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RU2017125758A - Антитела к с5 и способы их применения - Google Patents

Антитела к с5 и способы их применения Download PDF

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RU2017125758A
RU2017125758A RU2017125758A RU2017125758A RU2017125758A RU 2017125758 A RU2017125758 A RU 2017125758A RU 2017125758 A RU2017125758 A RU 2017125758A RU 2017125758 A RU2017125758 A RU 2017125758A RU 2017125758 A RU2017125758 A RU 2017125758A
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Ёсинао Руике
Дзендзиро Сампей
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Чугаи Сейяку Кабусики Кайся
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Claims (45)

1. Выделенное антитело, которое связывается с С5, где антитело связывается с эпитопом в бета-цепи С5 с более высокой аффинностью при нейтральном рН, чем при кислом рН.
2. Антитело по п. 1, где антитело обладает характеристикой, выбранное из группы, которая состоит из следующих характеристик:
(а) антитело контактирует с аминокислотами D51 и K109 С5 (SEQ ID NO: 39);
(б) аффинность антитела к С5 (SEQ ID NO: 39) больше, чем аффинность антитела к мутант С5, содержащему замену Е48А в SEQ ID NO: 39; и
(в) антитело связывается с белком С5, состоящим из аминокислотной последовательности SEQ ID NO: 39, при рН 7,4, но не связывается с белком С5, состоящим из аминокислотной последовательности SEQ ID NO: 39 с заменой H72Y, при рН 7,4.
3. Антитело по п. 1, где антитело конкурирует за связывание с С5 с антителом, которое содержит пару VH и VL, выбранную из:
(а) VH, имеющей SEQ ID NO: 1, и VL, имеющей SEQ ID NO: 11;
(б) VH, имеющей SEQ ID NO: 5, и VL, имеющей SEQ ID NO: 15;
(в) VH, имеющей SEQ ID NO: 4, и VL, имеющей SEQ ID NO: 14;
(г) VH, имеющей SEQ ID NO: 6, и VL, имеющей SEQ ID NO: 16;
(д) VH, имеющей SEQ ID NO: 2, и VL, имеющей SEQ ID NO: 12;
(е) VH, имеющей SEQ ID NO: 3, и VL, имеющей SEQ ID NO: 13;
(ж) VH, имеющей SEQ ID NO: 9, и VL, имеющей SEQ ID NO: 19;
(з) VH, имеющей SEQ ID NO: 7, и VL, имеющей SEQ ID NO: 17;
(и) VH, имеющей SEQ ID NO: 8, и VL, имеющей SEQ ID NO: 18, и
(к) VH, имеющей SEQ ID NO: 10, и VL, имеющей SEQ ID NO: 20.
4. Антитело по одному из пп. 1-3, где антитело связывается с эпитопом в домене MG1-MG2 бета-цепи С5.
5. Антитело по одному из пп. 1-4, где антитело связывается с эпитопом во фрагменте, состоящем из аминокислот 33-124 бета-цепи (SEQ ID NO: 40) С5.
6. Антитело по одному из пп. 1-5, где антитело связывается с эпитопом в бета-цепи (SEQ ID NO: 40) С5, который содержит по меньшей мере один фрагмент выбранный из группы, которая состоит из аминокислот 47-57, 70-76 и 107-110.
7. Антитело по одному из пп. 1-6, где антитело связывается с эпитопом во фрагменте бета-цепи (SEQ ID NO: 40) С5, который содержит по меньшей мере одну аминокислоту, выбранную из группы, которая состоит из Glu48, Asp51, His70, His72, Lys109 и His110.
8. Антитело по одному из пп. 1-7, которое связывается с тем же эпитопом, что и антитело, описанное в таблицах 2, 7 или 8.
9. Антитело по одному из пп. 1-8, которое ингибирует активацию С5.
10. Антитело по одному из пп. 1-9, которое ингибирует активацию R885H-варианта С5.
11. Антитело по одному из пп. 1-10, которое представляет собой моноклональное антитело.
12. Антитело по одному из пп. 1-11, которое представляет собой человеческое, гуманизированное или химерное антитело.
13. Антитело по одному из пп. 1-12, которое представляет собой фрагмент антитела, который связывается с С5.
14. Антитело по одному из пп. 1-13, где антитело содержит (a) HVR-H3, включающий аминокислотную последовательность DX1GYX2X3PTHAMX4X5, в которой X1 обозначает G или А, Х2 обозначает V, Q или D, Х3 обозначает Т или Y, X4 обозначает Y или H, X5 обозначает L или Y (SEQ ID NO: 128), (б) a HVR-L3, включающий аминокислотную последовательность QX1TX2VGSSYGNX3, в которой X1 обозначает S, С, N или Т, Х2 обозначает F или K, Х3 обозначает А, Т или Н (SEQ ID NO: 131), и (в) HVR-H2, включающий аминокислотную последовательность X1IX2TGSGAX3YX4AX5WX6KG, в которой X1 обозначает С, А или G, Х2 обозначает Y или F, Х3 обозначает Т, D или Е, Х4 обозначает Y, K или Q, Х5 обозначает S, D или Е, Х6 обозначает А или V (SEQ ID NO: 127).
15. Антитело по одному из пп. 1-13, где антитело содержит (a) HVR-H1, включающий аминокислотную последовательность SSYYX1X2, в которой X1 обозначает М или V, Х2 обозначает С или A (SEQ ID NO: 126), (б) HVR-H2, включающий аминокислотную последовательность X1IX2TGSGAX3YX4AX5WX6KG, в которой X1 обозначает С, А или G, Х2 обозначает Y или F, Х3 обозначает Т, D или Е, Х4 обозначает Y, K или Q, Х5 обозначает S, D или Е, Х6 обозначает А или V (SEQ ID NO: 127), и (в) HVR-H3, включающий аминокислотную последовательность DX1GYX2X3PTHAMX4X5, в которой X1 обозначает G или А, Х2 обозначает V, Q или D, Х3 обозначает Т или Y, Х4 обозначает Y или Н, Х5 обозначает L или Y (SEQ ID NO: 128).
