JP7297734B2 - 可変ドメインと定常ドメインとの間のエルボー領域に機能的ドメインを有する抗体 - Google Patents
可変ドメインと定常ドメインとの間のエルボー領域に機能的ドメインを有する抗体 Download PDFInfo
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Description
第1の態様において、本発明は、第1のポリペプチド鎖と第2のポリペプチド鎖とを含む抗体又はその抗原結合断片であって、
前記第1のポリペプチド鎖は、NからC末端方向に、
(i)1つ以上の可変ドメイン;
(ii)(追加の)機能的ドメイン;及び
(iii)1つ以上の定常ドメインを含み、
前記第2のポリペプチド鎖は、NからC末端方向に、
(iv)前記第1のポリペプチド鎖の前記1つ以上の可変ドメイン(i)と抗原結合部位を形成する1つ以上の可変ドメイン;
(v)任意に、(追加の)機能的ドメイン;及び
(vi)1つ以上の定常ドメインを含み、
前記第1のポリペプチド鎖の前記(追加の)機能的ドメイン(ii)が前記第2のポリペプチド鎖の断片を含まず、前記第2のポリペプチド鎖の前記任意の(追加の)機能的ドメイン(v)が前記第1のポリペプチド鎖の断片を含まない抗体又はその抗原結合断片を提供する。
タ1糖タンパク質11)、PSG11s’(妊娠特異的ベータ1糖タンパク質11s’)、PTGFRN(プロスタグランジンF2受容体阻害剤)、PTK7(タンパク質チロシンキナーゼ7(不活性))、PTPRD(タンパク質チロシンホスファターゼ、受容体タイプD)、PTPRK(タンパク質チロシンホスファターゼ、受容体タイプK)、PTPRM(タンパク質チロシンホスファターゼ、受容体タイプM)、PTPRSタンパク質チロシンホスファターゼ、受容体タイプS)、PTPRT(タンパク質チロシンホスファターゼ、受容体タイプT)、PTPシグマ、PUNC、PVR(ポリオウイルス受容体)、PVRL1、PVRL2、PVRL4、NECTIN1(ネクチン細胞接着分子1)、NECTIN2(ネクチン細胞接着分子2)、NECTIN3(ネクチン細胞接着分子3)、RAGE、ROBO3(ラウンドアバウト誘導受容体3)、SCN1B(ナトリウム電圧ゲートチャネルベータサブユニット1)、SDK1(サイドキック細胞接着分子1)、SDK2(サイドキック細胞接着分子2)、SEMA3A(セマフォリン3A)、SEMA3B(セマフォリン3B)、SEMA3E(セマフォリン3E)、SEMA3F(セマフォリン3F)、SEMA3G(セマフォリン3G)、SEMA4C(セマフォリン4C)、SEMA4D(セマフォリン4D)、SEMA4G(セマフォリン4G)、SEMA7A 8セマフォリン7A(ジョンミルトンハーゲン血液型))、SIGIRR(単一IgとTIRドメイン含有)、SIGLEC1(シアル酸結合Ig様レクチン1)、SIGLEC5(シアル酸結合Ig様レクチン5)、SIGLEC6(シアル酸結合Ig様レクチン6)、SIGLEC7(シアル酸結合Ig様レクチン7)、SIGLEC8(シアル酸結合Ig様レクチン8)、SIGLEC9(シアル酸結合Ig様レクチン9)、SIGLEC10(シアル酸結合Ig様レクチン10)、SIGLEC11(シアル酸結合Ig様レクチン11)、SIGLEC12(シアル酸結合Ig様レクチン12)(遺伝子/偽遺伝子))、SIGLEC14(シアル酸結合Ig様レクチン14)、SIGLEC15(シアル酸結合Ig様レクチン15)、SLAMF1(シグナル伝達リンパ球活性化分子ファミリーメンバー1)、SLAMF6(SLAMファミリーメンバー6)、SLAMF8(SLAMファミリーメンバー8)、SIRPG;TARM1(T細胞相互作用骨髄細胞活性化受容体1)、TEK(TEK受容体チロシンキナーゼ)、THY1 