US 12,331,114 B2
Humanized anti-CD19 chimeric antigen receptor, its nucleic acid sequence and its preparation
Rahul Purwar, Mumbai (IN); Alka Dwivedi, Mumbai (IN); Atharva Karulkar, Mumbai (IN); and Srisathya Srinivasan, Mumbai (IN)
Assigned to INDIAN INSTITUTE OF TECHNOLOGY BOMBAY, Mumbai (IN)
Appl. No. 16/969,788
Filed by INDIAN INSTITUTE OF TECHNOLOGY BOMBAY, Mumbai (IN)
PCT Filed Feb. 12, 2019, PCT No. PCT/IN2019/050111
§ 371(c)(1), (2) Date Aug. 13, 2020,
PCT Pub. No. WO2019/159193, PCT Pub. Date Aug. 22, 2019.
Claims priority of application No. 201821005458 (IN), filed on Feb. 13, 2018.
Prior Publication US 2021/0002366 A1, Jan. 7, 2021
Int. Cl. C07K 16/28 (2006.01); A61K 40/11 (2025.01); A61K 40/31 (2025.01); A61K 40/42 (2025.01); A61P 35/00 (2006.01); C07K 14/705 (2006.01); C07K 14/725 (2006.01); C12N 15/86 (2006.01)
CPC C07K 16/2803 (2013.01) [A61K 40/11 (2025.01); A61K 40/31 (2025.01); A61K 40/4211 (2025.01); A61P 35/00 (2018.01); C07K 14/7051 (2013.01); C07K 14/70517 (2013.01); C07K 14/70578 (2013.01); C12N 15/86 (2013.01); A61K 2239/48 (2023.05); C07K 2317/24 (2013.01); C07K 2317/53 (2013.01); C07K 2317/56 (2013.01); C07K 2317/565 (2013.01); C07K 2317/567 (2013.01); C07K 2317/622 (2013.01); C07K 2319/02 (2013.01); C07K 2319/03 (2013.01)] 16 Claims
 
1. A recombinant nucleic acid molecule encoding a humanized chimeric antigen receptor (CAR), wherein the humanized CAR comprises:
(a) a single chain antibody or single chain antibody fragments comprising a humanized anti-CD 19 binding domain;
(b) a hinge region wherein said hinge region comprises the nucleic acid sequences of SEQ ID No. 38;
(c) a transmembrane domain wherein said transmembrane domain comprises the nucleotide sequence selected from SEQ ID No. 39, SEQ ID No. 40 and SEQ ID No. 45; and
(d) a cytoplasmic domain comprising a human 4-1BB costimulatory signaling domain wherein said costimulatory signaling domain comprises the nucleotide sequence selected from SEQ ID No. 41 and SEQ ID No. 47 and a CD33 signaling domain wherein said signaling domain comprises the nucleotide sequence selected from SEQ ID No. 42, SEQ ID No. 48 and SEQ ID No. 49;
wherein said humanized anti-CD 19 binding domain is scFv and comprises a heavy chain framework region 1 (HFR1) of SEQ ID No: 4 or SEQ ID No: 5, a heavy chain framework region 2 (HFR2) of SEQ ID No: 7, a heavy chain framework region 3 (HFR3) of SEQ ID No: 9 or SEQ ID No: 10, a heavy chain framework region 4 (HFR4) of SEQ ID No: 12 and a light chain framework region 1 (LFR1) of SEQ ID No: 13, a light chain framework region 2 (LFR2) of SEQ ID No: 15, a light chain framework region 3 (LFR3) of SEQ ID No: 17 and a light chain framework region 4 (LFR4) of SEQ ID No: 19;
wherein said nucleic acid molecule encodes a heavy chain complementary determining region 1 (HC CDR1) of SEQ ID No: 6, a heavy chain complementary determining region 2 (HC CDR2) of SEQ ID No: 8, a heavy chain complementary determining region 3 (HC CDR3) of SEQ ID No: 11 and a light chain complementary determining region 1 (LC CDR1) of SEQ ID No: 14, a light chain complementary determining region 2 (LC CDR2) of SEQ ID No: 16, and a light chain complementary determining region 3 (LC CDR3) of SEQ ID No: 18.