD8-92(すなわち、HMSL10094-101-1);BIRB796;ドラマピモド(すなわち、HMSL10169-101-1);スニチニブリンゴ酸塩(すなわち、SU11248;ステント;HMSL10175-106-1);GDC-0879(すなわち、HMSL10181-101-1);XMD8-85(すなわち、HMSL10093-101-1);AMN-107(すなわち、ニロチニブ;HMSL10099-101-1);Y39983(すなわち、HMSL10149-102-1);SB203580(すなわち、RWJ64809;PB203580;HMSL10167-101-1);VX-745(すなわち、HMSL10168-101-1);pseudoXL765(すなわち、HMSL10173-101-1);Y-27632(すなわち、HMSL10176-101-1);PH-797804(すなわち、HMSL10439-101);VX-702(すなわち、HMSL10440-101);NG25(すなわち、HMSL10419-101);SB202190(すなわち、HMSL10441-101);
BI-D1870(すなわち、HMSL10423-101);BIX02565(すなわち、HMSL10434-101);URMC-099(すなわち、HMSL10453-101);スタウロスポリンアグリコン(すなわち、K252C;HMSL10454-101);ラリメチニブ(すなわち、LY2228820;HMSL10438-103);BMX-IN-1(すなわち、HMSL10427-101);PF3644022(すなわち、HMSL10476-101);NVP-BHG712(すなわち、KIN001-265;HMSL10200-101);ボスチニブ(すなわち、SKI-606;HMSL10189-101);NVP-TAE226(すなわち、CHIR-265;HMSL10207-101);RAD001(すなわち、エベロリムス;HMSL10235-101);CC-401(すなわち、HMSL10185-101);CGP74514A(すなわち、HMSL10355-101);KIN001-269(すなわち、HMSL10195-101);RAF265(すなわち、HMSL10206-101);OTSSP167(すなわち、HMSL10337-102);ドルソモルフィン(すなわち、Compound C;BML275;HMSL10399-102);ロスマピモド(すなわち、GSK-AHAB;SB856553;GW856553X;HMSL10402-101);AZD5363(すなわち、HMSL10370-101);RO31-8220(すなわち、ビスインドリルマレイミドIX;HMSL10407-103);ソトラスタウリン(すなわち、AEB071;HMSL10408-101);TAK-632(すなわち、HMSL10409-101);FRAX597(すなわち、HMSL10400-101);GW2580(すなわち、HMSL10401-101);アリセルチブ(すなわち、MLN8237;HMSL10391-101)、表5に列挙されているキナーゼ阻害剤、またはその誘導体、塩、代謝産物、プロドラッグ、および立体異性体を含む。
73. 上述のパラグラフ1~41、51~57、59~60、66、および67~68のいずれかに記載の一部の実施形態では、エピジェネティック修飾因子は、HDAC修飾因子(例えば、HDAC1、HDAC3、および/またはHDAC6修飾因子)、BRD修飾因子(例えば、BRD2および/またはBRD4修飾因子)、またはEGLN1修飾因子を含む。
73. 上述のパラグラフ1~41、51~57、59~60、66、および67~68のいずれかに記載の一部の実施形態では、エピジェネティック修飾因子が、(+)-JQ1;S)-JQ1;ベリノスタット(すなわち、PXD101);MS-275(すなわち、エンチノスタット;MS-27-275);ボリノスタット(すなわち、スベロイルアニリドヒドロキサム酸(SAHA);ゾリンザ);モセチノスタット(すなわち、MGCD0103);I-BET(すなわち、GSK525762A);SB939(すなわち、プラシノスタット);PFI-1);ロシリノスタット(すなわち、ACY-1215);I-BET151(すなわち、GSK1210151A);IOX2;またはその誘導体、塩、代謝産物、プロドラッグ、および立体異性体を含む。
