RU2021114137A - Онколитические вирусные векторы и их применение - Google Patents
Онколитические вирусные векторы и их применение Download PDFInfo
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- RU2021114137A RU2021114137A RU2021114137A RU2021114137A RU2021114137A RU 2021114137 A RU2021114137 A RU 2021114137A RU 2021114137 A RU2021114137 A RU 2021114137A RU 2021114137 A RU2021114137 A RU 2021114137A RU 2021114137 A RU2021114137 A RU 2021114137A
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- recombinant oncolytic
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Classifications
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Claims (35)
1. Рекомбинантный онколитический вирус простого герпеса (HSV), содержащий одну или более копий двух или более целевых последовательностей опухолесупрессирующих микроРНК (миР), вставленных в два или более локусов вирусных генов, причем первый локус из двух или более локусов вирусных генов являются локусом UL1, UL5, UL6, UL7, UL8, UL9, UL11, UL12, UL14, UL15, UL17, UL18, UL19, UL20, UL22, UL25, UL26, UL26.5, UL27, UL28, UL29, UL30, UL31, UL32, UL33, UL34, UL35, UL36, UL37, UL38, UL39, UL40, UL42, UL48, UL49, UL52, UL53, UL54, ICP0, ICP4, ICP22, ICP27, ICP47, гамма-34.5, US3, US4, US5, US6, US7, US8, US9, US10, US11 или US12.
2. Рекомбинантный онколитический HSV по п.1, в котором каждая из двух или более целевых последовательностей опухолесупрессирующих миР вставлены только в один из двух или более локусов вирусных генов.
3. Рекомбинантный онколитический HSV по п.1, в котором по меньшей мере одна из целевых последовательностей опухолесупрессирующих миР вставлена в по меньшей мере два локуса вирусных генов.
4. Рекомбинантный онколитический HSV по любому из пп. 1-3, в котором первый локус представляет собой локус ICP4, ICP22, ICP27, ICP34.5, UL5, UL8, UL9, UL30, UL39, UL40 или UL42.
5. Рекомбинантный онколитический HSV по любому из пп. 1-3, в котором второй локус из двух или более локусов вирусных генов представляет собой локус ICP4, ICP22, ICP27, ICP34.5, UL5, UL8, UL9, UL30, UL39, UL40 или UL42, и при этом второй локус отличается от первого локуса.
6. Рекомбинантный онколитический HSV по любому из пп. 1-5, в котором две или более целевых последовательностей опухолесупрессирующих миР представляют собой целевые последовательности для миР, выбранных из таблицы 3.
7. Рекомбинантный онколитический HSV по п.6, в котором по меньшей мере одна из двух или более целевых последовательностей опухолесупрессирующих миР представляют собой целевые последовательности миР для миР-124, миР-122, миР-1, миР-143, миР-128, миР-204, миР-137, миР-126 или миР-217.
8. Рекомбинантный онколитический HSV по любому из пп. 1-7, в котором две или более целевых последовательностей опухолесупрессирующих миР встроены в 5'-нетранслируемую область (НТО) или 3'-НТО двух или более локусов вирусных генов.
9. Рекомбинантный онколитический HSV по любому из пп. 1-8, в котором первый локус вирусного гена содержит по меньшей мере две из целевых последовательностей миР.
10. Рекомбинантный онколитический HSV по любому из пп. 1-9, в котором второй локус вирусного гена содержит по меньшей мере две из целевых последовательностей миР.
11. Рекомбинантный онколитический HSV по любому из пп. 1-10, в котором одна или более копий целевых последовательностей опухолесупрессирующих миР представляют собой две, три, четыре или более копий.
12. Рекомбинантный онколитический HSV по любому из пп. 1-11, дополнительно включающий делецию во внутренней повторяющейся соединительной области, так что рекомбинантный геном HSV содержит по одной копии каждого из генов ICP0, γ34.5, LAT и ICP4.
13. Рекомбинантный онколитический HSV по любому из пп. 1-12, в котором вирусный геном рекомбинантного онколитического HSV дополнительно содержит одну или более гетерологичных полинуклеотидных последовательностей, кодирующих один или более белков и/или олигонуклеотидов.
14. Рекомбинантный онколитический HSV по п.13, в котором белок представляет собой нуклеазу, биспецифический агент, взаимодействующий с Т-клетками (BiTE), антииммуносупрессивный белок или иммуногенный антиген.
