JPWO2019165116A5

JPWO2019165116A5 -

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Publication number: JPWO2019165116A5
Application number: JP2020544661A
Authority: JP
Publication date: 2022-03-01

Description

したがって、がんを処置するための養子細胞療法における新しい戦略が必要である。ここで開示された実施形態は、これらの必要性に取り組むものであり、他の関連の利点を提供する。
本発明は、例えば以下の項目を提供する。
(項目１)
（ａ）標的抗原に特異的に結合する結合ドメインを含む細胞外成分；
（ｂ）改変された機能的なＣＤ２８共刺激シグナル伝達ドメインを含む細胞内成分であって、前記改変された機能的なＣＤ２８共刺激シグナル伝達ドメインは、少なくとも１つのアミノ酸置換を含む、細胞内成分；および
（ｃ）前記細胞外成分と前記細胞内成分との間に配置された疎水性部分
を含む融合タンパク質であって、
前記融合タンパク質は、野生型ＣＤ２８共刺激シグナル伝達ドメインを含む融合タンパク質とは異なる１つまたは複数の機能的な活性を有する、融合タンパク質。
(項目２)
前記ＣＤ２８共刺激シグナル伝達ドメイン中の少なくとも１つのチロシン残基が、異なるアミノ酸残基で置換されている、項目１に記載の融合タンパク質。
(項目３)
前記少なくとも１つのチロシン残基が、１９１、２０６、２０９および２１８位のいずれか１つから選択される、項目２に記載の融合タンパク質。
(項目４)
１９１、２０６、２０９、および２１８位のいずれかから選択される少なくとも２つのチロシン残基がそれぞれ、異なるアミノ酸残基で置換されている、項目２に記載の融合タンパク質。
(項目５)
１９１、２０６、２０９、および２１８位のいずれかから選択される少なくとも３つのチロシン残基がそれぞれ、異なるアミノ酸残基で置換されている、項目２に記載の融合タンパク質。
(項目６)
１９１、２０６、２０９、および２１８位における４つのチロシン残基が、異なるアミノ酸残基で置換されている、項目２に記載の融合タンパク質。
(項目７)
各チロシン残基が、独立して、トリプトファン残基またはフェニルアラニン残基で置換されている、項目２から６のいずれか一項に記載の融合タンパク質。
(項目８)
各チロシン残基が、フェニルアラニン残基で置換されている、項目２から６のいずれか一項に記載の融合タンパク質。
(項目９)
各チロシン残基が、トリプトファン残基で置換されている、項目２から６のいずれか一項に記載の融合タンパク質。
(項目１０)
前記改変されたＣＤ２８共刺激シグナル伝達ドメインが、Ｙ２１８Ｆ置換を含む、項目２から７のいずれか一項に記載の融合タンパク質。
(項目１１)
前記改変されたＣＤ２８共刺激シグナル伝達ドメインが、Ｙ２０６Ｆ、Ｙ２０９Ｆ、およびＹ２１８Ｆ置換を含む、項目２から７のいずれか一項に記載の融合タンパク質。
(項目１２)
前記改変されたＣＤ２８共刺激シグナル伝達ドメインが、Ｙ１９１Ｆ、Ｙ２０６Ｆ、Ｙ２０９Ｆ、およびＹ２１８Ｆ置換を含む、項目２から７のいずれか一項に記載の融合タンパク質。
(項目１３)
前記ＣＤ２８共刺激シグナル伝達ドメイン中の少なくとも１つのプロリン残基が、異なるアミノ酸残基で置換されている、項目１から１２のいずれか一項に記載の融合タンパク質。
(項目１４)
前記少なくとも１つのプロリン残基が、１９６、１９９、２０８、および２１１位のいずれか１つから選択される、項目１３に記載の融合タンパク質。
(項目１５)
１９６、１９９、２０８、および２１１位のいずれかから選択される少なくとも２つのプロリン残基がそれぞれ、異なるアミノ酸残基で置換されている、項目１３に記載の融合タンパク質。
(項目１６)
１９６、１９９、２０８、および２１１位のいずれかから選択される少なくとも３つのプロリン残基がそれぞれ、異なるアミノ酸残基で置換されている、項目１３に記載の融合タンパク質。
(項目１７)
１９６、１９９、２０８、および２１１位における４つのプロリン残基が、異なるアミノ酸残基で置換されている、項目１３に記載の融合タンパク質。
(項目１８)
各プロリン残基が、アラニン残基で置換されている、項目１３から１７のいずれか一項に記載の融合タンパク質。
(項目１９)
前記改変されたＣＤ２８共刺激シグナル伝達ドメインが、Ｌ１８６およびＬ１８７置換をさらに含む、項目１から１８のいずれか一項に記載の融合タンパク質。
(項目２０)
前記Ｌ１８６置換が、Ｌ１８６Ｇであり、前記Ｌ１８７置換が、Ｌ１８７Ｇである、項目１９に記載の融合タンパク質。
(項目２１)
前記改変されたＣＤ２８共刺激シグナル伝達ドメインが、Ｙ１９１、Ｐ２０８、Ｐ２１１、またはそれらのあらゆる組合せにおける置換を含まない、項目１から１１、１３から１５、および１８から２０のいずれか一項に記載の融合タンパク質。
(項目２２)
前記結合ドメインが、ｓｃＦｖ、ｓｃＴＣＲ、受容体の細胞外ドメイン、またはリガンドである、項目１から２１のいずれか一項に記載の融合タンパク質。
(項目２３)
前記結合ドメインが、ＣＤ８の細胞外結合ドメインまたは部分も、機能的なＩｇＶ様ドメインを含むそれらのいずれの部分も含まない、項目１から２２のいずれか一項に記載の融合タンパク質。
(項目２４)
前記結合ドメインが、ＣＤ８α鎖由来の結合ドメインも、ＣＤ８β鎖由来の結合ドメインも、ＣＤ８αホモ二量体由来の結合ドメインも、ＣＤ８αβヘテロ二量体由来の結合ドメインも含まない、項目２３に記載の融合タンパク質。
(項目２５)
前記結合ドメインが、キメラ、ヒト、またはヒト化されたものである、項目１から２４のいずれか一項に記載の融合タンパク質。
(項目２６)
前記細胞外成分が、前記結合ドメインと前記疎水性部分との間に配置されたスペーサーをさらに含む、項目１から２５のいずれか一項に記載の融合タンパク質。
(項目２７)
前記スペーサーが、免疫グロブリンのヒンジ領域、ＣＨ２ドメイン、ＣＨ３ドメイン、またはそれらのあらゆる組合せを含む、項目２６に記載の融合タンパク質。
(項目２８)
前記免疫グロブリンのヒンジ領域が、ＩｇＧ４のヒンジ領域である、項目２７に記載の融合タンパク質。
(項目２９)
前記細胞外成分が、前記結合ドメインと前記疎水性部分との間に配置されたタグをさらに含む、項目１から２８のいずれか一項に記載の融合タンパク質。
(項目３０)
前記タグが、Ｔｒｐ－Ｓｅｒ－Ｈｉｓ－Ｐｒｏ－Ｇｌｎ－Ｐｈｅ－Ｇｌｕ－Ｌｙｓ（配列番号４０）のアミノ酸配列を有する、項目２９に記載の融合タンパク質。
(項目３１)
前記細胞内成分が、ＩＴＡＭ含有Ｔ細胞活性化ドメインをさらに含む、項目１から３０のいずれか一項に記載の融合タンパク質。
(項目３２)
前記ＩＴＡＭ含有Ｔ細胞活性化ドメインが、ＣＤ３γ、ＣＤ３δ、ＣＤ３ε、ＣＤ３ζ、ＦｃεＲＩのガンマ鎖、またはＦｃγＲＩのガンマ鎖の細胞内シグナル伝達ドメインを含む、項目３１に記載の融合タンパク質。
(項目３３)
前記細胞内成分が、ＣＤ３ζ細胞内シグナル伝達ドメインをさらに含む、項目３２に記載の融合タンパク質。
(項目３４)
前記細胞内成分が、少なくとも１つの追加の共刺激シグナル伝達ドメインをさらに含む、項目１から３３のいずれか一項に記載の融合タンパク質。
(項目３５)
前記少なくとも１つの追加の共刺激シグナル伝達ドメインが、ＣＤ２７、ＣＤ４０Ｌ、ＧＩＴＲ、ＮＫＧ２Ｃ、ＣＡＲＤ１、ＣＤ２、ＣＤ７、ＣＤ２７、ＣＤ３０、ＣＤ４０、ＣＤ５４（ＩＣＡＭ）、ＣＤ８３、ＣＤ１３４（ＯＸ－４０）、ＣＤ１３７（４－１ＢＢ）、ＣＤ１５０（ＳＬＡＭＦ１）、ＣＤ１５２（ＣＴＬＡ４）、ＣＤ２２３（ＬＡＧ３）、ＣＤ２７０（ＨＶＥＭ）、ＣＤ２７３（ＰＤ－Ｌ２）、ＣＤ２７４（ＰＤ－Ｌ１）、ＣＤ２７８（ＩＣＯＳ）、ＤＡＰ１０、ＬＡＴ、ＮＫＤ２ＣＳＬＰ７６、ＴＲＩＭ、ＺＡＰ７０、ＣＤ５、ＢＡＦＦ－Ｒ、ＳＬＡＭＦ７、ＮＫｐ８０、ＣＤ１６０、Ｂ７－Ｈ３、ＣＤ８３と特異的に結合するリガンド、またはそれらの組合せから選択される、項目３４に記載の融合タンパク質
(項目３６)
前記疎水性部分が、膜貫通ドメインである、項目１から３５のいずれか一項に記載の融合タンパク質。
(項目３７)
前記膜貫通ドメインが、ＣＤ２８、ＣＤ２、ＣＤ３ε、ＣＤ３δ、ＣＤ３ζ、ＣＤ２５、ＣＤ２７、ＣＤ４０、ＣＤ７９Ａ、ＣＤ７９Ｂ、ＣＤ８０、ＣＤ８６、ＣＤ９５（Ｆａｓ）、ＣＤ１３４（ＯＸ４０）、ＣＤ１３７（４－１ＢＢ）、ＣＤ１５０（ＳＬＡＭＦ１）、ＣＤ１５２（ＣＴＬＡ４）、ＣＤ２００Ｒ、ＣＤ２２３（ＬＡＧ３）、ＣＤ２７０（ＨＶＥＭ）、ＣＤ２７２（ＢＴＬＡ）、ＣＤ２７３（ＰＤ－Ｌ２）、ＣＤ２７４（ＰＤ－Ｌ１）、ＣＤ２７８（ＩＣＯＳ）、ＣＤ２７９（ＰＤ－１）、ＣＤ３００、ＣＤ３５７（ＧＩＴＲ）、Ａ２ａＲ、ＤＡＰ１０、ＦｃＲα、ＦｃＲβ、ＦｃＲγ、Ｆｙｎ、ＧＡＬ９、ＫＩＲ、Ｌｃｋ、ＬＡＴ、ＬＲＰ、ＮＫＧ２Ｄ、ＮＯＴＣＨ１、ＮＯＴＣＨ２、ＮＯＴＣＨ３、ＮＯＴＣＨ４、ＰＴＣＨ２、ＲＯＲ２、Ｒｙｋ、Ｓｌｐ７６、ＳＩＲＰα、ｐＴα、ＴＣＲα、ＴＣＲβ、ＴＩＭ３、ＴＲＩＭ、ＬＰＡ５、またはＺａｐ７０の膜貫通ドメインを含む、項目３６に記載の融合タンパク質。
(項目３８)
前記標的抗原が、がん抗原、ウイルス抗原、細菌抗原、または自己抗原である、項目１から３７のいずれか一項に記載の融合タンパク質。
(項目３９)
前記標的抗原が、ＢＣＭＡ、ＣＤ３、ＣＥＡＣＡＭ６、ｃ－Ｍｅｔ、ＥＧＦＲ、ＥＧＦＲｖＩＩＩ、ＥｒｂＢ２、ＥｒｂＢ３、ＥｒｂＢ４、ＥｐｈＡ２、ＩＧＦ１Ｒ、ＧＤ２、Ｏ－アセチルＧＤ２、Ｏ－アセチルＧＤ３、ＧＨＲＨＲ、ＧＨＲ、ＦＬＴ１、ＫＤＲ、ＦＬＴ４、ＣＤ４４ｖ６、ＣＤ１５１、ＣＡ１２５、ＣＥＡ、ＣＴＬＡ－４、ＧＩＴＲ、ＢＴＬＡ、ＴＧＦＢＲ２、ＴＧＦＢＲ１、ＩＬ６Ｒ、ｇｐ１３０、ルイスＡ、ルイスＹ、ＴＮＦＲ１、ＴＮＦＲ２、ＰＤ１、ＰＤ－Ｌ１、ＰＤ－Ｌ２、ＨＶＥＭ、ＭＡＧＥ－Ａ、メソテリン、ＮＹ－ＥＳＯ－１、ＰＳＭＡ、ＲＡＮＫ、ＲＯＲ１、ＴＮＦＲＳＦ４、ＣＤ４０、ＣＤ１３７、ＴＷＥＡＫ－Ｒ、ＨＬＡ、ＨＬＡに結合した腫瘍または病原体関連のペプチド、ＨＬＡに結合したｈＴＥＲＴペプチド、ＨＬＡに結合したチロシナーゼペプチド、ＨＬＡに結合したＷＴ－１ペプチド、ＬＴβＲ、ＬＩＦＲβ、ＬＲＰ５、ＭＵＣ１、ＯＳＭＲβ、ＴＣＲα、ＴＣＲβ、ＣＤ１９、ＣＤ２０、ＣＤ２２、ＣＤ２５、ＣＤ２８、ＣＤ３０、ＣＤ３３、ＣＤ５２、ＣＤ５６、ＣＤ７９ａ、ＣＤ７９ｂ、ＣＤ８０、ＣＤ８１、ＣＤ８６、ＣＤ１２３、ＣＤ１７１、ＣＤ２７６、Ｂ７Ｈ４、ＴＬＲ７、ＴＬＲ９、ＰＴＣＨ１、ＷＴ－１、ＨＡ ^１－Ｈ、Ｒｏｂｏ１、α－フェトプロテイン（ＡＦＰ）、Ｆｒｉｚｚｌｅｄ、ＯＸ４０、ＰＲＡＭＥ、およびＳＳＸ－２から選択されるがん抗原である、項目３６に記載の融合タンパク質。
