JP2019528312A - mRNA媒介性の免疫化方法 - Google Patents
mRNA媒介性の免疫化方法 Download PDFInfo
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Abstract
Description
本出願は、ASCII形式で電子的手段により提出された配列表を含み、参照によりその全体が本明細書に組み込まれる。2017年8月3日に作成された前記配列表のASCII複製は、PAT057169−WO−PCT_SL.txtと命名され、サイズは146,992バイトである。
(i)Gタンパク質共役型受容体(GPCR);
(ii)1回膜貫通型タンパク質受容体;
(iii)腫瘍壊死因子受容体スーパーファミリー(TNFRSF)メンバー;
(iv)インターロイキン(IL)受容体;
(v)イオンチャネル;
(vi)溶質輸送体;
(vii)免疫受容体;および
(viii)複数回膜貫通タンパク質
から選択される、態様3の方法。
ABRB1、GABRB2、GABRB3、GABRD、GABRE、GABRG1、GABRG2、GABRG3、GABRP、GABRQ、GABRR1、GABRR2、GABRR3、GRIA1、GRIA2、GRIA3、GRIA4、GRID1、GRID2、GRIK1、GRIK2、GRIK3、GRIK4、GRIK5、GRIN1、GRIN2A、GRIN2B、GRIN2C、GRIN2D、GRIN3A、GRIN3B、GLRA1、GLRA2、GLRA3、GLRA4、P2RX1、P2RX2、P2RX3、P2RX4、P2RX5、P2RX6、P2RX7、ZACN、ASIC1、ASIC2、ASIC3、ASIC4、AQP1、AQP2、AQP3、AQP4、AQP5、AQP6、AQP7、AQP8、AQP9、AQP10、AQP11、AQP12A、AQP12B、MIP、CLCN1、CLCN2、CLCN3、CLCN4、CLCN5、CLCN6、CLCN7、CLCNKA、CLCNKB、嚢胞性線維症膜コンダクタンス制御因子(CFTR)、ANO1、ANO2、ANO3、ANO4、ANO5、ANO6、ANO7、ANO8、ANO9、ANO10、BEST1、BEST2、BEST3、BEST4、CLIC1、CLIC2、CLIC3、CLIC4、CLIC5、CLIC6、GJA1、GJA3、GJA4、GJA5、GJA6P、GJA8、GJA9、GJA10、GJB1、GJB2、GJB3、GJB4、GJB5、GJB6、GJB7、GJC1、GJC2、GJC3、GJD2、GJD3、GJD4、GJE1、ITPR1、ITPR2、ITPR3、PANX1、PANX2、PANX3、RYR1、RYR2、RYR3、NALCN、SCNN1A、SCNN1B、SCNN1D、SCNN1G、TEM16A、ADAMTS7、ANGPTL3、ANGPTL4、ANGPTL8、LPL、GDF15、ガレクチン−1、ガレクチン−2、ガレクチン−3、ガレクチン−4、ガレクチン−7、ガレクチン−8、ガレクチン−9、ガレクチン−10、ガレクチン−12、ガレクチン−13、マトリックスglaタンパク質(MGP)、PRNP、DGAT1、GPAT3、DMC1、BLM、BRCA2、ヒト内在性レトロウイルスタイプK(HERV−K)ファミリーのメンバー、エクトヌクレオシド三リン酸ジホスホヒドロラーゼ1(ENTPD1)、エクトヌクレオシド三リン酸ジホスホヒドロラーゼ2(ENTPD2)、SLC1A1、SLC1A2、SLC1A3、SLC1A4、SLC1A5、SLC1A6、SLC1A7、SLC2A1、SLC2A2、SLC2A3、SLC2A4、SLC2A5、SLC2A6、SLC2A7、SLC2A8、SLC2A9、SLC2A10、SLC2A11、SLC2A12、SLC2A13、SLC2A14、SLC3A1、SLC3A2、SLC4A1、SLC4A2、SLC4A3、SLC4A4、SLC4A5、SLC4A6、SLC4A7、SLC4A8、SLC4A9、SLC4A10、SLC4A11、SLC5A1、SLC5A2、SLC5A3、SLC5A4、SLC5A5、SLC5A6、SLC5A7、SLC5A8、SLC5A9、SLC5A10、SLC5A11、SLC5A12、SLC6A1、SLC6A2、SLC6A3、SLC6A4、SLC6A5、SLC6A6、SLC6A7、SLC6A8、SLC6A9、SLC6A10、SLC6A11、SLC6A12、SLC6A13、SLC6A14、SLC6A15、SLC6A16、SLC6A17、SLC6A18、SLC6A19、SLC6A20、SLC7A5、SLC7A6、SLC7A7、SLC7A8、SLC7A9、SLC7A10、SLC7A11、SLC7A13、SLC7A14、SLC8A1、SLC8A2、SLC8A3、SLC9A1、SLC9A2、SLC9A3、SLC9A4、SLC9A5、SLC9A6、SLC9A7、SLC9A8、SLC9A9、SLC9A10、SLC9A11、SLC9B1、SLC9B2、SLC10A1、SLC10A2、SLC10A3、SLC10A4、SLC10A5、SLC10A6、SLC10A7、SLC11A1、SLC11A2、SLC12A1、SLC12A2、SLC12A3、SLC12A4、SLC12A5、SLC12A6、SLC12A7、SLC12A8、SLC12A9、SLC13A1、SLC13A2、SLC13A3、SLC13A4、SLC13A5、SLC14A1、SLC14A2、SLC15A1、SLC15A2、SLC15A3、SLC15A4、SLC16A1、SLC16A2、SLC16A3、SLC16A4、SLC16A5、SLC16A6、SLC16A7、SLC16A8、SLC16A9、SLC16A10、SLC16A11、SLC16A12、SLC16A13、SLC16A14、SLC17A1、SLC17A2、SLC17A3、SLC17A4、SLC17A5、SLC17A6、SLC17A7、SLC17A8、SLC17A9、SLC18A1、SLC18A2、SLC18A3、SLC19A1、SLC19A2、SLC19A3、SLC20A1、SLC20A2、SLCO1A2、SLCO1B1、SLCO1B3、SLCO1C1、SLCO2A1、SLCO2B1、SLCO3A1、SLCO4A1、SLCO4C1、SLCO5A1、SLCO6A1、SLC22A1、SLC22A2、SLC22A3、SLC22A4、SLC22A5、SLC22A6、SLC22A7、SLC22A8、SLC22A9、SLC22A10、SLC22A11、SLC22A12、SLC22A13、SLC22A14、SLC22A15、SLC22A16、SLC22A17、SLC22A18、SLC22A18AS,SLC22A19、SLC22A20、SLC22A23、SLC22A24、SLC22A25、SLC22A31、SLC23A1、SLC23A2、SLC23A3、SLC23A4、SLC24A1、SLC24A2、SLC24A3、SLC24A4、SLC24A5、SLC24A6、SLC25A1、SLC25A2、SLC25A3、SLC25A4、SLC25A5、SLC25A6、SLC25A7、SLC25A8、SLC25A9、SLC25A10、SLC25A11、SLC25A12、SLC25A13、SLC25A14、SLC25A15、SLC25A16、SLC25A17、SLC25A18、SLC25A19,SLC25A20、SLC25A21、SLC25A22、SLC25A23、SLC25A24、SLC25A25、SLC25A26、SLC25A27、SLC25A28、SLC25A29、SLC25A30、SLC25A31、SLC25A32、SLC25A33、SLC25A34、SLC25A35、SLC25A36、SLC25A37,SLC25A38、SLC25A39、SLC25A40、SLC25A41、SLC25A42、SLC25A43、SLC25A44、SLC25A45、SLC25A46、SLC26A1、SLC26A2、SLC26A3、SLC26A4、SLC26A5、SLC26A6、SLC26A7、SLC26A8、SLC26A9、SLC26A10、SLC26A11、SLC27A1、SLC27A2、SLC27A3、SLC27A4、SLC27A5、SLC27A6、SLC28A1、SLC28A2、SLC28A3、SLC29A1、SLC29A2、SLC29A3、SLC29A4、SLC30A1、SLC30A2、SLC30A3、SLC30A4、SLC30A5、SLC30A6、SLC30A7、SLC30A8、SLC30A9、SLC30A10、SLC31A1、SLC31A2、SLC32A1、SLC33A1、SLC34A1、SLC34A2、SLC34A3、SLC35A1、SLC35A2、SLC35A3、SLC35A4、SLC35A5、SLC35B1、SLC35B2、SLC35B3、SLC35B4、SLC35C1、SLC35C2、SLC35D1、SLC35D2、SLC35D3、SLC35E1、SLC35E2、SLC35E3、SLC35E4、SLC35F1、SLC35F2、SLC35F3、SLC35F4、SLC35F5、SLC35G1、SLC35G3、SLC35G4、SLC35G5、SLC35G6、SLC36A1、SLC36A2、SLC36A3、SLC36A4、SLC37A1、SLC37A2、SLC37A3、SLC37A4、SLC38A1、SLC38A2、SLC38A3、SLC38A4、SLC38A5、SLC38A6、SLC38A7、SLC38A8、SLC38A9、SLC38A10、SLC38A11、SLC39A1、SLC39A2、SLC39A3、SLC39A4、SLC39A5、SLC39A6、SLC39A7、SLC39A8、SLC39A9、SLC39A10、SLC39A11、SLC39A12、SLC39A13、SLC39A14、SLC40A1、SLC41A1、SLC41A2、SLC41A3、RhAG、RhBG、RhCG、SLC43A1、SLC43A2、SLC43A3、SLC44A1、SLC44A2、SLC44A3、SLC44A4、SLC44A5、SLC45A1、SLC45A2、SLC45A3、SLC45A4、SLC46A1、SLC46A2、SLC46A3、SLC47A1、SLC47A2、HCP−1、MFSD5、MFSD10、SLC50A1、OSTα、OSTβ、SLC52A1、SLC52A2、ならびにSLC52A3から選択される、態様1または2の方法。
ABRB1、GABRB2、GABRB3、GABRD、GABRE、GABRG1、GABRG2、GABRG3、GABRP、GABRQ、GABRR1、GABRR2、GABRR3、GRIA1、GRIA2、GRIA3、GRIA4、GRID1、GRID2、GRIK1、GRIK2、GRIK3、GRIK4、GRIK5、GRIN1、GRIN2A、GRIN2B、GRIN2C、GRIN2D、GRIN3A、GRIN3B、GLRA1、GLRA2、GLRA3、GLRA4、P2RX1、P2RX2、P2RX3、P2RX4、P2RX5、P2RX6、P2RX7、ZACN、ASIC1、ASIC2、ASIC3、ASIC4、AQP1、AQP2、AQP3、AQP4、AQP5、AQP6、AQP7、AQP8、AQP9、AQP10、AQP11、AQP12A、AQP12B、MIP、CLCN1、CLCN2、CLCN3、CLCN4、CLCN5、CLCN6、CLCN7、CLCNKA、CLCNKB、嚢胞性線維症膜コンダクタンス制御因子(CFTR)、ANO1、ANO2、ANO3、ANO4、ANO5、ANO6、ANO7、ANO8、ANO9、ANO10、BEST1、BEST2、BEST3、BEST4、CLIC1、CLIC2、CLIC3、CLIC4、CLIC5、CLIC6、GJA1、GJA3、GJA4、GJA5、GJA6P、GJA8、GJA9、GJA10、GJB1、GJB2、GJB3、GJB4、GJB5、GJB6、GJB7、GJC1、GJC2、GJC3、GJD2、GJD3、GJD4、GJE1、ITPR1、ITPR2、ITPR3、PANX1、PANX2、PANX3、RYR1、RYR2、RYR3、NALCN、SCNN1A、SCNN1B、SCNN1D、SCNN1G、TEM16A、ADAMTS7、ANGPTL3、ANGPTL4、ANGPTL8、LPL、GDF15、ガレクチン−1、ガレクチン−2、ガレクチン−3、ガレクチン−4、ガレクチン−7、ガレクチン−8、ガレクチン−9、ガレクチン−10、ガレクチン−12、ガレクチン−13、マトリックスglaタンパク質(MGP)、PRNP、DGAT1、GPAT3、DMC1、BLM、BRCA2、ヒト内在性レトロウイルスタイプK(HERV−K)ファミリーのメンバー、エクトヌクレオシド三リン酸ジホスホヒドロラーゼ1(ENTPD1)、エクトヌクレオシド三リン酸ジホスホヒドロラーゼ2(ENTPD2)、SLC1A1、SLC1A2、SLC1A3、SLC1A4、SLC1A5、SLC1A6、SLC1A7、SLC2A1、SLC2A2、SLC2A3、SLC2A4、SLC2A5、SLC2A6、SLC2A7、SLC2A8、SLC2A9、SLC2A10、SLC2A11、SLC2A12、SLC2A13、SLC2A14、SLC3A1、SLC3A2、SLC4A1、SLC4A2、SLC4A3、SLC4A4、SLC4A5、SLC4A6、SLC4A7、SLC4A8、SLC4A9、SLC4A10、SLC4A11、SLC5A1、SLC5A2、SLC5A3、SLC5A4、SLC5A5、SLC5A6、SLC5A7、SLC5A8、SLC5A9、SLC5A10、SLC5A11、SLC5A12、SLC6A1、SLC6A2、SLC6A3、SLC6A4、SLC6A5、SLC6A6、SLC6A7、SLC6A8、SLC6A9、SLC6A10、SLC6A11、SLC6A12、SLC6A13、SLC6A14、SLC6A15、SLC6A16、SLC6A17、SLC6A18、SLC6A19、SLC6A20、SLC7A5、SLC7A6、SLC7A7、SLC7A8、SLC7A9、SLC7A10、SLC7A11、SLC7A13、SLC7A14、SLC8A1、SLC8A2、SLC8A3、SLC9A1、SLC9A2、SLC9A3、SLC9A4、SLC9A5、SLC9A6、SLC9A7、SLC9A8、SLC9A9、SLC9A10、SLC9A11、SLC9B1、SLC9B2、SLC10A1、SLC10A2、SLC10A3、SLC10A4、SLC10A5、SLC10A6、SLC10A7、SLC11A1、SLC11A2、SLC12A1、SLC12A2、SLC12A3、SLC12A4、SLC12A5、SLC12A6、SLC12A7、SLC12A8、SLC12A9、SLC13A1、SLC13A2、SLC13A3、SLC13A4、SLC13A5、SLC14A1、SLC14A2、SLC15A1、SLC15A2、SLC15A3、SLC15A4、SLC16A1、SLC16A2、SLC16A3、SLC16A4、SLC16A5、SLC16A6、SLC16A7、SLC16A8、SLC16A9、SLC16A10、SLC16A11、SLC16A12、SLC16A13、SLC16A14、SLC17A1、SLC17A2、SLC17A3、SLC17A4、SLC17A5、SLC17A6、SLC17A7、SLC17A8、SLC17A9、SLC18A1、SLC18A2、SLC18A3、SLC19A1、SLC19A2、SLC19A3、SLC20A1、SLC20A2、SLCO1A2、SLCO1B1、SLCO1B3、SLCO1C1、SLCO2A1、SLCO2B1、SLCO3A1、SLCO4A1、SLCO4C1、SLCO5A1、SLCO6A1、SLC22A1、SLC22A2、SLC22A3、SLC22A4、SLC22A5、SLC22A6、SLC22A7、SLC22A8、SLC22A9、SLC22A10、SLC22A11、SLC22A12、SLC22A13、SLC22A14、SLC22A15、SLC22A16、SLC22A17、SLC22A18、SLC22A18AS,SLC22A19、SLC22A20、SLC22A23、SLC22A24、SLC22A25、SLC22A31、SLC23A1、SLC23A2、SLC23A3、SLC23A4、SLC24A1、SLC24A2、SLC24A3、SLC24A4、SLC24A5、SLC24A6、SLC25A1、SLC25A2、SLC25A3、SLC25A4、SLC25A5、SLC25A6、SLC25A7、SLC25A8、SLC25A9、SLC25A10、SLC25A11、SLC25A12、SLC25A13、SLC25A14、SLC25A15、SLC25A16、SLC25A17、SLC25A18、SLC25A19,SLC25A20、SLC25A21、SLC25A22