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RU2012112829A - Анти-gitr-антитела - Google Patents

Анти-gitr-антитела Download PDF

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RU2012112829A
RU2012112829A RU2012112829/10A RU2012112829A RU2012112829A RU 2012112829 A RU2012112829 A RU 2012112829A RU 2012112829/10 A RU2012112829/10 A RU 2012112829/10A RU 2012112829 A RU2012112829 A RU 2012112829A RU 2012112829 A RU2012112829 A RU 2012112829A
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seq
variable domain
sequence
heavy chain
amino acid
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RU2012112829/10A
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RU2595409C2 (ru
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Сяо Минь ШИБАЙ
Григорий П. ЕРМАКОВ
Дуглас Дж. ХОДЖЕС
Леонард Г. ПРЕСТА
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Шеринг Корпорейшн
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Abstract

1. Связывающее соединение, которое связывается с GITR человека, содержащее:a) вариабельный домен легкой цепи антитела или его антигенсвязывающий фрагмент, имеющий три последовательности CDR, выбранные из группы, состоящей из последовательностей SEQ ID NO: 56-88, иb) вариабельный домен тяжелой цепи антитела или его антигенсвязывающий фрагмент, имеющий три последовательности CDR, выбранные из группы, состоящей из последовательностей SEQ ID NO: 23-55.2. Связывающее соединение по п.1, в котором CDR содержат:a) по меньшей мере одну CDRL1 из группы, состоящей из последовательностей SEQ ID NO: 56-66; по меньшей мере одну CDRL2 из группы, состоящей из последовательностей SEQ ID NO: 67-77; и по меньшей мере одну CDRL3 из группы, состоящей из последовательностей SEQ ID NO: 78-88; иb) по меньшей мере одну CDRH1 из группы, состоящей из последовательностей SEQ ID NO: 23-33; по меньшей мере одну CDRH2 из группы, состоящей из последовательностей SEQ ID NO: 34-44; и по меньшей мере одну CDRH3 из группы, состоящей из последовательностей SEQ ID NO: 45-55.3. Связывающее соединение по п.1, в котором:a) аминокислотная последовательность вариабельного домена тяжелой цепи выбрана из группы, состоящей из последовательностей SEQ ID NO: 90, 92, 94, 96, 98, 100, 102, 104, 106, 108 и 110; иb) аминокислотная последовательность вариабельного домена легкой цепи выбрана из группы, состоящей из последовательностей 91, 93, 95, 97, 99, 101, 103, 105, 107, 109 и 111.4. Связывающее соединение по п.3, в котором:a) аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 90, и последовательностью вариабельного домена легкой цепи является последовательность SEQ ID NO: 91;b) аминокислотной последовательностью вариабельного домена тяжелой ц

Claims (22)