(項目４０)
項目１から３９のいずれか一項に記載の融合タンパク質をコードするポリヌクレオチド。
(項目４１)
核酸分子が、コドン最適化されている、項目４０に記載のポリヌクレオチド。
(項目４２)
配列番号１８、２０、２２、２４、２６、２８、３０、３２、３４、３６、４８、５０、５２、５４、５６、または５８のいずれか１つに記載の融合タンパク質をコードするヌクレオチド配列に対して、少なくとも約７５％の同一性を有するポリヌクレオチドを含む、項目４０または４１に記載のポリヌクレオチド。
(項目４３)
形質導入マーカー、自殺遺伝子、またはその両方をコードするポリヌクレオチドをさらに含む、項目４０から４２のいずれか一項に記載のポリヌクレオチド。
(項目４４)
前記形質導入マーカーが、トランケート型ＥＧＦＲ分子である、項目４３に記載のポリヌクレオチド。
(項目４５)
配列番号１８、２０、２２、２４、２６、２８、３０、３２、３４、３６、４８、５０、５２、５４、５６、または５８のいずれか１つに対して少なくとも約７５％の同一性を有するポリヌクレオチドを含むかまたはそれからなる、項目４０から４４のいずれか一項に記載のポリヌクレオチド。
(項目４６)
項目４０から４５のいずれか一項に記載のポリヌクレオチドを含むベクター。
(項目４７)
ウイルスベクターである、項目４６に記載のベクター。
(項目４８)
前記ウイルスベクターが、レンチウイルスまたはレトロウイルスベクターである、項目４７に記載のベクター。
(項目４９)
項目１から３９のいずれか一項に記載の融合タンパク質、項目４０から４５のいずれか一項に記載のポリヌクレオチド、または項目４６から４８のいずれか一項に記載のベクターを含む宿主細胞。
(項目５０)
免疫系細胞である、項目４９に記載の宿主細胞。
(項目５１)
前記免疫系細胞が、Ｔ細胞である、項目５０に記載の宿主細胞。
(項目５２)
前記Ｔ細胞が、ＣＤ４ ^＋Ｔ細胞またはＣＤ８ ^＋Ｔ細胞である、項目５１に記載の宿主細胞。
(項目５３)
前記Ｔ細胞が、メモリーＴ細胞である、項目５１または５２に記載の宿主細胞。
(項目５４)
ヒト細胞である、項目４９から５３のいずれか一項に記載の宿主細胞。
(項目５５)
内因性遺伝子の発現が、前記宿主細胞中で阻害され、阻害される前記内因性遺伝子は、ＴＣＲ遺伝子、ＨＬＡ遺伝子、免疫抑制成分の遺伝子、またはそれらのあらゆる組合せから選択される、項目４９から５４のいずれか一項に記載の宿主細胞。
(項目５６)
前記ＴＣＲ遺伝子が、Ｔ細胞受容体α定常（ＴＲＡＣ）遺伝子、Ｔ細胞受容体β定常（ＴＲＢＣ）遺伝子、またはその両方である、項目５５に記載の宿主細胞。
(項目５７)
項目４０から４５のいずれか一項に記載のポリヌクレオチド、または項目４６から４８のいずれか一項に記載のベクターが、エンドヌクレアーゼ系を介して、前記ＴＣＲ遺伝子またはＨＬＡ遺伝子の遺伝子座に標的化されている、項目５５または５６に記載の宿主細胞。
(項目５８)
前記エンドヌクレアーゼ系が、ＣＲＩＳＰＲ／Ｃａｓヌクレアーゼ系、ジンクフィンガーヌクレアーゼ（ＺＦＮ）系、または転写活性化因子様エフェクターヌクレアーゼ（ＴＡＬＥＮ）系である、項目５７に記載の宿主細胞。
(項目５９)
前記融合タンパク質の持続的なリン酸化の減少、前記融合タンパク質のシグナル伝達の減少、サイトカイン発現の減少、存続の強化、抗原特異的な細胞溶解活性の強化、またはそれらのあらゆる組合せを呈示する、項目４９から５８のいずれか一項に記載の宿主細胞。
(項目６０)
野生型ＣＤ２８共刺激シグナル伝達ドメインを含む融合タンパク質を含む宿主細胞と比較して減少したレベルの標的抗原によって誘導されたＩＬ－２、ＴＮＦ－α、またはその両方を発現する、項目４９から５９のいずれか一項に記載の宿主細胞。
(項目６１)
項目４０から４５のいずれか一項に記載のポリヌクレオチド、項目４６から４８のいずれか一項に記載のベクター、または項目４９から６０のいずれか一項に記載の宿主細胞、および薬学的に許容される担体、希釈剤、または賦形剤を含む、医薬組成物。
(項目６２)
被験体における疾患を処置する方法であって、項目４９から６０のいずれか一項に記載の宿主細胞または項目６１に記載の医薬組成物を前記被験体に投与することを含み、前記疾患は、融合タンパク質と結合した抗原の存在に関連する、方法。
(項目６３)
前記疾患が、ウイルス感染、細菌感染、がん、炎症性疾患、または自己免疫疾患である、項目６２に記載の方法。
(項目６４)
前記被験体が、ヒトである、項目６２または６３に記載の方法。
(項目６５)
前記宿主細胞が、前記被験体にとって同種または自己である、項目６２から６４のいずれか一項に記載の方法。
(項目６６)
前記被験体が、前記処置の後、低いレベルまたは軽度のサイトカイン放出症候群、低いレベルまたは軽度の細胞関連脳症症候群、またはその両方を有するか、および／あるいは参照宿主細胞または組成物が投与された参照被験体と比較して、低減されたサイトカイン放出症候群、低減された細胞関連脳症症候群、またはその両方を有し、前記融合タンパク質は、野生型ＣＤ２８共刺激シグナル伝達ドメインを含む、項目６２から６５のいずれか一項に記載の方法。
(項目６７)
前記疾患が、がんである、項目６２から６６のいずれか一項に記載の方法。
(項目６８)
前記がんが、固形腫瘍、黒色腫、非小細胞肺がん、腎細胞癌、腎臓がん、血液がん、前立腺がん、去勢抵抗性前立腺がん、結腸がん、直腸がん、胃がん、食道がん、膀胱がん、頭頸部がん、甲状腺がん、乳がん、トリプルネガティブ乳がん、卵巣がん、子宮頸がん、肺がん、尿路上皮がん、膵臓がん、膠芽腫、肝細胞がん、骨髄腫、多発性骨髄腫、白血病、ホジキンリンパ腫、非ホジキンリンパ腫、骨髄異形成症候群、脳がん、ＣＮＳがん、または悪性神経膠腫である、項目６７に記載の方法。
(項目６９)
化学療法剤または免疫抑制成分の阻害剤を投与することをさらに含む、項目６２から６８のいずれか一項に記載の方法。
(項目７０)
（ａ）項目４６から４８のいずれか一項に記載のベクターまたは発現構築物、および前記ベクターまたは前記発現構築物を宿主細胞に形質導入するための必要に応じた試薬；
（ｂ）項目４０から４５のいずれか一項に記載の単離されたポリヌクレオチド、および前記ポリヌクレオチドを宿主細胞に形質導入するための必要に応じた試薬；ならびに／あるいは
（ｃ）項目４９から６０のいずれか一項に記載の宿主細胞
を含む、キット。 Therefore, new strategies in adoptive cell therapy to treat cancer are needed. The embodiments disclosed herein address these needs and provide other relevant benefits.
The present invention provides, for example, the following items.
(Item 1)
(A) Extracellular components containing a binding domain that specifically binds to the target antigen;
(B) An intracellular component comprising a modified functional CD28 co-stimulation signaling domain, wherein the modified functional CD28 co-stimulating signaling domain comprises at least one amino acid substitution. ;and
(C) A hydrophobic moiety arranged between the extracellular component and the intracellular component.
Is a fusion protein containing
The fusion protein is a fusion protein having one or more functional activities different from the fusion protein containing the wild-type CD28 co-stimulating signaling domain.
(Item 2)
The fusion protein of item 1, wherein at least one tyrosine residue in the CD28 co-stimulation signaling domain is replaced with a different amino acid residue.
(Item 3)
The fusion protein of item 2, wherein the at least one tyrosine residue is selected from any one of positions 191, 206, 209 and 218.
(Item 4)
Item 2. The fusion protein of item 2, wherein at least two tyrosine residues selected from any of positions 191, 206, 209, and 218 are each substituted with different amino acid residues.
(Item 5)