16. Антитело по п. 15, которое дополнительно содержит (a) HVR-L1, включающий аминокислотную последовательность X1ASQX2IX3SX4LA, в которой X1 обозначает Q или R, Х2 обозначает N, Q или G, Х3 обозначает G или S, Х4 обозначает D, K или S (SEQ ID NO: 129); (б) HVR-L2, включающий аминокислотную последовательность GASX1X2X3S, в которой X1 обозначает K, Е или Т, Х2 обозначает L или Т, Х3 обозначает А, Н, Е или Q (SEQ ID NO: 130); и (в) HVR-L3, включающий аминокислотную последовательность QX1TX2VGSSYGNX3, в которой X1 обозначает S, С, N или Т, Х2 обозначает F или K, Х3 обозначает А, Т или Н (SEQ ID NO: 131).
17. Антитело по одному из пп. 1-13, которое содержит (a) HVR-L1, включающий аминокислотную последовательность X1ASQX2IX3SX4LA, в которой X1 обозначает Q или R, Х2 обозначает N, Q или G, Х3 обозначает G или S, Х4 обозначает D, K или S (SEQ ID NO: 129); (б) HVR-L2, включающий аминокислотную последовательность GASX1X2X3S, в которой X1 обозначает K, Е или Т, Х2 обозначает L или Т, Х3 обозначает А, Н, Е или Q (SEQ ID NO: 130); и (в) HVR-L3, включающий аминокислотную последовательность QX1TX2VGSSYGNX3, в которой X1 обозначает S, С, N или Т, Х2 обозначает F или K, Х3 обозначает А, Т или Н (SEQ ID NO: 131).
18. Антитело по п. 15, которое дополнительно содержит каркасный участок вариабельного домена тяжелой цепи FR1, включающий одну из аминокислотных последовательностей SEQ ID NO: 132-134; FR2, включающий одну из аминокислотных последовательностей SEQ ID NO: 135-136; и FR3, включающий одну из аминокислотных последовательностей SEQ ID NO: 137-139; и FR4, включающий одну из аминокислотных последовательностей SEQ ID NO: 140-141.
19. Антитело по п. 17, которое дополнительно содержит каркасный участок вариабельного домена легкой цепи FR1, включающий одну из аминокислотных последовательностей SEQ ID NO: 142-143; FR2, включающий одну из аминокислотных последовательностей SEQ ID NO: 144-145; FR3, включающий одну из аминокислотных последовательностей SEQ ID NO: 146-147; и FR4, включающий аминокислотную последовательность SEQ ID NO: 148.
20. Антитело по одному из пп. 1-13, которое содержит (а) последовательность VH, идентичную по меньшей мере на 95% одной из аминокислотных последовательностей SEQ ID NO: 10, 106-110; (б) последовательность VL, идентичную по меньшей мере на 95% одной из аминокислотных последовательностей SEQ ID NO: 20, 111-113; или (в) последовательность VH, указанную в подпункте (а), и последовательность VL, указанную в подпункте (б).
21. Антитело по п. 20, содержащее последовательность VH, представленную в одной из SEQ ID NO: 10, 106-110.
22. Антитело по п. 20, содержащее последовательность VL, представленную в одной из SEQ ID NO: 20, 111-113.
23. Антитело, содержащее последовательность VH, представленную в одной из SEQ ID NO: 10, 106-110, и последовательность VL, представленную в одной из SEQ ID NO: 20, 111-113.
24. Антитело по одному из пп. 1-23, которое представляет собой полноразмерное антитело IgG1-класса или IgG4-класса.
25. Выделенная нуклеиновая кислота, кодирующая антитело по одному из пп. 1-24.
26. Клетка-хозяин, содержащая нуклеиновую кислоту по п. 25.
27. Способ получения антитела, включающий культивирование клетки-хозяина по п. 26, в результате чего получают антитело.
28. Способ получения антитела к С5, включающий иммунизацию животного полипептидом, где полипептид содержит домен MG1-MG2 (SEQ ID NO: 43) бета-цепи С5.
29. Способ получения антитела к С5, включающий иммунизацию животного полипептидом, где полипептид содержит область, соответствующую аминокислотам в положениях 33-124 бета-цепи (SEQ ID NO: 40) С5.
30. Способ получения антитела к С5, включающий иммунизацию животного полипептидом, где полипептид содержит по меньшей мере один фрагмент, выбранный из аминокислот 47-57, 70-76 и 107-110 бета-цепи (SEQ ID NO: 40) С5.
31. Способ получения антитела к С5, включающий иммунизацию животного полипептидом, где полипептид содержит фрагмент бета-цепи (SEQ ID NO: 40) С5, который содержит по меньшей мере одну аминокислоту, выбранную из Glu48, Asp51, His70, His72, Lys109 и His110.
32. Фармацевтическая композиция, содержащая антитело по одному из пп. 1-24 и фармацевтически приемлемый носитель.
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