1. Связывающее соединение, которое связывается с GITR человека, содержащее:
a) вариабельный домен легкой цепи антитела или его антигенсвязывающий фрагмент, имеющий три последовательности CDR, выбранные из группы, состоящей из последовательностей SEQ ID NO: 56-88, и
b) вариабельный домен тяжелой цепи антитела или его антигенсвязывающий фрагмент, имеющий три последовательности CDR, выбранные из группы, состоящей из последовательностей SEQ ID NO: 23-55.
2. Связывающее соединение по п.1, в котором CDR содержат:
a) по меньшей мере одну CDRL1 из группы, состоящей из последовательностей SEQ ID NO: 56-66; по меньшей мере одну CDRL2 из группы, состоящей из последовательностей SEQ ID NO: 67-77; и по меньшей мере одну CDRL3 из группы, состоящей из последовательностей SEQ ID NO: 78-88; и
b) по меньшей мере одну CDRH1 из группы, состоящей из последовательностей SEQ ID NO: 23-33; по меньшей мере одну CDRH2 из группы, состоящей из последовательностей SEQ ID NO: 34-44; и по меньшей мере одну CDRH3 из группы, состоящей из последовательностей SEQ ID NO: 45-55.
3. Связывающее соединение по п.1, в котором:
a) аминокислотная последовательность вариабельного домена тяжелой цепи выбрана из группы, состоящей из последовательностей SEQ ID NO: 90, 92, 94, 96, 98, 100, 102, 104, 106, 108 и 110; и
b) аминокислотная последовательность вариабельного домена легкой цепи выбрана из группы, состоящей из последовательностей 91, 93, 95, 97, 99, 101, 103, 105, 107, 109 и 111.
4. Связывающее соединение по п.3, в котором:
a) аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 90, и последовательностью вариабельного домена легкой цепи является последовательность SEQ ID NO: 91;
b) аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 92, и аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 93;
c) аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 94, и аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 95;
d) аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 96, и аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 97;
e) аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 98, и аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 99;
f) аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 100, и аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 101;
g) аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 102, и аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 103;
h) аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 104, и аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 105;
i) аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 106, и аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 107;
j) аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 108, и аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 109; или
k) аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 110, и аминокислотной последовательностью вариабельного домена тяжелой цепи является последовательность SEQ ID NO: 111.
5. Антитело, которое способно блокировать связывание связывающего соединения по п.4 с GITR человека в анализе перекрестного блокирования.
6. Изолированная нуклеиновая кислота, кодирующая по меньшей мере один из вариабельного домена легкой цепи или вариабельного домена тяжелой цепи связывающего соединения по п.4.
7. Экспрессирующий вектор, содержащий нуклеиновую кислоту по п.6, функционально связанную с регуляторными последовательностями, которые распознает клетка-хозяин, когда клетка-хозяин трансфицирована вектором.
8. Клетка-хозяин, содержащая экспрессирующий вектор по п.7.
9. Способ получения полипептида, включающий
культивирование клетки-хозяина по п.8 в культуральной среде в условиях, в которых экспрессируется последовательность нуклеиновой кислоты, таким образом получая полипептиды, содержащие вариабельные домены легкой и тяжелой цепи; и
извлечение полипептидов из клетки-хозяина или культуральной среды.
10. Связывающее соединение по п.4, дополнительно содержащее константную область тяжелой цепи, содержащую константную область тяжелой цепи γ1 человека или ее вариант, при этом вариант константной области содержит до 20 консервативно модифицированных аминокислотных замен.
11. Связывающее соединение по п.4, дополнительно содержащее константную область тяжелой цепи, содержащую константную область тяжелой цепи γ4 человека или ее вариант, при этом вариант константной области содержит до 20 консервативно модифицированных аминокислотных замен.
12. Связывающее соединение по п.4, в котором связывающее соединение представляет собой фрагмент антитела, выбранный из группы, состоящей из Fab, Fab', Fab'-SH, Fv, scFv, F(ab')2 и диантитело.
13. Способ усиления иммунного ответа у человека, включающий введение человеку, нуждающемуся в этом, антитела, специфичного для GITR, или его антигенсвязывающего фрагмента в количестве, эффективном для усиления биологической активности GITR, при этом антителом является антитело по п.5.
14. Способ по п.13, в котором GITR-антитело или его антигенсвязывающий фрагмент вводят совместно с анти-TGFβ-антителом.
15. Способ по п.13, в котором GITR-антитело или его антигенсвязывающий фрагмент применяют совместно с местным облучением.
16. Способ по п.13, в котором иммунный ответ направлен против пролиферативного нарушения.
17. Способ по п.13, в котором иммунный ответ направлен против вирусной инфекции.
18. Фармацевтическая композиция, содержащая антитело по п.4 и фармацевтически приемлемый носитель.
19. Антитело или его антигенсвязывающий фрагмент, продуцированные гибридомой, депонированной в Американской коллекции типовых культур (ATCC), при этом гибридома выбрана из группы, состоящей из PTA-9889, PTA-9890, PTA-9891, PTA-9892, PTA-9893, PTA-10286, PTA-10287, PTA-10288, PTA-10289, PTA-10290 и PTA-10291.
20. Антитело или антигенсвязывающий фрагмент, который связывается с белком GITR человека, при этом антитело или антигенсвязывающий фрагмент узнает эпитоп, охватывающий модуль 3 и модуль 4 белка GITR человека (SEQ ID NO: 89).
21. Антитело или антигенсвязывающий фрагмент по п.20, в котором эпитоп содержит Gly57, Arg65, His67, Lys80, Phe81, Ser82 и Gln86.
22. Антитело по п.20, которое перекрестно блокирует по меньшей мере одно из антител или фрагментов антител, продуцируемых гибридомами, выбранными из группы, состоящей из PTA-9889, PTA-9890, PTA-9891, PTA-9892, PTA-9893, PTA-10286, PTA-10287, PTA-10288, PTA-10289, PTA-10290 и PTA-10291.
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