Item 2. The fusion protein of item 2, wherein at least three tyrosine residues selected from any of positions 191, 206, 209, and 218 are each substituted with different amino acid residues.
(Item 6)
The fusion protein of item 2, wherein the four tyrosine residues at positions 191, 206, 209, and 218 are replaced with different amino acid residues.
(Item 7)
Item 6. The fusion protein according to any one of items 2 to 6, wherein each tyrosine residue is independently substituted with a tryptophan residue or a phenylalanine residue.
(Item 8)
The fusion protein according to any one of items 2 to 6, wherein each tyrosine residue is replaced with a phenylalanine residue.
(Item 9)
The fusion protein according to any one of items 2 to 6, wherein each tyrosine residue is replaced with a tryptophan residue.
(Item 10)
The fusion protein according to any one of items 2 to 7, wherein the modified CD28 co-stimulation signaling domain comprises a Y218F substitution.
(Item 11)
The fusion protein according to any one of items 2 to 7, wherein the modified CD28 co-stimulation signaling domain comprises a Y206F, Y209F, and Y218F substitution.
(Item 12)
The fusion protein according to any one of items 2 to 7, wherein the modified CD28 co-stimulation signaling domain comprises Y191F, Y206F, Y209F, and Y218F substitutions.
(Item 13)
The fusion protein according to any one of items 1 to 12, wherein at least one proline residue in the CD28 co-stimulation signaling domain is replaced with a different amino acid residue.
(Item 14)
13. The fusion protein of item 13, wherein the at least one proline residue is selected from any one of positions 196, 199, 208, and 211.
(Item 15)
196. The fusion protein of item 13, wherein at least two proline residues selected from positions 196, 199, 208, and 211 are each substituted with different amino acid residues.
(Item 16)
196. The fusion protein of item 13, wherein at least three proline residues selected from positions 196, 199, 208, and 211 are each substituted with different amino acid residues.
(Item 17)
196. The fusion protein of item 13, wherein the four proline residues at positions 199, 208, and 211 are replaced with different amino acid residues.
(Item 18)
The fusion protein according to any one of items 13 to 17, wherein each proline residue is replaced with an alanine residue.
(Item 19)
The fusion protein according to any one of items 1 to 18, wherein the modified CD28 co-stimulation signaling domain further comprises L186 and L187 substitutions.
(Item 20)
19. The fusion protein of item 19, wherein the L186 substitution is L186G and the L187 substitution is L187G.
(Item 21)
The item any one of Items 1-11, 13-15, and 18-20, wherein the modified CD28 co-stimulation signaling domain does not include substitutions in Y191, P208, P211 or any combination thereof. Fusion protein.
(Item 22)
The fusion protein according to any one of items 1 to 21, wherein the binding domain is scFv, scTCR, an extracellular domain of a receptor, or a ligand.
(Item 23)
The fusion protein according to any one of items 1 to 22, wherein the binding domain does not contain an extracellular binding domain or portion of CD8, nor any portion thereof, including a functional IgV-like domain.
(Item 24)
23. Item 23, wherein the binding domain does not include a CD8α chain-derived binding domain, a CD8β chain-derived binding domain, a CD8α homodimer-derived binding domain, or a CD8αβ heterodimer-derived binding domain. Fusion protein.
(Item 25)
The fusion protein according to any one of items 1 to 24, wherein the binding domain is chimeric, human, or humanized.
(Item 26)
The fusion protein according to any one of items 1 to 25, wherein the extracellular component further comprises a spacer disposed between the binding domain and the hydrophobic moiety.
(Item 27)
26. The fusion protein of item 26, wherein the spacer comprises a hinge region of an immunoglobulin, a CH2 domain, a CH3 domain, or any combination thereof.
(Item 28)
27. The fusion protein according to item 27, wherein the hinge region of the immunoglobulin is the hinge region of IgG4.
(Item 29)
The fusion protein according to any one of items 1 to 28, wherein the extracellular component further comprises a tag placed between the binding domain and the hydrophobic moiety.
(Item 30)
29. The fusion protein of item 29, wherein the tag has the amino acid sequence of Trp-Ser-His-Pro-Gln-Phe-Glu-Lys (SEQ ID NO: 40).
(Item 31)
The fusion protein according to any one of items 1 to 30, wherein the intracellular component further comprises an ITAM-containing T cell activation domain.