、SLC25A23、SLC25A24、SLC25A25、SLC25A26、SLC25A27、SLC25A28、SLC25A29、SLC25A30、SLC25A31、SLC25A32、SLC25A33、SLC25A34、SLC25A35、SLC25A36、SLC25A37,SLC25A38、SLC25A39、SLC25A40、SLC25A41、SLC25A42、SLC25A43、SLC25A44、SLC25A45、SLC25A46、SLC26A1、SLC26A2、SLC26A3、SLC26A4、SLC26A5、SLC26A6、SLC26A7、SLC26A8、SLC26A9、SLC26A10、SLC26A11、SLC27A1、SLC27A2、SLC27A3、SLC27A4、SLC27A5、SLC27A6、SLC28A1、SLC28A2、SLC28A3、SLC29A1、SLC29A2、SLC29A3、SLC29A4、SLC30A1、SLC30A2、SLC30A3、SLC30A4、SLC30A5、SLC30A6、SLC30A7、SLC30A8、SLC30A9、SLC30A10、SLC31A1、SLC31A2、SLC32A1、SLC33A1、SLC34A1、SLC34A2、SLC34A3、SLC35A1、SLC35A2、SLC35A3、SLC35A4、SLC35A5、SLC35B1、SLC35B2、SLC35B3、SLC35B4、SLC35C1、SLC35C2、SLC35D1、SLC35D2、SLC35D3、SLC35E1、SLC35E2、SLC35E3、SLC35E4、SLC35F1、SLC35F2、SLC35F3、SLC35F4、SLC35F5、SLC35G1、SLC35G3、SLC35G4、SLC35G5、SLC35G6、SLC36A1、SLC36A2、SLC36A3、SLC36A4、SLC37A1、SLC37A2、SLC37A3、SLC37A4、SLC38A1、SLC38A2、SLC38A3、SLC38A4、SLC38A5、SLC38A6、SLC38A7、SLC38A8、SLC38A9、SLC38A10、SLC38A11、SLC39A1、SLC39A2、SLC39A3、SLC39A4、SLC39A5、SLC39A6、SLC39A7、SLC39A8、SLC39A9、SLC39A10、SLC39A11、SLC39A12、SLC39A13、SLC39A14、SLC40A1、SLC41A1、SLC41A2、SLC41A3、RhAG、RhBG、RhCG、SLC43A1、SLC43A2、SLC43A3、SLC44A1、SLC44A2、SLC44A3、SLC44A4、SLC44A5、SLC45A1、SLC45A2、SLC45A3、SLC45A4、SLC46A1、SLC46A2、SLC46A3、SLC47A1、SLC47A2、HCP−1、MFSD5、MFSD10、SLC50A1、OSTα、OSTβ、SLC52A1、SLC52A2、ならびにSLC52A3から選択される、態様51〜77のいずれか1つの方法。
(a)次のシチジンについての修飾ヌクレオチド:5−ホルミルシチジン、5−メチルシチジン、5−メトキシシチジン、5−ヒドロキシシチジン、および5−ヒドロキシメチルシチジンの1つ以上;
(b)次のウリジンについての修飾ヌクレオチド:5−ホルミルウリジン、5−メチルウリジン、5−メトキシウリジン、5−カルボキシメチルエステルウリジン、プソイドウリジン、およびN1−メチルプソイドウリジンの1つ以上;
(c)アデノシンについての修飾ヌクレオチドとしてN6−メチルアデノシン:および/または
(d)グアノシンについての修飾ヌクレオチドとしてチエノグアノシンを含む、態様1〜84のいずれか1つの方法。
「クローニングベクター」は、プラスミドもしくはファージDNAまたはその他のDNA配列を意味し、これは、宿主細胞において自律的に複製が可能であり、かつ1つまたは少数の制限エンドヌクレアーゼ認識部位により特徴づけられ、そこでは、そのようなDNA配列をベクターの必須の生物学的機能を失うことなく確定的に切断し得、その複製およびクローニングをもたらすためにそこにDNAがスプライスされ得る。クローニングベクターはさらに、クローニングベクターによりトランスフェクトされた細胞の同定における使用に適したマーカーを含有してもよい。マーカーは、例えば、テトラサイクリン抵抗性またはアンピシリン抵抗性をもたらす。
本明細書で提供されるのは、動物(例えば、非ヒト動物)における標的タンパク質(例えば、ヒト膜貫通タンパク質、例えば、ヒトGPCRなどのヒトタンパク質)またはその断片に対する免疫応答を誘導するための方法であって、その動物に、脂質および標的タンパク質またはその断片をコードするポリリボヌクレオチド(例えば、mRNA)などのポリヌクレオチドを含む複合体を含む組成物を投与する方法と、そのような標的タンパク質に対する抗体を作製する関連した方法である。本明細書で提供される方法は、目的の任意の標的タンパク質(例えば、膜貫通タンパク質)に対する抗体(例えば、モノクローナル抗体)を作製するか、またはそれに対する免疫応答を誘導するために有用である。特定の態様では、標的タンパク質は、ヒト標的タンパク質であり、本明細書に記載の方法において使用される免疫化された動物は、非ヒト動物である。
、CHRNA2、CHRNA3、CHRNA4、CHRNA5、CHRNA6、CHRNA7、CHRNA9、CHRNA10、CHRNB1、CHRNB2、CHRNB3、CHRNB4、CHRND、CHRNE、CHRNG、GABRA1、GABRA2、GABRA3、GABRA4、GABRA5、GABRA6、GABRB1、GABRB2、GABRB3、GABRD、GABRE、GABRG1、GABRG2、GABRG3、GABRP、GABRQ、GABRR1、GABRR2、GABRR3、GRIA1、GRIA2、GRIA3、GRIA4、GRID1、GRID2、GRIK1、GRIK2、GRIK3、GRIK4、GRIK5、GRIN1、GRIN2A、GRIN2B、GRIN2C、GRIN2D、GRIN3A、GRIN3B、GLRA1、GLRA2、GLRA3、GLRA4、P2RX1、P2RX2、P2RX3、P2RX4、P2RX5、P2RX6、P2RX7、ZACN、ASIC1、ASIC2、ASIC3、ASIC4、AQP1、AQP2、AQP3、AQP4、AQP5、AQP6、AQP7、AQP8、AQP9、AQP10、AQP11、AQP12A、AQP12B、MIP、CLCN1、CLCN2、CLCN3、CLCN4、CLCN5、CLCN6、CLCN7、CLCNKA、CLCNKB、嚢胞性線維症膜コンダクタンス制御因子(CFTR)、ANO1/TMEM16a、ANO2、ANO3、ANO4、ANO5、ANO6、ANO7、ANO8、ANO9、ANO10、BEST1、BEST2、BEST3、BEST4、CLIC1、CLIC2、CLIC3、CLIC4、CLIC5、CLIC6、GJA1、GJA3、GJA4、GJA5、GJA6P、GJA8、GJA9、GJA10、GJB1、GJB2、GJB3、GJB4、GJB5、GJB6、GJB7、GJC1、GJC2、GJC3、GJD2、GJD3、GJD4、GJE1、ITPR1、ITPR2、ITPR3、PANX1、PANX2、PANX3、RYR1、RYR2、RYR3、NALCN、SCNN1A、SCNN1B、SCNN1D、SCNN1G、ADAMTS7、ANGPTL3、ANGPTL4、ANGPTL8、LPL、GDF15、ガレクチン−1、ガレクチン−2、ガレクチン−3、ガレクチン−4、ガレクチン−7、ガレクチン−8、ガレクチン−9、ガレクチン−10、ガレクチン−12、ガレクチン−13、マトリックスglaタンパク質(MGP)、PRNP、DGAT1、GPAT3、DMC1、BLM、BRCA2、ヒト内在性レトロウイルスタイプK(HERV−K)ファミリーのメンバー、エクトヌクレオシド三リン酸ジホスホヒドロラーゼ1(ENTPD1)、エクトヌクレオシド三リン酸ジホスホヒドロラーゼ2(ENTPD2)、SLC1A1、SLC1A2、SLC1A3、SLC1A4、SLC1A5、SLC1A6、SLC1A7、SLC2A1、SLC2A2、SLC2A3、SLC2A4、SLC2A5、SLC2A6、SLC2A7、SLC2A8、SLC2A9、SLC2A10、SLC2A11、SLC2A12、SLC2A13、SLC2A14、SLC3A1、SLC3A2、SLC4A1、SLC4A2、SLC4A3、SLC4A4、SLC4A5、SLC4A6、SLC4A7、SLC4A8、SLC4A9、SLC4A10、SLC4A11、SLC5A1、SLC5A2、SLC5A3、SLC5A4、SLC5A5、SLC5A6、SLC5A7、SLC5A8、SLC5A9、SLC5A10、SLC5A11、SLC5A12、SLC6A1、SLC6A2、SLC6A3、SLC6A4、SLC6A5、SLC6A6、SLC6A7、SLC6A8、SLC6A9、SLC6A10、SLC6A11、SLC6A12、SLC6A13、SLC6A14、SLC6A15、SLC6A16、SLC6A17、SLC6A18、SLC6A19、SLC6A20、SLC7A5、SLC7A6、SLC7A7、SLC7A8、SLC7A9、SLC7A10、SLC7A11、SLC7A13、SLC7A14、SLC8A1、SLC8A2、SLC8A3、SLC9A1、SLC9A2、SLC9A3、SLC9A4、SLC9A5、SLC9A6、SLC9A7、SLC9A8、SLC9A9、SLC9A10、SLC9A11、SLC9B1、SLC9B2、SLC10A1、SLC10A2、SLC10A3、SLC10A4、SLC10A5、SLC10A6、SLC10A7、SLC11A1、SLC11A2、SLC12A1、SLC12A2、SLC12A3、SLC12A4、SLC12A5、SLC12A6、SLC12A7、SLC12A8、SLC12A9、SLC13A1、SLC13A2、SLC13A3、SLC13A4、SLC13A5、SLC14A1、SLC14A2、SLC15A1、SLC15A2、SLC15A3、SLC15A4、SLC16A1、SLC16A2、SLC16A3、SLC16A4、SLC16A5、SLC16A6、SLC16A7、SLC16A8、SLC16A9、SLC16A10、SLC16A11、SLC16A12、SLC16A13、SLC16A14、SLC17A1、SLC17A2、SLC17A3、SLC17A4、SLC17A5、SLC17A6、SLC17A7、SLC17A8、SLC17A9、SLC18A1、SLC18A2、SLC18A3、SLC19A1、SLC19A2、SLC19A3、SLC20A1、SLC20A2、SLCO1A2、SLCO1B1、SLCO1B3、SLCO1C1、SLCO2A1、SLCO2B1、SLCO3A1、SLCO4A1、SLCO4C1、SLCO5A1、SLCO6A1、SLC22A1、SLC22A2、SLC22A3、SLC22A4、SLC22A5、SLC22A6、SLC22A7、SLC22A8、SLC22A9、SLC22A10、SLC22A11、SLC22A12、SLC22A13、SLC22A14、SLC22A15、SLC22A16、SLC22A17、SLC22A18、SLC22A18AS,SLC22A19、SLC22A20、SLC22A23、SLC22A24、SLC22A25、SLC22A31、SLC23A1、SLC23A2、SLC23A3、SLC23A4、SLC24A1、SLC24A2、SLC24A3、SLC24A4、SLC24A5、SLC24A6、SLC25A1、SLC25A2、SLC25A3、SLC25A4、SLC25A5、SLC25A6、SLC25A7、SLC25A8、SLC25A9、SLC25A10、SLC25A11、SLC25A12、SLC25A13、SLC25A14、SLC25A15、SLC25A16、SLC25A17、SLC25A18、SLC25A19,SLC25A20、SLC25A21、SLC25A22、SLC25A23、SLC25A24、SLC25A25、SLC25A26、SLC25A27、SLC25A28、SLC25A29、SLC25A30、SLC25A31、SLC25A32、SLC25A33、SLC25A34、SLC25A35、SLC25A36、SLC25A37,SLC25A38、SLC25A39、SLC25A40、SLC25A41、SLC25A42、SLC25A43、SLC25A44、SLC25A45、SLC25A46、SLC26A1、SLC26A2、SLC26A3、SLC26A4、SLC26A5、SLC26A6、SLC26A7、SLC26A8、SLC26A9、SLC26A10、SLC26A11、SLC27A1、SLC27A2、SLC27A3、SLC27A4、SLC27A5、SLC27A6、SLC28A1、SLC28A2、SLC28A3、SLC29A1、SLC29A2、SLC29A3、SLC29A4、SLC30A1、SLC30A2、SLC30A3、SLC30A4、SLC30A5、SLC30A6、SLC30A7、SLC30A8、SLC30A9、SLC30A10、SLC31A1、SLC31A2、SLC32A1、SLC33A1、SLC34A1、SLC34A2、SLC34A3、SLC35A1、SLC35A2、SLC35A3、SLC35A4、SLC35A5、SLC35B1、SLC35B2、SLC35B3、SLC35B4、SLC35C1、SLC35C2、SLC35D1、SLC35D2、SLC35D3、SLC35E1、SLC35E2、SLC35E3、SLC35E4、SLC35F1、SLC35F2、SLC35F3、SLC35F4、SLC35F5、SLC35G1、SLC35G3、SLC35G4、SLC35G5、SLC35G6、SLC36A1、SLC36A2、SLC36A3、SLC36A4、SLC37A1、SLC37A2、SLC37A3、SLC37A4、SLC38A1、SLC38A2、SLC38A3、SLC38A4、SLC38A5、SLC38A6、SLC38A7、SLC38A8、SLC38A9、SLC38A10、SLC38A11、SLC39A1、SLC39A2、SLC39A3、SLC39A4、SLC39A5、SLC39A6、SLC39A7、SLC39A8、SLC39A9、SLC39A10、SLC39A11、SLC39A12、SLC39A13、SLC39A14、SLC40A1、SLC41A1、SLC41A2、SLC41A3、RhAG、RhBG、RhCG、SLC43A1、SLC43A2、SLC43A3、SLC44A1、SLC44A2、SLC44A3、SLC44A4、SLC44A5、SLC45A1、SLC45A2、SLC45A3、SLC45A4、SLC46A1、SLC46A2、SLC46A3、SLC47A1、SLC47A2、HCP−1、MFSD5、MFSD10、SLC50A1、OSTα、OSTβ、SLC52A1、SLC52A2、ならびにSLC52A3から選択される。
特定の態様では、本明細書で提供される方法における使用のための複合体は、脂質および標的タンパク質をコードするポリヌクレオチド(例えば、mRNA)を含み、標的タンパク質(例えば、ヒト標的タンパク質)は、GPCR(例えば、ヒトGPCR)である。7回膜貫通(7TM)ドメイン受容体およびGタンパク質結合受容体(GPLR)としても知られるGPCR。GPCRに関係する2つの主要なシグナル伝達経路は、cAMPシグナル経路およびホスファチジルイノシトールシグナル経路である。
特定の態様では、本明細書で提供される方法における使用のための複合体は、脂質および標的タンパク質をコードするポリヌクレオチド(例えば、mRNA)を含み、標的タンパク質は、パターン認識受容体(PRR)、トール様受容体(TLR)、キラー活性化およびキラー抑制受容体(KARおよびKIR)、補体受容体、Fc受容体、B細胞受容体、およびT細胞受容体(例えば、TCR−α、TCR−β、CD3、ζ−鎖アクセサリー、CD4、およびCD8)ならびに主要組織適合複合体を含む。
