JP2014500886A - 低親和性血液脳関門受容体抗体及びその使用 - Google Patents
低親和性血液脳関門受容体抗体及びその使用 Download PDFInfo
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Abstract
Description
本出願は、2010年11月30日に出願された米国仮出願第61/418,223号の利益を主張し、それは、参照によりその全体が本明細書に援用される。
本発明は、血液脳関門受容体(BBB−R)に結合する抗体及びその同一物を使用する方法に関する。
大きな分子の薬物の脳透過性は大部分が不透過性である血液脳関門(BBB)によって厳しく制限される。この障害を克服するための多くの戦略の中で、脳毛細血管内皮で発現される内因性受容体のトランスサイトーシストラフィッキング経路を利用することが挙げられる。モノクローナル抗体などの組換えタンパク質が、脳への大きな分子の受容体を介した送達を可能にするために、これらの受容体に対して設計されている。しかし、脳の取り込みを最大にしつつ、一方でもとの血液へ戻す逆トランスサイトーシスを最小限とする戦略、及び治療的投与後の蓄積の程度は未だ探求されぬままである。更に、BBBを通過する抗体が薬力学的に機能性であるかどうかは未知である。
定義
「血液脳関門」又は「BBB」は、脳毛細血管内皮原形質膜内の密着結合によって形成され、脳内への分子の輸送を、尿素(60ダルトン)などの非常に小さな分子さえも、制限する強固な障壁を作りだす末梢循環と脳及び脊髄の間の生理的障壁を指す。脳内の血液脳関門、脊髄内の血液脊髄関門、及び網膜内の血液網膜関門は、CNS内における連続的な毛細血管障壁であり、ここに総称して血液脳関門又はBBBと呼ばれる。BBBはまた、障壁が毛細血管内皮細胞よりむしろ上衣細胞で構成される、血液脳脊髄液関門を(脈絡叢)をも包含する。
本明細書における用語「抗体」は、最も広範な意味で用いられ、具体的には、モノクローナル抗体、ポリクローナル抗体、少なくとも二つのインタクトな抗体から形成される多重特異性抗体(たとえば二重特異性抗体)、所望の生物学的活性を示す限りにおいて抗体断片を網羅する。
ネイキッド抗体は、細胞傷害性部分、ポリマー、又は放射性標識などの異種分子にコンジュゲートされていない抗体(本明細書で定義される)である。
II.抗BBB−R抗体とそのコンジュゲートの産生
本発明の方法及び製造品は、BBB−Rに結合する抗体を用いるか又は組み込む。抗体の産生又はスクリーニングに使用されるBBB−R抗原は、所望のエピトープを含む可溶性形態またはその一部(例えば、細胞外ドメイン)であってもよい。あるいは、又は追加として、それらの細胞表面でBBB−Rを発現する細胞が、抗体を生成する又はスクリーニングするために使用することができる。抗体を生成するために有用なBBB−Rの他の形態は当業者には明らかであろう。本明細書におけるBBB−Rの別は、トランスフェリン受容体(TfR)、インスリン受容体、インスリン様増殖因子受容体(IGF−R)、低密度リポタンパク質受容体関連タンパク質1(LRP1)及びLRP8など、及びヘパリン結合上皮細胞増殖因子様増殖因子(HB−EGF)を含む。
本明細書に従って使用される抗体の治療製剤は、所望の程度の純度を有する抗体と任意の薬学的に許容される担体、賦形剤又は安定剤(Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed.: Williams and Wilkins PA, USA (1980))とを、凍結乾燥製剤または水性溶液の形態で混合することによって調製される。