RU2018108259A - Векторы для экспрессии простатоассоциированных антигенов - Google Patents
Векторы для экспрессии простатоассоциированных антигенов Download PDFInfo
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- RU2018108259A RU2018108259A RU2018108259A RU2018108259A RU2018108259A RU 2018108259 A RU2018108259 A RU 2018108259A RU 2018108259 A RU2018108259 A RU 2018108259A RU 2018108259 A RU2018108259 A RU 2018108259A RU 2018108259 A RU2018108259 A RU 2018108259A
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- 239000013598 vector Substances 0.000 title claims 18
- 239000000427 antigen Substances 0.000 title claims 2
- 102000036639 antigens Human genes 0.000 title claims 2
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- 239000002773 nucleotide Substances 0.000 claims 49
- 125000003729 nucleotide group Chemical group 0.000 claims 49
- 108090000765 processed proteins & peptides Proteins 0.000 claims 30
- 102000004196 processed proteins & peptides Human genes 0.000 claims 23
- 229920001184 polypeptide Polymers 0.000 claims 22
- 230000002163 immunogen Effects 0.000 claims 13
- 210000004027 cell Anatomy 0.000 claims 9
- 150000001413 amino acids Chemical class 0.000 claims 8
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- 238000000034 method Methods 0.000 claims 7
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- LBWFXVZLPYTWQI-IPOVEDGCSA-N n-[2-(diethylamino)ethyl]-5-[(z)-(5-fluoro-2-oxo-1h-indol-3-ylidene)methyl]-2,4-dimethyl-1h-pyrrole-3-carboxamide;(2s)-2-hydroxybutanedioic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O.CCN(CC)CCNC(=O)C1=C(C)NC(\C=C/2C3=CC(F)=CC=C3NC\2=O)=C1C LBWFXVZLPYTWQI-IPOVEDGCSA-N 0.000 claims 3
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
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Claims (46)
1. Вектор экспрессии C68, содержащий
(а) нуклеотидную последовательность C68; и
(б) мультиантигенную конструкцию ДНК, которая содержит (1) первую кодирующую нуклеотидную последовательность, кодирующую иммуногенный полипептид PSA (простатоспецифический антиген), и (2) вторую кодирующую нуклеотидную последовательность, кодирующую иммуногенный полипептид PSCA (антиген стволовых клеток простаты) или кодирующую иммуногенный полипептид PSMA (простатоспецифический мембранный антиген), где иммуногенный полипептид PSA содержит аминокислоты 27-263 из SEQ ID NO: 15, аминокислоты 4-240 из SEQ ID NO: 17 или аминокислотную последовательность с SEQ ID NO: 17.
2. Вектор C68 по п. 1, где нуклеотидная последовательность C68 представляет собой последовательность с SEQ ID NO: 57, в которой отсутствует по меньшей мере один ген, выбранный из группы, состоящей из генов E1A, E1B, E2A, E2B, E3, E4, L1, L2, L3, L4 и L5, где иммуногенный полипептид PSCA содержит аминокислотную последовательность с SEQ ID NO: 21 или аминокислоты 4-125 из SEQ ID NO: 21 и где иммуногенный полипептид PSMA выбран из группы, состоящей из:
1) полипептида, содержащего аминокислоты 15-750 из SEQ ID NO: 1;
2) полипептида, содержащего аминокислотную последовательность с SEQ ID NO: 3;
3) полипептида, содержащего аминокислотную последовательность с SEQ ID NO: 5;
4) полипептида, содержащего аминокислотную последовательность с SEQ ID NO: 7;
5) полипептида, содержащего аминокислоты 4-739 из SEQ ID NO: 9;
6) полипептида, содержащего аминокислоты 4-739 из SEQ ID NO: 3;
7) полипептида, содержащего аминокислоты 4-739 из SEQ ID NO: 5;
8) полипептида, содержащего аминокислоты 4-739 из SEQ ID NO: 7; и
9) полипептида, содержащего аминокислотную последовательность с SEQ ID NO: 9.
3. Вектор C68 по п. 2, где нуклеотидная последовательность C68 представляет собой последовательность с SEQ ID NO: 57, в которой отсутствуют гены E1A, E1B и E3, и где мультиантигенная конструкция дополнительно содержит разделяющую последовательность между первой и второй кодирующими нуклеотидными последовательностями.
