Nucleic acid molecules encoding chimeric antigen receptors comprising a CD20 binding domain

Patent metadata

Field	Value
Publication number	US-10525083-B2
Application number	US-201715727402-A
Country	US
Kind code	B2
Filing date	Oct 6, 2017
Priority date	Oct 7, 2016
Publication date	Jan 7, 2020
Grant date	Jan 7, 2020

Abstract

The invention provides compositions and methods for treating diseases associated with expression of CD20 or CD22. The invention also relates to chimeric antigen receptor (CAR) specific to CD20 or CD22, vectors encoding the same, and recombinant T or natural killer (NK) cells comprising the CD20 CAR or CD22 CAR. The invention also includes methods of administering a genetically modified T cell or NK cell expressing a CAR that comprises a CD20 or CD22 binding domain.

First claim

We claim:

1. An isolated nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the CAR comprises a CD20 binding domain, a transmembrane domain, and an intracellular signaling domain, wherein said CD20 binding domain comprises a light chain complementarity determining region 1 (LCDR1), light chain complementarity determining region 2 (LCDR2), light chain complementarity determining region 3 (LCDR3) heavy chain complementarity determining region 1 (HCDR1), heavy chain complementarity determining region 2 (HCDR2), and heavy chain complementarity determining region 3 (HCDR3), wherein the LCDR1, LCDR2, LCDR3, HCDR1, HCDR2, and HCDR3 comprise:
(i) SEQ ID NOs: 147, 148, 149, 136, 137, and 138, respectively;
(ii) SEQ ID NOs: 150, 151, 152, 139, 140, and 141, respectively;
(iii) SEQ ID NOs: 153, 154, 155, 142, 143, and 144, respectively; or
(iv) SEQ ID NOs: 929, 930, 931, 926, 927, and 928, respectively.

2. The isolated nucleic acid molecule of claim 1 , wherein the CD20 binding domain comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein:
(i) the VL comprises the amino acid sequence of SEQ ID NO: 156, 129, or 439, or an amino acid sequence with at least 95% identity thereto;
(ii) the VH comprises the amino acid sequence of SEQ ID NO: 145, 118, or 437, or an amino acid sequence with at least 95% identity thereto; or
(iii) the VL and VH comprise:
(a) SEQ ID NOs: 156 and 145, respectively;
(b) SEQ ID NOs: 129 and 118, respectively; or
(c) SEQ ID NOs: 439 and 437, respectively.

3. The isolated nucleic acid molecule of claim 1 , wherein:
(i) the nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 157, 130, or 440, or a nucleotide sequence with at least 95% identity thereto;
(ii) the nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 146, 119, or 438, or a nucleotide sequence with at least 95% identity thereto; or
(iii) the nucleic acid molecule comprises:
(a) SEQ ID NOs: 157 and 146;
(b) SEQ ID NOs: 130 and 119; or
(c) SEQ ID NOs: 440 and 438.

4. The isolated nucleic acid molecule of claim 1 , wherein the CD20 binding domain comprises the amino acid sequence of SEQ ID NO: 159 or 132, or an amino acid sequence with at least 95% identity thereto.

5. The isolated nucleic acid molecule of claim 1 , wherein the nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 160 or 133, or a nucleotide sequence with at least 95% identity thereto.

6. The isolated nucleic acid molecule of claim 1 , wherein the transmembrane domain comprises a transmembrane domain of a protein selected from the group consisting of the alpha, beta or zeta chain of T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 and CD154.

7. The isolated nucleic acid molecule of claim 1 , wherein:
(i) the transmembrane domain comprises the amino acid sequence of SEQ ID NO: 801, or an amino acid sequence with at least 95% identity thereto; or
(ii) the nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 802 or 1072, or a nucleotide sequence with at least 95% identity thereto.

8. The isolated nucleic acid molecule of claim 1 , wherein the CD20 binding domain is connected to the transmembrane domain by a hinge region.

9. The isolated nucleic acid molecule of claim 8 , wherein:
(i) the hinge region comprises the amino acid sequence of SEQ ID NO: 799 or SEQ ID NO: 814, or an amino acid sequence with at least 95% identity thereto; or
(ii) the nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 800 or SEQ ID NO: 815, or a nucleotide sequence with at least 95% identity thereto.

