RU2016143388A - Экспрессия трансгенов, регулируемая лекарственным средством - Google Patents
Экспрессия трансгенов, регулируемая лекарственным средством Download PDFInfo
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- RU2016143388A RU2016143388A RU2016143388A RU2016143388A RU2016143388A RU 2016143388 A RU2016143388 A RU 2016143388A RU 2016143388 A RU2016143388 A RU 2016143388A RU 2016143388 A RU2016143388 A RU 2016143388A RU 2016143388 A RU2016143388 A RU 2016143388A
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Claims (54)
1. Система для индуцируемой экспрессии химерного антигенного рецептора, содержащая:
a) первую нуклеиновую кислоту, содержащую первый промотор, индуцируемый лекарственным средством, причем указанная первая нуклеиновая кислота функционально соединена с полинуклеотидом, кодирующим химерный антигенный рецептор, содержащий лигандсвязывающий домен, причем указанный лигандсвязывающий домен специфичен в отношении лиганда, при этом лиганд представляет собой опухолеспецифичную молекулу, вирусную молекулу или любую другую молекулу, экспрессируемую на популяции клеток-мишеней, при этом указанный лиганд может индуцировать распознавание, модулирование, ингибирование и/или устранение лимфоцитом; полинуклеотид, кодирующий полипептидный спейсер, причем указанный спейсер оптимизирован; полинуклеотид, кодирующий трансмембранный домен; и полинуклеотид, кодирующий внутриклеточный сигнальный домен; и
b) вторую нуклеиновую кислоту, содержащую второй промотор, функционально соединенный с нуклеиновой кислотой, кодирующей активатор транскрипции для индуцируемого промотора.
2. Система по п. 1, отличающаяся тем, что указанное лекарственное средство представляет собой тамоксифен и/или его метаболиты.
3. Система по п. 2, отличающаяся тем, что указанный первый промотор содержит последовательность нуклеиновой кислоты, представленную в SEQ ID NO: 41.
4. Система по любому из пп. 1-3, отличающаяся тем, что указанный второй промотор представляет собой конститутивный промотор.
5. Система по п. 4, отличающаяся тем, что указанный второй промотор представляет собой EFlαp.
6. Система по любому из пп. 1-3, 5, отличающаяся тем, что указанный активатор транскрипции содержит последовательность SEQ ID NO: 40.
7. Система по любому из пп. 1-3, 5, отличающаяся тем, что указанная первая нуклеиновая кислота дополнительно включает первый вектор, и вторая нуклеиновая кислота дополнительно включает второй вектор.
8. Система по п. 7, отличающаяся тем, что оба вектора упакованы в вирусный вектор.
9. Система по п. 8, отличающаяся тем, что указанный вирусный вектор представляет собой лентивирус.
10. Система по любому из пп. 1-3, 5, отличающаяся тем, что указанные первая и вторая нуклеиновые кислоты включают вектор.
11. Система по любому из пп. 1-3, 5, 8-9, отличающаяся тем, что указанная первая нуклеиновая кислота дополнительно содержит последовательность нуклеиновой кислоты, кодирующую селективный маркер.
12. Система по любому из пп. 1-3, 5, 8-9, отличающаяся тем, что указанная вторая нуклеиновая кислота дополнительно содержит нуклеиновую кислоту, кодирующую селективный маркер.
13. Система по любому из пп. 1, отличающаяся тем, что указанная вторая нуклеиновая кислота дополнительно содержит полинуклеотид, кодирующий химерный антигенный рецептор, содержащий лигандсвязывающий домен, при этом лиганд представляет собой опухолеспецифичную молекулу, вирусную молекулу или любую другую молекулу, экспрессируемую на популяции клеток-мишеней, при этом указанный лиганд может индуцировать распознавание, модулирование, ингибирование и/или устранение лимфоцитом; полинуклеотид, кодирующий полипептидный спейсер, причем указанный спейсер оптимизирован; полинуклеотид, кодирующий трансмембранный домен; и полинуклеотид, кодирующий внутриклеточный сигнальный домен.
14. Система по п. 1, отличающаяся тем, что указанный первый промотор находится в противоположной ориентации по отношению ко второму промотору.
15. Система по п. 1, отличающаяся тем, что указанный лигандсвязывающий домен представляет собой фрагмент антитела.