許容される担体、賦形剤又は安定剤は、使用される投薬量および濃度でレシピエントに毒性でなく、リン酸塩、クエン酸塩および他の有機酸のような緩衝液;アスコルビン酸およびメチオニンを含む抗酸化剤;防腐剤(例えば、オクタデシルジメチオルベンジルアンモニウムクロライド;ヘキサメトニウムクロライド;塩化ベンザルコニウム、塩化ベンゼトニウム、フェノール、ブチルまたはベンジルアルコール;アルキルパラベン、例えば、メチルまたはプロピルパラベン;カテコール;レゾルシノール;シクロヘキサノール;3−ペンタノール;およびm−クレゾール);低分子量(約10残基未満)ポリペプチド;タンパク質、例えば、血清アルブミン、ゼラチン、または免疫グロブリン;親水性ポリマー、例えば、ポリビニルピロリドン;アミノ酸、例えば、グリシン、グルタミン、アスパラギン、ヒスチジン、アルギニンまたはリジン;マンノサッカライド、ジサッカライド、およびグルコース、マンノースまたはデキストリンを含む他の炭水化物;キレート剤、例えば、EDTA;糖、例えば、スクロース、マンニトール、トレハロースまたはソルビトール;塩形成対イオン、例えば、ナトリウム、金属錯体(例えば、Zn−タンパク質錯体);及び/又はTWEEN、PLURONICS又はポリエチレングリコール(PEG)等の非イオン性界面活性剤が挙げられる。
抗BBB−R抗体(それらを構成する多重特異性抗体を含む)は、様々なインビトロの方法で利用することができる。例えば、本発明は、抗体がBBBを通過してそれに結合した治療化合物を輸送するように、治療化合物に結合した抗BBB−R抗体(BBB−Rと脳抗原の両方を結合する多重特異性抗体など)をBBBへ曝露することを含む、血液脳関門を通過する治療用化合物を輸送する方法を提供する。別の実施例において、本発明は、抗体がBBBを通過してそれに結合した神経障害薬を輸送するように、神経障害薬に結合した抗BBB−R抗体(BBB−Rと脳抗原の両方を結合する多重特異性抗体など)をBBBへ曝露することを含む、血液脳関門を通過する神経障害薬を輸送する方法を提供する。一実施態様において、本明細書においてBBBは、哺乳動物(例えばヒト)、例えば神経障害を有するものであり、限定されないが、アルツハイマー病(AD)、脳卒中、認知症、筋ジストロフィー(MD)、多発性硬化症(MS)、筋萎縮性側索硬化症(ALS)、嚢胞性線維症、アンジェルマン症候群、リドル症候群、パーキンソン病、ピック病、パジェット病、癌、外傷性脳損傷などを含む。
本発明の他の実施態様において、上述した障害の治療、予防、及び/又は診断に有用な物質を含む製造品が提供される。製造品は、容器とラベルまたは容器上にあるまたは容器に付属するパッケージ挿入物を含む。好適な容器は、例としてボトル、バイアル、シリンジ、IV輸液バッグ等を含む。容器はガラス又はプラスチックなどの様々な物質から形成されうる。容器は、疾患の治療、予防、及び/又は診断に有効である、それ自体か、又はその他の組成物と併用される化合物を収容し、無菌のアクセスポートを有し得る(例えば、容器は皮下注射針による穴あきストッパーを有する静脈内溶液バッグ又はバイアルであってよい)。組成物中の少なくとも一の活性剤は本発明の抗体である。ラベルまたはパッケージ挿入物は、組成物が特定の症状の治療のために使用されることを示している。更に、製造品は、(a)組成物が本発明の抗体を包含する組成物を含む第一の容器;および(b)組成物が更なる細胞障害性又はその他の治療的薬剤を包含する組成物を含む第2の容器を含み得る。本発明の本実施態様における製造品は、組成物が特定の疾患を治療することに用いることができることを示すパッケージ挿入物をさらに含んでいてもよい。別法として、または加えて、製造品は、薬学的に許容されるバッファー、例えば注射用静菌水(BWFI)、リン酸緩衝化塩水、リンガー溶液およびデキストロース溶液を含む第二(または第三)の容器をさらに含んでもよい。これは、他のバッファー、希釈剤、フィルター、針、およびシリンジを含む、商業的およびユーザーの立場から望まれる他の物質のさらに含んでもよい。
Claims (55)
- 抗体がそれに結合した化合物を血液脳関門を通過して輸送するように、化合物に結合して血液脳関門受容体(BBB−R)に低親和性で結合する抗体を血液脳関門に曝すことを含む、血液脳関門を横切る化合物を輸送する方法。
- 化合物へのCNSの曝露を増やす方法であって、該化合物がBBB−Rに低親和性で結合する抗体に結合し、これによってCNSの化合物への曝露を増加させる方法。