便宜的に、本明細書、明細書、実施例および添付の特許請求の範囲で用いられる特定の用語をここに集める。他の点で述べられるか、または文脈で示されない限り、次の用語および語句は、下記に示す意味を含む。他の点で明確に述べられるか、または文脈から明らかでない限り、下記の用語および語句は、それが関係する分野で、その用語または語句が獲得している意味を除外するものではない。定義は、特定の実施形態の説明を補助するために示されているものであって、請求している発明を制限することを意図したものではない。なぜなら、本発明の範囲は、特許請求の範囲によってのみ制限されるからである。さらに、文脈によって他の点で必要とされない限り、単数形は複数形を含むこととし、複数形の用語は、単数形を含むこととする。
衛星細胞をCAG-B-アクチン-GFPマウス、2~4カ月齢から、Sherwoodら、2004年およびCerlettiら、2008年に概説されているFACS単離法により採取した。簡単に述べると、すべての四肢筋肉および腹部筋肉を動物から採取し、初めは0.2%コラゲナーゼ溶液中で、次いで、0.0125%コラゲナーゼ/0.05%ディスパーゼ溶液中で消化した。濾過した溶液を、細胞表面マーカーについて染色し、FACSに掛けた。Mac1、Sca1およびCD45については陰性で、かつベータ-インテグリン1およびCXCR4については陽性だった細胞をスクリーニングアッセイのために使用した。細胞を、FACS機器から直接、50細胞/ウェルで、10ug/mLラミニンでコーティングされた96ウェルプレートにプレーティングした(37℃で4~6時間、次いで、部分的に除去した)。スクリーニングのための培地は、10%熱不活化ウマ血清、1×ペニシリン/ストレプトマイシンおよび1×L-グルタミンを補充されたHam F-10であった。塩基性FGF(bFGF)を陽性対照ウェルにのみ添加し;他のウェルはすべて、培地のみを与えられた。プレーティングの日を0日目と呼んだ。プレーティングの翌日、1日目に、化合物を添加した。化合物はすべて、DMSOに溶解し、初めは、10uM、1uMまたは0.1uMで2連で試験した。各プレートのための陰性対照は、同じ濃度のDMSOであった(化合物は溶解していない)。化合物を、培地を換えずに4日目まで細胞と共にインキュベートした。bFGFを、陽性対照ウェルに毎日加えた。4日目に、プレートを4%パラホルムアルデヒド中で固定し、リン酸緩衝食塩水で洗浄した。プレートをOpera Confocalイメージャーで直接的に画像化した。なぜなら、細胞は、CAG-EGFP-ベータ-アクチン導入遺伝子から容易に見ることができたからである。アカペラスクリプトを使用して、各ウェル内の細胞の数を計数した。ウェルを、細胞数に基づき、増殖についてスコアリングした(DMSO処置ウェルは通常、アッセイの終了時に約50個の細胞を有し、bFGF処置ウェルは通常、終了時に150~250個の細胞を有した)。発明者らは、化合物がDMSO対照から1(または、スクリーニングの第2のセッションでは、3)標準偏差(複数可)よりも大きいか、またはそれに等しい細胞数を有した場合に、その化合物をヒットとして選択した。次いで、そうして選択された化合物を、初期用量曲線で、通常は、ヒットとしてフラグを立てた濃度に対応する20nM~30uMの間の濃度で試験した。本発明者らは、化合物が、DMSO陰性対照に対して少なくとも2標準偏差の細胞数まで、増殖させることができた場合に、その化合物を活性と呼んだ。用量曲線において活性であると見出された任意の化合物について、元の供給業者から再供給を受けた。再供給された化合物を、増殖能について用量曲線で再び試験した。次いで、依然として活性であると見出された化合物を、最適化および特徴づけのための列に入れた。