(Item 32)
31. The fusion protein of item 31, wherein the ITM-containing T cell activation domain comprises an intracellular signaling domain of the gamma chain of CD3γ, CD3δ, CD3ε, CD3ζ, FcεRI, or the gamma chain of FcγRI.
(Item 33)
32. The fusion protein of item 32, wherein the intracellular component further comprises a CD3ζ intracellular signaling domain.
(Item 34)
The fusion protein according to any one of items 1 to 33, wherein the intracellular component further comprises at least one additional co-stimulation signaling domain.
(Item 35)
The at least one additional co-stimulation signaling domain is CD27, CD40L, GITR, NKG2C, CARD1, CD2, CD7, CD27, CD30, CD40, CD54 (ICAM), CD83, CD134 (OX-40), CD137 (4). -1BB), CD150 (SLAMF1), CD152 (CTLA4), CD223 (LAG3), CD270 (HVEM), CD273 (PD-L2), CD274 (PD-L1), CD278 (ICOS), DAP10, LAT, NKD2C SLP76, 34. The fusion protein of item 34, selected from a ligand that specifically binds to TRIM, ZAP70, CD5, BAFF-R, SLAMF7, NKp80, CD160, B7-H3, CD83, or a combination thereof.
(Item 36)
The fusion protein according to any one of items 1 to 35, wherein the hydrophobic portion is a transmembrane domain.
(Item 37)
The transmembrane domains are CD28, CD2, CD3ε, CD3δ, CD3ζ, CD25, CD27, CD40, CD79A, CD79B, CD80, CD86, CD95 (Fas), CD134 (OX40), CD137 (4-1BB), CD150 (SLAMF1). ), CD152 (CTLA4), CD200R, CD223 (LAG3), CD270 (HVEM), CD272 (BTLA), CD273 (PD-L2), CD274 (PD-L1), CD278 (ICOS), CD279 (PD-1), CD300, CD357 (GITR), A2aR, DAP10, FcRα, FcRβ, FcRγ, Fyn, GAL9, KIR, Lck, LAT, LRP, NKG2D, NOTCH1, NOTCH2, NOTCH3, NOTCH4, PTCH2, ROR2, Ryk, Slp76 36. The fusion protein of item 36, comprising the transmembrane domain of TCRα, TCRβ, TIM3, TRIM, LPA5, or Zap70.
(Item 38)
The fusion protein according to any one of items 1 to 37, wherein the target antigen is a cancer antigen, a viral antigen, a bacterial antigen, or a self-antigen.
(Item 39)
The target antigens are BCMA, CD3, CEACAM6, c-Met, EGFR, EGFRvIII, ErbB2, ErbB3, ErbB4, EphA2, IGF1R, GD2, O-acetylGD2, O-acetylGD3, GHRHR, GHR, FLT1, KDR, L , CD44v6, CD151, CA125, CEA, CTLA-4, GITR, BTLA, TGFBR2, TGFBR1, IL6R, gp130, Lewis A, Lewis Y, TNFR1, TNFR2, PD1, PD-L1, PD-L2, HVEM, MAGE-A , Mesoterin, NY-ESO-1, PSMA, RANK, ROR1, TNFRSF4, CD40, CD137, TWEAK-R, HLA, HLA-bound tumor or pathogen-related peptide, HLA-bound hTERT peptide, HLA-bound tyrosinase. Peptide, WT-1 peptide bound to HLA, LTβR, LIFRβ, LRP5, MUC1, OSMRβ, TCRα, TCRβ, CD19, CD20, CD22, CD25, CD28, CD30, CD33, CD52, CD56, CD79a, CD79b, CD80, CD81 , CD86, CD123, CD171, CD276, B7H4, TLR7, TLR9, PTCH1, WT-1, HA ¹ - H, Robo1, α-fet protein (AFP), Frizzled, OX40, PRAME, and SSX-2. 36. The fusion protein according to item 36, which is an antigen.
(Item 40)
The polynucleotide encoding the fusion protein according to any one of items 1 to 39.
(Item 41)
40. The polynucleotide of item 40, wherein the nucleic acid molecule is codon-optimized.
(Item 42)
To the nucleotide sequence encoding the fusion protein set forth in any one of SEQ ID NOs: 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 48, 50, 52, 54, 56, or 58. The polynucleotide according to item 40 or 41, comprising a polynucleotide having at least about 75% identity relative to it.
(Item 43)
The polynucleotide according to any one of items 40 to 42, further comprising a polynucleotide encoding a transduction marker, a suicide gene, or both.
(Item 44)
The polynucleotide according to item 43, wherein the transduction marker is a truncated EGFR molecule.
(Item 45)
At least about 75% identity to any one of SEQ ID NOs: 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 48, 50, 52, 54, 56, or 58. The polynucleotide according to any one of items 40 to 44, which comprises or comprises the polynucleotide having.
(Item 46)
A vector comprising the polynucleotide according to any one of items 40 to 45.
(Item 47)
The vector according to item 46, which is a viral vector.
(Item 48)
47. The vector according to item 47, wherein the viral vector is a lentivirus or a retrovirus vector.
(Item 49)
A host cell comprising the fusion protein according to any one of items 1 to 39, the polynucleotide according to any one of items 40 to 45, or the vector according to any one of items 46 to 48.
(Item 50)
The host cell according to item 49, which is an immune system cell.
(Item 51)
The host cell according to item 50, wherein the immune system cell is a T cell.
(Item 52)
51. The host cell of item 51, wherein the T cell is a CD4 ⁺ T cell or a CD8 ⁺ T cell.
