RU2010112935A - Комплексы на основе рнк и катионных пептидов для трансфекции и иммуностимуляции - Google Patents

Комплексы на основе рнк и катионных пептидов для трансфекции и иммуностимуляции Download PDF

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RU2010112935A
RU2010112935A RU2010112935/10A RU2010112935A RU2010112935A RU 2010112935 A RU2010112935 A RU 2010112935A RU 2010112935/10 A RU2010112935/10 A RU 2010112935/10A RU 2010112935 A RU2010112935 A RU 2010112935A RU 2010112935 A RU2010112935 A RU 2010112935A
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rna
arg
orn
lys
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Мариола ФОТИНМЛЕЧЕК (DE)
Мариола ФОТИНМЛЕЧЕК
Патрик БАУМХОФ (DE)
Патрик БАУМХОФ
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Куревак Гмбх (De)
Куревак Гмбх
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Abstract

1. РНК в составе комплекса, включающего по крайней мере одну РНК в виде комплекса с одним или более олигопептидами, при этом олигопептид содержит от 8 до 15 аминокислотных остатков, и характеризуется следующей общей формулой: ! ! где l+m+n+o+x=8-15 и l, m, n или о независимо друг от друга равны числу, выбранному из 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 или 15, при условии, что общее содержание Arg, Lys, His и Orn составляет по крайней мере 50% от всех аминокислотных остатков, входящих в состав олигопептида, и ! Хаа обозначает любую аминокислоту, выбранную из природных или неприродных аминокислот, за исключением Arg, Lys, His или Orn, и ! х равен любому числу, выбранному из 0, 1, 2, 3, 4, 5, 6, 7 или 8, при условии, что общее содержание Хаа не превышает 50% от всех аминокислотных остатков, входящих в состав олигопептида. ! 2. РНК в составе комплекса по п.1, где по крайней мере одной РНК является мРНК. ! 3. РНК в составе комплекса по п.1, где олигопептид включает от 8 до 14, от 8 до 13, от 8 до 12, или от 9 до 12, или от 9 до 11 аминокислотных остатков. ! 4. РНК в составе комплекса по п.1, где общее содержание Arg, Lys, His или Orn составляет по крайней мере 60%, 70%, 80%, 90% или 95% от всех аминокислотных остатков, входящих в состав олигопептида, образующего комплекс с РНК. ! 5. РНК в составе комплекса по любому из пп.1-4, где Хаа в общей формуле (Arg)1(Lys)m(His)n(Orn)o(Xaa)x выбран из аминокислот, содержащих нейтральные боковые цепи, включая аминокислоты, содержащие нейтральную гидрофобную боковую цепь, и аминокислоты, содержащие нейтральную полярную боковую цепь. ! 6. РНК в составе комплекса по любому из пп.1-4, где олигопептид общей формулы (Arg)1(Lys)m(His)n(Orn)o(Xaa)x в одном или обоих концевых фрагментах не содержит аминокислот, содержа�

Claims (23)

