Affinity-enhanced monomeric streptavidin chimeric antigen receptor (CAR)

Patent metadata

Field	Value
Publication number	US-11117936-B2
Application number	US-201816183579-A
Country	US
Kind code	B2
Filing date	Nov 7, 2018
Priority date	Nov 10, 2017
Publication date	Sep 14, 2021
Grant date	Sep 14, 2021

Abstract

A chimeric antigen receptor is disclosed that includes: (a) an extracellular high affinity streptavidin;(b) a hinge domain from CD8; (c) a CD28 transmembrane domain; (d) an intracellular 4-1BB and/or CD28 signaling domain; and (e) an intracellular CD3 zeta signaling domain, wherein (a)-(e) are in N-terminal to C-terminal order. Nucleic acids encoding this chimeric antigen receptor, and T and natural killer (NK) cells transformed with this chimeric antigen receptor are also disclosed. The use of this chimeric antigen receptor for the treatment of tumors is also disclosed.

First claim

I claim:

1. A method for treating a subject with a tumor, comprising
administering to the subject a therapeutically effective amount of a monoclonal antibody that specifically binds a tumor associated antigen expressed by the tumor, wherein the monoclonal antibody is biotinylated, and
administering to the subject a pharmaceutical composition, thereby treating the tumor in the subject,
wherein the pharmaceutical composition comprises i) an effective amount of an expression vector comprising a promoter operably linked to a nucleic acid molecule encoding a chimeric antigen receptor comprising amino acids 1-369 of SEQ ID NO: 11 or amino acids 1-371 of SEQ ID NO: 12 or ii) an effective amount of CD3+T cells and/or natural killer cells transduced with the expression vector of (i), and a pharmaceutically acceptable carrier, thereby treating the tumor in the subject.

2. The method of claim 1 , wherein the pharmaceutical composition comprises the CD3+T cells, and wherein the T cells are autologous to the subject.

3. The method of claim 1 , wherein
a) the pharmaceutical composition comprises the CD3+ T cells, wherein the CD3+ T cells are CD3 + CD4 + T cells and/or CD3 + CD8 + T cells; and/or
b) the tumor associated antigen is MUC-1, CD19 or CD20.

4. The method of claim 1 , wherein the subject is human.

5. The method of claim 1 , wherein a) the tumor is a lymphoma or leukemia; or b) the tumor is a solid tumor.

6. The method of claim 1 , wherein the chimeric antigen receptor comprises amino acids 1-371 of SEQ ID NO: 12.

7. The method of claim 1 , wherein the expression vector comprises the nucleic acid sequence of SEQ ID NO: 13.

8. The method of claim 1 , wherein the expression vector comprises the nucleic acid sequence of SEQ ID NO: 14.

9. The method of claim 1 , wherein the expression vector comprises (a) the nucleic acid sequence of SEQ ID NO: 15; and/or (b) the nucleic acid of SEQ ID NO: 16.

10. The method of claim 1 , wherein the expression vector comprises the nucleic acid sequence of SEQ ID NO: 17 or SEQ ID NO: 18.

11. The method of claim 1 , wherein the expression vector comprises the nucleic acid sequence of SEQ ID NO: 19.

12. The method of claim 1 , wherein the expression vector is codon-optimized for expression in human cells.

13. The method of claim 1 , wherein the expression vector comprises the nucleic acid sequence of SEQ ID NO: 20 or SEQ ID NO: 21.

14. The method of claim 1 , wherein the expression vector is a viral vector.

15. The method of claim 14 , wherein the viral vector is a lentiviral vector or a gamma retroviral vector.

16. The method of claim 1 , wherein the tumor associated antigen is EGFR or HER2.

17. A method for treating a subject with a tumor, comprising
transducing CD3+ T cells and/or natural killer cells from the subject with an expression vector comprising a promoter operably linked to a nucleic acid molecule encoding a chimeric antigen receptor comprising amino acids 1-369 of SEQ ID NO: 11 or amino acids 1-371 of SEQ ID NO: 12 to produce autologous transduced cells that express the chimeric antigen receptor:
administering to the subject a therapeutically effective amount of a monoclonal antibody that specifically binds a tumor associated antigen expressed by the tumor, wherein the monoclonal antibody is biotinylated; and
administering to the subject a therapeutically effective amount of the autologous transduced cells that express the chimeric antigen receptor, thereby treating the tumor in the subject.

18. The method of claim 17 , wherein the tumor associated antigen is MUC-1, CD19 or CD20.

19. The method of claim 17 , wherein the expression vector is a gamma retroviral vector.

20. The method of claim 17 , wherein the tumor associated antigen is EGFR or HER2.

Assignees

• Univ Of Pittsburgh—Of The Commonwealth System Of Higher Education
• Univ Of Pittsburg—Of The Commonwealth System Of Higher Education

Inventors

• Lohmueller Jason

Classifications

• A61K40/4211

  CD19 or B4 · CPC title

• A61K40/31

  Chimeric antigen receptors [CAR] · CPC title

• A61K40/11

  T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells · CPC title

• A61K35/17

  Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes (when activated by a specific antigen A61K39/00) · CPC title

• C07K14/70517

  CD8 · CPC title