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RU2018145738A - Экзосомы, содержащие терапевтические полипептиды - Google Patents

Экзосомы, содержащие терапевтические полипептиды Download PDF

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Publication number
RU2018145738A
RU2018145738A RU2018145738A RU2018145738A RU2018145738A RU 2018145738 A RU2018145738 A RU 2018145738A RU 2018145738 A RU2018145738 A RU 2018145738A RU 2018145738 A RU2018145738 A RU 2018145738A RU 2018145738 A RU2018145738 A RU 2018145738A
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Russia
Prior art keywords
polypeptide
pol
exosomal
construct
polynucleotide
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RU2018145738A
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RU2018145738A3 (ru
RU2737732C2 (ru
Inventor
Джастин ХИН
Имре МЕЙДЖЕР
Меттью ВУД
Самир ЭЛЬ АНДАЛУССИ
Оскар ВИКЛАНДЕР
Джоэл НОРДИН
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Ивокс Терапьютикс Лтд
Охфорд Юниверсити Инновейшн Лимитед
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Publication of RU2018145738A3 publication Critical patent/RU2018145738A3/ru
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Claims (27)

1. Внеклеточная везикула (ВВ), содержащая по меньшей мере одну полипептидную конструкцию, содержащую
(i) по меньшей мере один представляющий интерес полипептид (Pol);
(ii) по меньшей мере один экзосомальный полипептид; и
(iii) систему высвобождения на основе полипептида,
где система высвобождения на основе полипептида освобождает по меньшей мере один Pol от по меньшей мере одного экзосомального полипептида при эндогенной активации.
2. ВВ по п. 1, где систему высвобождения на основе полипептида выбирают из цис-расщепляющей системы высвобождения или системы высвобождения на основе комплекса сигнал ядерной локализации (NLS) - сигнал ядерной локализации-связывающий белок (NLSBP) (NLS-NLSBP).
3. ВВ по любому из предшествующих пунктов, где Pol представляет собой терапевтический полипептид, выбранный из группы, содержащей антитела, интратела, одноцепочечные вариабельные фрагменты, аффитела, ферменты для, например, ферментозаместительной терапии или редактирования генов, онкосупрессоры, вирусные или бактериальные ингибиторы, белки клеточных компонентов, ДНК и/или РНК-связывающие белки, ингибиторы репарации ДНК, нуклеазы, протеиназы, интегразы, факторы транскрипции, факторы роста, ингибиторы и индукторы апоптоза, токсины, структурные белки, нейротрофические факторы, мембранные транспортеры, нуклеотид-связывающие белки, белки теплового шока, CRISPR-ассоциированные белки и любую их комбинацию.
4. ВВ по любому из предшествующих пунктов, где экзосомальный полипептид выбирают из группы, содержащей CD9, CD53, CD63, CD81, CD54, CD50, FLOT1, FLOT2, CD49d, CD71, CD133, CD138, CD235a, ALIX, синтенин-1, синтенин-2, Lamp2b, TSPAN8, TSPAN14, CD37, CD82, CD151, CD231, CD102, NOTCH1, NOTCH2, NOTCH3, NOTCH4, DLL1, DLL4, JAG1, JAG2, CD49d/ITGA4, ITGB5, ITGB6, ITGB7, CD11a, CD11b, CD11c, CD18/ITGB2, CD41, CD49b, CD49c, CD49e, CD51, CD61, CD104, тетраспанины, Fc-рецепторы, рецепторы интерлейкина, иммуноглобулины, компоненты MHC-I или MHC-II, CD2, CD3-эпсилон, CD3-дзета, CD13, CD18, CD19, CD30, CD34, CD36, CD40, CD40L, CD44, CD45, CD45RA, CD47, CD86, CD110, CD111, CD115, CD117, CD125, CD135, CD184, CD200, CD279, CD273, CD274, CD362, COL6A1, AGRN, EGFR, GAPDH, GLUR2, GLUR3, HLA-DM, HSPG2, L1CAM, LAMB1, LAMC1, LFA-1, LGALS3BP, Mac-1 альфа, Mac-1 бета, MFGE8, SLIT2, STX3, TCRA, TCRB, TCRD, TCRG, VTI1A, VTI1B и любые другие экзосомальные полипептиды.
5. ВВ по любому из предшествующих пунктов, где ВВ дополнительно содержит по меньшей мере один представляющий интерес полинуклеотид.
6. ВВ по п. 5, где транспорт представляющего интерес полинуклеотида в ВВ осуществляют через взаимодействие с РНК-связывающим Pol или ДНК-связывающим Pol.
7. ВВ, содержащая по меньшей мере один Pol, где освобождение Pol от по меньшей мере одного экзосомального полипептида осуществляют через эндогенную активацию системы высвобождения на основе полипептида.
8. ВВ по п. 7, где ВВ дополнительно содержит представляющий интерес полинуклеотид.
9. ВВ по любому из предшествующих пунктов, где ВВ дополнительно содержит по меньшей один нацеливающий фрагмент.
10. Способ продуцирования ВВ по любому из предшествующих пунктов, содержащий
i. введение в продуцирующую ВВ клетку по меньшей мере одной полинуклеотидной конструкции, кодирующей по меньшей мере одну полипептидную конструкцию, содержащую по меньшей мере один Pol, эндогенно активируемую систему высвобождения на основе полипептида и по меньшей мере один экзосомальный полипептид;
ii. экспрессию продуцирующей ВВ клеткой по меньшей мере одной полипептидной конструкции, кодируемой по меньшей мере одной полинуклеотидной конструкцией; и iii. сбор из продуцирующей ВВ клетки содержащих Pol ВВ.
11. Полинуклеотидная конструкция, кодирующая:
i. по меньшей мере один Pol;
ii. по меньшей мере одну эндогенно активируемую систему на основе полипептида; и iii) по меньшей мере один экзосомальный полипептид.
12. Полипептидная конструкция, содержащая
i. по меньшей мере один Pol;
ii. по меньшей мере одну эндогенно активируемую систему на основе полипептида; и iii) по меньшей мере один экзосомальный полипептид.
13. ВВ, содержащая: (i) по меньшей мере одну полинуклеотидную конструкцию по п. 11 и/или (ii) по меньшей мере одну полипептидную конструкцию по п. 12.
14. Клетка, содержащая: (i) по меньшей мере одну полинуклеотидную конструкцию по п. 11 и/или (ii) по меньшей мере одну полипептидную конструкцию по п. 12, и/или (iii) по меньшей мере одну ВВ по любому из пп. 1-9 и/или 13.
15. In vitro способ внутриклеточной доставки по меньшей мере одного Pol, включающий контактирование целевой клетки с по меньшей мере одной ВВ по любому из пп. 1-9 и/или по меньшей мере одной ВВ по любому из пп. 1-9 или 13.
16. Фармацевтическая композиция, содержащая: (i) по меньшей мере одну полинуклеотидную конструкцию по п. 11 и/или (ii) по меньшей мере одну полипептидную конструкцию по п. 12, и/или (iii) по меньшей мере одну ВВ по любому из пп. 1-9 и/или 13, и/или (iv) клетку по п. 14.
17. Применение ВВ по любому из пп. 1-9 или 13 в медицине.
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