Thy-1細胞表面抗原)、TIE1(免疫グロブリン様及びEGF様ドメイン1を有するチロシンキナーゼ)、TMEM81(膜貫通タンパク質81)、TMIGD1(膜貫通及び免疫グロブリンドメイン含有1)、TMIGD2(膜貫通及び免疫グロブリンドメイン含有2)、TTN(チチン)、TYRO3(TYRO3タンパク質チロシンキナーゼ)、UNC5D、VCAM1(血管細胞接着分子1)、VSIG1(Vセット及び免疫グロブリンドメイン含有1)、VSIG2(Vセット及び免疫グロブリンドメイン含有2)、VSIG4(Vセット及び免疫グロブリンドメイン含有4)、VSIG10(Vセット及び免疫グロブリンドメイン含有10)、VSIG10L(Vセット及び免疫グロブリンドメイン含有10様)、VSTM1(Vセット及び膜貫通ドメイン含有1)、VTCN1(Vセットドメイン含有T細胞活性化阻害剤1)、ZPBP(透明帯結合タンパク質)、又はZPBP2(透明帯結合タンパク質2)。
前記第1のポリペプチド鎖が、NからC末端方向に、
(i)1つ以上の可変ドメイン;
(ii)(追加の)機能的ドメイン;及び
(iii)1つ以上の定常ドメインを含み、
前記第2のポリペプチド鎖が、NからC末端方向に、
(iv)第1のポリペプチド鎖の1つ以上の可変ドメインと抗原結合部位を形成する1つ以上の可変ドメイン;及び
(v)1つ以上の定常ドメインを含み、
前記第2のポリペプチド鎖の最もC末端側の可変ドメインのC末端が、第2のポリペプチド鎖の最もN末端側の定常ドメインのN末端に直接結合する抗体又はその抗原結合断片を提供する。
Vは、可変ドメイン(i)であり、
Dは、(追加の)機能的ドメイン(ii)であり、
CH1は、CH1定常ドメイン(iii)である。
V-D-CH1は、前記したとおりであり、
CH2とCH3はそれぞれ、CH2定常ドメインとCH3定常ドメインである。
Aは、1~5、好ましくは1~4、より好ましくは1~3、更により好ましくは1又は2の整数であり、
可変ドメインVは、直接又はリンカーを介して互いに結合されてもよい。
Vは、可変ドメイン(iv)であり、
CLは定常ドメイン(vi)である。
V及びCLは、前記したようにリンカーを介して連結されてもよく、又は互いに直接連結されてもよい。
V-CLは、前記した通りであり、
Aは、1~5、好ましくは1~4、より好ましくは1~3、更により好ましくは1又は2の整数であり、
可変ドメインVは、直接又はリンカーを介して互いに結合されてもよい。
別の態様では、本発明はまた、本発明に係る抗体又はその抗原結合断片の第1のポリペプチド鎖をコードする第1のポリヌクレオチド、及び/又は本発明に係る抗体又はその抗原結合断片の第2のポリペプチド鎖をコードする第2のポリペプチドを含む核酸分子を提供する。
更に、本発明に係る核酸分子を含むベクター、例えば発現ベクターを提供も本発明の範囲に含まれる。好ましくは、ベクターは、前記の核酸分子を含む。
更なる態様では、本発明は、また、本発明に係る抗体又はその抗原結合断片を発現する、及び/又は本発明に係る核酸分子又はベクターを含む細胞を提供する。
本発明はまた、以下のうちの1つ以上を含む組成物を提供する。
(i)本発明に係る抗体又はその抗体断片;
(ii)本発明に係る核酸分子;
(iii)本発明に係る核酸を含むベクター;及び/又は
(iv)本発明に係る抗体を発現する、又は本発明に係るベクター又は核酸分子を含む細胞。
本発明に係る抗体は、当技術分野において公知の任意の方法によって作製することができる。例えば、足場抗体を提供し、エルボー領域で遺伝子操作することができる。例えば、足場抗体を得るために、例えば、ハイブリドーマ技術を使用してモノクローナル抗体を作製するための一般的な方法論は、周知である(Kohler,G.and Milstein,C,.1975;Kozbar et al.1983)。特に、国際公開第2004/076677号に記載される代替のEBV不死化方法を使用することができる。
第1のポリペプチド鎖と第2のポリペプチド鎖をコードする前記した1つ以上の核酸分子、例えば、前記した本発明に係るベクターを、(例えば、前記のように)組み込むことにより真核宿主細胞を形質転換すること;