16. Система по п. 15, отличающаяся тем, что указанный лигандсвязывающий домен представляет собой одноцепочечный вариабельный фрагмент.
17. Система по п. 1, отличающаяся тем, что указанная опухолеспецифичная молекула выбрана из группы, состоящей из CD19, CD20, CD22, CD23, CD123, CS-1, ROR1, СЕ7, EGFR, hB7H3, мезотелина, c-Met, PSMA, Her2, GD-2, MAGE A3 TCR и их комбинаций.
18. Система для индуцируемой экспрессии химерного антигенного рецептора, содержащая: а) первую нуклеиновую кислоту, содержащую первый промотор, индуцируемый лекарственным средством, причем указанная первая нуклеиновая кислота функционально соединена с полинуклеотидом, кодирующим цитокин, рецептор хемокина, полипептид, который регулирует апоптоз, или полипептид, который модулирует передачу сигналов с участием контрольных точек; и
b) вторую нуклеиновую кислоту, содержащую второй промотор, функционально соединенный с нуклеиновой кислотой, кодирующей активатор транскрипции для индуцируемого промотора.
19. Система по п. 13, отличающаяся тем, что указанная вторая нуклеиновая кислота дополнительно содержит полинуклеотид, кодирующий химерный антигенный рецептор, содержащий лигандсвязывающий домен, при этом лиганд представляет собой опухолеспецифичную молекулу, вирусную молекулу или любую другую молекулу, экспрессируемую на популяции клеток-мишеней, при этом указанный лиганд может индуцировать распознавание, модулирование, ингибирование и/или устранение лимфоцитом; полинуклеотид, кодирующий полипептидный спейсер, причем указанный спейсер оптимизирован; полинуклеотид, кодирующий трансмембранный домен; и полинуклеотид, кодирующий внутриклеточный сигнальный домен.
20. Система по п. 13, отличающаяся тем, что указанный первый промотор находится в противоположной ориентации по отношению ко второму промотору.
21. Система по п. 13, отличающаяся тем, что указанный лигандсвязывающий домен представляет собой фрагмент антитела.
22. Система по п. 13, отличающаяся тем, что указанная опухолеспецифичная молекула выбрана из группы, состоящей из CD19, CD20, CD22, CD23, CD123, CS-1, ROR1, СЕ7, EGFR, hb7Н3, мезотелина, c-Met, PSMA, Her2, GD-2, MAGE A3 TCR и их комбинаций.
23. Система по любому из пп. 1-3, 5, 8-9, 13-17, отличающаяся тем, что указанный второй промотор представляет собой индуцируемый промотор.
24. Система по любому из пп. 1-3, 5, 8-9, 13-17, отличающаяся тем, что указанный спейсер оптимизирован для увеличения пролиферации Т-клеток и/или выработки цитокинов в ответ на лиганд по сравнению с эталонным химерным рецептором.
25. Система по любому из пп. 18-22, отличающаяся тем, что указанный второй промотор представляет собой индуцируемый промотор.
26. Система по любому из пп. 18-22, отличающаяся тем, что указанный второй промотор представляет собой конститутивный промотор.
27. Система по любому из пп. 18-22, отличающаяся тем, что указанный спейсер оптимизирован для увеличения пролиферации Т-клеток и/или выработки цитокинов в ответ на лиганд по сравнению с эталонным химерным рецептором.
28. Полипептид химерного рецептора, кодируемый первой и/или второй нуклеиновой кислотой системы по любому из пп. 1-27.
29. Клетка-хозяин, содержащая систему по любому из пп. 1-27.
30. Клетка-хозяин по п. 29, отличающаяся тем, что указанная клетка-хозяин представляет собой цитотоксический CD8+ Т-лимфоцит, выбранный из группы, состоящей из наивных CD8+ Т-клеток, CD8+ Т-клеток центральной памяти, CD8+ Т-клеток эффекторной памяти и общей популяции CD8+ Т-клеток.
31. Клетка-хозяин по п. 30, отличающаяся тем, что указанная клетка-хозяин представляет собой хелперный CD4+ Т-лимфоцит, который выбран из группы, состоящей из наивных CD4+ Т-клеток, CD4+ Т-клеток центральной памяти, CD4+ Т-клеток эффекторной памяти и общей популяции CD4+ Т-клеток.