衛星細胞をCAG-B-アクチン-GFPマウス、2~4カ月齢から、実施例1において概説されているとおりにFACS単離法により採取した。簡単に述べると、すべての四肢筋肉および腹部筋肉を動物から採取し、初めは0.2%コラゲナーゼ溶液中で、次いで、0.0125%コラゲナーゼ/0.05%ディスパーゼ溶液中で消化した。濾過した溶液を、細胞表面マーカーについて染色し、FACSに掛けた。Mac1、Sea1およびCD45については陰性で、かつベータ-インテグリン1およびCXCR4については陽性だった細胞をスクリーニングアッセイのために使用した。細胞を、FACS機器から直接、細胞50個/ウェルで、10ug/mLラミニンでコーティングされた96ウェルプレートにプレーティングした(37℃で4~6時間、次いで、部分的に除去した)。スクリーニングのための培地は、10%熱不活化ウマ血清、1×ペニシリン/ストレプトマイシンおよび1×L-グルタミンを補充されたHam F-10であった。塩基性FGF(bFGF)を陽性対照ウェルにのみ添加し;他のウェルはすべて、培地のみを与えられた。プレーティングの日を0日目と呼んだ。
この実施例は、in vitroで衛星細胞の増殖を促進する化合物を同定するように設計されたスクリーニングからの一次ヒットのリストを含む。衛星細胞は、生後の筋成長および再生を担う骨格筋幹細胞である。in vitroで衛星細胞を増殖させる化合物は、筋肉再生のための重要な関連を有する。なぜなら、それらが、in vivoで同等に有効であり、細胞補充療法のために移植可能な細胞を増殖させる可能性を有するからである。試験された4つの化合物ライブラリ、LINCS1、2、3および4は、Harvard Medical SchoolのSorger研究室により、本発明者らに提供された。Sorger研究室は、治療に関連する細胞経路のさらなる調査のために公的データを作成することを目的とするNIHイニシアティブであるLINCS(Library of Integrated Network-Based Cellular Signatures)Consortiumのメンバーである。LINCS1、2、および4はキナーゼ阻害剤を含む一方で、LINCS3はエピジェネティック修飾因子を含む。スクリーニングの結果を表5に示す。
衛星細胞の単離および化学物質スクリーニング。
衛星細胞をインタクトな四肢筋肉から単離し、以前に記載されたとおり(Rocheteauら、2012年)、蛍光活性化細胞選別(FACS)のために調製した。FACSは、Harvard UniversityのBauer Flow Cytometry Core Facilityで、Beckman Coulter MoFlo Legacy(Beckman Coulter)により行われた。衛星細胞をエッペンドルフ管に選別し、手動で96ウェルプレートにプレーティングした(Greiner)。プレートを、PBS中で希釈された10ug/uLラミニンで37℃で4時間、予めコーティングした。選別された細胞を、0.05ng/mL塩基性線維芽細胞成長因子(bFGF、Life Technologies)を補充されたStemSpan SFEM
II基本培地(Stem Cell Technologies)中で培養した。陽性対照のためのウェルを600nM Jak3阻害剤IV(EMD Millipore)で処置した。スクリーニングおよび用量応答フォローアップのために、細胞を細胞150個/ウェルでプレーティングした。
細胞をプレーティングした日を、0日目とみなした。化合物を、1日目に添加した。スクリーニングおよび用量応答フォローアップのために、4日目まで培地を換えることなく、細胞をインキュベートし、プレートを画像化のために調製した。すべての化合物をジメチルスルホキシド(DMSO、Sigma Aldrich)に懸濁し;0.1%DMSOを、すべてのアッセイで陰性対照として添加した。
本発明の実施形態の例として以下の項目が挙げられる。
(項目1)
衛星細胞の増殖を増加させる方法であって、衛星細胞を、キナーゼ阻害剤、Gタンパク質共役受容体(GPCR)モジュレーター、ヒストンデアセチラーゼ(HDAC)モジュレーター、エピジェネティック修飾因子、ヘッジホッグシグナル伝達経路モジュレーター、神経ペプチド、ドーパミン受容体モジュレーター、セロトニン受容体モジュレーター、ヒスタミン受容体モジュレーター、アデノシン受容体アゴニスト、イオノフォア、イオンチャネルモジュレーター、ガンマ-セクレターゼモジュレーター、コルチコステロイド、およびその任意の組み合わせからなる群より選択される化合物と接触させるステップを含む、方法。