(Item 53)
The host cell according to item 51 or 52, wherein the T cell is a memory T cell.
(Item 54)
The host cell according to any one of items 49 to 53, which is a human cell.
(Item 55)
Expression of the endogenous gene is inhibited in the host cell, and the endogenous gene is selected from the TCR gene, the HLA gene, the immunosuppressive component gene, or any combination thereof, items 49-54. The host cell according to any one of the above.
(Item 56)
55. The host cell of item 55, wherein the TCR gene is a T cell receptor α constant (TRAC) gene, a T cell receptor β constant (TRBC) gene, or both.
(Item 57)
The polynucleotide according to any one of items 40 to 45, or the vector according to any one of items 46 to 48, is targeted to the locus of the TCR gene or HLA gene via an endonuclease system. 55. The host cell according to item 55 or 56.
(Item 58)
58. The host cell according to item 57, wherein the endonuclease system is a CRISPR / Cas nuclease system, a zinc finger nuclease (ZFN) system, or a transcriptional activator-like effector nuclease (TALEN) system.
(Item 59)
Items that present a decrease in sustained phosphorylation of the fusion protein, a decrease in signaling of the fusion protein, a decrease in cytokine expression, an enhancement of survival, an enhancement of antigen-specific cytolytic activity, or any combination thereof. The host cell according to any one of 49 to 58.
(Item 60)
Items 49-59 expressing IL-2, TNF-α, or both induced by target antigens at reduced levels compared to host cells containing a fusion protein containing a wild-type CD28 co-stimulating signaling domain. The host cell according to any one of the above.
(Item 61)
The polynucleotide according to any one of items 40 to 45, the vector according to any one of items 46 to 48, or the host cell according to any one of items 49 to 60, and pharmaceutically acceptable. A pharmaceutical composition comprising a carrier, diluent, or excipient to be used.
(Item 62)
A method of treating a disease in a subject comprising administering to the subject the host cell according to any one of items 49 to 60 or the pharmaceutical composition according to item 61. A method associated with the presence of an antigen bound to a fusion protein.
(Item 63)
62. The method of item 62, wherein the disease is a viral infection, a bacterial infection, a cancer, an inflammatory disease, or an autoimmune disease.
(Item 64)
62. The method of item 62 or 63, wherein the subject is a human.
(Item 65)
62. The method of any one of items 62-64, wherein the host cell is allogeneic or self to the subject.
(Item 66)
References to which the subject had low or mild cytokine release syndrome, low or mild cell-related encephalopathy syndrome, or both after the treatment and / or was administered a reference host cell or composition. The fusion protein has reduced cytokine release syndrome, reduced cell-related encephalopathy syndrome, or both as compared to the subject, wherein the fusion protein comprises a wild-type CD28 costimulatory signaling domain, items 62-65. The method described in any one of the items.
(Item 67)
The method according to any one of items 62 to 66, wherein the disease is cancer.
(Item 68)
The cancers are solid tumor, melanoma, non-small cell lung cancer, renal cell cancer, kidney cancer, blood cancer, prostate cancer, castration-resistant prostate cancer, colon cancer, rectal cancer, gastric cancer, esophagus. Cancer, bladder cancer, head and neck cancer, thyroid cancer, breast cancer, triple negative breast cancer, ovarian cancer, cervical cancer, lung cancer, urinary tract epithelial cancer, pancreatic cancer, glioblastoma, hepatocellular carcinoma 67. The method of item 67, which is marrow, myeloma, multiple myeloma, leukemia, hodgkin lymphoma, non-hodgkin lymphoma, myelodystrophy syndrome, brain cancer, CNS cancer, or malignant glioma.
(Item 69)
The method of any one of items 62-68, further comprising administering a chemotherapeutic agent or an inhibitor of an immunosuppressive component.
(Item 70)
(A) The vector or expression construct according to any one of items 46 to 48, and the necessary reagents for transducing the vector or the expression construct into a host cell;
(B) The isolated polynucleotide according to any one of items 40 to 45, and an optional reagent for transducing the polynucleotide into a host cell; and / or
(C) The host cell according to any one of items 49 to 60.
Including, kit.