32. Клетка-хозяин по п. 29, отличающаяся тем, что указанная клетка-хозяин представляет собой клетку-предшественника Т-лимфоцитов.
33. Клетка-хозяин по п. 32, отличающаяся тем, что указанная клетка-предшественник Т-лимфоцитов представляет собой гемопоэтическую стволовую клетку.
34. Композиция, содержащая клетку-хозяина по любому из пп. 29-33 в фармацевтически приемлемом вспомогательном веществе.
35. Композиция по п. 34, содержащая клетку-хозяина по п. 30 и клетку-хозяина по п. 31.
36. Композиция по п. 34, содержащая клетку-хозяина по п. 30 или 31 и клетку-хозяина по п. 32 или 33.
37. Способ для получения клетки-хозяина по любому из пп. 29-33 в условиях in vitro, включающий:
a) обеспечение системы по любому из пп. 1-27; и
b) введение указанной системы в отдельную выделенную популяцию Т-лимфоцитов и размножение каждой популяции Т-лимфоцитов в условиях in vitro.
38. Способ по п. 37, отличающийся тем, что указанные Т-лимфоциты размножают, причем указанный способ дополнительно включает культивирование клеток в присутствии анти-CD3 и/или анти-CD28, и по меньшей мере одного гомеостатического цитокина до получения количества клеток, которое пригодно для применения в виде инфузии.
39. Способ по любому из пп. 37 или 38, отличающийся тем, что указанный лимфоцит представляет собой CD8+ или CD4+ клетку.
40. Применение клетки-хозяина по любому из пп. 29-33 или композиции по пп. 34-35 или 36 в комбинации с лекарственным средством, которое индуцирует экспрессию трансгена в клетке-хозяине или композиции для лечения рака или вирусной инфекции.
41. Применение по п. 40, отличающееся тем, что указанный рак представляет собой плотную опухоль или гематологическое злокачественное новообразование.
42. Применение по п. 41, отличающееся тем, что указанная плотная опухоль выбрана из группы, состоящей из рака молочной железы, рака мозга, рака легких, рака толстой кишки, рака почек, рака поджелудочной железы, рака предстательной железы и рака яичников.
43. Способ осуществления клеточной иммунотерапии у субъекта, имеющего рак или вирусную инфекцию, включающий: введение композиции по любому из пп. 34-35 или клетки-хозяина по любому из пп.29-33 субъекту и введение лекарственного средства, который индуцирует экспрессию трансгена в композиции или клетках-хозяевах.
44. Способ по п. 43, отличающийся тем, что указанный рак выбран из плотной опухоли или гематологического злокачественного новообразования.
45. Способ по п. 44, отличающийся тем, что указанная плотная опухоль выбрана из группы, состоящей из рака молочной железы, рака мозга, рака легких, рака толстой кишки, рака почек, рака поджелудочной железы, рака предстательной железы и рака яичников.
46. Способ по п. 37 или 38, отличающийся тем, что указанная популяция выделенных Т-лимфоцитов содержит клетки-предшественники Т-лимфоцитов.
47. Способ по п.46, отличающийся тем, что указанные клетки-предшественники Т-лимфоцитов представляют собой гемопоэтические стволовые клетки.
48. Способ по п. 37, отличающийся тем, что введение указанного лекарственного средства осуществляют при введении указанной композиции или клеток-хозяев.
49. Способ по п. 43, отличающийся тем, что введение указанного лекарственного средства осуществляют после введения композиции или клеток-хозяев, причем введение осуществляют 1 день, 2 дня, 3 дня, 4 дня, 5 дней, 6 дней, 7 дней, 2 недели, 4 недели или два месяца, или в течение любого периода времени, величина которого находится в диапазоне, определенном любыми двумя перечисленными периодами времени.
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Publication number | Priority date | Publication date | Assignee | Title |
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WO2023239267A1 (en) * | 2022-06-10 | 2023-12-14 | Joint Stock Company "Biocad" | Nucleic acid having promoter activity and use thereof |
RU2818112C2 (ru) * | 2022-06-10 | 2024-04-24 | Акционерное общество "БИОКАД" | Нуклеиновая кислота, имеющая промоторную активность, и ее применение |
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