4. Вектор C68 по п. 3, где разделяющая последовательность выбрана из последовательностей 2A-пептидов и участков внутренней посадки рибосомы (IRES).
5. Вектор C68 по п. 4, где последовательность 2A-пептида выбрана из группы, состоящей из последовательности 2A-пептида из вируса ящура (FMDV), вируса A, вызывающего ринит у лошадей (ERAV), вируса 1 болезни Тешена у свиней (PTV1), вируса энцефаломиокардита, варианта B (EMC-B), вируса энцефаломиокардита (EMCV), вируса мышиного энцефаломиелита Тейлора GD7 (TME-GD7), вируса B, вызывающего ринит у лошадей (ERBV), вируса Thosea asigna (TaV), вируса С дрозофилы (DrosC), вируса паралича сверчка (CrPV), вируса острого паралича пчел (ABPV), вируса инфекционного заболевания тутового шелкопряда (IFV), ротавируса свиней, ротавируса человека, TSR1 T. brucei и апуриновой/апиримидиновой (AP) эндонуклеазы T. cruzi; и где IRES (участок внутренней посадки рибосомы) представляет собой IRES из EMCV.
6. Вектор C68 по п. 5, где вторая кодирующая нуклеотидная последовательность кодирует иммуногенный полипептид PSCA.
7. Вектор C68 по п. 5, где вторая кодирующая нуклеотидная последовательность кодирует иммуногенный полипептид PSMA.
8. Вектор C68 по п. 6 или п. 7, где разделяющая последовательность кодирует последовательность 2A-пептида, выбранную из последовательности 2A-пептида FMDV или последовательности 2A-пептида TaV.
9. Вектор C68 по п. 1, где:
(1) нуклеотидная последовательность, кодирующая иммуногенный полипептид PSA, выбрана из группы, состоящей из: (а) нуклеотидной последовательности с SEQ ID NO: 18; (б) нуклеотидной последовательности с SEQ ID NO: 20; (в) нуклеотидной последовательности, содержащей нуклеотиды 10-720 из SEQ ID NO: 18; (г) нуклеотидной последовательности, содержащей нуклеотиды 1115-1825 из SEQ ID NO: 58; и (д) нуклеотидной последовательности, содержащей нуклеотиды 1106-1825 из SEQ ID NO: 58;
(2) нуклеотидная последовательность, кодирующая иммуногенный полипептид PSCA, выбрана из группы, состоящей из: (а) нуклеотидной последовательности с SEQ ID NO: 22; (б) нуклеотидной последовательности, содержащей нуклеотиды 10-372 из SEQ ID NO: 22; (в) нуклеотидной последовательности, содержащей нуклеотиды 1892-2257 из SEQ ID NO: 58; и (г) нуклеотидной последовательности, содержащей нуклеотиды 1886-2257 из SEQ ID NO: 58; и
(3) нуклеотидная последовательность, кодирующая иммуногенный полипептид PSMA, выбрана из группы, состоящей из: (а) нуклеотидной последовательности, содержащей нуклеотиды 43-2250 из SEQ ID NO: 2; (б) нуклеотидной последовательности с SEQ ID NO: 4; (в) нуклеотидной последовательности с SEQ ID NO: 6; (г) нуклеотидной последовательности с SEQ ID NO: 8; (д) нуклеотидной последовательности с SEQ ID NO: 10; (е) нуклеотидной последовательности, содержащей нуклеотиды 10-2217 из SEQ ID NO: 4; (ж) нуклеотидной последовательности, содержащей нуклеотиды 10-2217 из SEQ ID NO: 6; (з) нуклеотидной последовательности, содержащей нуклеотиды 10-2217 из SEQ ID NO: 8; (и) нуклеотидной последовательности, содержащей нуклеотиды 10-2217 из SEQ ID NO: 10; (к) нуклеотидной последовательности, содержащей нуклеотиды 2333-4543 из SEQ ID NO: 58; и (л) нуклеотидной последовательности, содержащей нуклеотиды 2324-4543 из SEQ ID NO: 58.
