RU2015122371A - Слитые белки для применения в качестве иммуногенных усиливающих агентов для индуцирования антигенспецифического т-клеточного ответа - Google Patents

Слитые белки для применения в качестве иммуногенных усиливающих агентов для индуцирования антигенспецифического т-клеточного ответа Download PDF

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RU2015122371A
RU2015122371A RU2015122371A RU2015122371A RU2015122371A RU 2015122371 A RU2015122371 A RU 2015122371A RU 2015122371 A RU2015122371 A RU 2015122371A RU 2015122371 A RU2015122371 A RU 2015122371A RU 2015122371 A RU2015122371 A RU 2015122371A
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fusion protein
protein according
amino acid
peptide
binding domain
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RU2015122371A
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RU2631002C2 (ru
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Чиа-Мао У
Сиу-Кан ЧАН
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Тевакс Дженетикс Вэксин Ко., Лтд.
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Claims (33)

1. Слитый белок, содержащий:
(a) домен связывания с антигенпрезентирующей клеткой (АПК) или домен связывания с рецептором CD91, расположенный на N-конце слитого белка;
(b) домен трансдукции белка, расположенный на С-конце домена связывания с АПК или домена связывания с рецептором CD91, причем домен трансдукции белка выбран из группы, состоящей из:
(i) слитого полипептида, содержащего пептид, трансдуцирующий сигнал, сенсибилизирующий Т-клетки, линкер и транслокационный пептид, причем:
(1) пептид, трансдуцирующий сигнал, сенсибилизирующий Т-клетки, расположен на N-конце слитого полипептида;
(2) линкер содержит последовательность SEQ ID NO: 15, связывающую пептид, трансдуцирующий сигнал, сенсибилизирующий Т-клетки, и транслокационный пептид; и
(3) транслокационный пептид имеет длину от 34 до 112 аминокислотных остатков и содержит аминокислотную последовательность, которая по меньшей мере на 90% идентична SEQ ID NO: 3, 20 или 4;
(ii) пептида, трансдуцирующего сигнал, сенсибилизирующий Т-клетки; и
(iii) транслокационного пептида, длина которого равна 34-46 аминокислотным остаткам, содержащего аминокислотную последовательность, которая по меньшей мере на 90% идентична SEQ ID NO: 3 или 20; и
(c) антиген патогена, расположенный на С-конце домена трансдукции белка,
причем:
- пептид, трансдуцирующий сигнал, сенсибилизирующий Т-клетки, имеет длину от 28 до 53 аминокислотных остатков и содержит аминокислотную последовательность SEQ ID NO: 31, в которой Хаа8 соответствует I или L; Хаа10 соответствует V, F или А, Хаа11 соответствует М или L, Хаа17 соответствует L или I; и
- домен связывания с антигенпрезентирующей клеткой (АПК) или домен связывания с рецептором CD91 не содержит аминокислотной последовательности домена I связывания экзотоксина A Pseudomonas (РЕ; от англ. Pseudomonas exotoxin), если домен трансдукции белка является транслокационным пептидом (biii).
2. Слитый белок по п. 1, отличающийся тем, что домен связывания с АПК или домен связывания с рецептором CD91 является полипептидом, содержащим аминокислотную последовательность, которая по меньшей мере на 90% идентична последовательности, выбранной из группы, состоящей из последовательностей SEQ ID NO: 5, 9, 6, 7 и 8.
3. Слитый белок по п. 1, отличающийся тем, что он дополнительно содержит последовательность, обеспечивающую удержание в эндоплазматическом ретикулуме, расположенную на С-конце слитого белка.
4. Слитый белок по п. 3, отличающийся тем, что последовательность, обеспечивающая удержание в эндоплазматическом ретикулуме, содержит аминокислотную последовательность Lys-Asp-Glu-Leu (SEQ ID NO: 14).
5. Слитый белок по п. 1, отличающийся тем, что он не содержит последовательности, обеспечивающей удержание в эндоплазматическом ретикулуме, на своем С-конце, если антиген содержит 10 или более эпитопов.
6. Слитый белок по п. 1, отличающийся тем, что домен трансдукции белка является слитым полипептидом (bi).
7. Слитый белок по п. 1, отличающийся тем, что домен трансдукции белка является пептидом, трансдуцирующим сигнал, сенсибилизирующий Т-клетки (bii).
8. Слитый белок по п. 7, отличающийся тем, что он содержит дополнительный линкер между доменом трансдукции белка и антигеном, причем дополнительный линкер содержит последовательность SEQ ID NO: 15.
9. Слитый белок по п. 1, отличающийся тем, что домен трансдукции белка является транслокационным пептидом (biii).
10. Слитый белок по п. 9, отличающийся тем, что он содержит дополнительный линкер между доменом связывания АПК или доменом связывания рецептора CD91 и транслокационным пептидом, причем дополнительный линкер содержит последовательность SEQ ID NO: 15.
11. Слитый белок по п. 1, отличающийся тем, что домен трансдукции белка содержит последовательность SEQ ID NO: 30.
12. Композиция вакцины, содержащая:
(a) терапевтически эффективное количество слитого белка по п. 1; и
(b) адъювант.
13. Слитый белок по п. 1, отличающийся тем, что домен связывания АПК или домен связывания рецептора CD91 является полипептидом, содержащим аминокислотную последовательность, выбранную из группы, состоящей из последовательностей SEQ ID NO: 5, 9, 6, 7 и 8.
14. Слитый белок по п. 1, отличающийся тем, что пептид, трансдуцирующий сигнал, сенсибилизирующий Т-клетки, содержит аминокислотную последовательность, выбранную из группы, состоящей из последовательностей SEQ ID NO: 1 и 2.
15. Слитый белок по п. 1, отличающийся тем, что транслокационный пептид содержит аминокислотную последовательность SEQ ID NO: 3.
16. Способ индуцирования усиленных антигенспецифических Т-клеточных ответов против патогенов, включающий введение субъекту, нуждающемуся в этом, терапевтически эффективного количества слитого белка по п. 1.
17. Способ уничтожения болезненной клетки, презентирующей антиген через молекулы класса 1 МНС на мембране болезненной клетки, включающий введение субъекту, нуждающемуся в этом, терапевтически эффективного количества слитого белка по п. 1.
18. Способ по п. 17, где болезненная клетка является раковой клеткой.
19. Способ профилактики или лечения инфекции, вызванной патогеном, и/или минимизации симптомов, вызванных инфекцией, включающий введение субъекту, нуждающемуся в этом, терапевтически эффективного количества слитого белка по п. 1.
RU2015122371A 2012-12-05 2013-12-03 Слитые белки для применения в качестве иммуногенных усиливающих агентов для индуцирования антигенспецифического т-клеточного ответа RU2631002C2 (ru)

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