(項目2)
前記化合物が、小さな有機または無機分子;サッカリン;オリゴ糖;多糖;ペプチド、タンパク質、ペプチド類似体および誘導体;抗体、抗体断片、ペプチド模倣物質;核酸;核酸類似体および誘導体;生物材料から作製される抽出物;天然に存在する組成物または合成組成物;ならびにその任意の組み合わせからなる群より選択される、項目1に記載の方法。
(項目3)
前記化合物が、B-Raf阻害剤、JAK3阻害剤、p38MAPK阻害剤、C-Raf1阻害剤、Akt阻害剤、ERK阻害剤、BMK1/ERK5阻害剤、p38MAPK阻害剤、RTK阻害剤、ERK5阻害剤、Bcr-Abl阻害剤、RhoK阻害剤、p38阻害剤、p110阻害剤、FAK阻害剤、ATP競合JNK阻害剤、MELK阻害剤、表5に特定されている経路の阻害剤、Flt3キナーゼ阻害剤、PDGFR/EGFR阻害剤、Bcr-abl阻害剤、Jak3阻害剤、SRCキナーゼ阻害剤、HDAC1修飾因子、HDA31修飾因子、HDAC6修飾因子、BRD2修飾因子、BRD2修飾因子、EGLN1修飾因子、またはその誘導体、塩、代謝産物、プロドラッグ、もしくは立体異性体のうちの1種または複数である、項目1から2のいずれかに記載の方法。
(項目4)
前記化合物が、プロテインキナーゼ阻害剤および受容体キナーゼ阻害剤からなる群より選択される、項目1から3のいずれかに記載の方法。
(項目5)
前記化合物が、
(項目6)
前記化合物を、前記衛星細胞と約0.01nM~約100μMの濃度で接触させる、項目1から5のいずれかに記載の方法。
(項目7)
前記接触させるステップが少なくとも1時間にわたる、項目1から6のいずれかに記載の方法。
(項目8)
前記接触させるステップが1~7日間にわたる、項目1から7のいずれかに記載の方法。
(項目9)
前記接触がin vitroである、項目1から8のいずれかに記載の方法。
(項目10)
前記接触がex vivoである、項目1から9のいずれかに記載の方法。
(項目11)
前記接触がin vivoである、項目1から10のいずれかに記載の方法。
(項目12)
in vivoでの接触が哺乳動物における接触である、項目11に記載の方法。
(項目13)
in vivoでの接触がヒトにおける接触である、項目11または12に記載の方法。
(項目14)
前記in vivoでの接触が被験体における接触であり、前記被験体が損傷筋組織のための処置を必要としている、項目11から13のいずれかに記載の方法。
(項目15)
前記損傷筋組織が、物理的傷害または事故、疾患、感染、濫用、血液循環の喪失、または筋萎縮もしくは消耗の結果である、項目14に記載の方法。
(項目16)
前記損傷筋組織が、ジストロフィーの筋肉または老化している筋肉である、項目14または15に記載の方法。
(項目17)
前記損傷筋組織が筋萎縮/消耗の結果である、項目14から16のいずれかに記載の方法。
(項目18)
被験体において筋肉を修復または再生するための方法であって、治療有効量の化合物を被験体に投与するステップを含み、前記被験体が損傷筋組織を有し、前記化合物が、キナーゼ阻害剤、Gタンパク質共役受容体(GPCR)モジュレーター、ヒストンデアセチラーゼ(HDAC)モジュレーター、エピジェネティック修飾因子、ヘッジホッグシグナル伝達経路モジュレーター、神経ペプチド、ドーパミン受容体モジュレーター、セロトニン受容体モジュレーター、ヒスタミン受容体モジュレーター、イオノフォア、イオンチャネルモジュレーター、ガンマ-セクレターゼモジュレーター、およびその任意の組み合わせからなる群より選択される、方法。
(項目19)
前記損傷筋組織が、物理的傷害または事故、疾患、感染、濫用、血液循環の喪失、または筋萎縮もしくは消耗の結果である、項目18に記載の方法。
(項目20)
前記損傷筋組織が、ジストロフィーの筋肉または老化している筋肉である、項目18から19のいずれかに記載の方法。
(項目21)
前記損傷筋組織が筋萎縮/消耗の結果である、項目18から20のいずれかに記載の方法。
(項目22)
前記被験体が哺乳動物である、項目18から21のいずれかに記載の方法。