Claims

(A) An extracellular component containing a binding domain that specifically binds to a target antigen;
(B) An intracellular component comprising a modified functional CD28 co-stimulation signaling domain, wherein the modified functional CD28 co-stimulating signaling domain comprises at least one amino acid substitution. And (c) a fusion protein containing a hydrophobic moiety disposed between the extracellular component and the intracellular component.
The fusion protein is a fusion protein having one or more functional activities different from the fusion protein containing the wild-type CD28 co-stimulating signaling domain.

The fusion protein of claim 1, wherein at least one tyrosine residue in the CD28 co-stimulation signaling domain is replaced with a different amino acid residue.

(A) Whether the at least one tyrosine residue is selected from any one of positions 191, 206, 209 and 218.
(B) Whether at least two tyrosine residues selected from any of positions 191, 206, 209, and 218 are substituted with different amino acid residues, respectively.
(C) At least three tyrosine residues selected from any of positions 191, 206, 209 and 218 are each substituted with different amino acid residues or
(D) The fusion protein of claim 2, wherein the four tyrosine residues at positions 191, 206, 209, and 218 are replaced with different amino acid residues .

Each tyrosine residue is
(A) Independently substituted with tryptophan or phenylalanine residues ,
(B) Substituted with a phenylalanine residue or
(C) The fusion protein according to claim 2 or 3 , which is substituted with a tryptophan residue .

The modified CD28 co-stimulation signaling domain
(A) Including Y218F substitution or
(B) Contains or contains Y206F, Y209F, and Y218F substitutions
(C) The fusion protein according to any one of claims 2 to 4 , comprising Y191F, Y206F, Y209F, and Y218F substitutions .

The fusion protein according to any one of claims 1 to 5 , wherein at least one proline residue in the CD28 co-stimulation signaling domain is replaced with a different amino acid residue.

(A) Whether the at least one proline residue is selected from any one of positions 196, 199, 208, and 211.
(B) Whether at least two proline residues selected from any of positions 196, 199, 208, and 211 are substituted with different amino acid residues, respectively.
( C) At least three proline residues selected from any of positions 196, 199, 208, and 211 are each substituted with different amino acid residues, or
(D) The fusion protein of claim 6 , wherein the four proline residues at positions 196, 199, 208, and 211 are replaced with different amino acid residues .

The fusion protein of claim 6 or 7 , wherein each of the at least one, at least two, at least three, or four proline residues is substituted with an alanine residue.

The modified CD28 co-stimulation signaling domain further comprises L186 and L187 substitutions, wherein the L186 substitution is L186G and the L187 substitution is L187G, as needed, from claim 1. 8. The fusion protein according to any one of 8.

