Regulating chimeric antigen receptors

Patent metadata

Field	Value
Publication number	US-11311609-B2
Application number	US-201816483204-A
Country	US
Kind code	B2
Filing date	Feb 8, 2018
Priority date	Feb 8, 2017
Publication date	Apr 26, 2022
Grant date	Apr 26, 2022

Abstract

This invention is in the area of compositions and methods for regulating chimeric antigen receptor immune effector cell, for example T-cell (CAR-T), therapy to modulate associated adverse inflammatory responses, for example, cytokine release syndrome and tumor lysis syndrome, using targeted protein degradation.

First claim

What is claimed is:

1. A T cell or natural killer (NK) cell comprising:
a. a chimeric antigen receptor polypeptide, wherein the chimeric antigen receptor polypeptide comprises:
i. an extracellular ligand binding domain comprising a single chain variable fragment (scFv) antibody that binds human CD19, BCMA, or CD38;
ii. a CD8, CD28, or CD137 transmembrane domain; and
iii. a cytoplasmic domain comprising at least one intracellular signaling domain, wherein the at least one intracellular signaling domain comprises CD3; and

b. a cytoplasmic costimulatory polypeptide, wherein the cytoplasmic costimulatory polypeptide comprises:
i. at least one intracellular signaling domain selected from a CD28, 4-1BB, OX40, CD27, ICOS, DAP-10, and DAP-12 signaling domain; and
ii. a heterobifunctional compound targeting protein, wherein the heterobifunctional compound targeting protein comprises the amino acid sequence selected from SEQ ID NO: 1 and 2 and wherein the heterobifunctional compound targeting protein binds to a heterobifunctional compound selected from the group consisting of dFKBP-1 to dFKBP-13;

wherein the binding of the heterobifunctional compound to the heterobifunctional compound targeting protein brings the cytoplasmic costimulatory polypeptide into proximity of a ubiquitin ligase;
and wherein the cytoplasmic costimulatory polypeptide is ubiquitinated and then degraded by a proteasome.

2. The T cell or natural killer (NK) cell of claim 1 , wherein the heterobifunctional compound targeting protein comprises the amino acid sequence of SEQ ID NO: 2.

3. The T cell or natural killer (NK) cell of claim 1 , wherein the T cell or natural killer (NK) cell is a T cell.

4. The T cell or natural killer (NK) cell of claim 1 , wherein the chimeric antigen receptor polypeptide further comprises a CD8α hinge domain.

5. The T cell or natural killer (NK) cell of claim 1 , wherein the at least one intracellular signaling domain of the cytoplasmic costimulatory polypeptide is CD28.

6. A therapeutic system, comprising:
a. the T cell or natural killer (NK) cell of claim 1 ; and
b. a heterobifunctional compound comprising any one of dFKBP-1 to dFKBP-13.

7. A method of reducing an adverse immune response in a subject caused by an activated T cell or natural killer (NK) cell that expresses a chimeric antigen receptor polypeptide and a cytoplasmic costimulatory polypeptide of claim 1 comprising:
administering to the subject experiencing an adverse immune response an effective amount of a heterobifunctional compound comprising any one of dFKBP-1 to dFKBP-13;
wherein the subject had previously been administered the T cell or natural killer (NK) cell of claim 1 .

8. A T cell or natural killer (NK) cell comprising a chimeric antigen receptor polypeptide, wherein the chimeric antigen receptor polypeptide comprises:
a. an extracellular ligand binding domain comprising a single chain variable fragment (scFv) antibody that binds human CD19, BCMA, or CD38;
b. a CD8, CD28, or CD137 transmembrane domain; and
c. a cytoplasmic domain, wherein the cytoplasmic domain comprises:
i. at least one intracellular signaling domain, wherein the at least one intracellular signaling domain comprises CD3; and
ii. a heterobifunctional compound targeting protein, wherein the heterobifunctional compound targeting protein comprises an amino acid sequence selected from SEQ ID NO: 1 and 2 and wherein the heterobifunctional compound targeting protein binds to a heterobifunctional compound;

wherein the binding of the heterobifunctional compound to the heterobifunctional compound targeting protein brings the chimeric antigen receptor polypeptide into proximity of a ubiquitin ligase;
and wherein the chimeric antigen receptor polypeptide is ubiquitinated and then degraded by a proteasome.