(項目23)
前記被験体がヒトである、項目18から22のいずれかに記載の方法。
(項目24)
前記化合物を治療剤と共投与する、項目18から23のいずれかに記載の方法。
(項目25)
前記化合物および前記治療剤を同じ製剤中で投与する、項目24に記載の方法。
(項目26)
前記化合物を1μg/kg~150mg/kgの投薬量で投与する、項目18から25のいずれかに記載の方法。
(項目27)
前記投与するステップが、注射、注入、点滴注入、吸入、または摂取による、項目18から26のいずれかに記載の方法。
(項目28)
前記投与するステップが1日1回である、項目18から27のいずれかに記載の方法。
(項目29)
前記被験体を筋修復または再生のために処置する前に、前記被験体を筋損傷または筋萎縮/消耗について診断するステップをさらに含む、項目18から28のいずれかに記載の方法。
(項目30)
前記化合物が、
(項目31)
衛星細胞の増殖を誘導する、刺激する、増強する、または増加させる化合物をスクリーニングするためのハイスループットアッセイであって、
(a)衛星細胞を試験化合物と接触させるステップと;
(b)衛星細胞の増殖を評価するステップと;
(c)衛星細胞の複製または成長を誘導する、刺激する、増強する、または増加させる化合物を選択するステップと
を含む、ハイスループットアッセイ。
(項目32)
衛星細胞の増殖を評価する前記ステップが、細胞マーカーを検出することを含む、項目31に記載のアッセイ。
(項目33)
前記細胞マーカーが、CXCR4、β1-インテグリン、Sca-1、Mac-1、CD45、PAX7、PAX3、Myf5、MyoD、デスミン、およびその任意の組み合わせからなる群より選択される、項目32に記載のアッセイ。
(項目34)
前記試験化合物が0.1nM~1000mMの範囲の濃度を有する、項目31から33のいずれかに記載のアッセイ。
(項目35)
前記アッセイを約15℃~約55℃の範囲の温度で行う、項目31から34のいずれかに記載のアッセイ。
(項目36)
前記試験化合物を、膵臓細胞と1時間から7日間にわたって接触させる、項目31から35のいずれかに記載のアッセイ。
(項目37)
前記試験化合物が、衛星細胞の増殖を、非処置対照に対して少なくとも5%、10%、20%、30%、40%、50%、50%、70%、80%、90%、1倍、1.1倍、1.5倍、2倍、3倍、4倍、5倍、10倍、50倍、100倍またはそれより多く増加させる、項目31から36のいずれかに記載のアッセイ。
(項目38)
前記衛星細胞を哺乳動物から単離する、項目31から37のいずれかに記載のアッセイ。
(項目39)
前記衛星細胞を被験体から単離し、前記被験体が損傷筋組織のための処置を必要としている、項目31から38のいずれかに記載のアッセイ。
(項目40)
前記損傷筋組織が、物理的傷害または事故、疾患、感染、濫用、血液循環の喪失、または筋萎縮もしくは消耗の結果である、項目39に記載の方法。
(項目41)
前記損傷筋組織が、ジストロフィーの筋肉または老化している筋肉である、項目39または40に記載の方法。
(項目42)
前記損傷筋組織が筋萎縮/消耗の結果である、項目39から41のいずれかに記載の方法。
Claims (15)
- 細胞の増殖を増加させる方法において使用するためのp38MAPK阻害剤を含む組成物であって、前記方法は、Pax7およびMyf5を発現する細胞を、SB239063(トランス-1-(4-ヒドロキシシクロヘキシル)-4-(4-フルオロフェニル)-5-(2-メトキシピリジミジン-4-イル)イミダゾール;HMSL10036-101-1)、ドラマピモド(BIRB796;HMSL10169-101-1)、SB203580(RWJ64809;PB203580;HMSL10167-101-1)、ネフラマピモド(VX-745;HMSL10168-101-1)、PH-797804(HMSL10439-101)、VX-702(HMSL10440-101)、SB202190(HMSL10441-101)、ラリメチニブ(LY2228820;HMSL10438-103)、ロスマピモド(GSK-AHAB;SB856553;GW856553X;HMSL10402-101)、およびその任意の組み合わせからなる群より選択される前記p38MAPK阻害剤と接触させることを含む、組成物。