特定の態様では、本明細書で提供される方法における使用のための複合体は、脂質および標的タンパク質をコードするポリヌクレオチド(例えば、mRNA)を含み、標的タンパク質は、サイトカイン受容体(例えば、インターロイキン(IL)受容体または線維芽細胞増殖因子(FGF)受容体)である。サイトカイン受容体の非限定的な例は、神経成長因子(NGF)、ミオスタチン(GDF−8)、増殖分化因子(GDF)、顆粒球マクロファージコロニー刺激因子(GM−CSF)、顆粒球コロニー刺激因子(G−CSF)、血小板由来増殖因子(PDGF)、エリスロポエチン(EPO)、トロンボポチエン(TPO)、上皮増殖因子(EGF)、線維芽細胞増殖因子(FGF)、血管内皮増殖因子(VEGF)、組織メタロプロテアーゼ阻害物質(TIMP)、マトリックスメタロプロテアーゼ(MMP)、マクロファージ刺激因子(MSF)、毛様体神経栄養因子(CNTF)、カルジオトロフィン、オンコスタチンM、白血病抑制因子(LIF)、形質転換増殖因子(TGF)−αおよび−β、インターフェロン(IFN)−βおよび−γ、ならびに腫瘍壊死因子(TNF)αに対する受容体を含む。ある特定の実施形態では、本明細書で提供される方法における使用のための複合体は、脂質および標的タンパク質をコードするポリヌクレオチド(例えば、mRNA)を含み、標的タンパク質はgp130であり、これは、IL−6、IL−11、IL−27、白血病抑制因子(LIF)、オンコスタチンM(OSM)、毛様体神経栄養因子(CNTF)、カルジオトロフィン1(CT−1)、およびカルジオトロフィン様サイトカイン(CLC)を含むいくつかの関連するサイトカインによって利用される共有のレセプターである。
特定の態様では、本明細書で提供される方法における使用のための複合体は、脂質および標的タンパク質をコードするポリヌクレオチド(例えば、mRNA)を含み、標的タンパク質は、イオンチャネルである。
特定の態様では、本明細書で提供される方法における使用のための複合体は、脂質および標的タンパク質をコードするポリヌクレオチド(例えば、mRNA)を含み、標的タンパク質は、溶質輸送体である。溶質輸送体タンパク質は、脂質膜の一方から他方に溶質を輸送する能力によって特徴づけられる膜内在性タンパク質である。このタンパク質の群は、溶質を電気化学的勾配に逆らって移動させる二次性能動輸送体、および溶質をそれらの電気化学的勾配の方向に移動させる促進性輸送体を含む。溶質輸送体は、300種を超えるタンパク質を包含する52種のファミリーにまとめられる。52種のファミリーおよびその非限定的な例となるメンバーを以下に示す:
(1)高親和性グルタミン酸および中性アミノ酸輸送体ファミリーであって、非限定的な例は、SLC1A1、SLC1A2、SLC1A3、SLC1A4、SLC1A5、SLC1A6、およびSLC1A7を含み;
(2)促進性グルコース(GLUT)輸送体ファミリーであって、非限定的な例は、SLC2A1、SLC2A2、SLC2A3、SLC2A4、SLC2A5、SLC2A6、SLC2A7、SLC2A8、SLC2A9、SLC2A10、SLC2A11、SLC2A12、SLC2A13、およびSLC2A14を含み;
(3)ヘテロ二量体アミノ酸ファミリーの大サブユニットであって、非限定的な例は、SLC3A1およびSLC3A2を含み;
(4)重炭酸塩ファミリーであって、例は、SLC4A1、SLC4A2、SLC4A3、SLC4A4、SLC4A5、SLC4A6、SLC4A7、SLC4A8、SLC4A9、SLC4A10、およびSLC4A11を含み;
(5)ナトリウム・グルコース共輸送体ファミリーであって、例は、SLC5A1、SLC5A2、SLC5A3、SLC5A4、SLC5A5、SLC5A6、SLC5A7、SLC5A8、SLC5A9、SLC5A10、SLC5A11、およびSLC5A12を含み;
(6)ナトリウムおよびクロライド依存性ナトリウム:神経伝達物質共輸送体ファミリーであって、例は、SLC6A1、SLC6A2、SLC6A3、SLC6A4、SLC6A5、SLC6A6、SLC6A7、SLC6A8、SLC6A9、SLC6A10、SLC6A11、SLC6A12、SLC6A13、SLC6A14、SLC6A15、SLC6A16、SLC6A17、SLC6A18、SLC6A19、およびSLC6A20を含み;
(7)カチオン性アミノ酸輸送体/糖タンパク質関連ファミリーであって、非限定的な例は、(i)カチオン性アミノ酸輸送体(SLC7A1、SLC7A2、SLC7A3、SLC7A4)および(ii)ヘテロ二量体アミノ酸輸送体の糖タンパク質関連/小または触媒サブユニット(SLC7A5、SLC7A6、SLC7A7、SLC7A8、SLC7A9、SLC7A10、SLC7A11、SLC7A13、SLC7A14)を含み;
(8)Na+/Ca2+交換輸送体ファミリーであって、非限定的な例は、SLC8A1、SLC8A2、およびSLC8A3を含み;
(9)Na+/H+交換輸送体ファミリーであって、非限定的な例は、SLC9A1、SLC9A2、SLC9A3、SLC9A4、SLC9A5、SLC9A6、SLC9A7、SLC9A8、SLC9A9、SLC9A10、SLC9A11、SLC9B1、およびSLC9B2を含み;
(10)ナトリウム胆汁酸塩共輸送ファミリーであって、非限定的な例は、SLC10A1、SLC10A2、SLC10A3、SLC10A4、SLC10A5、SLC10A6、およびSLC10A7を含み;
(11)プロトン共役型金属イオン輸送体ファミリーであって、非限定的な例は、SLC11A1およびSLC11A2を含み;
(12)電気的中性カチオン−Cl共輸送体ファミリーであって、非限定的な例は、SLC12A1、SLC12A1、SLC12A2、SLC12A3、SLC12A4、SLC12A5、SLC12A6、SLC12A7、SLC12A8、およびSLC12A9を含み;
(13)Na+−SO42−/カルボン酸共輸送体ファミリーであって、非限定的な例は、SLC13A1、SLC13A2、SLC13A3、SLC13A4、およびSLC13A5を含み;
(14)尿素輸送体ファミリーであって、非限定的な例は、SLC14A1およびSLC14A2を含み;
(15)プロトンオリゴペプチド共輸送体ファミリーであって、非限定的な例は、SLC15A1、SLC15A2、SLC15A3、およびSLC15A4を含み;
(16)モノカルボン酸輸送体ファミリーであって、非限定的な例は、SLC16A1、SLC16A2、SLC16A3、SLC16A4、SLC16A5、SLC16A6、SLC16A7、SLC16A8、SLC16A9、SLC16A10、SLC16A11、SLC16A12、SLC16A13、およびSLC16A14を含み;
(17)小胞グルタミン酸輸送体ファミリーであって、非限定的な例は、SLC17A1、SLC17A2、SLC17A3、SLC17A4、SLC17A5、SLC17A6、SLC17A7、SLC17A8、およびSLC17A9を含み;
(18)小胞アミン輸送体ファミリーであって、非限定的な例は、SLC18A1、SLC18A2、およびSLC18A3を含み;
(19)葉酸/チアミン輸送体ファミリーであって、非限定的な例は、SLC19A1、SLC19A2、およびSLC19A3を含み;
(20)III型Na+−リン酸共輸送体ファミリーであって、非限定的な例は、SLC20A1およびSLC20A2を含み;
(21)有機アニオン輸送体ファミリーであって、非限定的な例は、(i)サブファミリー1 SLCO1A2、SLCO1B1、SLCO1B3、およびSLCO1C1;(ii)サブファミリー2、SLCO2A1およびSLCO2B1を含み;(iii)サブファミリー3、SLCO3A1;(iv)サブファミリー4、SLCO4A1、SLCO4C1;(v)サブファミリー5、SLCO5A1;ならびに(vi)サブファミリー6、SLCO6A1を含み;
(22)有機カチオン/アニオン/双性イオン輸送体ファミリーであって、非限定的な例は、SLC22A1、SLC22A2、SLC22A3、SLC22A4、SLC22A5、SLC22A6、SLC22A7、SLC22A8、SLC22A9、SLC22A10、SLC22A11、SLC22A12、SLC22A13、SLC22A14、SLC22A15、SLC22A16、SLC22A17、SLC22A18、SLC22A18AS,SLC22A19、SLC22A20、SLC22A23、SLC22A24、SLC22A25、およびSLC22A31を含み;
(23)Na+依存性アスコルビン酸輸送体ファミリーであって、非限定的な例は、SLC23A1、SLC23A2、SLC23A3、およびSLC23A4を含み;
(24)Na+/(Ca2+−K+)交換輸送体ファミリーであって、非限定的な例は、SLC24A1、SLC24A2、SLC24A3、SLC24A4、SLC24A5、およびSLC24A6を含み;
(25)ミトコンドリアキャリアファミリーであって、非限定的な例は、SLC25A1、SLC25A2、SLC25A3、SLC25A4、SLC25A5、SLC25A6、SLC25A7、SLC25A8、SLC25A9、SLC25A10、SLC25A11、SLC25A12、SLC25A13、SLC25A14、SLC25A15、SLC25A16、SLC25A17、SLC25A18、SLC25A19,SLC25A20、SLC25A21、SLC25A22、SLC25A23、SLC25A24、SLC25A25、SLC25A26、SLC25A27、SLC25A28、SLC25A29、SLC25A30、SLC25A31、SLC25A32、SLC25A33、SLC25A34、SLC25A35、SLC25A36、SLC25A37,SLC25A38、SLC25A39、SLC25A40、SLC25A41、SLC25A42、SLC25A43、SLC25A44、SLC25A45、およびSLC25A46を含み;
(26)多機能性アニオン交換輸送体ファミリーであって、非限定的な例は、SLC26A1、SLC26A2、SLC26A3、SLC26A4、SLC26A5、SLC26A6、SLC26A7、SLC26A8、SLC26A9、SLC26A10、およびSLC26A11を含み;
(27)脂肪酸輸送タンパク質ファミリーであって、非限定的な例は、SLC27A1、SLC27A2、SLC27A3、SLC27A4、SLC27A5、およびSLC27A6を含み;
(28)Na+共役型ヌクレオシド輸送ファミリーであって、非限定的な例は、SLC28A1、SLC28A2、およびSLC28A3を含み;
(29)促進性ヌクレオシド輸送体ファミリーであって、非限定的な例は、SLC29A1、SLC29A2、SLC29A3、およびSLC29A4を含み;
(30)亜鉛排出ファミリーであって、非限定的な例は、SLC30A1、SLC30A2、SLC30A3、SLC30A4、SLC30A5、SLC30A6、SLC30A7、SLC30A8、SLC30A9、およびSLC30A10を含み;
(31)銅輸送体ファミリーであって、非限定的な例は、SLC31A1およびSLC31A2を含み;
(32)小胞抑制性アミノ酸輸送体ファミリーであって、非限定的な例は、SLC32A1を含み;
(33)アセチル−CoA輸送体ファミリーであって、非限定的な例は、SLC33A1を含み;
(34)II型Na+−リン酸共輸送体ファミリーであって、非限定的な例は、SLC34A1、SLC34A2、およびSLC34A3を含み;
(35)ヌクレオシド−糖輸送体ファミリーであって、非限定的な例は、(i)サブファミリーA、SLC35A1、SLC35A2、SLC35A3、SLC35A4、SLC35A5;(ii)サブファミリーB、SLC35B1、SLC35B2、SLC35B3、SLC35B4;(iii)サブファミリーC、SLC35C1、SLC35C2;(iv)サブファミリーD、SLC35D1、SLC35D2、SLC35D3;(v)サブファミリーE、SLC35E1、SLC35E2、SLC35E3、SLC35E4;(vi)サブファミリーF、SLC35F1、SLC35F2、SLC35F3、SLC35F4、SLC35F5;(vii)サブファミリーG、SLC35G1、SLC35G3、SLC35G4、SLC35G5、SLC35G6を含み;
(36)プロトン共役型アミノ酸輸送体ファミリーであって、非限定的な例は、SLC36A1、SLC36A2、SLC36A3、およびSLC36A4を含み;
(37)糖−リン酸/リン酸交換輸送体ファミリーであって、非限定的な例は、SLC37A1、SLC37A2、SLC37A3、およびSLC37A4を含み;
(38)システムAおよびNナトリウム共役型中性アミノ酸輸送体ファミリーであって、非限定的な例は、SLC38A1、SLC38A2、SLC38A3、SLC38A4、SLC38A5、SLC38A6、SLC38A7、SLC38A8、SLC38A9、SLC38A10、およびSLC38A11を含み;
(39)金属イオン輸送体ファミリーであって、非限定的な例は、SLC39A1、SLC39A2、SLC39A3、SLC39A4、SLC39A5、SLC39A6、SLC39A7、SLC39A8、SLC39A9、SLC39A10、SLC39A11、SLC39A12、SLC39A13、およびSLC39A14を含み;
(40)側底側鉄輸送体ファミリーであって、非限定的な例は、SLC40A1を含み;
(41)MgtE様マグネシウム輸送体ファミリーであって、非限定的な例は、SLC41A1、SLC41A2、およびSLC41A3を含み;
(42)アンモニア輸送体ファミリーであって、非限定的な例は、RhAG、RhBGおよびRhCGを含み;
(43)Na+依存性システムL様アミノ酸輸送体ファミリーであって、非限定的な例は、SLC43A1、SLC43A2、およびSLC43A3を含み;
(44)コリン様輸送体ファミリーであって、非限定的な例は、SLC44A1、SLC44A2、SLC44A3、SLC44A4、およびSLC44A5を含み;
(45)推定上の糖輸送体ファミリーであって、非限定的な例は、SLC45A1、SLC45A2、SLC45A3、およびSLC45A4を含み;
(46)葉酸輸送体ファミリーであって、非限定的な例は、SLC46A1、SLC46A2、およびSLC46A3を含み;
(47)多剤および毒素排出ファミリーであって、非限定的な例は、SLC47A1およびSLC47A2を含み;
(48)ヘム輸送体ファミリーであって、非限定的な例は、HCP−1を含み;
(49)主要促進因子の輸送体スーパーファミリーであって、非限定的な例は、MFSD5およびMFSD10を含み;
(50)糖排出輸送体のSWEETファミリーであって、非限定的な例は、SLC50A1を含み;
(51)ステロイド由来の分子の輸送体であって、非限定的な例は、OSTαおよびOSTβを含み;
(52)リボフラビン輸送体ファミリーRFVT/SLC52であって、非限定的な例は、SLC52A1、SLC52A2、およびSLC52A3を含む。
特定の態様では、抗体を作製するための従来の方法、例えば、精製された組換え体タンパク質により動物を免疫化することは、発現が困難な標的タンパク質について効果的ではない場合がある。多くの要因が、過剰発現/精製をさせないタンパク質の宿主産生細胞における細胞毒性、本質的に不十分な生物物理学的特性(例えば、サイズ、溶解性、コンフォメーション、翻訳後修飾(糖鎖付加など)などの、発現の問題に寄与し得る。発現が困難なタンパク質の特徴の非限定的な例は、大きなタンパク質(例えば、分子量≧150kDaを有するタンパク質)、膜貫通タンパク質、特有な翻訳後修飾を有するタンパク質または溶解性の低いタンパク質、不安定なタンパク質、シグナルペプチドを含有しない分泌タンパク質、膜結合タンパク質、天然変性タンパク質、および半減期の短いタンパク質を含むが、これらに限定されない。
従来技術において知られるカチオン性脂質のいずれかを、特許請求された発明の実施に際して採用してもよい。例えば、Felgner et al.(Proc.Natl.Acad.Sci.U.S.A.84:7413−7417(1987));Felgner et al.(Focus 11:21−25(1989));Felgner(“Cationic Liposome−Mediated Transfection with Lipofectin(商標) Reagent,”in Gene Transfer and Expression Protocols Vol.7,Murray,E.J.,Ed.,Humana Press,New Jersey,pp.81−89(1991));国際公開第91/17424号パンフレット;国際公開第91/16024号パンフレット;米国特許第4,897,355号明細書;米国特許第4,946,787号明細書;米国特許第5,049,386号明細書;米国特許第5,208,036号明細書;Behr et al.(Proc.Natl.Acad.Sci.USA 86:6982−6986(1989);EPO Publication 0 394 111);Gao et al.(Biochim.Biophys.Res.Comm.179:280−285(1991));Zhou et al.,(Biochim.Biophys.Res.Comm.165:8−14(1991));およびGebeychu et al.(共有の米国特許出願第07/937,508号明細書;1992年8月28日出願)を参照されたい。これらの内容は参照により全面的に組み込まれる。