10. The isolated nucleic acid molecule of claim 1 , wherein the intracellular signaling domain comprises a costimulatory domain, wherein the costimulatory domain is a functional signaling domain obtained from a protein selected from the group consisting of OX40, CD2, CD27, CD28, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278) and 4-1BB (CD137).

11. The isolated nucleic acid molecule of claim 1 , wherein the intracellular signaling domain comprises a costimulatory domain, wherein:
(i) the costimulatory domain comprises the amino acid sequence of SEQ ID NO: 803, or an amino acid sequence with at least 95% identity thereto; or
(ii) the nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 804, or a nucleotide sequence with at least 95% identity thereto.

12. The isolated nucleic acid molecule of claim 1 , wherein the intracellular signaling domain comprises a primary signaling domain, wherein the primary signaling domain comprises a functional signaling domain of CD3 zeta.

13. The isolated nucleic acid molecule of claim 1 , wherein the intracellular signaling domain comprises a primary signaling domain, wherein:
(i) the primary signaling domain comprises the amino acid sequence of SEQ ID NO: 807 or SEQ ID NO: 805, or an amino acid sequence with at least 95% identity thereto; or
(ii) the nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 893, 808, 806, or 1074, or a nucleotide sequence with at least 95% identity thereto.

14. The isolated nucleic acid molecule of claim 1 , wherein the intracellular signaling domain comprises a functional signaling domain of 4-1BB and/or a functional signaling domain of CD3 zeta.

15. The isolated nucleic acid molecule of claim 1 , wherein:
(i) the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO: 803, or an amino acid sequence with at least 95% identity thereto, and the amino acid sequence of SEQ ID NO: 807 or SEQ ID NO: 805, or an amino acid sequence with at least 95% identity thereto;
(ii) the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO: 803 and the amino acid sequence of SEQ ID NO: 807 or SEQ ID NO: 805;
(iii) the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO: 803 and the amino acid sequence of SEQ ID NO: 807 or SEQ ID NO: 805, wherein the sequences comprising the intracellular signaling domain are expressed in the same frame and as a single polypeptide chain, or
(iv) the nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 804, or a nucleotide sequence with at least 95% identity thereto, and/or the nucleotide sequence of SEQ ID NO: 893, 808, 806, or 1074, or a nucleotide sequence with at least 95% identity thereto.

16. The isolated nucleic acid molecule of claim 1 , further comprising a leader sequence, wherein the leader sequence encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 797, or an amino acid sequence with at least 95% identity thereto.

17. The isolated nucleic acid molecule of claim 1 , wherein the nucleic acid molecule encodes a CAR comprising the amino acid sequence of SEQ ID NO: 161 or 134, or an amino acid sequence with at least 95% identity thereto, wherein the CAR comprises or does not comprise a signal peptide comprising the amino acid sequence of SEQ ID NO: 797.

18. The isolated nucleic acid molecule of claim 1 , wherein the nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 162 or 135, or a nucleotide sequence with at least 95% identity thereto, wherein the nucleic acid molecule comprises or does not comprise the nucleotide sequence of SEQ ID NO: 798.

19. A nucleic acid comprising:
(i) a first nucleic acid comprising the nucleic acid molecule of claim 1 , and
(ii) a second nucleic acid encoding a CAR molecule that binds a B-cell antigen.

Assignees

• Novartis Ag
• Univ Pennsylvania

Inventors

• Brannetti Barbara
• Brogdon Jennifer
• Engels Boris
• Granda Brian
• Huang Lu
• Lei Ming
• Li Na
• Zhang Jimin
• Guimaraes Carla

Classifications

• C07K16/2803 Primary

  against the immunoglobulin superfamily · CPC title

• C07K2317/622

  Single chain antibody (scFv) · CPC title

• A61K2039/505

  comprising antibodies · CPC title

• C07K2319/33

  fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies · CPC title

• C07K2319/03

  containing a transmembrane segment · CPC title