前記核酸分子が発現されるように適切な条件下で宿主細胞を培養すること;
前記第1及び第2のポリペプチド鎖を組み合わせて、抗体又はその抗原結合断片を形成すること;及び
任意に、培養培地から前記抗体又はその抗原結合断片を精製すること。
更なる態様において、本発明は、医学における使用のための、本発明に係る抗体又はその抗原結合断片、本発明に係る核酸、本発明に係るベクター、本発明に係る細胞、又は本発明に係る(医薬)組成物の使用を提供する。
抗原の多価抗体への結合を可能にする条件下で、本発明に係る抗体又はその抗原結合断片と共に前記抗原をインキュベートすること;及び
抗原抗体結合を検出すること。
特に定義しない限り、本明細書で使用する全ての技術用語及び科学用語は、本発明が属する分野の当業者に一般的に理解されている意味と同じ意味を有する。本明細書に記載するものと類似又は等価な方法及び材料を本発明の実施又は試験で用いることができるが、好適な方法及び材料を以下に記載する。本明細書に記載する全ての刊行物、特許出願、特許、及び他の参照文献は、その全体が参照によって援用される。矛盾する場合、定義を含む本明細書が優先する。更に、材料、方法、及び実施例は、単なる例示であり、限定を意図するものではない。
抗体の特異性に対する抗体のエルボー領域に挿入された異なる(追加の)機能的ドメインの効果を調べるために、非変異LAIR1(配列番号12)、変異LAIR1(配列番号13)、又は他のIg様ドメインを足場として使用した抗体のエルボー領域に挿入した7種の構築物(「C2~C8」と命名)を設計した。構築物C2~C3は、構築物C1と同一の完全な重鎖定常領域を有する(VH:配列番号5、VL:配列番号6、重鎖定常領域:配列番号3、軽鎖定常領域:配列番号7)。構築物C4~C6は、抗体GCE536と同一の完全な重鎖定常領域を有する(VH:配列番号1、VL:配列番号2、重鎖定常領域:配列番号3、カッパ軽鎖定常領域:配列番号:4;Piccoli, L., et al. Neutralization and clearance of GM-CSF by autoantibodies in pulmonary alveolar proteinosis. Nature communications 6, 7375 (2015))。構築物C7~C8は、構築物C1bと同一の完全な重鎖定常領域を有する(VH:配列番号8、VL:配列番号9、重鎖定常領域:配列番号3、軽鎖定常領域:配列番号4)。構築物の軽鎖は、足場抗体と比較して改変しなかった。いずれの構築物も、最終的にモノクローナル抗体(重鎖及び軽鎖)として発現させた。
1.「C1」(VH:配列番号5、VL:配列番号6、重鎖定常領域:配列番号3、軽鎖ラムダ定常領域:配列番号7)は、対照目的の組換え単一特異性抗体である。C1は、以下によって(NからC末端にこの順で)形成される:重鎖可変ドメインのV(可変)遺伝子セグメントの発現産物(「VH3-30」);重鎖可変ドメインのD(多様性)遺伝子セグメントの発現産物(「D」);重鎖可変ドメインのJ(結合)遺伝子セグメントエレメントの発現産物(「JH6」);重鎖定常領域のC(定常)遺伝子セグメントの発現産物(IgG1アイソタイプ);及び別の鎖上に、軽鎖可変ドメインのV(可変)遺伝子セグメントの発現産物及び軽鎖可変ドメインのJ(結合)遺伝子セグメントエレメントの発現産物;軽鎖ラムダ定常領域のC(定常)遺伝子セグメントの発現産物。
実施例1に記載した8つの抗体構築物を、一過性トランスフェクションによる組換えにより産生した。この目的のために、抗体の重鎖及び軽鎖をヒトIgG1、Igκ、及びIgλ発現ベクターにクローニングし、ポリエチレンイミン(PEI)を使用したExpi293F細胞(ThermoFisher Scientific)の一過性トランスフェクションにより発現させた。細胞株は、マイコプラズマ汚染について汎用法により試験した。