2−(10−ドデシル−3−エチル−8,14−ジオキソ−7,9,13−トリオキサ−3−アザオクタデカン−18−イル)プロパン−1,3−ジイルジオクタノアート;
2−(9−ドデシル−2−メチル−7,13−ジオキソ−6,8,12−トリオキサ−2−アザヘプタデカン−17−イル)プロパン−1,3−ジイルジオクタノアート;
2−(9−ドデシル−2−メチル−7,13−ジオキソ−6,8,12−トリオキサ−2−アザペンタデカン−15−イル)プロパン−1,3−ジイルジオクタノアート;
2−(10−ドデシル−3−エチル−8,14−ジオキソ−7,9,13−トリオキサ−3−アザヘキサデカン−16−イル)プロパン−1,3−ジイルジオクタノアート;
2−(8−ドデシル−2−メチル−6,12−ジオキソ−5,7,11−トリオキサ−2−アザヘプタデカン−17−イル)プロパン−1,3−ジイルジオクタノアート;
2−(10−ドデシル−3−エチル−8,14−ジオキソ−7,9,13−トリオキサ−3−アザノナデカン−19−イル)プロパン−1,3−ジイルジオクタノアート;
2−(9−ドデシル−2−メチル−7,13−ジオキソ−6,8,12−トリオキサ−2−アザオクタデカン−18−イル)プロパン−1,3−ジイルジオクタノアート;
2−(8−ドデシル−2−メチル−6,12−ジオキソ−5,7,11−トリオキサ−2−アザオクタデカン−18−イル)プロパン−1,3−ジイルジオクタノアート;
2−(10−ドデシル−3−エチル−8,14−ジオキソ−7,9,13−トリオキサ−3−アザイコサン−20−イル)プロパン−1,3−ジイルジオクタノアート;
2−(9−ドデシル−2−メチル−7,13−ジオキソ−6,8,12−トリオキサ−2−アザノナデカン−19−イル)プロパン−1,3−ジイルジオクタノアート;
3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル4,4−ビス(オクチルオキシ)ブタノアート;
3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル4,4−ビス((2−エチルヘキシル)オキシ)ブタノアート;
3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル4,4−ビス((2−プロピルペンチル)オキシ)ブタノアート;
3−(((3−(エチル(メチル)アミノ)プロポキシ)カルボニル)オキシ)ペンタデシル4,4−ビス((2−プロピルペンチル)オキシ)ブタノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル4,4−ビス((2−プロピルペンチル)オキシ)ブタノアート;
3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル6,6−ビス(オクチルオキシ)ヘキサノアート;
3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル6,6−ビス(ヘキシルオキシ)ヘキサノアート;
3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル6,6−ビス((2−エチルヘキシル)オキシ)ヘキサノアート;
3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル8,8−ビス(ヘキシルオキシ)オクタノアート;
3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル8,8−ジブトキシオクタノアート;
3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル8,8−ビス((2−プロピルペンチル)オキシ)オクタノアート;
3−(((3−(エチル(メチル)アミノ)プロポキシ)カルボニル)オキシ)ペンタデシル8,8−ビス((2−プロピルペンチル)オキシ)オクタノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル8,8−ビス((2−プロピルペンチル)オキシ)オクタノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル3−オクチルウンデカノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル3−オクチルウンデカ−2−エノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル7−ヘキシルトリデカ−6−エノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル9−ペンチルテトラデカノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル9−ペンチルテトラデカ−8−エノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル5−ヘプチルドデカノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)トリデシル5−ヘプチルドデカノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ウンデシル5−ヘプチルドデカノアート;
1,3−ビス(オクタノイルオキシ)プロパン−2−イル(3−(((2−(ジメチルアミノ)エトキシ)カルボニル)オキシ)ペンタデシル)サクシナート;
1,3−ビス(オクタノイルオキシ)プロパン−2−イル(3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル)サクシナート;
1−(3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル)10−オクチルデカンジオアート;
1−(3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル)10−オクチルデカンジオアート;
1−(3−(((3−(エチル(メチル)アミノ)プロポキシ)カルボニル)オキシ)ペンタデシル)10−オクチルデカンジオアート;
1−(3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル)10−(2−エチルヘキシル)デカンジオアート;
1−(3−(((3−(エチル(メチル)アミノ)プロポキシ)カルボニル)オキシ)ペンタデシル)10−(2−エチルヘキシル)デカンジオアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル10−(オクタノイルオキシ)デカノアート;
8−ドデシル−2−メチル−6,12−ジオキソ−5,7,11−トリオキサ−2−アザノナデカン−19−イルデカノアート;
3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル10−(オクタノイルオキシ)デカノアート;
3−(((3−(エチル(メチル)アミノ)プロポキシ)カルボニル)オキシ)ペンタデシル10−(オクタノイルオキシ)デカノアート;
(9Z,12Z)−3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシルオクタデカ−9,12−ジエノアート;
(9Z,12Z)−3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシルオクタデカ−9,12−ジエノアート;
(9Z,12Z)−3−(((3−(エチル(メチル)アミノ)プロポキシ)カルボニル)オキシ)ペンタデシルオクタデカ−9,12−ジエノアート;
(9Z,12Z)−3−(((2−(ジメチルアミノ)エトキシ)カルボニル)オキシ)ペンタデシルオクタデカ−9,12−ジエノアート;
1−((9Z,12Z)−オクタデカ−9,12−ジエノイルオキシ)ペンタデカン−3−イル1,4−ジメチルピペリジン−4−カルボキシラート;
2−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)テトラデシル4,4−ビス((2−エチルヘキシル)オキシ)ブタノアート;
(9Z,12Z)−(12Z,15Z)−3−((3−(ジメチルアミノ)プロパノイル)オキシ)ヘンイコサ−12,15−ジエン−1−イルオクタデカ−9,12−ジエノアート;
(12Z,15Z)−3−((4−(ジメチルアミノ)ブタノイル)オキシ)ヘンイコサ−12,15−ジエン−1−イル3−オクチルウンデカノアート;
(12Z,15Z)−3−((4−(ジメチルアミノ)ブタノイル)オキシ)ヘンイコサ−12,15−ジエン−1−イル5−ヘプチルドデカノアート;
(12Z,15Z)−3−((4−(ジメチルアミノ)ブタノイル)オキシ)ヘンイコサ−12,15−ジエン−1−イル7−ヘキシルトリデカノアート;
(12Z,15Z)−3−((4−(ジメチルアミノ)ブタノイル)オキシ)ヘンイコサ−12,15−ジエン−1−イル9−ペンチルテトラデカノアート;
(12Z,15Z)−1−((((9Z,12Z)−オクタデカ−9,12−ジエン−1−イルオキシ)カルボニル)オキシ)ヘンイコサ−12,15−ジエン−3−イル3−(ジメチルアミノ)プロパノアート;
(13Z,16Z)−4−(((2−(ジメチルアミノ)エトキシ)カルボニル)オキシ)ドコサ−13,16−ジエン−1−イル2,2−ビス(ヘプチルオキシ)アセタート;
(13Z,16Z)−4−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ドコサ−13,16−ジエン−1−イル2,2−ビス(ヘプチルオキシ)アセタート;
2,2−ビス(ヘプチルオキシ)エチル3−((3−エチル−10−((9Z,12Z)−オクタデカ−9,12−ジエン−1−イル)−8,15−ジオキソ−7,9,14−トリオキサ−3−アザヘプタデカン−17−イル)ジスルファニル)プロパノアート;
(13Z,16Z)−4−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ドコサ−13,16−ジエン−1−イルヘプタデカン−9−イルサクシナート;
(9Z,12Z)−2−(((11Z,14Z)−2−((3−(ジメチルアミノ)プロパノイル)オキシ)イコサ−11,14−ジエン−1−イル)オキシ)エチルオクタデカ−9,12−ジエノアート;
(9Z,12Z)−3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−13−(オクタノイルオキシ)トリデシルオクタデカ−9,12−ジエノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−13−(オクタノイルオキシ)トリデシル3−オクチルウンデカノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−13−ヒドロキシトリデシル5−ヘプチルドデカノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−13−(オクタノイルオキシ)トリデシル5−ヘプチルドデカノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−13−(オクタノイルオキシ)トリデシル7−ヘキシルトリデカノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−13−ヒドロキシトリデシル9−ペンチルテトラデカノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−13−(オクタノイルオキシ)トリデシル9−ペンチルテトラデカノアート;
1−(3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−13−(オクタノイルオキシ)トリデシル)10−オクチルデカンジオアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−13−(オクタノイルオキシ)トリデシル10−(オクタノイルオキシ)デカノアート;
(9Z,12Z)−3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−5−オクチルトリデシルオクタデカ−9,12−ジエノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−5−オクチルトリデシルデカノアート;
5−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−7−オクチルペンタデシルオクタノアート;
(9Z,12Z)−5−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−7−オクチルペンタデシルオクタデカ−9,12−ジエノアート;
9−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−11−オクチルノナデシルオクタノアート;
9−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)−11−オクチルノナデシルデカノアート;
(9Z,12Z)−9−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ノナデシルオクタデカ−9,12−ジエノアート;9−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ノナデシルヘキサノアート;
9−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ノナデシル3−オクチルウンデカノアート;
9−((4−(ジメチルアミノ)ブタノイル)オキシ)ノナデシルヘキサノアート;
9−((4−(ジメチルアミノ)ブタノイル)オキシ)ノナデシル3−オクチルウンデカノアート;
(9Z,9’Z,12Z,12’Z)−2−((4−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノイル)オキシ)プロパン−1,3−ジイルビス(オクタデカ−9,12−ジエノアート);
(9Z,9’Z,12Z,12’Z)−2−((4−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノイル)オキシ)プロパン−1,3−ジイルビス(オクタデカ−9,12−ジエノアート);
(9Z,9’Z,12Z,12’Z,15Z,15’Z)−2−((4−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノイル)オキシ)プロパン−1,3−ジイルビス(オクタデカ−9,12,15−トリエノアート);
(Z)−2−((4−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノイル)オキシ)プロパン−1,3−ジイルジオレアート;
2−((4−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノイル)オキシ)プロパン−1,3−ジイルジテトラデカノアート;
2−((4−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノイル)オキシ)プロパン−1,3−ジイルジテトラデカノアート;
2−((4−(((3−(エチル(メチル)アミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノイル)オキシ)プロパン−1,3−ジイルジテトラデカノアート;
2−((4−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノイル)オキシ)プロパン−1,3−ジイルジドデカノアート;
2−((4−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノイル)オキシ)プロパン−1,3−ジイルジドデカノアート;
2−((4−(((3−(エチル(メチル)アミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノイル)オキシ)プロパン−1,3−ジイルジドデカノアート;
2−((4−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノイル)オキシ)プロパン−1,3−ジイルビス(デカノアート);