9. The T cell or natural killer (NK) cell of claim 8 , wherein the T cell or natural killer (NK) cell is a T cell.

10. The T cell or natural killer (NK) cell of claim 1 , wherein the chimeric antigen receptor polypeptide further comprises a CD8α hinge domain.

11. A therapeutic system for downregulating activation of a T cell or natural killer (NK) cell expressing a chimeric antigen receptor polypeptide, the system comprising:
a. the T cell or natural killer (NK) cell of claim 8 ; and
b. a heterobifunctional compound comprising any one of dFKBP-1 to dFKBP-13.

12. A method of reducing an adverse immune response in a subject caused by an activated T cell or natural killer (NK) cell that expresses a chimeric antigen receptor polypeptide of claim 8 comprising:
administering to the subject experiencing an adverse immune response an effective amount of a heterobifunctional compound comprising any one of dFKBP-1 to dFKBP-13;
wherein the subject had previously been administered the T cell or natural killer (NK) cell of claim 8 .

13. The T cell or natural killer (NK) cell of claim 1 , wherein the extracellular ligand binding domain binds CD19.

14. The T cell or natural killer (NK) cell of claim 8 , wherein the extracellular ligand binding domain binds CD19.

15. The T cell or natural killer (NK) cell of claim 1 , wherein the extracellular ligand binding domain that binds CD19 comprises scFv comprising the amino acid sequence of SEQ ID NO: 10 or 89.

16. The T cell or natural killer (NK) cell of claim 1 , wherein the extracellular ligand binding domain that binds BCMA comprises a variable light chain (VL) and a variable heavy chain (VH) wherein the VL comprises the amino acid sequence of SEQ ID NO: 68 or SEQ ID NO: 78 and wherein the VH comprises the amino acid sequence of SEQ ID NO: 70 or SEQ ID NO: 79.

17. The T cell or natural killer (NK) cell of claim 1 , wherein the extracellular ligand binding domain that binds CD38 comprises a variable light chain (VL) and a variable heavy chain (VH) wherein the VL comprises the amino acid sequence of SEQ ID NO: 81 or SEQ ID NO: 86 and the VH comprises the amino acid sequence of SEQ ID NO: 82 or SEQ ID NO: 87.

18. The T cell or natural killer (NK) cell of claim 1 , wherein the CD28 transmembrane domain comprises the amino acid sequence of SEQ ID NO: 16.

19. The T cell or natural killer (NK) cell of claim 1 , wherein the CD8 transmembrane domain comprises a CD8 alpha hinge comprising the amino acid sequence of SEQ ID NO: 15.

20. The T cell or natural killer (NK) cell of claim 1 , wherein the CD3 intracellular signaling domain comprises the amino acid sequence of SEQ ID NO: 17.

21. The T cell or natural killer (NK) cell of claim 8 , wherein the extracellular ligand binding domain that binds CD19 comprises scFv comprising the amino acid sequence of SEQ ID NO: 10.

22. The T cell or natural killer (NK) cell of claim 8 , wherein the extracellular ligand binding domain that binds BCMA comprises a variable light chain (VL) and a variable heavy chain (VH) wherein the VL comprises the amino acid sequence of SEQ ID NO: 68 or SEQ ID NO: 78 and the VH comprises the amino acid sequence of SEQ ID NO: 70 or SEQ ID NO: 79.

23. The T cell or natural killer (NK) cell of claim 8 , wherein the extracellular ligand binding domain that binds CD38 comprises a variable light chain (VL) and a variable heavy chain (VH) wherein the VL comprises the amino acid sequence of SEQ ID NO: 81 or SEQ ID NO: 86 and the VH comprises the amino acid sequence of SEQ ID NO: 82 or SEQ ID NO: 87.

24. The T cell or natural killer (NK) cell of claim 8 , wherein the CD28 transmembrane domain com

Assignees

• Dana Farber Cancer Inst Inc

Inventors

• Bradner James
• Roberts Justin
• Nabet Behnam
• Winter Georg
• Phillips Andrew J
• Heffernan Timothy P
• Buckley Dennis

Classifications

• A61K40/4222

  CD38 not IgG · CPC title

• A61K40/4215

  Receptors for tumor necrosis factors [TNF], e.g. lymphotoxin receptor [LTR], CD30 · CPC title

• A61K40/4211

  CD19 or B4 · CPC title

• A61K40/4205

  Her-2/neu/ErbB2, Her-3/ErbB3 or Her 4/ ErbB4 · CPC title

• A61K40/31

  Chimeric antigen receptors [CAR] · CPC title