1. РНК в составе комплекса, включающего по крайней мере одну РНК в виде комплекса с одним или более олигопептидами, при этом олигопептид содержит от 8 до 15 аминокислотных остатков, и характеризуется следующей общей формулой:
Figure 00000001
где l+m+n+o+x=8-15 и l, m, n или о независимо друг от друга равны числу, выбранному из 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 или 15, при условии, что общее содержание Arg, Lys, His и Orn составляет по крайней мере 50% от всех аминокислотных остатков, входящих в состав олигопептида, и
Хаа обозначает любую аминокислоту, выбранную из природных или неприродных аминокислот, за исключением Arg, Lys, His или Orn, и
х равен любому числу, выбранному из 0, 1, 2, 3, 4, 5, 6, 7 или 8, при условии, что общее содержание Хаа не превышает 50% от всех аминокислотных остатков, входящих в состав олигопептида.
2. РНК в составе комплекса по п.1, где по крайней мере одной РНК является мРНК.
3. РНК в составе комплекса по п.1, где олигопептид включает от 8 до 14, от 8 до 13, от 8 до 12, или от 9 до 12, или от 9 до 11 аминокислотных остатков.
4. РНК в составе комплекса по п.1, где общее содержание Arg, Lys, His или Orn составляет по крайней мере 60%, 70%, 80%, 90% или 95% от всех аминокислотных остатков, входящих в состав олигопептида, образующего комплекс с РНК.
5. РНК в составе комплекса по любому из пп.1-4, где Хаа в общей формуле (Arg)1(Lys)m(His)n(Orn)o(Xaa)x выбран из аминокислот, содержащих нейтральные боковые цепи, включая аминокислоты, содержащие нейтральную гидрофобную боковую цепь, и аминокислоты, содержащие нейтральную полярную боковую цепь.
6. РНК в составе комплекса по любому из пп.1-4, где олигопептид общей формулы (Arg)1(Lys)m(His)n(Orn)o(Xaa)x в одном или обоих концевых фрагментах не содержит аминокислот, содержащих боковые кислотные цепи.
7. РНК в составе комплекса по любому из пп.1-4, где олигопептид общей формулы (Arg)1(Lys)m(His)n(Orn)o(Xaa)x в одном или обоих концевых фрагментах содержит нейтральные или основные аминокислотные остатки или в одном или обоих концевых фрагментах содержит основные аминокислотные остатки, или по крайней мере один, предпочтительно два, более предпочтительно три основных аминокислотных остатка в обоих концевых фрагментах.
8. РНК в составе комплекса по любому из пп.1-4, где олигопептид общей формулы (Arg)1(Lys)m(His)n(Orn)o(Xaa)x включает в своей последовательности участок, содержащий по крайней мере 3 последовательно расположенных основных аминокислотных остатков.
9. РНК в составе комплекса по любому из пп.1-4, где олигопептид общей формулы (Arg)1(Lys)m(His)n(Orn)o(Xaa)x включает по крайней мере 1, 2 или 3 некатионных аминокислотных остатка в одном или обоих концевых фрагментах, а некатионную аминокислоту предпочтительно выбирают из гиститидина.
10. РНК в составе комплекса по любому из пп.1-4, где по крайней мере одной РНК в составе комплекса, содержащего РНК, является мРНК, при этом мРНК кодирует терапевтически эффективный белок или пептид, иммуностимулирующий белок или пептид, опухолевый антиген или антитело.
11. РНК в составе комплекса по любому из пп.1-4, где РНК является модифицированная РНК, прежде всего стабилизированная РНК, по сравнению с РНК дикого типа.
12. РНК в составе комплекса по п.11, где содержание G/C в кодирующей области модифицированной РНК увеличено по сравнению с содержанием G/C в кодирующей области РНК дикого типа, при этом кодируемая аминокислотная последовательность предпочтительно не модифицирована по сравнению с аминокислотной последовательностью, кодируемой РНК дикого типа.
13. РНК в составе комплекса по любому из пп.1-4, где массовое соотношение по крайней мере одной РНК в составе комплекса, содержащего РНК, и одного или более олигопептидов находится в диапазоне приблизительно от 1:100 до приблизительно 1:0,5, приблизительно от 1:50 до приблизительно 1:1, или составляет приблизительно 1:100, приблизительно 1:90, приблизительно 1:80, приблизительно 1:70, приблизительно 1:60, приблизительно 1:50, приблизительно 1:45, приблизительно 1:40, приблизительно 1:35, приблизительно 1:30, приблизительно 1:25, приблизительно 1:20, приблизительно 1:15, приблизительно 1:10, приблизительно 1:5, приблизительно 1:4, приблизительно 1:3, приблизительно 1:2, приблизительно 1:1 или приблизительно 1:0,5.
14. РНК в составе комплекса по любому из пп.1-4, где молярное соотношение по крайней мере одной РНК в составе комплекса, содержащего РНК, и одного или более олигопептидов находится в диапазоне приблизительно от 1:20000 до приблизительно 1:500 или 1:250, или в диапазоне приблизительно от 1:10000 до приблизительно 1:1000, или составляет приблизительно 1:9500, приблизительно 1:9000, приблизительно 1:8500, приблизительно 1:8000, приблизительно 1:7500, приблизительно 1:7000, приблизительно 1:6500, приблизительно 1:6000, приблизительно 1:5500, приблизительно 1:5000, приблизительно 1:4500, приблизительно 1:4000, приблизительно 1:3500, приблизительно 1:3000, приблизительно 1:2500, приблизительно 1:2000, приблизительно 1:1500, приблизительно 1:1000, приблизительно 1:500, приблизительно 1:450, приблизительно 1:400, приблизительно 1:350, приблизительно 1:300 или приблизительно 1:250.
15. РНК в составе комплекса по любому из пп.1-4, где соотношение азот/фосфат (N/P-соотношение) по крайней мере в одной РНК в составе комплекса, содержащего РНК, и одного или более олигопептидов, находится в диапазоне приблизительно 0,2-50, в диапазоне приблизительно 0,5-50 или в диапазоне приблизительно 0,75-25.
16. РНК в составе комплекса по любому пп.1-4, где олигопептид выбран из группы, включающей:
Arg9His3, His3Arg9His3, TyrSerSerArg9SerSerTyr, His3Arg9SerSerTyr, (ArgLysHis)4, Tyr(ArgLysHis)2Arg,
или из
Arg8, Arg9, Arg10, Arg11, Arg12, Arg13, Arg14, Arg15 (SEQ ID №1-8),
Lys8, Lys9, Lys10, Lys11, Lys12, Lys13, Lys14, Lys15 (SEQ ID №9-16),
His8, His9, His10, His11, His12, His13, His14, His15 (SEQ ID №17-24) и
Orn8, Orn9, Orn10, Orn11, Orn12, Orn13, Orn14, Orn15, (SEQ ID №25-32).
17. Фармацевтическая композиция, включающая РНК в составе комплекса по любому из пп.1-16 и необязательно фармацевтически приемлемый носитель.
18. Способ трансфекции клеток или организма, включающий следующие стадии:
а. необязательно конструирование и/или получение РНК в составе комплекса по пп.1-16, включающего по крайней мере одну РНК в составе комплекса и один или более олигопептидов общей формулы (Arg)1(Lys)m(His)n(Orn)o(Xaa)x,
б. трансфекцию клеток, ткани или организма с использованием РНК в составе комплекса, полученного на стадии а.
19. Способ по п.18, где организмом является млекопитающее, выбранное из группы, включающей человека, козу, крупный рогатый скот, свинью, собаку, кошку, осла, обезьяну, приматов или грызунов, включая мышь, хомяка и кролика.
20. Применение РНК в составе комплекса по любому из пп.1-16 для трансфекции клеток или организма.
21. Применение РНК в составе комплекса по любому из пп.1-16 для получения агента, предназначенного для лечения заболевания, выбранного из опухолевого заболевания или рака, сердечно-сосудистого заболевания, инфекционного заболевания, (инфекционного) вирусного заболевания, аутоиммунного заболевания, (моно-)генетического заболевания и/или аллергии.
22. Применение по п.21, где трансфекция с использованием РНК в составе комплекса индуцирует иммунный ответ в организме.
23. Набор, включающий фармацевтическую композицию по п.17, цитокин или композицию, содержащую цитокин, и/или по крайней мере один дополнительный адъювант или композицию, включающую по крайней мере один дополнительный адъювант.
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