エルボー領域に結合部位を有する二重特異性構築物が両方の特異性で同時に結合できるかどうかを試験するために、同時結合を表面プラズモン共鳴(SPR)で調べた。この目的のために、GM-CSFに特異的なV(D)J領域を有し、エルボーに非変異LAIR1(コラーゲンの結合部位)を有する二重特異性構築物C4を、表面プラズモン共鳴(SPR)によって、GM-CSFとコラーゲンへの同時結合について試験した。
抗体構築物C5b及びC6bは、C5及びC6構築物のエルボーに挿入された同一ドメイン(PD1又はSLAM)を含み、JHとドメインとの間及びドメインとCH1との間に2つの追加15-merアミノ酸リンカーが挿入されている。構築物を、ELISAにより組換えヒトGM-CSF、抗PD1、及び抗SLAM抗体への結合について試験した。簡潔に説明すると、Certified Reference Material 470(ERMs-DA470、Sigma-Aldrich)を標準として使用し、ヤギ抗ヒトIgG(SouthernBiotech、2040-01)でコーティングした96ウェルMaxiSorpプレート(Nunc)を使用して、総IgGを定量した。抗体構築物の特異的結合を試験するために、ELISAプレートを1μgml-1の組換えヒトGM-CSF(Gentaur)、2μgml-1の抗PD1又は抗SLAM抗体(R&D Systems、AF1086及びAF164)でコーティングした。プレートを1%ウシ血清アルブミン(BSA)でブロッキングし、滴定抗体と共にインキュベートした後、APコンジュゲートヤギ抗ヒトIgG、Fcγ断片特異的(Jackson Immuno Research、109-056-098)とインキュベートした。次いで、プレートを洗浄し、基質(p-NPP、Sigma)を加え、プレートを405nmで読み取った。
抗体検出のために抗体のエルボー領域に挿入されたタグを使用する可能性について調べるために、ツインStrepタグを足場抗体C1b(実施例1参照)の重鎖のエルボー領域に挿入した1つの構築物(「C9」と名付ける)。構築物の軽鎖は、足場抗体と比較して変更しなかった。構築物は、最終的にモノクローナル抗体(重鎖及び軽鎖)として発現した。
実施例5に記載した抗体構築物「C9」を、一過性トランスフェクションによる組換えにより産生した。この目的のために、抗体の重鎖及び軽鎖をヒトIgG1及びIgκ発現ベクターにクローニングし、ポリエチレンイミン(PEI)を使用したExpi293F細胞(ThermoFisher Scientific)の一過性トランスフェクションにより発現させた。細胞株は、マイコプラズマ汚染について汎用法により試験した。
Ig様ドメイン以外の(追加の)機能的ドメインのエルボー領域への挿入が機能的多重特異性抗体をもたらすかどうかを調べるために、4種の新たな構築物を設計した(C9~C12)。4種の新たな構築物C9~C12では、破傷風トキソイド(TT)又は呼吸器合胞体ウイルス(RSV)の融合(F)タンパク質に特異的な、単鎖抗体可変ドメイン(VHH)又は単鎖可変断片(ScFv)それぞれを、インフルエンザH1ヘマグルチニンに特異的な抗体(FI174;Pappas, L., et al. Rapid development of broadly influenza neutralizing antibodies through redundant mutations. Nature 516, 418-422 (2014);VH:配列番号10、VL:配列番号11、重鎖定常領域:配列番号3、軽鎖カッパ定常領域:配列番号12/配列番号4)のエルボー領域に挿入した。構築物の軽鎖は、足場抗体FI174と比較して変更しなかった。いずれもの築物も、最終的にモノクローナル抗体(重鎖及び軽鎖)として発現させた。