- 被験体に投与された化合物のクリアランスを減らす方法であって、該化合物がBBB−Rに低親和性で結合する抗体に結合し、これによって化合物のクリアランスが減少する方法。
- 被験体に投与された化合物のCNSにおける保持を増やす方法であって、該化合物がBBB−Rに低親和性で結合する抗体に結合し、これによって化合物のCNSにおける保持が増加する方法。
- 被検体のCNSにおいて有効であるべき化合物の薬物動態及び/又は薬力学を最適化する方法であって、該化合物は、BBB−Rに低親和性で結合する抗体に結合されており、その抗体は、化合物に結合後にBBB−Rに対するその親和性が、BBBを通過する化合物に結合した抗体の搬送量を生じるように選択され、CNSにおける化合物の薬物動態及び/又は薬力学を最適化する方法。
- BBB−Rに結合し、かつ化合物に結合している抗体により哺乳動物を治療することを含む哺乳動物における神経障害を治療する方法であって、該抗体は、BBB−Rに対して低い親和性を有するように選択され、それによって抗体及び結合した化合物のCNSへの取り込みを改善する方法。
- 化合物が神経障害薬又は造影剤である、請求項1から6の何れかに記載の方法。
- 増加又は減少がBBB−Rに対する低下した親和性を持たない典型的抗体に対して測定される、請求項2から4の何れかに記載の方法。
- 抗体が一以上のその天然リガンドに対するBBB−Rの結合を阻害しない、請求項1から6の何れかに記載の方法。
- 血液脳関門が哺乳動物にある、請求項1から6の何れかに記載の方法。
- 哺乳動物が神経障害を有する、請求項10に記載の方法。
- 神経障害が、アルツハイマー病(AD)、脳卒中、認知症、筋ジストロフィー(MD)、多発性硬化症(MS)、筋萎縮性側索硬化症(ALS)、嚢胞性線維症、アンジェルマン症候群、リドル症候群、パーキンソン病、ピック病、パジェット病、癌、および外傷性脳損傷からなる群から選択される、請求項11に記載の方法。
- 哺乳動物がヒトである、請求項10に記載の方法。
- 抗体が、約1nMから約100μMのBBB−Rに対するIC50を有する、請求項1から6の何れかに記載の方法。
- IC50が約5nMから約100μMである、請求項8に記載の方法。
- IC50が約50nMから約100μMである、請求項8に記載の方法。
- IC50が約100nMから約100μMである、請求項8に記載の方法。
- 抗体が、約5nMから約10μMのBBB−Rに対する親和性を有する、請求項1から6の何れかに記載の方法。
- 化合物に結合した抗体が、約30nMから約1μMのBBB−Rに対する親和性を有する、請求項1から6の何れかに記載の方法。
- BBB−Rが、トランスフェリン受容体(TfR)、インスリン受容体、インスリン様増殖因子受容体(IGF受容体)、低密度リポタンパク質受容体関連タンパク質8(LRP8)、低密度リポタンパク質受容体関連タンパク質1(LRP1)、及びヘパリン結合上皮細胞増殖因子様増殖因子(HB−EGF)からなる群から選択される、請求項1から6の何れかに記載の方法。
- BBB−Rがトランスフェリン受容体である、請求項20に記載の方法。
- 抗体がトランスフェリンに対するTfRの結合を阻害しない、請求項21に記載の方法。
- 化合物に結合した抗体が治療的投与量で投与される、請求項1から6の何れかに記載の方法。
- 治療的投与量がBBB−R飽和である、請求項23に記載の方法。
- 抗体が多重特異性抗体であり、化合物が多重特異性抗体の一部を任意で形成する、請求項1から6の何れかに記載の方法。
- 多重特異抗体がBBB−Rに結合する第一の抗原結合部位と脳の抗原に結合する第二の抗原結合部位を含む、請求項25に記載の方法。
