Compositions and methods for TCR reprogramming using fusion proteins

• US 10,208,285 B2
• Filed: 02/05/2018
• Issued: 02/19/2019
• Est. Priority Date: 10/07/2016
• Status: Active Grant

• Current Assignee: TCR2 Therapeutics Inc.
• Original Assignee: TCR2 Therapeutics Inc.
• Inventors: Baeuerle, Patrick, Sieczkiewicz, Gregory, Hofmeister, Robert
• Primary Examiner(s): Skelding, Zachary S
• Application Number:
  US15/888,897
• Publication Number:
  US 20180230429A1
• Time in Patent Office: 379 Days
• Field of Search: None
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/5156 expressing foreign proteins
• CPC Class Codes: A61K 2039/5158 Antigen-pulsed cells, e.g. T-cells
• CPC Class Codes: A61K 2039/572 cytotoxic response
• CPC Class Codes: A61K 2039/585 wherein the target is cancer
• CPC Class Codes: A61K 2039/6006 Cells recombinantly expressing antigens A61K2039/5156, A61K2039/523
• CPC Class Codes: A61K 39/0011 Cancer antigens
• CPC Class Codes: A61K 39/001168 Mesothelin [MSLN]
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4631 Chimeric Antigen Receptors [CAR]
• CPC Class Codes: A61K 39/464429 Molecules with a "CD" designation not provided for elsewhere
• CPC Class Codes: A61K 39/46444 Interleukins [IL]
• CPC Class Codes: A61K 39/464441 Interferons [IFN]
• CPC Class Codes: A61K 39/464468 Mesothelin [MSLN]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/4748 Tumour specific antigens; Tumour rejection antigen precursors [TRAP], e.g. MAGE
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 2317/70 characterized by effect upon binding to a cell or to an antigen
• CPC Class Codes: C07K 2319/02 : containing a signal sequence
• CPC Class Codes: C07K 2319/03 : containing a transmembrane segment
• CPC Class Codes: C07K 2319/30 : Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/33 : fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C07K 2319/70 : containing domain for protein-protein interaction
• CPC Class Codes: C12N 5/0638 : Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK]
• CPC Class Codes: C12N 5/0646 : Natural killers cells [NK], NKT cells

Abstract

Provided herein are T-cell receptor (TCR) fusion proteins (TFPs), T-cells engineered to express one or more TFPs, and methods of use thereof for the treatment of diseases, including cancer.

Claims

1. A recombinant nucleic acid molecule encoding a T cell receptor (TCR) fusion protein (TFP) comprising

(a) a TCR subunit comprising

(i) at least a portion of a TCR extracellular domain, and

(ii) a TCR intracellular domain comprising a stimulatory domain from an intracellular signaling domain of CD3 epsilon or CD3 gamma;

and

(b) a human or humanized antibody domain comprising an antigen binding domain that is a sdAb or V_HH anti-mesothelin binding domain comprising the sequence of SEQ ID NO;

58 or SEQ ID NO;

59,

wherein the TCR subunit and the antibody domain are operatively linked, and

wherein the TFP incorporates into a functional TCR when expressed in a T cell.

2. The recombinant nucleic acid molecule of claim 1, wherein the encoded antigen binding domain is connected to the TCR extracellular domain by a linker sequence.

3. The recombinant nucleic acid molecule of claim 2, wherein the encoded linker sequence comprises (G₄S)_n, wherein n=1 to 4.

4. The recombinant nucleic acid molecule of claim 1, wherein the TCR subunit comprises (i) a TCR extracellular domain, (ii) a TCR transmembrane domain, and (iii) a TCR intracellular domain, wherein at least two of (i), (ii), and (iii) are from the same TCR subunit.

5. The recombinant nucleic acid molecule of claim 1, wherein the TCR subunit comprises a TCR intracellular domain comprising a stimulatory domain selected from an intracellular signaling domain of CD3 epsilon, CD3 gamma, or an amino acid sequence having at least one, two or three modifications thereto.

6. The recombinant nucleic acid molecule of claim 1, wherein the antigen binding domain is a V_HH domain.

7. The recombinant nucleic acid molecule of claim 1, wherein the anti-mesothelin binding domain comprises the sequence of SEQ ID NO:

58.

8. A human T cell comprising the recombinant nucleic acid molecule of claim 1.

9. The T cell of claim 8, wherein the T cell is a CD8+ T cell, a CD4+ T cell, a gamma-delta T cell, or a NKT cell.

10. The T cell of claim 8, wherein the T cell exhibits increased cytotoxicity to a human cell expressing mesothelin compared to a T cell not containing the TFP.

11. The T cell of claim 8, wherein production of IL-2 by the T cell is increased in the presence of a human cell expressing mesothelin compared to a T cell not containing the TFP.

12. The T cell of claim 8, wherein the T cell is from a human subject.

13. The T cell of claim 8, wherein the T cell or its progeny persists in a human subject administered a pharmaceutical composition comprising the T cell for at least four months.

14. The T cell of claim 8, wherein the T cell is from a mammal having a disease associated with expression of mesothelin.

15. The recombinant nucleic acid molecule of claim 1, wherein the TCR intracellular domain comprises a stimulatory domain from an intracellular signaling domain of CD3 epsilon.

16. The T cell of claim 14, wherein the disease associated with expression of mesothelin is mesothelioma.

17. The T cell of claim 8, wherein production of a cytokine by the T cell is lower than production of the cytokine by a T cell comprising a nucleic acid encoding a CAR comprising the antigen binding domain operatively linked to at least a portion of a CD28 extracellular domain, a CD28 transmembrane domain, at least a portion of a CD28 intracellular domain, and a CD3 zeta intracellular domain.

18. The T cell of claim 17, wherein the cytokine is IL-2 or IFNγ

.

19. A pharmaceutical composition comprising the T cell of claim 8.

20. The recombinant nucleic acid molecule of claim 1, wherein the anti-mesothelin binding domain comprises the sequence of SEQ ID NO:

59.