15. Рекомбинантный онколитический HSV по п.14, в котором нуклеаза выбрана из эндонуклеазы, ассоциированной с короткими палиндромными повторами, регулярно расположенными кластерами (CRISPR), нуклеазы с цинковыми пальцами (ZFN) или эффекторной нуклеазы, подобной активатору транскрипции (TALEN).
16. Рекомбинантный онколитический HSV по п.13, в котором один или более белков содержат одно или более антител или их антигенсвязывающих фрагментов, которые связываются с рецепторами иммунных контрольных точек, один или более провоспалительных цитокинов или их комбинацию.
17. Рекомбинантный онколитический HSV по п.16, в котором рецепторы иммунных контрольных точек выбраны из группы, состоящей из CTLA4, LAG3, PD1 и PDL1.
18. Рекомбинантный онколитический HSV по п.16, в котором провоспалительные цитокины выбраны из группы, состоящей из IFNγ, IFNα, IFNβ, TNFα, IL-12, IL-2, IL-6, IL-8 и GM-CSF.
19. Рекомбинантный онколитический HSV по п.18, в котором провоспалительный цитокин представляет собой человеческий IL-12.
20. Молекула нуклеиновой кислоты, кодирующая рекомбинантный онколитический HSV по любому из пп. 1-19.
21. Вирусный сток, содержащий рекомбинантный онколитический HSV по любому из пп. 1-19, где вирусный сток предназначен для введения субъекту.
22. Композиция, содержащая рекомбинантный онколитический HSV по любому из пп. 1-19 и фармацевтически приемлемый носитель, причем композиция предназначена для введения субъекту.
23. Способ уничтожения раковой клетки, включающий воздействие на раковую клетку рекомбинантного онколитического HSV по любому из пп. 1-19 или его композиций в условиях, достаточных для инфицирования и репликации рекомбинантного онколитического HSV в указанной раковой клетке, и при этом репликация рекомбинантный онколитический HSV в раковой клетке приводит к гибели клетки.
24. Способ по п.23, в котором раковая клетка имеет пониженную экспрессию опухолесупрессирующих миР, способных связываться с одной из двух или более целевых последовательностей опухолесупрессирующих миР по сравнению с экспрессией опухолесупрессирующих миР в нераковой клетке.
25. Способ по п.24, в котором уровень экспрессии опухолесупрессирующих миР в раковой клетке по меньшей мере на 5% ниже, чем уровень экспрессии опухолесупрессирующих миР в нераковой клетке.
26. Способ по любому из пп. 23-25, в котором репликация рекомбинантного онколитического HSV увеличивается или поддерживается в раковых клетках со сниженной экспрессией опухолесупрессирующих миР, способных связываться с одной из двух или более целевых последовательностей опухолесупрессирующих миР.
27. Способ по п.26, в котором репликация вируса в раковых клетках по меньшей мере на 5% больше по сравнению с репликацией вируса в нераковых клетках.
28. Способ по любому из пп. 23-27, в котором клетка находится в условиях in vivo.
29. Способ по п.28, в котором клетка находится внутри опухоли.
30. Способ лечения рака у субъекта, нуждающегося в этом, включающий введение субъекту рекомбинантного онколитического HSV по любому из пп. 1-19 или его композиций.
31. Способ по любому из пп. 23-30, в котором субъектом является мышь, крыса, кролик, кошка, собака, лошадь, примат, не являющийся человеком, или человек.
32. Способ по п.30 или 31, в котором рекомбинантный онколитический HSV или его композиции вводят внутривенно, подкожно, внутриопухолево, внутримышечно или интраназально.
33. Способ по любому из пп. 30-32, в котором рак выбран из рака легкого, рака молочной железы, рака яичников, рака шейки матки, рака предстательной железы, рака яичка, колоректального рака, рака толстой кишки, рака поджелудочной железы, рака печени, рака желудка, рака головы и шеи, рака щитовидной железы, злокачественной глиомы, глиобластомы, меланомы, В-клеточного хронического лимфоцитарного лейкоза, диффузной В-крупноклеточной лимфомы (ДВКЛ) и лимфомы из клеток маргинальной зоны (ЛМЗ).
34. Способ по п.33, в котором рак легкого представляет собой мелкоклеточный рак легкого или немелкоклеточный рак легкого.
35. Способ по п.33, в котором рак печени представляет собой гепатоцеллюлярную карциному (HCC).
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