- 脳の抗原が、β−セクレターゼ1(BACE1)、Abeta、上皮増殖因子受容体(EGFR)、ヒト上皮増殖因子受容体2(HER2)、タウ、アポリポタンパク質E4(ApoE4)、α−シヌクレイン、CD20、ハンチンチン、プリオンタンパク質(PrP)、ロイシンリッチリピートキナーゼ2(LRRK2)、パーキン、プレセニリン1、プレセニリン2、ガンマセクレターゼ、細胞死受容体6(DR6)、アミロイド前駆体タンパク質(APP)、p75ニューロトロフィン受容体(p75NTR)、及びカスパーゼ6からなる群から選択される、請求項26に記載の方法。
- 多重特異性抗体がTfRとBACE1の両方に結合する、請求項26に記載の方法。
- 多重特異性抗体がTfRとAbetaの両方に結合する、請求項26に記載の方法。
- 抗体がBBB−Rに対して望ましく低い親和性を有するため、血液脳関門受容体(BBB−R)に特異的な抗体を選択することを含む、BBBを通過して化合物を輸送するために有用な抗体を作成する方法。
- BBB−Rに対する親和性が約5nMから約10μMである、請求項30に記載の方法。
- 抗体が、約1nMから約100μMのBBB−Rに対するIC50を有する、請求項30に記載の方法。
- 抗体が、選択された抗体の親和性に基づいて抗体のパネルから選択される、請求項30に記載の方法。
- 抗体が親和性を有するように操作される、請求項30に記載の方法。
- 抗体を治療用化合物と結合させることを含む、請求項30に記載の方法。
- 治療用化合物が神経障害薬である、請求項35に記載の方法。
- BBB−Rに結合する第一の抗原結合部位と脳の抗原に結合する第二の抗原結合部位を含む多重特異性抗体を作成することを含む、請求項30に記載の方法。
- BBB−Rに対する抗体の親和性が、約5nMから約10μMである、BBB−Rに結合する抗体。
- 親和性が約20nMから約1μMである、請求項38に記載の抗体。
- BBB−Rが、トランスフェリン受容体(TfR)、インスリン受容体、インスリン様増殖因子受容体(IGF受容体)、低密度リポタンパク質受容体関連タンパク質8(LRP8)、低密度リポタンパク質受容体関連タンパク質38(LRP1)、及びヘパリン結合上皮細胞増殖因子様増殖因子(HB−EGF)からなる群から選択される、請求項38に記載の抗体。
- BBB−Rがトランスフェリン受容体(TfR)である、請求項40に記載の抗体。
- BBB−Rがインスリン受容体である、請求項40に記載の抗体。
- BBB−RがヒトBBB−Rである、請求項38に記載の抗体。
- 神経障害薬と結合した、請求項38に記載の抗体。
- BBB−Rに結合する第一の抗原結合部位と脳の抗原に結合する第二の抗原結合部位を含む多重特異性抗体である、請求項44に記載の抗体。
- 脳の抗原が、β−セクレターゼ1(BACE1)、Abeta、上皮増殖因子受容体(EGFR)、ヒト上皮増殖因子受容体2(HER2)、タウ、アポリポタンパク質E4(ApoE4)、α−シヌクレイン、CD20、ハンチンチン、プリオンタンパク質(PrP)、ロイシンリッチリピートキナーゼ2(LRRK2)、パーキン、プレセニリン1、プレセニリン2、ガンマセクレターゼ、細胞死受容体6(DR6)、アミロイド前駆体タンパク質(APP)、p75ニューロトロフィン受容体(p75NTR)、及びカスパーゼ6からなる群から選択される、請求項45に記載の抗体。
- 脳の抗原がBACE1である、請求項46に記載の抗体。
- 脳の抗原がAbetaである、請求項46に記載の抗体。
- BBB−Rに結合する抗原結合領域を持つ抗体断片である、請求項38に記載の抗体。
- Fab断片である、請求項49に記載の抗体断片。
- 完全長抗体又は抗体断片である、請求項45に記載の多重特異性抗体。
- 神経障害を治療するための医薬の製造のために、BBB−Rに対して低親和性で結合する抗体の使用。
- 神経障害を治療するための医薬の製造のための、請求項38から51の何れかに記載の抗体の使用。
- 神経障害の治療において使用のための、BBB−Rに対して低親和性で結合する抗体。
- 神経障害の治療において使用のための、請求項38から51の何れかに記載の抗体。
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