US 11,655,452 B2
Chimeric antigen receptors (CARs), compositions and methods of use thereof
Yupo Ma, Stony Brook, NY (US); Kevin Pinz, Stony Brook, NY (US); Xun Jiang, Stony Brook, NY (US); Masayuki Wada, Stony Brook, NY (US); and Kevin Chen, Stony Brook, NY (US)
Assigned to iCell Gene Therapeutics Inc., Stony Brook, NY (US)
Appl. No. 15/739,596
Filed by iCell Gene Therapeutics LLC, Stony Brook, NY (US)
PCT Filed Jun. 24, 2016, PCT No. PCT/US2016/039306
§ 371(c)(1), (2) Date Dec. 22, 2017,
PCT Pub. No. WO2016/210293, PCT Pub. Date Dec. 29, 2016.
Claims priority of provisional application 62/244,435, filed on Oct. 21, 2015.
Claims priority of provisional application 62/235,840, filed on Oct. 1, 2015.
Claims priority of provisional application 62/184,321, filed on Jun. 25, 2015.
Prior Publication US 2018/0187149 A1, Jul. 5, 2018
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/0783 (2010.01); C07K 14/725 (2006.01); C07K 14/705 (2006.01); C07K 16/28 (2006.01)
CPC C12N 5/0636 (2013.01) [C07K 14/7051 (2013.01); C07K 14/70517 (2013.01); C07K 14/70521 (2013.01); C07K 14/70578 (2013.01); C07K 16/289 (2013.01); C12N 5/0646 (2013.01); C07K 2319/00 (2013.01); C07K 2319/02 (2013.01); C07K 2319/03 (2013.01); C07K 2319/50 (2013.01); C12N 2510/00 (2013.01)] 4 Claims
 
1. An ex vivo engineered T cell or NK cell co-expressing two distinct chimeric antigen receptor (CAR) units at the cell surface, wherein the engineered T cell or NK cell comprises a nucleotide sequence comprising from 5′ to 3′ a single promoter selected from human elongation factor-1 alpha (EF-1α) or spleen focus forming virus (SFFV), a first polynucleotide encoding a first chimeric antigen receptor polypeptide (CAR), a nucleotide encoding a viral self-cleavage peptide, a second polynucleotide encoding a second chimeric antigen receptor polypeptide (CAR), and an enhancer selected from the group consisting of secreted IL-15/IL-15sushi, Il-15/Il-15 sushi anchor, PD-1, PD-L1, CSF1R, CTLA-4, TIM-3, TGFR-beta, IL-2, Il-7, IL-12, IL-15, IL-15RA, IL-21 or a functional fragment thereof, or a combination thereof, wherein said enhancer is separated from the first CAR and second CAR by a second cleavage site that flanks either end of the two distinct CAR units, wherein:
the first CAR comprises a first signal peptide, a first antibody binding domain, a first hinge region, a first transmembrane domain, a first co-stimulatory domain, and a first signaling domain; and
the second CAR comprises a second signal peptide, a second antibody binding domain, a second hinge region, a second transmembrane domain, a second co-stimulatory domain, and a second signaling domain; and
wherein the first antibody binding domain and the second antibody binding domain are different and each bind to a different target, wherein the targets of the first and second antibody binding domains irrespective of order are CD19 and CD20, or CD123 and CD33, or B cell maturation antigen (BCMA) (CD269) and CD19, or BCMA (CD269) and CD38, or and BCMA (CD269) and CS1, wherein the first and second co-stimulatory domains are intracellular, and wherein the cleavage site is selected from the group consisting of porcine teschovirus-1 2A (P2A), thoseaasigna virus 2A (T2A), equine rhinitis A virus (ERAV) 2A (E2A), and FMDV 2A (F2A).