10. Вектор C68 по п. 3, где вторая кодирующая нуклеотидная последовательность кодирует иммуногенный полипептид PSCA и где разделяющая последовательность кодирует 2A-пептид FMDV.
11. Вектор C68 по п. 9, где вторая кодирующая нуклеотидная последовательность кодирует иммуногенный полипептид PSMA и где разделяющая последовательность кодирует 2A-пептид TaV.
12. Вектор C68 по п. 1, где мультиантигенная конструкция содержит:
(1) нуклеотидную последовательность с SEQ ID NO: 29; или
(2) вырожденный вариант нуклеотидной последовательности с SEQ ID NO: 29.
13. Вектор C68 по п. 1, дополнительно содержащий промотор CMV.
14. Вектор C68 по п. 1, где мультиантигенная конструкция содержит:
(1) нуклеотидную последовательность с SEQ ID NO: 32; или
(2) вырожденный вариант нуклеотидной последовательности с SEQ ID NO: 32.
15. Композиция для вызова иммунного ответа на простатоассоциированный антиген, содержащая эффективное количество вектора C68 по любому из пп. 1-14.
16. Клетка для экспрессии вектора по п. 1, содержащая вектор C68 по любому из пп. 1-14.
17. Фармацевтическая композиция для лечения или предупреждения рака предстательной железы, содержащая эффективное количество вектора по любому из пп. 1-14 и фармацевтически приемлемый эксципиент.
18. Способ лечения рака предстательной железы у человека, включающий введение человеку эффективного количества фармацевтической композиции по п. 17.
19. Способ по п. 18, дополнительно включающий введение человеку эффективного количества иммуномодулятора.
20. Способ по п. 18, дополнительно включающий введение человеку (а) эффективного количества по меньшей мере одного ингибитора супрессорных клеток иммунной системы и (б) эффективного количества по меньшей мере одного усилителя эффекторных клеток иммунной системы.
21. Способ по п. 20, где ингибитор супрессорных клеток иммунной системы выбран из группы, состоящей из ингибитора протеинкиназы, ингибитора циклооксигеназы-2 (COX-2) и ингибитора фосфодиэстеразы типа 5 (PDE5), и где усилитель эффекторных клеток иммунной системы выбран из группы, состоящей из ингибитора цитотоксического T-лимфоцитарного антигена-4 (CTLA-4), агониста CD40, агониста toll-подобного рецептора (TLR), агониста 4-1BB, агониста OX40, агониста индуцированного глюкокортикоидами рецептора фактора некроза опухолей (GITR), антагониста белка 1 программируемой клеточной смерти (PD-1) и антагониста лиганда 1 PD-1 (PD-L1).
22. Способ по п. 21, где:
(1) ингибитор протеинкиназы выбран из группы, состоящей из иматиниба, сорафениба, лапатиниба, зактима, MP-412, дазатиниба, лестауртиниба, сунитиниба малата, акситиниба, эрлотиниба, гефитиниба, босутиниба, темсиролимуса и нилотиниба;
(2) ингибитор CTLA-4 выбран из группы, состоящей из ипилимумаба и тремелимумаба;
(3) агонист CD40 представляет собой антитело к CD40, выбранное из группы, состоящей из G28-5, mAb89, EA-5, S2C6, CP870893 и дацетузумаба; и
(4) агонист TLR представляет собой CpG-олигонуклеотид, выбранный из группы, состоящей из CpG 24555, CpG 10103, CpG7909 и CpG1018.
23. Способ по п. 22, где ингибитор супрессорных клеток иммунной системы представляет собой ингибитор протеинкиназы, выбранный из группы, состоящей из сорафениба, дазатиниба, иматиниба, акситиниба и сунитиниба малата, и где усилитель эффекторных клеток иммунной системы представляет собой тремелимумаб.
24. Способ по п. 22, где ингибитор супрессорных клеток иммунной системы представляет собой ингибитор протеинкиназы, выбранный из группы, состоящей из сорафениба, дазатиниба, иматиниба, акситиниба и сунитиниба малата, и где усилитель эффекторных клеток иммунной системы представляет собой CpG-олигонуклеотид, выбранный из группы, состоящей из CpG24555, CpG10103, CpG7909 и CpG1018.
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