1.構築物「C10」では、抗TT VHH(「T3-VHH」;Rossotti, M.A., et al. Increasing the potency of neutralizing single-domain antibodies by functionalization with a CD11b/CD18 binding domain. mAbs 7, 820-828 (2015);配列番号16)をFI174のエルボー領域に挿入した。構築物「C10」(完全な重鎖:配列番号66、完全な軽鎖:配列番号67)は、以下によって(NからC末端にこの順で)形成される:FI174の重鎖可変ドメインのV(可変)遺伝子セグメントの発現産物(「VH」);FI174の重鎖可変ドメインのD(多様性)遺伝子セグメントの発現産物(「D」);FI174の重鎖可変ドメインのJ(結合)遺伝子セグメントエレメントの発現産物(「JH」);抗TT VHH(「T3-VHH」;Rossotti, M.A., et al. Increasing the potency of neutralizing single-domain antibodies by functionalization with a CD11b/CD18 binding domain. mAbs 7, 820-828 (2015)の発現産物;重鎖定常領域のC(定常)遺伝子セグメントの発現産物(IgG1アイソタイプ);及び別の鎖上に、FI174の軽鎖可変ドメインのV(可変)遺伝子セグメントの発現産物及びFI174の軽鎖可変ドメインのJ(結合)遺伝子セグメントエレメントの発現産物;軽鎖定常領域のC(定常)遺伝子セグメントの発現産物。
実施例4に記載した4種の新たな抗体構築物を、一過性トランスフェクションにより組換えにより産生した。この目的のために、抗体の重鎖及び軽鎖をヒトIgG1、Igκ、及びIgλ発現ベクターにクローニングし、ポリエチレンイミン(PEI)を使用したExpi293F細胞(ThermoFisher Scientific)の一過性トランスフェクションにより発現させた。構築物を、表面プラズモン共鳴(SPR)により、(i)H1及び(ii)TT又はRSV Fタンパク質への二重の結合について試験した。
Claims (26)
- 第1のポリペプチド鎖と第2のポリペプチド鎖とを含む抗体又はその抗原結合断片であって、
前記第1のポリペプチド鎖が、NからC末端方向に、
(i)1つ以上の可変ドメイン;
(ii)機能的ドメイン;及び
(iii)1つ以上の定常ドメインを含み、
前記第2のポリペプチド鎖が、NからC末端方向に、
(iv)前記第1のポリペプチド鎖の前記1つ以上の可変ドメインと抗原結合部位を形成する1つ以上の可変ドメイン;
(v)機能的ドメイン;及び
(vi)1つ以上の定常ドメインを含み、
前記第1のポリペプチド鎖の前記機能的ドメイン(ii)が前記第2のポリペプチド鎖の断片を含まず、前記第2のポリペプチド鎖の前記機能的ドメイン(v)が前記第1のポリペプチド鎖の断片を含まず;前記第2のポリペプチド鎖又はその断片は、前記第1のポリペプチド鎖の前記機能的ドメイン(ii)の機能性に必要ではなく、関与もせず、前記第1のポリペプチド鎖又はその断片は、前記第2のポリペプチド鎖の前記機能的ドメイン(v)の機能性に必要ではなく、関与もせず;前記第1のポリペプチド鎖の前記機能的ドメイン(ii)の機能性に必要な又は関与する全てのアミノ酸は、前記第1のポリペプチド鎖自体の前記機能的ドメイン(ii)に含まれ、前記第2のポリペプチド鎖の前記機能的ドメイン(v)の機能性に必要な又は関与する全てのアミノ酸は、前記第2のポリペプチド鎖自体の前記機能的ドメイン(v)に含まれ、
(a)前記第1のポリペプチド鎖の前記1つ以上の定常ドメインが少なくともCH1定常ドメインを含む重鎖定常ドメインであり、前記第1のポリペプチド鎖の前記1つ以上の可変ドメインが重鎖可変ドメインVHであり、前記第2のポリペプチド鎖の前記定常ドメインが軽鎖定常ドメインCLであり、前記第2のポリペプチド鎖の前記1つ以上の可変ドメインが軽鎖可変ドメインVLである、又は