(I) Whether the at least one tyrosine residue and / or the at least one proline residue has been replaced with a conservative amino acid.
(Ii) The modified CD28 co-stimulation signaling domain is Y191, Y206, Y209, Y218, Y191 / Y206, Y191 / Y209, Y191 / Y218, Y206 / Y209, Y206 / Y218, Y209 / Y218, Y191 / Y206. Includes / Y209, Y191 / Y206 / Y218, Y191 / Y209 / Y218, Y206 / Y209 / Y218, or Y191 / Y206 / Y209 / Y218 substitutions, or
(Iii) The modified CD28 co-stimulation signaling domain is P196, P199, P208, P211, P196 / P199, P196 / P208, P196 / P211, P199 / P208, P199 / P211, P208 / P211, P196 / P199. Includes / P208, P196 / P199 / P211, P196 / P208 / P211, P199 / P208 / P211 or P196 / P199 / P208 / P211 substitutions, or
(Iv) The modified CD28 co-stimulation signaling domain is Y191, Y206, Y209, Y218, Y191 / Y206, Y191 / Y209, Y191 / Y218, Y206 / Y209, Y206 / Y218, Y209 / Y218, Y191 / Y206. Contains at least one tyrosine substitution selected from any one of / Y209, Y191 / Y206 / Y218, Y191 / Y209 / Y218, Y206 / Y209 / Y218, or Y191 / Y206 / Y209 / Y218, and P196. , P199, P208, P211, P196 / P199, P196 / P208, P196 / P211, P199 / P208, P199 / P211, P208 / P211, P196 / P199 / P208, P196 / P199 / P211, P196 / P208 / P211, P199. The modified CD28 co-stimulation signaling domain comprises the substitution of at least one proline selected from any one of / P208 / P211 or P196 / P199 / P208 / P211. Includes Y218 / P208 / P211 substitution or Y206 / P208 / Y209 / P211 / Y218 substitution,
(V) The modified CD28 co-stimulation signaling domain is Y191F, Y206F, Y209F, Y218F, Y191F / Y206F, Y191F / Y209F, Y191F / Y218F, Y206F / Y209F, Y206F / Y218F, Y209F / Y218F, Y191F. Includes / Y209F, Y191F / Y206F / Y218F, Y191F / Y209F / Y218F, Y206F / Y209F / Y218F, or Y191F / Y206F / Y209F / Y218F substitutions, or
(Vi) The modified CD28 co-stimulation signaling domains are P196A, P199A, P208A, P211A, P196A / P199A, P196A / P208A, P196A / P211A, P199A / P208A, P199A / P211A, P208A / P211A, P196A / P199. Includes / P208A, P196A / P199A / P211A, P196A / P208A / P211A, P199A / P208A / P211A, or P196A / P199A / P208A / P211A substitutions, or
(Vii) The modified CD28 co-stimulation signaling domain is Y191F, Y206F, Y209F, Y218F, Y191F / Y206F, Y191F / Y209F, Y191F / Y218F, Y206F / Y209F, Y206F / Y218F, Y209F / Y218F, Y19F. Contains at least one tyrosine substitution selected from any one of / Y209F, Y191F / Y206F / Y218F, Y191F / Y209F / Y218F, Y206F / Y209F / Y218F, or Y191F / Y206F / Y209F / Y218F, and P196A. , P199A, P208A, P211A, P196A / P199A, P196A / P208A, P196A / P211A, P199A / P208A, P199A / P211A, P208A / P211A, P196A / P199A / P208A, P196A / P199A / P211A, P196A / P199A / P211A Containing a substitution of at least one proline selected from any one of / P208A / P211A or P196A / P199A / P208A / P211A, wherein the modified CD28 co-stimulation signaling domain, if desired, comprises the above-mentioned modified CD28 co-stimulation signaling domain. Includes Y218F / P208A / P211A substitution or Y206F / P208A / Y209F / P211A / Y218F.
The fusion protein according to any one of claims 2 to 9.

(A) The modified CD28 co-stimulation signaling domain does not contain substitutions in Y191, P208, P211 or any combination thereof and / or .
(B) The binding domain is a scFv, scTCR, extracellular domain of a receptor, or ligand and / or
(C) The binding domain contains neither the extracellular binding domain of CD8 nor any portion thereof, nor any portion thereof, including a functional IgV-like domain, wherein the binding domain is optionally the CD8α chain. 17 . Fusion protein.

(I) The binding domain is chimeric, human, or humanized and / or
(Ii) The extracellular component further comprises a spacer disposed between the binding domain and the hydrophobic moiety, wherein the spacer, if necessary, is an immunoglobulin hinge region, CH2 domain, CH3. Includes domains, or any combination thereof, wherein the immunoglobulin hinge region is, optionally, the IgG4 hinge region and / or.
(Iii) The extracellular component further comprises a tag placed between the binding domain and the hydrophobic moiety, wherein the tag is optionally Trp-Ser-His-Pro-Gln-. The fusion protein according to any one of claims 1 to 11 , which has an amino acid sequence of Phe-Glu-Lys (SEQ ID NO: 40) .

The intracellular component further comprises an ITAM-containing T cell activation domain, wherein the ITAM-containing T cell activation domain is, optionally, a gamma chain of CD3γ, CD3δ, CD3ε, CD3ζ, FcεRI, or FcγRI. 13 . Fusion protein.

(A) The intracellular component further comprises at least one additional co-stimulation signaling domain, wherein the at least one additional co-stimulating signaling domain is optionally included in CD27, CD40L, GITR. NKG2C, CARD1, CD2, CD7, CD27, CD30, CD40, CD54 (ICAM), CD83, CD134 (OX-40), CD137 (4-1BB), CD150 (SLAMF1), CD152 (CTLA4), CD223 (LAG3), CD270 (HVEM), CD273 (PD-L2), CD274 (PD-L1), CD278 (ICOS), DAP10, LAT, NKD2C SLP76, TRIM, ZAP70, CD5, BAFF-R, SLAMF7, NKp80, CD160, B7-H3 , A ligand that specifically binds to CD83, or a combination thereof and / or
(B) The hydrophobic moiety is a transmembrane domain, where, if necessary, the transmembrane domain is CD28, CD2, CD3ε, CD3δ, CD3ζ, CD25, CD27, CD40, CD79A, CD79B, CD80, CD86. , CD95 (Fas), CD134 (OX40), CD137 (4-1BB), CD150 (SLAMF1), CD152 (CTLA4), CD200R, CD223 (LAG3), CD270 (HVEM), CD272 (BTLA), CD273 (PD-L2). ), CD274 (PD-L1), CD278 (ICOS), CD279 (PD-1), CD300, CD357 (GITR), A2aR, DAP10, FcRα, FcRβ, FcRγ, Fyn, GAL9, KIR, Lck, LAT, LRP, 1 . The fusion protein according to one item.