2−((4−(((3−(エチル(メチル)アミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノイル)オキシ)プロパン−1,3−ジイルビス(デカノアート);
2−((4−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノイル)オキシ)プロパン−1,3−ジイルジオクタノアート;
2−((4−(((3−(エチル(メチル)アミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノイル)オキシ)プロパン−1,3−ジイルジオクタノアート;
2−(((13Z,16Z)−4−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ドコサ−13,16−ジエノイル)オキシ)プロパン−1,3−ジイルジオクタノアート;
2−(((13Z,16Z)−4−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ドコサ−13,16−ジエノイル)オキシ)プロパン−1,3−ジイルジオクタノアート;
(9Z,9’Z,12Z,12’Z)−2−((2−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)テトラデカノイル)オキシ)プロパン−1,3−ジイルビス(オクタデカ−9,12−ジエノアート);
(9Z,9’Z,12Z,12’Z)−2−((2−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ドデカノイル)オキシ)プロパン−1,3−ジイルビス(オクタデカ−9,12−ジエノアート);
(9Z,9’Z,12Z,12’Z)−2−((2−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)テトラデカノイル)オキシ)プロパン−1,3−ジイルビス(オクタデカ−9,12−ジエノアート);
(9Z,9’Z,12Z,12’Z)−2−((2−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ドデカノイル)オキシ)プロパン−1,3−ジイルビス(オクタデカ−9,12−ジエノアート);
2−((2−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)テトラデカノイル)オキシ)プロパン−1,3−ジイルジオクタノアート;
4.4−ビス(オクチルオキシ)ブチル4−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ヘキサデカノアート;
4,4−ビス(オクチルオキシ)ブチル2−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ドデカノアート;
(9Z,12Z)−10−ドデシル−3−エチル−14−(2−((9Z,12Z)−オクタデカ−9,12−ジエノイルオキシ)エチル)−8,13−ジオキソ−7,9−ジオキサ−3,14−ジアザヘキサデカン−16−イルオクタデカ−9,12−ジエノアート;
2−((4−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)−11−(オクタノイルオキシ)ウンデカノイル)オキシ)プロパン−1,3−ジイルジオクタノアート;
(9Z,9’Z,12Z,12’Z)−2−(9−ドデシル−2−メチル−7,12−ジオキソ−6,8,13−トリオキサ−2−アザテトラデカン−14−イル)プロパン−1,3−ジイルビス(オクタデカ−9,12−ジエノアート);
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル4,4−ビス(オクチルオキシ)ブタノアート;
3−(((3−(ピペリジン−1−イル)プロポキシ)カルボニル)オキシ)ペンタデシル6,6−ビス(オクチルオキシ)ヘキサノアート;
3−(((3−(ピペラジン−1−イル)プロポキシ)カルボニル)オキシ)ペンタデシル6,6−ビス(オクチルオキシ)ヘキサノアート;
3−(((4−(ジエチルアミノ)ブトキシ)カルボニル)オキシ)ペンタデシル6,6−ビス(オクチルオキシ)ヘキサノアート;
3−(((3−(4−メチルピペラジン−1−イル)プロポキシ)カルボニル)オキシ)ペンタデシル6,6−ビス(オクチルオキシ)ヘキサノアート;
3−((((1−メチルピペリジン−4−イル)メトキシ)カルボニル)オキシ)ペンタデシル6,6−ビス(オクチルオキシ)ヘキサノアート;
3−(((3−モルホリノポロポキシ)カルボニル)オキシ)ペンタデシル6,6−ビス(オクチルオキシ)ヘキサノアート;
3−(((2−(ジエチルアミノ)エトキシ)カルボニル)オキシ)ペンタデシル6,6−ビス(オクチルオキシ)ヘキサノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル6,6−ビス(オクチルオキシ)ヘキサノアート;
3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル6,6−ビス((2−プロピルペンチル)オキシ)ヘキサノアート;
3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル6,6−ビス((2−プロピルペンチル)オキシ)ヘキサノアート LXR420:3−(((3−(ジメチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル6,6−ビス((3−エチルペンチル)オキシ)ヘキサノアート;
(2R)−1−((6,6−ビス(オクチルオキシ)ヘキサノイル)オキシ)ペンタデカン−3−イル1−メチルピロリジン−2−カルボキシラート;
(2S)−1−((6,6−ビス(オクチルオキシ)ヘキサノイル)オキシ)ペンタデカン−3−イル1−メチルピロリジン−2−カルボキシラート;
(2R)−1−((6,6−ビス(オクチルオキシ)ヘキサノイル)オキシ)ペンタデカン−3−イルピロリジン−2−カルボキシラート;
1−((6,6−ビス(オクチルオキシ)ヘキサノイル)オキシ)ペンタデカン−3−イル1,3−ジメチルピロリジン−3−カルボキシラート;
3−((3−(1−メチルピペリジン−4−イル)プロパノイル)オキシ)ペンタデシル6,6−ビス(オクチルオキシ)ヘキサノアート;
1−((6,6−ビス(オクチルオキシ)ヘキサノイル)オキシ)ペンタデカン−3−イル1,4−ジメチルピペリジン−4−カルボキシラート;
3−((5−(ジエチルアミノ)ペンタノイル)オキシ)ペンタデシル6,6−ビス(オクチルオキシ)ヘキサノアート;
3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ペンタデシル5−(4,6−ジヘプチル−1,3−ジオキサン−2−イル)ペンタノアート;
3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ウンデシル6,6−ビス(オクチルオキシ)ヘキサノアート;
3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)トリデシル6,6−ビス(オクチルオキシ)ヘキサノアート;
(12Z,15Z)−3−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ヘンイコサ−12,15−ジエン−1−イル6,6−ビス(オクチルオキシ)ヘキサノアート;
6−((6,6−ビス(オクチルオキシ)ヘキサノイル)オキシ)−4−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ヘキシルオクタノアート;
4,4−ビス(オクチルオキシ)ブチル5−(((3−(ジエチルアミノ)プロポキシ)カルボニル)オキシ)ヘプタデカノアート;
4,4−ビス(オクチルオキシ)ブチル(3−(ジエチルアミノ)プロピル)ペンタデカン−1,3−ジイルジカルボナート;
2−(5−((4−((1,4−ジメチルピペリジン−4−カルボニル)オキシ)ヘキサデシル)オキシ)−5−オキソペンチル)プロパン−1,3−ジイルジオクタノアート;
2−(5−((4−((1,3−ジメチルピロリジン−3−カルボニル)オキシ)ヘキサデシル)オキシ)−5−オキソペンチル)プロパン−1,3−ジイルジオクタノアート;
2−(5−オキソ−5−((4−(((S)−ピロリジン−2−カルボニル)オキシ)ヘキサデシル)オキシ)ペンチル)プロパン−1,3−ジイルジオクタノアート;
2−(5−((4−(((((S)−1−メチルピロリジン−3−イル)オキシ)カルボニル)オキシ)ヘキサデシル)オキシ)−5−オキソペンチル)プロパン−1,3−ジイルジオクタノアート;
2−(5−((4−(((((R)−1−メチルピロリジン−3−イル)オキシ)カルボニル)オキシ)ヘキサデシル)オキシ)−5−オキソペンチル)プロパン−1,3−ジイルジオクタノアート;
2−(5−((4−((((1−エチルピペリジン−3−イル)メトキシ)カルボニル)オキシ)ヘキサデシル)オキシ)−5−オキソペンチル)プロパン−1,3−ジイルジオクタノアート;
2−(5−((4−((((1−メチルピペリジン−4−イル)オキシ)カルボニル)オキシ)ヘキサデシル)オキシ)−5−オキソペンチル)プロパン−1,3−ジイルジオクタノアート;
2−(10−ドデシル−3−エチル−8,15−ジオキソ−7,9,14−トリオキサ−3−アザノナデカン−19−イル)プロパン−1,3−ジイルジオクタノアート;
2−(11−ドデシル−3−エチル−9,15−ジオキソ−8,10,14−トリオキサ−3−アザノナデカン−19−イル)プロパン−1,3−ジイルジオクタノアート;
2−(5−((3−(((3−(1H−イミダゾール−イル)プロポキシ)カルボニル)オキシ)ペンタデシル)オキシ)−5−オキソペンチル)プロパン−1,3−ジイルジオクタノアート;
2−(5−オキソ−5−((3−(((3−(ピペリジン−1−イル)プロポキシ)カルボニル)オキシ)ペンタデシル)オキシ)ペンチル)プロパン−1,3−ジイルジオクタノアート;および2−(12−ドデシル−3−エチル−8,14−ジオキソ−7,9,13−トリオキサ−3−アザオクタデカン−18−イル)プロパン−1,3−ジイルジオクタノアート。
ある特定の実施形態では、本発明の免疫化方法のために使用されるポリヌクレオチドは、ポリリボヌクレオチドをベースにした、例えば、mRNAをベースにしたものである。このmRNA分子は、抗体応答が望まれる標的タンパク質の翻訳を直接にコードし、かつ促進することができなければならない。そのため、その分子はいくつかの成分を含有しなければならない。特定の態様では、第1の成分は、タンパク質、例えば、ヒト標的タンパク質またはその断片のアミノ酸配列に対応するオープンリーディングフレームである。天然のコドン配列を使用してもよく、あるいは、翻訳効率および根本的に標的のタンパク質発現レベルを上昇させるために、宿主種(マウスまたはウサギなど)に関してコドン最適化を実施してもよい。さらなる改変をオープンリーディングフレームに対して行い、タンパク質発現/輸送を亢進してもよい。分泌タンパク質または膜タンパク質については、このことは、インターロイキン2(IL−2)からの分泌シグナルなどの異種のシグナルペプチドの使用を含み得る。分泌タンパク質についての特定の例では、mRNA分子は、ヒトIL−2またはIgGカッパのシグナルペプチドなどの異種のシグナルペプチドを含んでもよい。
本開示によれば、特定の態様では、脂質/ポリヌクレオチド複合体を使用して、インビボトランスフェクションを行う。トランスフェクトされた細胞は、ポリヌクレオチド(例えば、mRNAなどのポリリボヌクレオチド)によってコードされたタンパク質を発現し、外来タンパク質を、例えば、細胞表面上に発現または提示し得る。その結果、宿主動物(例えば、非ヒト宿主動物)は、外来タンパク質、すなわち免疫原に対する免疫応答を開始する。
本明細書に記載の組成物および方法のためのmRNAなどのポリリボヌクレオチドは、エンドヌクレアーゼおよびエキソヌクレアーゼによる迅速な分解を予防し、かつRNAに対する細胞の自然免疫またはインターフェロン応答を回避または低減するための修飾を含むことができる。修飾には、例えば、(a)末端の修飾、例えば、5’末端修飾(リン酸化、脱リン酸化、コンジュゲーション、逆方向結合など)、3’末端修飾(コンジュゲーション、DNAヌクレオチド、逆方向結合など)、(b)塩基修飾、例えば、修飾された塩基、安定化塩基、不安定化塩基、もしくはパートナーの広範なレパートリーと塩基対合する塩基による置換、またはコンジュゲートされた塩基、(c)糖の修飾(例えば、2’位もしくは4’位での)または糖の置換、および(d)ホスホジエステル結合の修飾または置換を含むヌクレオシド間の結合の修飾を含むが、これらに限定されない。
RXFP1、すなわちリラキシン/インスリン様ファミリーペプチド受容体1は、ロイシンリッチ反復N末端細胞外ドメインを含有する757アミノ酸のクラスA Gタンパク質共役型受容体(GPCR)である。系統学的に、卵胞刺激ホルモン受容体、黄体形成ホルモン受容体、および甲状腺刺激ホルモン受容体を含む同一の受容体サブファミリーの一部である。RXFP1の内因性のリガンドは、タンパク質ホルモンのリラキシンである。RXFP1およびそのリガンドは、月経ならびに妊娠および分娩と関係した生理学的応答の一部の制御において意味づけられてきた。急性非代償性心不全に罹患した患者において、第III相臨床試験(RELAX−AHF)は、入院期間中の48時間の組換え体リラキシン注射により6ヶ月後の死亡率を有意に低減したことを示している。
SLC52A2(GPR172A)は、10個または11個の推定上の膜貫通ヘリックスを有すると予測される445アミノ酸の複数回膜貫通タンパク質である。それは、リボフラビンの細胞内取り込みを媒介することが示されており、ブタ内在性レトロウイルスサブグループAに対する受容体であることが報告されている。SLC52A2のある種の遺伝的変異体は、運動、感覚、および頭部ニューロン症と関連がある。
ANGPTL8は、脂質代謝に関与する分泌タンパク質であり、ヒト血漿においてng/mlの低い濃度で見出すことができる。しかし、このタンパク質は、異種系統において、その天然の可溶性形態での発現は困難である。さらに、このタンパク質の生化学的機能は記述されておらず、したがって、組換え技術によって生成されたタンパク質の活性を測定するために使用され得るインビトロ機能アッセイがない。作製の難しさ、および抗原として使用するための組換え体ANGPTL8の品質を検証する難しさを考慮すると、このタンパク質は、モノクローナル抗体を作製するmRNA媒介性の免疫化のための良好な候補であった。
甲状腺刺激ホルモン受容体(TSHR)は、甲状腺の発育および甲状腺ホルモン産生に必須のGタンパク質共役型受容体である。それはまた、グレーブス病における自己抗原でもある。TSHRに特異的な抗体によるTSHRの遷延性の活性化は、グレーブス病の根底にある主要な原因の1つである(Davies TF(2015)Expert Opin Ther Targets;19:835−47)。
APJ、アンジオテンシン様1受容体、アンジオテンシンII様1受容体などとも称されるアペリン受容体は、内因性リガンドのアペリンに対して同族である以前はオーファンであったGタンパク質共役型受容体(GPCR)である。アペリン/APJ経路は、心血管系において広範にわたって発現され、アペリンは、前臨床モデルにおいて、有益な主要な心血管効果を示している。ヒトにおける急性アペリン投与は、末梢および冠動脈の血管拡張を引き起こし、心拍出量を増加させる(Circulation.2010;121:1818−1827)。その結果、APJのアゴニズムは、心不全患者にとっての重要な治療標的として台頭している。アペリン受容体APJの活性化は、現行の療法の傾向を伴わずに、心収縮性を増大させ、心臓保護をもたらすと考えられている。
糖タンパク質130(GP130)は、918アミノ酸を含有するタンパク質であり、I型1回膜貫通型タンパク質受容体ファミリーのメンバーである。それは、インターロイキン6、インターロイキン11、毛様体神経栄養因子、白血病抑制因子、およびオンコスタチンMを含む多くのサイトカインによって用いられるシグナル伝達複合体の中心的な成分である。インターロイキン6(IL6)の場合、GP130はIL6/IL6R(α鎖)複合体に結合して、高親和性IL6結合部位の形成およびシグナル伝達の開始を引き起こす。GP130は、5つのIII型フィブロネクチンドメインおよび1つのIg様C2タイプドメインを含有する。