(b)前記第1のポリペプチド鎖の前記定常ドメインが軽鎖定常ドメインCLであり、前記第1のポリペプチド鎖の前記1つ以上の可変ドメインが軽鎖可変ドメインVLであり、前記第2のポリペプチド鎖の前記1つ以上の定常ドメインが少なくともCH1定常ドメインを含む重鎖定常ドメインであり、前記第2のポリペプチド鎖の前記1つ以上の可変ドメインが重鎖可変ドメインVHであり;
前記第1のポリペプチド鎖の前記機能的ドメイン(ii)及び前記第2のポリペプチド鎖の前記機能的ドメイン(v)が、独立して下記を含む又はからなることを特徴とする抗体又はその抗原結合断片:
- キャリアドメイン;
- レポータードメイン;
- タグ;
- 局在化ドメイン;
- (独立した)結合部位;
- 酵素若しくは酵素ドメイン;
- 受容体若しくはその機能的断片;又は
- リガンド若しくはその機能的断片。 - 第1のポリペプチド鎖と第2のポリペプチド鎖とを含む抗体又はその抗原結合断片であって、
前記第1のポリペプチド鎖が、NからC末端方向に、
(i)1つ以上の可変ドメイン;
(ii)機能的ドメイン;及び
(iii)1つ以上の定常ドメインを含み、
前記第2のポリペプチド鎖が、NからC末端方向に、
(iv)前記第1のポリペプチド鎖の前記1つ以上の可変ドメインと抗原結合部位を形成する1つ以上の可変ドメイン;及び
(vi)1つ以上の定常ドメインを含み、
前記第2のポリペプチド鎖が機能的ドメイン(v)を含まず、前記第2のポリペプチド鎖の最もC末端側の可変ドメインのC末端が前記第2のポリペプチド鎖の最もN末端側の定常ドメインのN末端に直接結合しており、
前記第1のポリペプチド鎖の前記機能的ドメイン(ii)が前記第2のポリペプチド鎖の断片を含まず;前記第2のポリペプチド鎖又はその断片は、前記第1のポリペプチド鎖の前記機能的ドメイン(ii)の機能性に必要ではなく、関与もせず;前記第1のポリペプチド鎖の前記機能的ドメイン(ii)の機能性に必要な又は関与する全てのアミノ酸は、前記第1のポリペプチド鎖自体の前記機能的ドメイン(ii)に含まれ、
(a)前記第1のポリペプチド鎖の前記1つ以上の定常ドメインが少なくともCH1定常ドメインを含む重鎖定常ドメインであり、前記第1のポリペプチド鎖の前記1つ以上の可変ドメインが重鎖可変ドメインVHであり、前記第2のポリペプチド鎖の前記定常ドメインが軽鎖定常ドメインCLであり、前記第2のポリペプチド鎖の前記1つ以上の可変ドメインが軽鎖可変ドメインVLである、又は
(b)前記第1のポリペプチド鎖の前記定常ドメインが軽鎖定常ドメインCLであり、前記第1のポリペプチド鎖の前記1つ以上の可変ドメインが軽鎖可変ドメインVLであり、前記第2のポリペプチド鎖の前記1つ以上の定常ドメインが少なくともCH1定常ドメインを含む重鎖定常ドメインであり、前記第2のポリペプチド鎖の前記1つ以上の可変ドメインが重鎖可変ドメインVHであり;
前記第1のポリペプチド鎖の前記機能的ドメイン(ii)が、下記を含む又はからなることを特徴とする抗体又はその抗原結合断片:
- キャリアドメイン;
- レポータードメイン;
- タグ;
- 局在化ドメイン;
- (独立した)結合部位;
- 酵素若しくは酵素ドメイン;
- 受容体若しくはその機能的断片;又は
- リガンド若しくはその機能的断片。 - 前記CH1ドメインが、以下のクラス:γ、α、μ、ε、及びδから選択される;及び/又は
前記CLドメインが、以下のクラス:κ及びλから選択される請求項1から2のいずれかに記載の抗体又はその抗原結合断片。 - 前記第1のポリペプチド鎖の前記機能的ドメイン(ii)及び/又は前記第2のポリペプチド鎖の前記機能的ドメイン(v)が、Ig様ドメインを含む又はからなる請求項1から3のいずれかに記載の抗体又はその抗原結合断片。
- 前記Ig様ドメインが、PD1、SLAM、CD2、CD3、CD4、CD8、CD19、CD22、CD33、CD80、又はCD86のIg様ドメインから選択される請求項4に記載の抗体又はその抗原結合断片。
- 前記キャリアドメインが、前記抗体又はその抗原結合断片の、薬剤又は造影剤へのコンジュゲーションを提供する請求項1から5のいずれかに記載の抗体又はその抗原結合断片。