The target antigen is a cancer antigen, a viral antigen, a bacterial antigen, or a self-antigen, and if necessary, the target antigen is BCMA, CD3, CEACAM6, c-Met, EGFR, EGFRvIII, ErbB2, ErbB3, ErbB4, EphA2, IGF1R, GD2, O-acetyl GD2, O-acetyl GD3, GHRHR, GHR, FLT1, KDR, FLT4, CD44v6, CD151, CA125, CEA, CTLA-4, GITR, BTLA, TBF IL6R, gp130, Lewis A, Lewis Y, TNFR1, TNFR2, PD1, PD-L1, PD-L2, HVEM, MAGE-A, Mesoterin, NY-ESO-1, PSMA, RANK, ROR1, TNFRSF4, CD40, CD137, TWEAK-R, HLA, HLA-bound tumor or pathogen-related peptide, HLA-bound hTERT peptide, HLA-bound tyrosinase peptide, HLA-bound WT-1 peptide, LTβR, LIFRβ, LRP5, MUC1, OSMRβ, TCRα, TCRβ, CD19, CD20, CD22, CD25, CD28, CD30, CD33, CD52, CD56, CD79a, CD79b, CD80, CD81, CD86, CD123, CD171, CD276, B7H4, TLR7, TLR9, PTCH1, WT-1, The fusion protein according to any one of claims 1 to 14 , which is a cancer antigen selected from HA ¹ -H, Robo1, α-fet protein (AFP), Frizzled, OX40, PRAME, and SSX-2. ..

A polynucleotide encoding the fusion protein according to any one of claims 1 to 15 , wherein as required.
(I) The polynucleotide is codon-optimized and / or
(Ii) The polynucleotide is described in any one of SEQ ID NOs: 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 48, 50, 52, 54, 56, or 58. Containing and / or a polynucleotide having at least about 75% identity to the nucleotide sequence encoding the fusion protein
(Iii) The polynucleotide further comprises a polynucleotide encoding a transduction marker, a suicide gene, or both, wherein the transduction marker is, optionally, a truncated EGFR molecule and / or.
(Iv) The polynucleotide is relative to any one of SEQ ID NOs: 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 48, 50, 52, 54, 56, or 58. A polynucleotide comprising or consisting of a polynucleotide having at least about 75% identity .

16. The vector comprising the polynucleotide according to claim 16 , wherein the vector is, if necessary, a viral vector, and here, further, if necessary, the viral vector is a lentivirus or a retroviral vector. There is a vector .

A host cell comprising the fusion protein according to any one of claims 1 to 15 , the polynucleotide according to claim 16 , or the vector according to claim 17 , as required herein.
(A) The host cell is a hematopoietic precursor cell or an immune system cell, and if necessary, the immune system cell is a T cell or a natural killer cell, and here, further, if necessary, the T. The cells are CD4 ⁺ T cells or CD8 ⁺ T cells and / or the T cells are memory T cells and / or
(B) The host cell is a human cell and / or
(C) The expression of an endogenous gene is inhibited and inhibited in the host cell. The endogenous gene is selected from a TCR gene, an HLA gene, an immunosuppressive component gene, or any combination thereof. Further, if necessary,
(C) (i) The TCR gene is a T cell receptor α constant (TRAC) gene, a T cell receptor β constant (TRBC) gene, or both.
(C) (ii) The polynucleotide according to claim 16, or the vector according to claim 17, is targeted to the locus of the TCR gene or the HLA gene via an endonuclease system, where. Further, if necessary, a host cell in which the endonuclease system is a CRISPR / Casnuclease system, a zinc finger nuclease (ZFN) system, or a transcriptional activator-like effector nuclease (TALEN) system .

Presenting a decrease in sustained phosphorylation of the fusion protein, a decrease in signal transduction of the fusion protein, a decrease in cytokine expression, an enhancement of survival, an enhancement of antigen-specific cytolytic activity, or any combination thereof. /or
18. Claim 18 , which expresses IL-2, TNF-α, or both induced by a reduced level of target antigen compared to a host cell comprising a fusion protein comprising a wild-type CD28 co-stimulating signaling domain. Host cell.

A pharmaceutical composition comprising the polynucleotide of claim 16 , the vector of claim 17 , or the host cell of claim 18 or 19 , and a pharmaceutically acceptable carrier, diluent, or excipient. thing.

The host cell of claim 18 or 19 or the pharmaceutical composition of claim 20 for use in a method of treating a disease in a subject , wherein the disease is the presence of an antigen bound to a fusion protein. In connection with, where the method comprises administering the host cell or pharmaceutical composition, wherein, as needed.
(A) The disease is a viral, bacterial, cancer, inflammatory, or autoimmune disease and / or
(B) The subject is human and / or
(C) The host cell is allogeneic or self to the subject and / or
(D) The subject has low or mild cytokine release syndrome, low or mild cell-related encephalopathy syndrome, or both after the treatment and / or is administered by a reference host cell or composition. Compared to the referenced subjects, they have reduced cytokine release syndrome, reduced cell-related encephalopathy syndrome, or both, wherein the fusion protein comprises a wild-type CD28 costimulatory signaling domain. And / or
(E) The disease is cancer, and if necessary, the cancer is a solid tumor, melanoma, non-small cell lung cancer, renal cell cancer, kidney cancer, blood cancer, prostate cancer. , Cast-resistant prostate cancer, colon cancer, rectal cancer, stomach cancer, esophageal cancer, bladder cancer, head and neck cancer, thyroid cancer, breast cancer, triple negative breast cancer, ovarian cancer, cervical cancer, Lung cancer, urinary tract epithelial cancer, pancreatic cancer, glioblastoma, hepatocellular carcinoma, myeloma, multiple myeloma, leukemia, Hodgkin lymphoma, non-Hodgkin lymphoma, myelodystrophy syndrome, brain cancer, CNS cancer , Or malignant glioma, and / or
(F) A composition, wherein the method further comprises administering a chemotherapeutic agent or an inhibitor of an immunosuppressive component .

(A) The vector according to claim 17 , and a reagent as required for transducing the vector into a host cell;
(B) The isolated polynucleotide according to claim 16 and the necessary reagent for transducing the polynucleotide into a host cell; and / or (c) the host according to claim 18 or 19 . A kit containing cells.

The method for producing a host cell according to claim 18 or 19, which comprises transforming or transfecting the host cell with the polynucleotide according to claim 16 or the vector according to claim 17, as required. A method for producing a host cell, which comprises culturing the host cell.