RU2021123419A - Химерные антигенные рецепторы и способы их получения - Google Patents

Химерные антигенные рецепторы и способы их получения Download PDF

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RU2021123419A
RU2021123419A RU2021123419A RU2021123419A RU2021123419A RU 2021123419 A RU2021123419 A RU 2021123419A RU 2021123419 A RU2021123419 A RU 2021123419A RU 2021123419 A RU2021123419 A RU 2021123419A RU 2021123419 A RU2021123419 A RU 2021123419A
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vectors
cd3ζ
separate
encode
cell
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RU2021123419A
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Лоуренс Дж. Н. КУПЕР
Ана Беатриц КОРНГОЛД
Брайан А. РАБИНОВИЧ
Харджеет СИНГХ
Саймон ОЛИВАРЕС
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Борд Оф Риджентс, Дзе Юниверсити Оф Техас Систем
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Claims (19)

1. Способ получения множества векторов, каждый из которых кодирует химерный антигенный рецептор (CAR), включающий:
(i) получение композиции, содержащей
(а) множество первых векторов, кодирующих один или несколько отдельных антигенсвязывающих доменов;
(b) множество вторых векторов, кодирующих один или несколько отдельных доменов шарнирной области; и
(c) множество третьих векторов, кодирующих один или несколько отдельных эндодоменов;
где по меньшей мере два из первого, второго и третьего векторов содержат множество из двух или более векторов, кодирующих различные антигенсвязывающие домены, домены шарнирной области и/или эндодомены, соответственно, и, кроме того, где векторы содержат сайты гомологичной рекомбинации, обеспечивающие получение четвертого вектора, кодирующего химерный антигенный рецептор (CAR); и
(ii) помещая композицию в условия, достаточные для рекомбинации различных антигенсвязывающих доменов, доменов шарнирной области и/или эндодоменов, кодируемых указанными векторами посредством гомологичной рекомбинации с получением множества четвертых векторов, где каждый из указанных четвертых векторов кодирует CAR.
2. Способ по п.1, где множество первых векторов кодирует множество отдельных антигенсвязывающих доменов, множество вторых векторов кодирует один домен шарнирной области, и множество третьих векторов кодирует множество отдельных эндодоменов.
3. Способ по п.1, где множество первых векторов кодирует множество отдельных антигенсвязывающих доменов, множество вторых векторов кодирует множество отдельных доменов шарнирного домена, и множество третьих векторов кодирует множество отдельных эндодоменов.
4. Способ по п.1, где множество первых векторов кодирует множество отдельных антигенсвязывающих доменов, множество вторых векторов кодирует множество доменов шарнирной области, и множество третьих векторов кодирует один эндодомен.
5. Способ по п.1, где множество первых векторов кодирует одни антигенсвязывающий домен, множество вторых векторов кодирует множество отдельных доменов шарнирного домена, и множество третьих векторов кодирует множество отдельных эндодоменов.
6. Способ по пп.1-5, где композиция дополнительно содержит множество пятых векторов, кодирующих один или несколько трансмембранных доменов; где первые векторы, вторые векторы, третьи векторы и пятые векторы содержат сайты гомологичной рекомбинации для получения четвертого вектора, кодирующего химерный антигенный рецептор (CAR).
7. Способ по пп.1-6, где антигенсвязывающий домен селективно связывается с CD19, универсальным антигеном (мышь), HER-3, GD2, Gp75, белок CS1, мезохелин, фосфатидилсерин, cMyc, CD22, CD4, CD44v6, CD45, CD28, CD3, CD3e, CD123, CD138, CD52, CD56, CD74, CD30, Gp75, CD38, CD33, CD20, слитая конструкция Her1/HER3, GD2, углевод, Aspergillus, ROR1, c-MET, EGFR, Дектин, Эбола, грибы, GP, HERV-K (HERVK), NY-ESO-1, VEGF-R2, TGF-b2R, IgG4, Биотин или O-AcGD2.
8. Способ по пп.1-7, где шарнирная область кодирует пептид 12 AA (GAGAGCAAGTACGCCCTCCCTGCCCCCCTTGCCCT, SEQ ID NO: 1), пептид t-20 AA, IgG4 Fc Δ EQ, IgG4 Fc Δ Q, (t-12AA + t-20AA), mKate, phiLov, dsRed, Venus, eGFP, CH3 HA, (CD8 α + t-20AA), двойной t-20 AA, (t-20AA + CD8α), (CD8α + основой белок 1 лейциновой застежки), (CD8α + кислый белок 1 лейциновой застежки), 2D3, CD8α или IgG4 Fc.
9. Способ по пп.1-8, где по меньшей мере один из эндодоменов содержит только (CD28 + CD3ζ), (CD28 + CD27 + CD3ζ), (CD28 + OX40 + CD3ζ), (CD28 + 4-1BB + CD3ζ), (CD28 + CD27 + OX40 + CD3ζ), (CD28 + 4-1BB + CD27 + CD3ζ), (CD28 + 4-1BB + OX40 + CD3ζ), (4-1BB + CD3ζ), (4-1BB + OX40 + CD3ζ ), (4-1BB + CD27 + CD3ζ), (CD27 + CD3ζ), (CD27 + OX 40 + CD3ζ), (CD28Δ + CD3ζ), (CD28Δ + CD27 + CD3ζ), (CD28Δ + OX40 + CD3ζ), ( CD28Δ + 4-1BB + CD3ζ), (CD28Δ + 4-1BB + OX40 + CD3ζ), (CD28Δ + CD27 + OX40 + CD3ζ), (CD28Δ + 4-1BB + CD27 + CD3ζ), (4-1BB + ICOS + CD3ζ), (CD28 + ICOS + CD3ζ), (ICOS + CD3ζ) CD3ζ или CD28.
10. Способ по пп.1-9, где способ дополнительно включает экспрессию CAR в клетке.
11. Способ по пп.1-10, где способ дополнительно включает тестирование CAR на активность, где активность включает способность CAR селективно связывать раковые клетки, селективно связывать патоген, селективно связывать клетку, участвующую в аутоиммунном заболевании, или усиливать активацию Т-клетки, разрушение Т-клетки, дифференцировку Т-клетки, пролиферацию Т-клетки, дедифференцировку Т-клетки, перемещение Т-клетки, продукцию цитокинов Т-клеткой или уничтожение Т-клеткой.
12. Способ по пп.1-11, где первые векторы, вторые векторы и/или третьи векторы кодируют транспозазу.
13. Способ по пп.1-12, где шестой вектор кодирует транспозазу, и где способ включает введение, электропорацию или трансфекцию одного или нескольких указанных четвертых векторов и указанного шестого вектора в клетку.
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