- 前記レポータードメインが、GFP、YFP、RFP、CFP、ルシフェラーゼ、ベータ-ガラクトシダーゼ、又はペルオキシダーゼをコードするアミノ酸配列を含む又はからなる請求項1から5のいずれかに記載の抗体又はその抗原結合断片。
- 前記タグが、親和性タグ、可溶化タグ、クロマトグラフィータグ、エピトープタグ、又は蛍光タグである請求項1から5のいずれかに記載の抗体又はその抗原結合断片。
- 前記酵素ドメインが、触媒ドメインである請求項1から5のいずれかに記載の抗体又はその抗原結合断片。
- 前記受容体の機能的断片が、受容体のIg様ドメインである請求項1から5のいずれかに記載の抗体又はその抗原結合断片。
- 前記機能的ドメインが、(i)PD1、SLAM、LAIR1、CTLA4、BTLA、TIM-3、TIGIT、CD200R1、2B4(CD244)、TLT2、LILRB4、KIR2DL2、ICOS、又はCD28からなる群から選択される受容体、又は(ii)その機能的断片を含む又はからなる請求項1から5及び10のいずれかに記載の抗体又はその抗原結合断片。
- 前記第1のポリペプチド鎖の前記機能的ドメイン(ii)及び/又は前記第2のポリペプチド鎖の前記機能的ドメイン(v)が、リガンド又はその機能的断片を含む又はからなり、
(1) 前記リガンドが、サイトカイン及び/又はPD1、SLAM、LAIR1、CTLA4、BTLA、TIM-3、TIGIT、CD200R1、2B4(CD244)、TLT2、LILRB4、KIR2DL2、ICOS、又はCD28のリガンドである;
(2) 前記リガンドが、PD-L1、PD-L2、B7-1、B7-2、B7-H4(B7ホモログ)、ガレクチン-9、ポリオウイルス受容体(PVR)、OX-2膜糖タンパク質、CD48、B7-H3(B7ホモログ)、MHCI、ICOS-Lからなる群から選択される;又は
(3) 前記リガンドが、サイトカインのSISファミリー、サイトカインのSIGファミリー、サイトカインのSCYファミリー、血小板因子4スーパーファミリー、インタークリン、CCケモカインリガンド(CCL)-1~-28、CXCL1~CXCL17、XCL1(リンホタクチン-α)、XCL2(リンホタクチン-β)、フラクタルカイン(又はCX 3 CL1);インターフェロン;インターロイキン;リンホカイン;腫瘍壊死因子;及びコロニー刺激因子からなる群から選択される請求項1から5のいずれかに記載の抗体又はその抗原結合断片。 - 前記(独立した)結合部位が、受容体又は結合部位を含むその機能的断片、リガンド又は結合部位を含むその機能的断片、CD分子又はその機能的断片、単鎖抗体又はその抗原結合断片、抗原又はその機能的断片、又は結合部位を含むタグを含む又はからなる請求項1から5のいずれかに記載の抗体又はその抗原結合断片。
- 前記機能的ドメインが、Ig様ドメイン、scFv、VHH、又はStrepタグを含む又はからなる請求項1から13のいずれかに記載の抗体又はその抗原結合断片。
- 前記機能的ドメインが、配列番号13~20のいずれかに示されるアミノ酸配列、又は少なくとも90%の配列同一性を有するその機能的配列変異体を含む又はからなる請求項14に記載の抗体又はその抗原結合断片。
- 前記第1のポリペプチド鎖が、前記機能的ドメイン(ii)のN末端に1つの単一の可変ドメイン(i)を含み、前記第2のポリペプチド鎖が、前記機能的ドメイン(v)のN末端に1つの単一の可変ドメイン(iv)を含み、前記第2のポリペプチド鎖の前記1つの単一の可変ドメイン(iv)が前記第1のポリペプチド鎖の前記可変ドメイン(i)と抗原結合部位を形成する;又は
前記第1のポリペプチド鎖が、前記機能的ドメイン(ii)のN末端に2つ以上の可変ドメイン(i)を含み、前記第2のポリペプチド鎖が、前記機能的ドメイン(v)のN末端に2つ以上の可変ドメイン(iv)を含み、前記第2のポリペプチド鎖の前記2つ以上の可変ドメイン(iv)が前記第1のポリペプチド鎖の前記2つ以上の可変ドメイン(i)と抗原結合部位を形成する請求項1及び3から15のいずれかに記載の抗体又はその抗原結合断片。 - 前記第2のポリペプチド鎖が、機能的ドメインを含まず、