ガレクチン−3は、26kDaのタンパク質であり、β−ガラクトシド−結合レクチンファミリーのメンバーである。それは、コラーゲン様N末端ドメイン、およびガレクチン−3の糖を結びつける能力を与えるC末端糖認識ドメインを含有する。そのN末端ドメインを介して、ガレクチン−3はより高次のオリゴマーを形成することができる。ガレクチン−3は、細胞接着、分化、転移、胚形成、炎症、および線維形成において役割を果たすことが示唆されている。
用語「脂質ナノ粒子」または「LNP」もしくは「LNPs」は、分子間力によって互いに物理的に会合(例えば、共有結合的にまたは非共有結合的に)した複数の(すなわち、2つ以上の)脂質分子を含む粒子を指す。脂質ナノ粒子は、例えば、ミクロスフィア(単層および多層の小胞を含む、例えば、リポソーム)、エマルション中で分散した相、ミセル、または懸濁液中の内相であってもよい。
下記方法を使用して、本明細書で提供される脂質ナノ粒子を作製することができる。脂質ナノ粒子を作製する非限定的な例が記載されており、例えば、PCT国際特許出願公開番号の国際公開第2016/010840号パンフレット、国際公開第2016/037053号パンフレット、国際公開第2015/095346号パンフレット、国際公開第2015/095340号パンフレット、国際公開第2014/136086号パンフレット、および国際公開第2011/076807号パンフレットを参照されたく、各々は、参照によりその全体が本明細書中に組み込まれる。粒子のサイズ縮小を達成するために、および/または粒子のサイズの一様性を増大させるために、当業者は、以下に提示する方法ステップを使用して、様々な組み合わせを実験してもよい。加えて、当業者は、リポソーム製剤で使用される超音波処理、濾過または他のサイジング手法を採用することができる。
免疫化に使用される宿主動物は、体液性(抗体)媒介性免疫応答を起こすことができる任意の種を包含する。免疫化に使用される非ヒト動物などの宿主動物の非限定的な例は、マウス、ラット、ウサギ、ヤギ、ヒツジ、ラクダ科動物、ウマ、ニワトリ、イヌ、ネコ、ブタ、ロバ、雌ウシ、サル、およびサメを含む。
0日目:免疫学的に無感作のマウスにおいてベースライン力価を規定するために採血する。
1日目(1回目の免疫化):5〜100μg、例えば、25〜50μgの被包性mRNAを、4匹のマウスの皮下および4匹のマウスの静脈内に注射する。
10日目:血清力価をモニターするために採血する。
21日目(2回目の免疫化):5〜100μg、例えば、25〜50μgの被包性mRNAを、4匹のマウスの皮下および4匹のマウスの静脈内に注射する。
31日目:血清力価をモニターするために採血する。
42日目(最後の免疫化):5〜100μg、例えば、25〜50μgの被包性mRNAを、マウスの静脈内に注射する。
45日目:脾細胞の単離およびハイブリドーマ融合のために脾臓を回収する。
モノクローナル抗体の生成
モノクローナル抗体(mAb)は、従来のモノクローナル抗体の方法論、例えば、KohlerおよびMilstein,1975 Nature 256:495の標準的な体細胞ハイブリダイゼーション技術を含む種々の手法により作製可能である。例えば、Bリンパ球のウイルス性形質転換または癌発生の形質転換など、モノクローナル抗体を作製するための多くの手法を採用することができる。
被包性mRNA作製のワークフロー
ステップ1:cDNAの設計およびインビトロ転写ベクターへのクローニング
cDNAコンストラクトの設計
天然のcDNA配列は、それが、サブクローニング計画において、または転写前の最終コンストラクトの線状化において使用される制限酵素のための任意のコンセンサス部位を含有しない場合に、サブクローニングの目的で使用され得る。この特定の例では、サブクローニングに使用される制限酵素はBamHIおよびRsrIIであり、線状化に使用される制限酵素はBspQIである。しかし、任意の好適な制限酵素および対応する制限酵素部位を使用することができる。例えば、目的の標的タンパク質をコードする特定のcDNA中に存在しないある種の制限酵素部位を、サブクローニング計画および線状化のために選択することができる。
標的タンパク質をコードするcDNA(例えば、表1〜7を参照のこと)を、T7 RNAポリメラーゼプロモーターからのRNAポリメラーゼ媒介性の転写を駆動するように設計されたベクターにクローン化した。T7プロモーターのすぐ下流は、タバコエッチ病ウイルス(Tobacco etch virus)(TEV)の5’非翻訳領域(UTR)をコードする配列である。このUTRは、真核細胞における翻訳効率を向上させることが示されている。TEVのUTRの下流に、標的タンパク質のcDNAが配置される。コザックコンセンサス配列(ccgccacc)を、翻訳を亢進するためにイニシエータメチオニン/開始コドンの上流に挿入した。2つの終止コドンを、RsrII制限酵素部位を後に伴うcDNAの末端に配置した。2つの直列のヒトβグロビン3’UTRが、そのcDNA配列に続く。このエレメントは、細胞内でのmRNAの安定性を高めることが示されている。ベクターの転写に関連する成分のC末端エレメントは、ポリAテールである。特定の実施形態では、標的タンパク質またはその断片をコードするmRNAのポリAテールは、約50bps〜120bpsまたは60bps〜120bpsである。特定の実施形態では、標的タンパク質またはその断片をコードするmRNAのポリAテールは、約60bpsまたは120bpsである。ある種の実施形態では、標的タンパク質またはその断片をコードするmRNAのポリAテールは、約70bps、80bps、90bps、100bps、または110bpsである。
制限酵素BamHI/RsrIIによりベクターとともにcDNAコンストラクトを消化することにより、アガロースゲル電気泳動によって精製された適合性の断片を生成し、続いて、精製されたcDNAを消化されたベクターに連結して、所望の転写ベクターコンストラクトを得た。
環状プラスミドDNAを、従来の方法に従って調製した。精製されたプラスミドDNAを、BspQI制限エンドヌクレアーゼにより消化した。プラスミドDNAを、適切な反応緩衝液(緩衝液4、New England Biolabs、10×原液)およびBspQI酵素(DNA1mgあたり1,250U)と混ぜ合わせた。反応を50℃で2時間インキュベートし、続いて氷上または4℃に置いた。少量の反応試料を、標準的なアガロース電気泳動上に流して、環状プラスミドDNAの完全な線状化を確認した。線状化DNA鋳型をエタノール沈殿により精製した。エタノール沈殿ステップのDNAペレットを、ヌクレアーゼフリー水を用いて溶解して、0.5mg/mlを超える濃度にした。
本実施例の修飾合成mRNAは、インビトロ転写(IVT)により作製され、塩化リチウム(LiCl)精製により精製され、New England Biolabs(登録商標)(Beverly、MA USA)の市販のキットを使用してキャップされた。材料および試薬を表9に示す。
1.上記の材料を30℃で2時間、温度をモニタリングしながらインキュベートした。DNA鋳型を、0.04U/μLのDNA分解酵素の添加により消化し、この反応混合物を37℃で30分間インキュベートした。
2.LiClを最終濃度2.81Mとなるように添加し、反応混合物を十分に混合し、−20℃で1時間を超えてインキュベートした。次に、混合物を4℃で15分間、約20,000×g(最大速度)の最大速度で遠心分離した。上清を除去し、ペレットを1mLの70%エタノールで洗浄した。調製物を、すぐ上に記載したように10分間遠心分離した。次に、上清を除去し、残留ペレットを上記のように1分未満の間再び遠心分離した。
3.残留エタノールを除去し、ペレットをヌクレアーゼフリー水中に再懸濁した。濃度を測定し、約1μg/μLに調整した。
4.調製物に、10%容量のpH5.5の3M酢酸ナトリウムを添加し、調製物を十分に混合した。次に、1容量の室温イソプロパノールを調製物に加え、よく混合した。調製物を−20℃で一晩インキュベートした。その後、調製物を4℃で15分間、約20,000×g(最大速度)の最大速度で遠心分離し、上清を除去し、残留ペレットを1mLの70%エタノールで洗浄した。再度、調製物をすぐ上に記載したように10分間遠心分離し、続いて上清を除去し、遠心分離ステップを、上記のように1分未満の間再び実施した。
5.残留エタノールを除去し、ペレットをヌクレアーゼフリー水中に再懸濁した。調製物の濃度を測定し、約4μg/μLに調整した。
最初のインビトロ転写反応の後および/またはキャッピング反応の後、Agilent 2100 Bioanalyzerを使用して、合成mRNAを、品質および完全性について分析した。Agilent 2100 BioAnalyzerは、DNA、RNAまたはタンパク質の少量試料のサイズ分画および定量を実施するナノ流体デバイスである。Agilent RNA 6000 Nano Kit(カタログ番号5067−1511)を使用して、分析を実施した。
ゲルマトリクス(550μL)をスピンフィルターにピペットで移し、室温にて10分間、1,500gで遠心分離し、続いて4℃で保存した(1ヶ月以内の使用用)。色素原液(1μl)を、濾過したゲルマトリクスの65μlアリコートに添加した。色素およびゲルマトリクスの混合物をボルテックスし、続いて室温にて10分間、13,000gで遠心分離した(1日以内の使用用)。mRNA試料ならびにラダーおよびキット標準を70℃で3〜5分間加熱変性させて、あらゆる高次構造を壊し、続いて分析前に氷上でクエンチした。
Bioanalyzerの電極を、350μlのRNaseZap電極洗浄剤およびヌクレアーゼフリー水で浄化した。
新しいチップをBioanalyzerのプライミングステーション(ポジションCでのプラットフォーム、トップポジションでのクリップ)に配置し、9μlのゲル/色素混合物をウェルに添加した。
5μl容量のマーカーをラダーウェルおよび試料ウェルに添加し、1μl容量のmRNA試料(<1μg)を試料ウェルに添加した。IKA Vortexerを使用して、2400rpmで1分間ボルテックスした。次に、試料を、mRNAアッセイ法を用いてAgilent 2100 Bioanalyzer上で測定した。アデニン、グアニン、シチジン、およびウリジンまたはプソイドウリジンのいずれかを用いて合成されたヒトRXFP1(コドン最適化およびコドン非最適化の両方)について調製されたmRNAの試料Bioanalyzerトレースについては、図4を参照されたい。
Thermo Fisher ScientificのLipofectamine(登録商標)2000試薬を使用したmRNAの培養哺乳動物細胞へのインビトロトランスフェクションによって、翻訳可能性を評価した。次に、トランスフェクトされた細胞を24〜48時間後に溶解し、溶解物中のタンパク質を、ポリアクリルアミドゲル電気泳動を用いて分離し、続いてmRNAによってコードされるタンパク質に特異的な抗体による免疫ブロットを行った(mRNA由来のヒトRXFP1発現の確証を示す試料のウェスタンブロットについては図5を参照のこと)。
本実施例で用いた材料は、以下の通りであった:
2.RNA分解酵素フリーの試薬、チューブ、チップおよび実施を用いて、表13に記載のように脂質ナノ粒子混合物試薬を計量し、バイアル中に混合した。
4.Amicon Ultra−15遠心分離デバイスにmRNA溶液を充填し、4℃にて4,000rpmで15分間遠心分離することによって、修飾合成mRNAを水からpH6.0緩衝液に交換した。濃縮したmRNAをpH6.0クエン酸緩衝液に再懸濁し、mRNA濃度を測定した。
5.pH6.0のクエン酸緩衝液中における修飾合成mRNAの0.5mg/mLの最終濃度を、すすがれたシンチレーションバイアルにおいて調製し(8mLに4mgのmRNA)、mRNA溶液の最終濃度を測定した。使用の準備ができるまで、mRNA希釈溶液を37℃でインキュベートした。
6.(a)脂質混合物;(b)mRNA溶液;(c)クエン酸緩衝液の各8mLにより、3つの10mlシリンジを調製した。シリンジ(a)および(b)を、T字型接合部のルアー嵌合部に取り付けた。要約すると、0.5mm内径のP727 T−混合機が、P652アダプタ(IDEX、Oak Harbor WA USA)に取り付けられた。シリンジ(a)および(b)は、P658ルアー嵌合部(IDEX)に取り付けた。シリンジ(a)および(b)を、P938xナットおよびフェルール(IDEX)を用いて、PTFE0.8mm内径チューブ(#3200068、Dolomite、Royston、UK)によって、T−混合機に接続した。シリンジ(c)は、P938xナットおよびフェルールによって最終の単一チューブに接続されたルアー嵌合部に取り付けた。チューブの末端は、撹拌棒入りの事前にすすいだビーカーの上に合わせて固定し、緩やかに撹拌した。
7.シリンジポンプ設定を、適切なシリンジ製造業者およびサイズにセットし、容量(8mL)および1.0mL/分の流速が入力された。ポンプを起動し、得られた材料を撹拌棒入りのRNA分解酵素フリーの50mLプラスチックビーカーに回収した。mRNAを含有する脂質ナノ粒子の懸濁液を、1バッグにつき2〜3mLで透析チューブに移し、4℃で一晩、リン酸緩衝生理食塩水(PBS)中に透析した。
8.分割した材料を、1本の15mLコニカルチューブにプールした。タンジェンシャルフロー濾過(TFF)を用いて、脂質ナノ粒子(LNP)懸濁液を濃縮した。新しいチューブを使用して新しい500Kの分子量カットオフカプセルをMinimateシステムに接続して、150rpmの流速により500mLのRNAフリー水ですすぐことによってTFFシステムを準備した。
9.脂質ナノ粒子/修飾合成mRNA懸濁液をTFFユニットリザーバーに充填し、75mL/分の流速で2〜3mlの最終容量に濃縮した。
10.修飾合成mRNAの被包パーセントは、Life Technologies(Grand Island NY USA)のQuant−iT Ribogreen RNAアッセイキットを用いて決定した。脂質ナノ粒子/修飾合成mRNA懸濁液を、緩衝液(粒子外のmRNA)および界面活性剤を加えた緩衝液(全mRNA)での蛍光測定によって分析した。提供されたリボソームRNAから1000ng/mL原液を調製し、これを使用してRibogreenアッセイのための標準曲線を作成した。アッセイのために、TE緩衝液またはTritonを加えたTE中に試料を調製し、蛍光試薬を各々に添加する。計算される差は、粒子内部のmRNAである。
12.試料蛍光を、蛍光マイクロプレートリーダーを用いて、480nmの励起、520nmの発光で測定した。各RNA試料の蛍光値から試薬ブランクの蛍光値を差し引いて、蛍光対RNA濃度の標準曲線を作成した。各試料の蛍光値から試薬ブランクの蛍光値を差し引き、標準曲線から試料のRNA濃度を決定した。試料の被包パーセントは、Tritonを加えた試料と試料だけの間の濃度の差を、Tritonを加えた試料の濃度によって割り算することによって決定した。脂質ナノ粒子/修飾合成懸濁液の6倍希釈溶液を作製し、Zetasizer Nano ZS機器(Malvern Instruments,Ltd、Worcestershire、UK)を用いて直径および多分散指数を決定した。
ヒトRXFP1などのGPCR標的タンパク質に対する免疫化戦略の概観を表16に示す。
ヒトANGPTL8などの発現が困難な標的タンパク質に対する免疫化戦略の概観を表17に示す。特異的な抗体を産生させるためのANGPTL8の組換え体発現に関連する問題の例としては、低収率、不十分な分泌、および凝集が挙げられる。例えば、標準的な発現プロトコルを使用すると、得られるタンパク質は、90%超が凝集しているように見えた。
ガレクチン−3などのレクチン結合タンパク質に対する免疫化戦略の概観を表18に示す。
表19は、選別された合計のハイブリドーマウェル(一般にこれらのウェルのうち約3分の1(1/3)がハイブリドーマを含有する)の標的タンパク質ごとの概要、および免疫原としての脂質に被包されたmRNAの使用後にそれらのハイブリドーマウェルから得られる確認された標的特異的な抗体の数を提供する。
特許、特許出願、論文、教科書などを含む、本明細書に引用される全ての参考文献およびそれらに引用される参考文献は、それらが既に引用されていない限り、本明細書に参照によりその全体が組み込まれる。