- 前記p38MAPK阻害剤が、前記細胞と約0.01nM~約100μMの濃度で接触させられることを特徴とする、請求項1に記載の組成物。
- 前記接触が少なくとも1時間にわたる、請求項1に記載の組成物。
- 前記接触が1~7日間にわたる、請求項1に記載の組成物。
- 前記接触が少なくとも2週間にわたる、請求項1に記載の組成物。
- 前記接触が少なくとも3週間にわたる、請求項1に記載の組成物。
- 前記接触がin vitroである、請求項1に記載の組成物。
- 前記接触がex vivoである、請求項1に記載の組成物。
- 前記接触がin vivoである、請求項1に記載の組成物。
- in vivoでの接触が哺乳動物における接触である、請求項9に記載の組成物。
- in vivoでの接触がヒトにおける接触である、請求項10に記載の組成物。
- 前記in vivoでの接触が被験体における接触であり、前記被験体が損傷筋組織のための処置を必要としている、請求項1に記載の組成物。
- 前記損傷筋組織が、物理的傷害または事故、疾患、感染、濫用、血液循環の喪失、または筋萎縮もしくは消耗の結果である、請求項12に記載の組成物。
- 前記損傷筋組織が、ジストロフィーの筋肉または老化している筋肉である、請求項12に記載の組成物。
- 前記損傷筋組織が筋萎縮/消耗の結果である、請求項12に記載の組成物。
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US9782417B2 (en) | 2011-06-16 | 2017-10-10 | Presidents And Fellows Of Harvard College | Methods of increasing satellite cell proliferation with kinase inhibitors |
US11963964B2 (en) | 2011-06-16 | 2024-04-23 | President And Fellows Of Harvard College | Small molecules for mouse satellite cell proliferation |
US11026952B2 (en) | 2011-06-16 | 2021-06-08 | President And Fellows Of Harvard College | Small molecules for mouse satellite cell proliferation |
WO2019071147A1 (en) | 2017-10-05 | 2019-04-11 | Fulcrum Therapeutics, Inc. | INHIBITORS OF KINASE P38 REDUCING EXPRESSION OF DUX4 GENE AND DOWNSTREAM GENES FOR THE TREATMENT OF FSHD |
US10342786B2 (en) | 2017-10-05 | 2019-07-09 | Fulcrum Therapeutics, Inc. | P38 kinase inhibitors reduce DUX4 and downstream gene expression for the treatment of FSHD |
WO2019143743A1 (en) * | 2018-01-16 | 2019-07-25 | President And Fellows Of Harvard College | Methods and agents that enhance myogenic progenitor cell engraftment |
CN108949677B (zh) * | 2018-07-05 | 2021-11-30 | 浙江大学 | 地黄苷c和丹酚酸a在促进体外培养骨髓间充质干细胞增殖和抑制复制性衰老中的应用 |
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