前記第1のポリペプチド鎖が、前記機能的ドメイン(ii)のN末端に1つの単一の可変ドメイン(i)を含み、前記第2のポリペプチド鎖が、前記1つ以上の定常ドメイン(vi)のN末端に1つの単一の可変ドメイン(iv)を含み、前記第2のポリペプチド鎖の前記1つの単一の可変ドメイン(iv)が前記第1のポリペプチド鎖の前記可変ドメイン(i)と抗原結合部位を形成する;又は
前記第1のポリペプチド鎖が、前記機能的ドメイン(ii)のN末端に2つ以上の可変ドメイン(i)を含み、前記第2のポリペプチド鎖が、前記1つ以上の定常ドメイン(vi)のN末端に2つ以上の可変ドメイン(iv)を含み、前記第2のポリペプチド鎖の前記2つ以上の可変ドメイン(iv)が前記第1のポリペプチド鎖の前記2つ以上の可変ドメイン(i)と抗原結合部位を形成する請求項2から15のいずれかに記載の抗体又はその抗原結合断片。 - 前記第1のポリペプチド鎖が、V-D-CH1を含む又はからなり、式中、
Vは、可変ドメイン(i)であり、
Dは、機能的ドメイン(ii)であり、
CH1は、CH1定常ドメイン(iii)である;及び/又は
前記第2のポリペプチド鎖が、V-CLを含む又はからなり、式中、
Vは、可変ドメイン(iv)であり、
CLは、定常ドメイン(vi)である請求項1から17のいずれかに記載の抗体又はその抗原結合断片。 - 前記第1のポリペプチド鎖の前記可変ドメイン(i)が、配列番号1、5、8、又は10のいずれかに示されるアミノ酸配列、又は少なくとも90%の配列同一性を有するその機能的配列変異体を含む又はからなる;及び/又は
前記第2のポリペプチド鎖の前記可変ドメイン(iv)が、配列番号2、6、9、又は11のいずれかに示されるアミノ酸配列、又は少なくとも90%の配列同一性を有するその機能的配列変異体を含む又はからなる請求項1から18のいずれかに記載の抗体又はその抗原結合断片。 - 前記抗体又はその抗原結合断片が、同一又は異なっていてもよい少なくとも2つの機能的ドメインを含む請求項1から19のいずれかに記載の抗体又はその抗原結合断片。
- 前記第1のポリペプチド鎖が、配列番号53、55、56、58、59、60、61、62、64、65、66、68、69、又は70のいずれかに示されるアミノ酸配列、又は少なくとも90%の配列同一性を有するその機能的配列変異体を含む又はからなる、及び/又は前記第2のポリペプチド鎖が、配列番号54、57、63、又は67のいずれかに示されるアミノ酸配列、又は少なくとも90%の配列同一性を有するその機能的配列変異体を含む又はからなる請求項1から20のいずれかに記載の抗体又はその抗原結合断片。
- 請求項1から21のいずれかに記載の抗体又はその抗原結合断片の第1のポリペプチド鎖及び第2のポリペプチド鎖をコードするポリヌクレオチドを含むことを特徴とする核酸分子。
- 請求項22に記載の核酸分子を含むことを特徴とするベクター。
- 請求項22に記載の核酸分子又は請求項23に記載のベクターを含むことを特徴とする宿主細胞。
- 請求項1から21のいずれかに記載の抗体又はその抗原結合断片、請求項22に記載の核酸分子、請求項23に記載のベクター、又は請求項24に記載の宿主細胞を含むことを特徴とする医薬組成物。
- 医学における使用のための、請求項1から21のいずれかに記載の抗体又はその抗原結合断片、請求項22に記載の核酸分子、請求項23に記載のベクター、請求項24に記載の宿主細胞、又は請求項25に記載の医薬組成物。
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BR112019028180A2 (pt) | 2020-07-07 |
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