上述した明細書で、当業者は本発明を十分に実施可能であると考えられる。上述の説明および実施例は、本発明のある種の実施形態を詳述している。しかし、上述の説明がどんなに詳細に本文中に記載されているとしても、本発明は様々な様式で実施することができ、本発明は添付の特許請求の範囲およびその任意の均等物に従い解釈されるべきであると理解されよう。
Claims (48)
- 標的タンパク質に対するモノクローナル抗体を作製するための方法であって、(a)少なくとも1つのカチオン性脂質を、前記標的タンパク質またはその断片をコードするメッセンジャーRNA(mRNA)などのポリリボヌクレオチドと混合し、それにより、カチオン性脂質−ポリリボヌクレオチド複合体を形成するステップと;(b)前記脂質−ポリリボヌクレオチド複合体を非ヒト動物に投与するステップと;(c)前記標的タンパク質に特異的に結合する抗体を前記動物から得るステップとを含む、方法。
- 標的タンパク質に対するモノクローナル抗体を作製するための方法であって、(a)脂質−ポリリボヌクレオチド複合体を非ヒト動物に投与し、それにより、前記標的タンパク質に対する免疫応答を誘導するステップであって、前記複合体が、前記標的タンパク質またはその断片をコードするmRNAなどのポリリボヌクレオチドを有する少なくとも1つのカチオン性脂質を含むステップと;(b)前記標的タンパク質に特異的に結合する抗体を前記動物から得るステップとを含む、方法。
- 非ヒト動物において標的タンパク質に対する免疫応答を誘発するための方法であって、脂質−ポリヌクレオチド複合体を前記動物に投与するステップを含み、前記脂質−ポリヌクレオチド複合体がカチオン性脂質および標的タンパク質をコードするmRNAを含み、前記標的タンパク質が前記動物と異なる種のタンパク質である、方法。
- 前記標的タンパク質が膜貫通タンパク質である、請求項1、2または3に記載の方法。
- 前記膜貫通タンパク質が、次の:
(i)Gタンパク質共役型受容体(GPCR);
(ii)1回膜貫通型タンパク質受容体;
(iii)腫瘍壊死因子受容体スーパーファミリー(TNFRSF)メンバー;
(iv)インターロイキン(IL)受容体;
(v)イオンチャネル;
(vi)溶質輸送体;
(vii)免疫受容体;および
(viii)複数回膜貫通タンパク質
から選択される、請求項4に記載の方法。 - 前記膜貫通タンパク質が、Gタンパク質共役型受容体(GPCR)である、請求項4または5に記載の方法。
- 前記GPCRが、RXFP1、TSHR、APJ、GPR40、GPR64、GPR4、またはGPR15である、請求項6に記載の方法。
- 前記膜貫通タンパク質が、GP130などの1回膜貫通型タンパク質受容体である、請求項4または5に記載の方法。
- 前記膜貫通タンパク質が、SLC52A2などの複数回膜貫通タンパク質である、請求項4または5に記載の方法。
- 前記膜貫通タンパク質が、IL−1受容体、IL−2受容体、IL−3受容体、IL−4受容体、IL−5受容体、IL−6受容体、IL−7受容体、IL−8受容体、IL−9受容体、IL−10受容体、IL−11受容体、IL−12受容体、IL−13受容体、IL−14受容体、IL−15受容体、IL−16受容体、IL−17受容体、IL−18受容体、IL−19受容体、IL−20受容体、IL−21受容体、IL−22受容体、IL−23受容体、IL−24受容体、IL−25受容体、IL−26受容体、IL−27受容体、IL−28受容体、IL−29受容体、IL−30受容体、IL−31受容体、IL−32受容体、IL−33受容体、IL−35受容体、またはIL−36受容体などのインターロイキン(IL)受容体である、請求項4または5に記載の方法。
- 前記膜貫通タンパク質が、次の:TNFRSF1A、TNFRSF1B、TNFRSF3、TNFRSF4、TNFRSF5、TNFRSF6、TNFRSF6B、TNFRSF7、TNFRSF8、TNFRSF9、TNFRSF10A、TNFRSF10B、TNFRSF10C、TNFRSF10D、TNFRSF11A、TNFRSF11B、TNFRSF12A、TNFRSF13B、TNFRSF13C、TNFRSF14、TNFRSF16、TNFRSF17、TNFRSF18、TNFRSF19、TNFRSF21、TNFRSF25、およびTNFRSF27からなる群から選択される腫瘍壊死因子受容体スーパーファミリー(TNFRSF)メンバーである、請求項4または5に記載の方法。
- 前記膜貫通タンパク質が、TMEM16Aなどのイオンチャネルである、請求項4または5に記載の方法。
- 前記膜貫通タンパク質が、溶質輸送体である、請求項4または5に記載の方法。
- 前記標的タンパク質が、次の:ACKR1、ACKR2、ACKR3、ACKR4、ADCYAP1R1、ADGRA1、ADGRA2、ADGRA3、ADGRB1、ADGRB2、ADGRB3、ADGRD1、ADGRD2、ADGRE1、ADGRE2、ADGRE3、ADGRE4P、ADGRE5、ADGRF1、ADGRF2、ADGRF3、ADGRF4、ADGRF5、ADGRG1、ADGRG2、ADGRG3、ADGRG4、ADGRG5、ADGRG6、ADGRG7、ADGRL1、ADGRL2、ADGRL3、ADGRL4、ADGRV1、ADORA1、ADORA2A、ADORA2B、ADORA3、ADRA1A、ADRA1B、ADRA1D、ADRA2A、ADRA2B、ADRA2C、ADRB1、ADRB2、ADRB3、AGTR1、AGTR2、APLNR/APJ、ASGR1、ASGR2、AVPR1A、AVPR1B、AVPR2、BDKRB1、BDKRB2、BRS3、BRS3、C3AR1、C5AR1、C5AR2、CALCR、CALCRL、CASR、CCKAR、CCKBR、CCR1、CCR10、CCR2、CCR3、CCR4、CCR5、CCR6、CCR7、CCR8、CCR9、CCRL2、CELSR1、CELSR2、CELSR3、CHRM1、CHRM2、CHRM3、CHRM4、CHRM5、CMKLR1、CNR1、CNR2、CRHR1、CRHR2、CX3CR1、CXCR1、CXCR2、CXCR3、CXCR4、CXCR5、CXCR6、CYSLTR1、CYSLTR2、DRD1、DRD2、DRD3、DRD4、DRD5、EDNRA、EDNRB、F2R、F2RL1、F2RL2、F2RL3、FFAR1、FFAR2、FFAR3、FFAR4、FPR1、FPR2、FPR2、FPR3、FSHR、FZD1、FZD10、FZD2、FZD3、FZD4、FZD5、FZD6、FZD7、FZD8、FZD9、GABBR1、GABBR2、GALR1、GALR2、GALR3、GCGR、GHRHR、GHSR、GIPR、GLP1R、GLP2R、GNRHR、GNRHR2、GPBAR1、GPER1、GPR1、GPR4、GPR12、GPR15、GPR17、GPR18、GPR19、GPR20、GPR21、GPR22、GPR25、GPR26、GPR27、GPR3、GPR31、GPR32、GPR33、GPR34、GPR35、GPR37、GPR37L1、GPR39、GPR40、GPR42、GPR42、GPR45、GPR50、GPR52、GPR55、GPR6、GPR61、GPR62、GPR63、GPR65、GPR68、GPR75、GPR78、GPR79、GPR82、GPR83、GPR84、GPR85、GPR87、GPR88、GPR101、GPR107、GPR132、GPR135、GPR137、GPR139、GPR141、GPR142、GPR143、GPR146、GPR148、GPR149、GPR15、GPR150、GPR151、GPR152、GPR153、GPR156、GPR157、GPR158、GPR160、GPR161、GPR162、GPR171、GPR173、GPR174、GPR176、GPR179、GPR182、GPR183、GPRC5A、GPRC5B、GPRC5C、GPRC5D、GPRC6A、GRM1、GRM2、GRM3、GRM4、GRM5、GRM6、GRM7、GRM8、GRPR、HCAR1、HCAR2、HCAR3、HCRTR1、HCRTR2、HRH1、HRH2、HRH3、HRH4、HTR1A、HTR1B、HTR1D、HTR1E、HTR1F、HTR2A、HTR2B、HTR2C、HTR4、HTR5A、HTR5BP、HTR6、HTR7、KISS1R、LGR3、LGR4、LGR5、LGR6、LHCGR、LPAR1、LPAR2、LPAR3、LPAR4、LPAR5、LPAR6、LTB4R、LTB4R2、MAS1、MAS1L、MC1R、MC2R、MC3R、MC4R、MC5R、MCHR1、MCHR2、MLNR、MRGPRD、MRGPRE、MRGPRF、MRGPRG、MRGPRX1、MRGPRX2、MRGPRX3、MRGPRX4、MTNR1A、MTNR1B、NMBR、NMUR1、NMUR2、NPBWR1、NPBWR2、NPFFR1、NPFFR2、NPSR1、NPY1R、NPY2R、NPY4R、NPY5R、NPY6R、NTSR1、NTSR2、OPN3、OPN4、OPN5、OPRD1、OPRK1、OPRL1、OPRM1、OR51E1、OXER1、OXGR1、OXTR、P2RY1、P2RY10、P2RY11、P2RY12、P2RY13、P2RY14、P2RY2、P2RY4、P2RY6、P2RY8、PRLHR、PROKR1、PROKR2、PTAFR、PTGDR、PTGDR2、PTGER1、PTGER2、PTGER3、PTGER4、PTGFR、PTGIR、PTH1R、PTH2R、QRFPR、RXFP1、RXFP2、RXFP3、RXFP4、S1PR1、S1PR2、S1PR3、S1PR4、S1PR5、SCTR、SMO、SSTR1、SSTR2、SSTR3、SSTR4、SSTR5、SUCNR1、TAAR1、TAAR2、TAAR3、TAAR4P、TAAR5、TAAR6、TAAR8、TAAR9、TACR1、TACR2、TACR3、TAS1R1、TAS1R2、TAS1R3、TAS2R1、TAS2R10、TAS2R13、TAS2R14、TAS2R16、TAS2R19、TAS2R20、TAS2R3、TAS2R30、TAS2R31、TAS2R38、TAS2R39、TAS2R4、TAS2R40、TAS2R41、TAS2R42、TAS2R43、TAS2R45、TAS2R46、TAS2R5、TAS2R50、TAS2R60、TAS2R7、TAS2R8、TAS2R9、TBXA2R、TPRA1、TRHR、TSHR、UTS2R、VIPR1、VIPR2、XCR1、TCR−α、TCR−β、CD3、ζ−鎖アクセサリー、CD4、CD8、SIGIRR、マンノース受容体(MR)、アシアロ糖タンパク質受容体ファミリー(例えば、アシアロ糖タンパク質受容体マクロファージガラクトース型レクチン(MGL))、DC−SIGN(CLEC4L)、ランゲリン(CLEC4K)、骨髄系DAP12会合レクチン(MDL)−1(CLEC5A)、デクチン1/CLEC7A、DNGR1/CLEC9A、骨髄系C型レクチン様受容体(MICL)(CLEC12A)、CLEC2(CLEC1Bとも呼ばれる)、CLEC12B、DCIR/CLEC4A、デクチン2/CLEC6A、血液DC抗原2(BDCA2)(CLEC4C)、マクロファージ誘導性C型レクチン(CLEC4E)、TLR1、TLR2、TLR3、TLR4、TLR5、TLR6、TLR7、TLR8、TLR9、TLR10、TLR11、TLR12、TLR13,FcγRI(CD64)、FcγRIIA(CD32)、FcγRIIB1(CD32)、FcγRIIB2(CD32)、FcγRIIIA(CD16a)、FcγRIIIB(CD16b)、FcεRI、FcεRII(CD23)、FcαR1(CD89)、Fcα/μR、FcRn、CD27、CD40、OX40、GITR、CD137、PD−1、CTLA−4、PD−L1、TIGIT、T細胞免疫グロブリンドメインおよびムチンドメイン3(TIM3)、T細胞活性化のV−ドメインIg抑制因子(VISTA)、CD28、CD122、ICOS、A2AR、B7−H3、B7−H4、BおよびTリンパ球アテニュエータ(BTLA)、インドールアミン2,3−ジオキシゲナーゼ(IDO)、キラー細胞免疫グロブリン様受容体(KIR)、リンパ球活性化遺伝子−3(LAG3)、FAM159B、HLA−A、HLA−B、HLA−C、HLA−DPA1、HLA−DPB1、HLA−DQA1、HLA−DQB1、HLA−DRA、HLA−DRB1、gp130、IL−1受容体、IL−2受容体、IL−3受容体、IL−4受容体、IL−5受容体、IL−6受容体、IL−7受容体、IL−8受容体、IL−9受容体、IL−10受容体、IL−11受容体、IL−12受容体、IL−13受容体、IL−14受容体、IL−15受容体、IL−16受容体、IL−17受容体、IL−18受容体、IL−19受容体、IL−20受容体、IL−21受容体、IL−22受容体、IL−23受容体、IL−24受容体、IL−25受容体、IL−26受容体、IL−27受容体、IL−28受容体、IL−29受容体、IL−30受容体、IL−31受容体、IL−32受容体、IL−33受容体、IL−35受容体、IL−36受容体、FGFR1、FGFR2、FGFR3、FGFR4、TNFRSF1A、TNFRSF1B、TNFRSF3、TNFRSF4、TNFRSF5、TNFRSF6、TNFRSF6B、TNFRSF7、TNFRSF8、TNFRSF9、TNFRSF10A、TNFRSF10B、TNFRSF10C、TNFRSF10D、TNFRSF11A、TNFRSF11B、TNFRSF12A、TNFRSF13B、TNFRSF13C、TNFRSF14、TNFRSF16、TNFRSF17、TNFRSF18、TNFRSF19、TNFRSF21、TNFRSF25、TNFRSF27、SCN1A、SCN1B、SCN2A、SCN2B、SCN3A、SCN3B、SCN4A、SCN5A、SCN7A、SCN8A、SCN9A、SCN10A、SCN11A、CACNA1A、CACNA1B、CACNA1C、CACNA1D、CACNA1E、CACNA1F、CACNA1G、CACNA1H、CACNA1I、CACNA1S、TRPA1、TRPC1、TRPC2、TRPC3、TRPC4、TRPC5、TRPC6、TRPC7、TRPM1、TRPM2、TRPM3、TRPM4、TRPM5、TRPM6、TRPM7、TRPM8、MCOLN1、MCOLN2、MCOLN3、PKD1、PKD2、PKD2L1、PKD2L2、TRPV1、TRPV2、TRPV3、TRPV4、TRPV5、TRPV6、CATSPER1、CATSPER2、CATSPER3、CATSPER4、TPCN1、TPCN2、CNGA1、CNGA2、CNGA3、CNGA4、CNGB1、CNGB3、HCN1、HCN2、HCN3、HCN4、KCNMA1、KCNN1、KCNN2、KCNN3、KCNN4、KCNT1、KCNT2、KCNU1、KCNA1、KCNA2、KCNA3、KCNA4、KCNA5、KCNA6、KCNA7、KCNA10、KCNB1、KCNB2、KCNC1、KCNC2、KCNC3、KCNC4、KCND1、KCND2、KCND3、KCNF1、KCNG1、KCNG2、KCNG3、KCNG4、KCNH1、KCNH2、KCNH3、KCNH4、KCNH5、KCNH6、KCNH7、KCNH8、KCNQ1、KCNQ2、KCNQ3、KCNQ4、KCNQ5、KCNS1、KCNS2、KCNS3、KCNV1、KCNV2、KCNJ1、KCNJ2、KCNJ3、KCNJ4、KCNJ5、KCNJ6、KCNJ8、KCNJ9、KCNJ10、KCNJ11、KCNJ12、KCNJ13、KCNJ14、KCNJ15、KCNJ16、KCNJ18、KCNK1、KCNK2、KCNK3、KCNK4、KCNK5、KCNK6、KCNK7、KCNK9、KCNK10、KCNK12、KCNK13、KCNK15、KCNK16、KCNK17、KCNK18、HVCN1、HTR3A、HTR3B、HTR3C、HTR3D、HTR3E、CHRNA1、CHRNA2、CHRNA3、CHRNA4、CHRNA5、CHRNA6、CHRNA7、CHRNA9、CHRNA10、CHRNB1、CHRNB2、CHRNB3、CHRNB4、CHRND、CHRNE、CHRNG、GABRA1、GABRA2、GABRA3、GABRA4、GABRA5、GABRA6、GABRB1、GABRB2、GABRB3、GA
