RU2016145958A - Химерные антигенные рецепторы с mnd-промотором - Google Patents

Химерные антигенные рецепторы с mnd-промотором Download PDF

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RU2016145958A
RU2016145958A RU2016145958A RU2016145958A RU2016145958A RU 2016145958 A RU2016145958 A RU 2016145958A RU 2016145958 A RU2016145958 A RU 2016145958A RU 2016145958 A RU2016145958 A RU 2016145958A RU 2016145958 A RU2016145958 A RU 2016145958A
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vector
ltr
virus
promoter
antibody
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RU2016145958A
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RU2708311C2 (ru
RU2016145958A3 (ru
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Ричард МОРГАН
Кевин ФРИДМАН
Пёун РЮ
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Блубёрд Био, Инк.
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    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/03—Fusion polypeptide containing a localisation/targetting motif containing a transmembrane segment
    • 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
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/15011—Lentivirus, not HIV, e.g. FIV, SIV
    • C12N2740/15041—Use of virus, viral particle or viral elements as a vector
    • C12N2740/15043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector

Claims (41)

1. Полинуклеотид, содержащий промотор (MND) с энхансером вируса миелопролиферативной саркомы, с удаленным участком отрицательного контроля, с сайтом связывания праймера, замещенным на последовательность из dl587rev, функционально связанный с нуклеиновой кислотой, кодирующей химерный антигенный рецептор (CAR).
2. Полинуклеотид по п. 1, где CAR содержит:
a) внеклеточный домен, который связывает антиген, выбранный из группы, включающей фолатный рецептор альфа, 5T4, αvβ6-интегрин, BCMA, B7-H3, B7-H6, CAIX, CD19, CD20, CD22, CD30, CD33, CD44, CD44v6, CD44v7/8, CD70, CD79a, CD79b, CD123, CD138, CD171, CEA, CSPG4, EGFR, семейство EGFR, в том числе ErbB2 (HER2), EGFRvIII, EGP2, EGP40, EPCAM, EphA2, EpCAM, FAP, фетальный AchR, FRα, GD2, GD3, 'глипикан-3 (GPC3), HLA-A1+MAGE1, HLA-A2+MAGE1, HLA-A3+MAGE1, HLA-A1+NY-ESO-1, HLA-A2+NY-ESO-1, HLA-A3+NY-ESO-1, IL-11Rα, IL-13Rα2, лямбда-цепь, Lewis-Y, каппа-цепь, мезотелин, Muc1, Muc16, NCAM, лиганды NKG2D, NY-ESO-1, PRAME, PSCA, PSMA, ROR1, SSX, сурвивин, TAG72, TEM и VEGFR2;
b) трансмембранный домен, полученный из полипептида, выбранного из группы, включающей CD8α; CD4, CD28, CD45, PD1 и CD152;
c) один или более внутриклеточных доменов передачи костимулирующего сигнала, выбранных из группы, включающей CD28, CD54 (ICAM), CD134 (OX40), CD137 (41BB), CD152 (CTLA4), CD273 (PD-L2), CD274 (PD-L1) и CD278 (ICOS); и
d) домен передачи первичного сигнала CD3ζ.
3. Полинуклеотид по п. 2, где:
a) внеклеточный домен содержит антитело или антиген-связывающий фрагмент, которые связывают антиген;
b) внеклеточный домен содержит антитело или антиген-связывающий фрагмент, которые связывают антиген, выбранные из группы, включающей верблюжий Ig, IgNAR, Fab-фрагменты, Fab'-фрагменты, F(ab)'2-фрагменты, F(ab)'3-фрагменты, Fv, одноцепочечное Fv-антитело («scFv»), бис-scFv, (scFv)2, минитело, диатело, триатело, тетратело, стабилизированный дисульфидными связями Fv-белок («dsFv») и однодоменное антитело (sdAb, нанотело); или
c) внеклеточный домен содержит антитело или антиген-связывающий фрагмент, которые связывают антиген, представляющие собой scFv; или
d) внеклеточный домен содержит человеческое антитело, мышиное антитело или гуманизированное антитело.
4. Полинуклеотид по любому из пп. 1-3, где
(a) трансмембранный домен получен из CD8α;
(b) один или более доменов передачи костимулирующего сигнала выбраны из группы, включающей: CD28, CD134 и CD137; или
(c) CAR содержит два или более доменов передачи костимулирующего сигнала, выбранных из группы, включающей: CD28, CD134 и CD137.
5. Полинуклеотид по любому из пп. 1-3, где CAR дополнительно содержит полипептид шарнирной области, спейсерную область или сигнальный пептид.
6. Полинуклеотид по п. 1, где полинуклеотид кодирует CAR, изложенный под любой из SEQ ID NO: 2-3.
7     Вектор, содержащий полинуклеотид по любому из пп. 1-6.
8. Вектор по п. 7, где:
a) вектор представляет собой вектор экспрессии;
b) вектор представляет собой вирусный вектор;
c) вектор представляет собой ретровирусный вектор;
d) вектор представляет собой лентивирусный вектор; или
e) вектор представляет собой лентивирусный вектор, выбранный из группы, включающей: вирус иммунодефицита человека (HIV); вирус висна-маэди (VMV); вирус артрита-энцефалита коз (CAEV); вирус инфекционной анемии лошадей (EIAV); вирус иммунодефицита кошек (FIV); вирус иммунодефицита крупного рогатого скота (BIV) и вирус иммунодефицита обезьян (SIV).
9. Вектор по любому из п. 7 или п. 8, содержащий левый (5') ретровирусный LTR, Psi (Ψ) сигнал упаковки, центральный полипуриновый тракт/ДНК-флэп (cPPT/FLAP), ретровирусный экспортный элемент; MND-промотор, функционально связанный с CAR по п. 1; и правый (3') ретровирусный LTR.
10. Вектор по п. 9, дополнительно содержащий:
a) гетерологичную последовательность полиаденилирования;
b) гетерологичную последовательность полиаденилирования, которая представляет собой последовательность полиаденилирования бычьего гормона роста или сигнальную последовательность полиаденилирования β-глобина кролика; или
c) посттранскрипционный регуляторный элемент вируса гепатита В (HPRE) или посттранскрипционный регуляторный элемент вируса гепатита сурков (WPRE).
11. Вектор по п. 10, где
(a) промотор 5' LTR замещен гетерологичным промотором;
(b) промотор 5' LTR замещен промотором цитомегаловируса (CMV), промотором вируса саркомы Рауса (RSV) или промотором вируса обезьян 40 (SV40);
(c) 5' LTR или 3' LTR представляет собой LTR лентивируса;
(d) 3' LTR содержит одну или более модификаций;
(e) 3' LTR содержит одну или более делеций; или
(f) 3' LTR представляет собой самоинактивирующийся (SIN) LTR.
12. Иммунная эффекторная клетка, содержащая вектор по любому из пп. 7-11.
13. Иммунная эффекторная клетка по п. 12, где иммунная эффекторная клетка представляет собой T-лимфоцит.
14. Композиция, содержащая иммунную эффекторную клетку по п. 12 или п. 13 и физиологически приемлемый наполнитель.
15. Способ лечения рака у субъекта, нуждающегося в этом, включающий введение субъекту терапевтически эффективного количества композиции по п. 14.
16. Способ лечения гемобластоза у субъекта, нуждающегося в этом, предусматривающий введение субъекту терапевтически эффективного количества композиции по п. 14.
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