BRD、GABRE、GABRG1、GABRG2、GABRG3、GABRP、GABRQ、GABRR1、GABRR2、GABRR3、GRIA1、GRIA2、GRIA3、GRIA4、GRID1、GRID2、GRIK1、GRIK2、GRIK3、GRIK4、GRIK5、GRIN1、GRIN2A、GRIN2B、GRIN2C、GRIN2D、GRIN3A、GRIN3B、GLRA1、GLRA2、GLRA3、GLRA4、P2RX1、P2RX2、P2RX3、P2RX4、P2RX5、P2RX6、P2RX7、ZACN、ASIC1、ASIC2、ASIC3、ASIC4、AQP1、AQP2、AQP3、AQP4、AQP5、AQP6、AQP7、AQP8、AQP9、AQP10、AQP11、AQP12A、AQP12B、MIP、CLCN1、CLCN2、CLCN3、CLCN4、CLCN5、CLCN6、CLCN7、CLCNKA、CLCNKB、嚢胞性線維症膜コンダクタンス制御因子(CFTR)、ANO1、ANO2、ANO3、ANO4、ANO5、ANO6、ANO7、ANO8、ANO9、ANO10、BEST1、BEST2、BEST3、BEST4、CLIC1、CLIC2、CLIC3、CLIC4、CLIC5、CLIC6、GJA1、GJA3、GJA4、GJA5、GJA6P、GJA8、GJA9、GJA10、GJB1、GJB2、GJB3、GJB4、GJB5、GJB6、GJB7、GJC1、GJC2、GJC3、GJD2、GJD3、GJD4、GJE1、ITPR1、ITPR2、ITPR3、PANX1、PANX2、PANX3、RYR1、RYR2、RYR3、NALCN、SCNN1A、SCNN1B、SCNN1D、SCNN1G、TEM16A、ADAMTS7、ANGPTL3、ANGPTL4、ANGPTL8、LPL、GDF15、ガレクチン−1、ガレクチン−2、ガレクチン−3、ガレクチン−4、ガレクチン−7、ガレクチン−8、ガレクチン−9、ガレクチン−10、ガレクチン−12、ガレクチン−13、マトリックスglaタンパク質(MGP)、PRNP、DGAT1、GPAT3、DMC1、BLM、BRCA2、ヒト内在性レトロウイルスタイプK(HERV−K)ファミリーのメンバー、エクトヌクレオシド三リン酸ジホスホヒドロラーゼ1(ENTPD1)、エクトヌクレオシド三リン酸ジホスホヒドロラーゼ2(ENTPD2)、SLC1A1、SLC1A2、SLC1A3、SLC1A4、SLC1A5、SLC1A6、SLC1A7、SLC2A1、SLC2A2、SLC2A3、SLC2A4、SLC2A5、SLC2A6、SLC2A7、SLC2A8、SLC2A9、SLC2A10、SLC2A11、SLC2A12、SLC2A13、SLC2A14、SLC3A1、SLC3A2、SLC4A1、SLC4A2、SLC4A3、SLC4A4、SLC4A5、SLC4A6、SLC4A7、SLC4A8、SLC4A9、SLC4A10、SLC4A11、SLC5A1、SLC5A2、SLC5A3、SLC5A4、SLC5A5、SLC5A6、SLC5A7、SLC5A8、SLC5A9、SLC5A10、SLC5A11、SLC5A12、SLC6A1、SLC6A2、SLC6A3、SLC6A4、SLC6A5、SLC6A6、SLC6A7、SLC6A8、SLC6A9、SLC6A10、SLC6A11、SLC6A12、SLC6A13、SLC6A14、SLC6A15、SLC6A16、SLC6A17、SLC6A18、SLC6A19、SLC6A20、SLC7A5、SLC7A6、SLC7A7、SLC7A8、SLC7A9、SLC7A10、SLC7A11、SLC7A13、SLC7A14、SLC8A1、SLC8A2、SLC8A3、SLC9A1、SLC9A2、SLC9A3、SLC9A4、SLC9A5、SLC9A6、SLC9A7、SLC9A8、SLC9A9、SLC9A10、SLC9A11、SLC9B1、SLC9B2、SLC10A1、SLC10A2、SLC10A3、SLC10A4、SLC10A5、SLC10A6、SLC10A7、SLC11A1、SLC11A2、SLC12A1、SLC12A2、SLC12A3、SLC12A4、SLC12A5、SLC12A6、SLC12A7、SLC12A8、SLC12A9、SLC13A1、SLC13A2、SLC13A3、SLC13A4、SLC13A5、SLC14A1、SLC14A2、SLC15A1、SLC15A2、SLC15A3、SLC15A4、SLC16A1、SLC16A2、SLC16A3、SLC16A4、SLC16A5、SLC16A6、SLC16A7、SLC16A8、SLC16A9、SLC16A10、SLC16A11、SLC16A12、SLC16A13、SLC16A14、SLC17A1、SLC17A2、SLC17A3、SLC17A4、SLC17A5、SLC17A6、SLC17A7、SLC17A8、SLC17A9、SLC18A1、SLC18A2、SLC18A3、SLC19A1、SLC19A2、SLC19A3、SLC20A1、SLC20A2、SLCO1A2、SLCO1B1、SLCO1B3、SLCO1C1、SLCO2A1、SLCO2B1、SLCO3A1、SLCO4A1、SLCO4C1、SLCO5A1、SLCO6A1、SLC22A1、SLC22A2、SLC22A3、SLC22A4、SLC22A5、SLC22A6、SLC22A7、SLC22A8、SLC22A9、SLC22A10、SLC22A11、SLC22A12、SLC22A13、SLC22A14、SLC22A15、SLC22A16、SLC22A17、SLC22A18、SLC22A18AS,SLC22A19、SLC22A20、SLC22A23、SLC22A24、SLC22A25、SLC22A31、SLC23A1、SLC23A2、SLC23A3、SLC23A4、SLC24A1、SLC24A2、SLC24A3、SLC24A4、SLC24A5、SLC24A6、SLC25A1、SLC25A2、SLC25A3、SLC25A4、SLC25A5、SLC25A6、SLC25A7、SLC25A8、SLC25A9、SLC25A10、SLC25A11、SLC25A12、SLC25A13、SLC25A14、SLC25A15、SLC25A16、SLC25A17、SLC25A18、SLC25A19,SLC25A20、SLC25A21、SLC25A22、SLC25A23、SLC25A24、SLC25A25、SLC25A26、SLC25A27、SLC25A28、SLC25A29、SLC25A30、SLC25A31、SLC25A32、SLC25A33、SLC25A34、SLC25A35、SLC25A36、SLC25A37,SLC25A38、SLC25A39、SLC25A40、SLC25A41、SLC25A42、SLC25A43、SLC25A44、SLC25A45、SLC25A46、SLC26A1、SLC26A2、SLC26A3、SLC26A4、SLC26A5、SLC26A6、SLC26A7、SLC26A8、SLC26A9、SLC26A10、SLC26A11、SLC27A1、SLC27A2、SLC27A3、SLC27A4、SLC27A5、SLC27A6、SLC28A1、SLC28A2、SLC28A3、SLC29A1、SLC29A2、SLC29A3、SLC29A4、SLC30A1、SLC30A2、SLC30A3、SLC30A4、SLC30A5、SLC30A6、SLC30A7、SLC30A8、SLC30A9、SLC30A10、SLC31A1、SLC31A2、SLC32A1、SLC33A1、SLC34A1、SLC34A2、SLC34A3、SLC35A1、SLC35A2、SLC35A3、SLC35A4、SLC35A5、SLC35B1、SLC35B2、SLC35B3、SLC35B4、SLC35C1、SLC35C2、SLC35D1、SLC35D2、SLC35D3、SLC35E1、SLC35E2、SLC35E3、SLC35E4、SLC35F1、SLC35F2、SLC35F3、SLC35F4、SLC35F5、SLC35G1、SLC35G3、SLC35G4、SLC35G5、SLC35G6、SLC36A1、SLC36A2、SLC36A3、SLC36A4、SLC37A1、SLC37A2、SLC37A3、SLC37A4、SLC38A1、SLC38A2、SLC38A3、SLC38A4、SLC38A5、SLC38A6、SLC38A7、SLC38A8、SLC38A9、SLC38A10、SLC38A11、SLC39A1、SLC39A2、SLC39A3、SLC39A4、SLC39A5、SLC39A6、SLC39A7、SLC39A8、SLC39A9、SLC39A10、SLC39A11、SLC39A12、SLC39A13、SLC39A14、SLC40A1、SLC41A1、SLC41A2、SLC41A3、RhAG、RhBG、RhCG、SLC43A1、SLC43A2、SLC43A3、SLC44A1、SLC44A2、SLC44A3、SLC44A4、SLC44A5、SLC45A1、SLC45A2、SLC45A3、SLC45A4、SLC46A1、SLC46A2、SLC46A3、SLC47A1、SLC47A2、HCP−1、MFSD5、MFSD10、SLC50A1、OSTα、OSTβ、SLC52A1、SLC52A2、ならびにSLC52A3から選択される、請求項1、2または3に記載の方法。 - 前記標的抗原が、発現させることが困難であるか、またはそれに対する抗体を産生させることが困難である、請求項1〜14のいずれか一項に記載の方法。
- 前記標的タンパク質の発現が、宿主産生細胞において細胞毒性をもたらす、請求項1〜15のいずれか一項に記載の方法。
- 前記標的タンパク質が、組換えにより発現されかつ/または精製される場合、不十分な収率、安定性、溶解性、および/または機能活性を示す、請求項1〜16のいずれか一項に記載の方法。
- 前記複合体の前記ポリヌクレオチドまたはmRNAが、次の:コンセンサスコザック配列;前記mRNAの5’末端上の7−メチルグアノシンキャップ;前記mRNA転写物の3’終端で見られるポリアデノシン(ポリA)テール;ならびに5’−および3’−非翻訳領域(UTR)の1つ以上を含む、請求項1〜17のいずれか一項に記載の方法。
- 前記投与が、非経口、静脈内、筋肉内、鼻腔内、または皮下である、請求項1〜18のいずれか一項に記載の方法。
- 前記標的タンパク質がRXFP1である、請求項1〜19のいずれか一項に記載の方法。
- 前記複合体が、配列番号4、または配列番号2、4および37のいずれか1つの核酸配列を含むポリリボヌクレオチドまたはmRNAを含む、請求項1〜20のいずれか一項に記載の方法。
- 前記標的タンパク質がSLC52A2である、請求項1〜21のいずれか一項に記載の方法。
- 前記複合体が、配列番号7、または配列番号5、7および40のいずれか1つの核酸配列を含むポリリボヌクレオチドまたはmRNAを含む、請求項1〜22のいずれか一項に記載の方法。
- 前記標的タンパク質がANGPTL8である、請求項1〜23のいずれか一項に記載の方法。
- 前記複合体が、配列番号10、または配列番号8、10および43のいずれか1つの核酸配列を含むポリリボヌクレオチドまたはmRNAを含む、請求項1〜24のいずれか一項に記載の方法。
- 前記標的タンパク質がTSHRである、請求項1〜25のいずれか一項に記載の方法。
- 前記複合体が、配列番号16、または配列番号14、16および46のいずれか1つの核酸配列を含むポリリボヌクレオチドまたはmRNAを含む、請求項1〜26のいずれか一項に記載の方法。
- 前記標的タンパク質がAPJである、請求項1〜27のいずれか一項に記載の方法。
- 前記複合体が、配列番号19、または配列番号17、19および49のいずれか1つの核酸配列を含むポリリボヌクレオチドまたはmRNAを含む、請求項1〜28のいずれか一項に記載の方法。
- 前記標的タンパク質がgp130である、請求項1〜29のいずれか一項に記載の方法。
- 前記複合体が、配列番号22、または配列番号20、22および52のいずれか1つの核酸配列を含むポリリボヌクレオチドまたはmRNAを含む、請求項1〜30のいずれか一項に記載の方法。
- 前記標的タンパク質がガレクチン3である、請求項1〜31のいずれか一項に記載の方法。
- 前記複合体が、配列番号55、または配列番号26、55および56のいずれか1つの核酸配列を含むポリリボヌクレオチドまたはmRNAを含む、請求項1〜32のいずれか一項に記載の方法。
- 前記複合体が、約30〜150nmの直径を有する、請求項1〜33のいずれか一項に記載の方法。
- 前記複合体がヘルパー脂質を含む、請求項1〜34のいずれか一項に記載の方法。
- 前記複合体が、(i)カチオン性脂質、(ii)ヘルパー脂質、例えば、コレステロール、(iii)中性脂質、例えば、DSPC、および(iv)ステルス脂質、例えば、S010、S024、S027、S031またはS033の任意の組み合わせを含む、請求項1〜35のいずれか一項に記載の方法。
- 前記動物が、10μg、12.5μg、20μg、25μg、30μg、40μg、50μg、60μg、70μg、80μg、90μg、100μg、110μg、120μg、130μg、140μg、または150μgのポリリボヌクレオチドを投与される、請求項1〜36のいずれか一項に記載の方法。
- 前記カチオン性脂質が、N,N−ジオレイル−N,N−ジメチルアンモニウムクロリド(DODAC)、N,N−ジステアリル−N,N−ジメチルアンモニウムブロミド(DDAB)、N−(1−(2,3−ジオレオイルオキシ)プロピル)−N,N,N−トリメチルアンモニウムクロリド(DOTAP)、1,2−ジオレオイル−3−ジメチルアンモニウム−プロパン(DODAP)、N−(1−(2,3−ジオレイルオキシ)プロピル)−N,N,N−トリメチルアンモニウムクロリド(DOTMA)、1,2−ジオレオイルカルバミル−3−ジメチルアンモニウム−プロパン(DOCDAP)、1,2−ジリネオイル−3−ジメチルアンモニウム−プロパン(DLINDAP)、ジラウリル(C12:0)トリメチルアンモニウムプロパン(DLTAP)、ジオクタデシルアミドグリシルスペルミン(DOGS)、DC−Chol、ジオレオイルオキシ−N−[2−スペルミンカルボキサミド)エチル}−N,N−ジメチル−1−プロパンアミニウムトリフルオロアセテート(DOSPA)、1,2−ジミリスチルオキシプロピル−3−ジメチル−ヒドロキシエチルアンモニウムブロミド(DMRIE)、3−ジメチルアミノ−2−(コレスト−5−エン−3−β−オキシブタン−4−オキシ)−1−(シス,シス−9,12−オクタデカジエノキシ)プロパン(CLinDMA)、N,N−ジメチル−2,3−ジオレイルオキシ)プロピルアミン(DODMA)、2−[5’−(コレスト−5−エン−3β−オキシ)−3’−オキサペントキシ)−3−ジメチル−1−(シス,シス−9’,12’−オクタデカジエノキシ)プロパン(CpLinDMA)およびN,N−ジメチル−3,4−ジオレイルオキシベンジルアミン(DMOBA)、ならびに1,2−N,N’−ジオレイルカルバミル−3−ジメチルアミノプロパン(DOcarbDAP)からなる群から選択される、請求項1〜37のいずれか一項に記載の方法。
- 前記カチオン性脂質がDOTAPまたはDLTAPである、請求項38に記載の方法。
- 前記標的タンパク質に特異的に結合する抗体を産生するハイブリドーマを作製するステップをさらに含む、請求項1〜39のいずれか一項に記載の方法。
- 前記標的タンパク質に特異的に結合する抗体を精製するステップをさらに含む、請求項1〜40のいずれか一項に記載の方法。
- 前記標的タンパク質に特異的に結合する精製された抗体に由来するキメラまたはヒト化抗体を作製するステップをさらに含む、請求項1〜41のいずれか一項に記載の方法。
- 前記方法が、cDNA、タンパク質もしくはペプチド、ウイルス粒子、または細胞全体による免疫化を含む方法と比較して、1回目の採血または2回目の採血由来の血清中においてより高い抗体力価を生じる、請求項1〜42のいずれか一項に記載の方法。
- 前記方法が、cDNA、タンパク質もしくはペプチド、ウイルス粒子、または細胞全体による免疫化を含む方法よりも、標的タンパク質に特異的な抗体を産生するハイブリドーマをより多く生じる、請求項1〜43のいずれか一項に記載の方法。
- 前記標的タンパク質がヒト標的タンパク質であり、非ヒト動物が、マウス、ラット、ウサギ、ヒツジ、ネコ、イヌ、ラクダ科動物、サメ、サル、ブタ、またはウマである、請求項1〜44のいずれか一項に記載の方法。
- 前記標的タンパク質に特異的に結合する抗体を産生するハイブリドーマであって、前記ハイブリドーマが請求項1〜45のいずれか1つの方法によって得られた、ハイブリドーマ。
- 前記標的タンパク質に特異的に結合するポリクローナル抗体の混合物であって、前記混合物が請求項1〜45のいずれか1つの方法によって得られた、混合物。
- 前記標的タンパク質に特異的に結合する単離されたモノクローナル抗体であって、前記モノクローナル抗体が請求項1〜45のいずれか1つの方法によって得られた、単離されたモノクローナル抗体。
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