TRIFUNCTIONAL T CELL-ANTIGEN COUPLER AND METHODS AND USES THEREOF

• US 20160368964A1
• Filed: 02/06/2015
• Published: 12/22/2016
• Est. Priority Date: 02/07/2014
• Status: Active Grant

• Current Assignee: Mcmaster University
• Original Assignee: Mcmaster University
• Inventors: Helsen, Christopher W., Giri, Rajanish, Mwawasi, Kenneth Anthony, Bramson, Jonathan, Denisova, Galina
• Granted Patent: US 10,435,453 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/5156 expressing foreign proteins
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• 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 39/4632 T-cell receptors [TCR]; antibody T-cell receptor constructs
• CPC Class Codes: A61K 39/464406 Her-2/neu/ErbB2, Her-3/ErbB3 or Her 4/ ErbB4
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 14/70514 CD4
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/2878 against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2318/20 Antigen-binding scaffold molecules wherein the scaffold is not an immunoglobulin variable region or antibody mimetics
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/03 containing a transmembrane segment
• 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: C12N 5/0636 : T lymphocytes

Abstract

A trifunctional molecule comprising a target-specific ligand, a ligand that binds a protein associated with the TCR complex and a T cell receptor signaling domain polypeptide is provided. Engineering T cells with this novel receptor engenders antigen specific activation of numerous T cell functions, including cytokine production, degranulation and cytolysis.

Claims

1. A nucleic acid comprising:

a. a first polynucleotide encoding a target-specific ligand;

b. a second polynucleotide encoding a ligand that binds a protein associated with the TCR complex; and

c. a third polynucleotide encoding a T cell receptor signaling domain polypeptide.

2. The nucleic acid of claim 1, wherein the target-specific ligand binds a tumor antigen.

3. The nucleic acid of claim 1, wherein the target-specific ligand is a designed ankyrin repeat (DARPin) polypeptide or scFv.

4. The nucleic acid of claim 1, wherein the protein associated with the TCR complex is CD3.

5. The nucleic acid of claim 2, wherein the ligand that binds a protein associated with the TCR complex is a single chain antibody.

6. The nucleic acid of claim 5, wherein the ligand that binds a protein associated with the TCR complex is UCHT1, or a variant thereof.

7. The nucleic acid of claim 1, wherein the T cell receptor signaling domain polypeptide comprises a cytosolic domain and a transmembrane domain.

8. The nucleic acid of claim 7, wherein the cytosolic domain is a CD4 cytosolic domain and the transmembrane domain is a CD4 transmembrane domain.

9. The nucleic acid of claim 1, wherein the first polynucleotide and third polynucleotide are fused to the second polynucleotide.

10. The nucleic acid of claim 1, wherein the second polynucleotide and third polynucleotide are fused to the first polynucleotide.

11. A polypeptide encoded by the nucleic acid of claim 1.

12. An expression vector comprising the nucleic acid of claim 1.

13. A T-cell expressing the nucleic acid of claim 1.

14. A vector construct comprising:

a. a first polynucleotide encoding a target-specific ligand;

b. a second polynucleotide encoding a ligand that binds a protein associated with the TCR complex; and

c. a third polynucleotide encoding a T cell receptor signaling domain polypeptide, and

d. a promoter functional in a mammalian cell.

15. The vector construct of claim 14, wherein the first polynucleotide and third polynucleotide are fused to the second polynucleotide to provide a T cell antigen coupler fusion and the coding sequence of the T cell antigen coupler fusion is operably connected to the promoter.

16. The vector construct of claim 14, wherein the second polynucleotide and third polynucleotide are fused to the first polynucleotide to provide a T cell antigen coupler fusion and the coding sequence of the T cell antigen coupler fusion is operably connected to the promoter.

17. The vector construct of claim 14, wherein the target-specific ligand binds a tumor antigen.

18. The vector construct of claim 17, wherein the target-specific ligand is a designed ankyrin repeat (DARPin) polypeptide or scFv.

19. The vector construct of claim 17, wherein the protein associated with the TCR complex is CD3.

20. The vector construct of claim 14, wherein the ligand that binds a protein associated with the TCR complex is a single chain antibody.

21. The vector construct of claim 14, wherein the ligand that binds a protein associated with the TCR complex is UCHT1, or a variant thereof.

22. The vector construct of claim 14, wherein the T cell receptor signaling domain polypeptide comprises a cytosolic domain and a transmembrane domain.

23. The vector construct of claim 22, wherein the cytosolic domain is a CD4 cytosolic domain and the transmembrane domain is a CD4 transmembrane domain.

24. The vector construct of claim 14, wherein the first polynucleotide and third polynucleotides are fused to the second polynucleotide.

25. An isolated T cell transfected with the vector construct of claim 14.

26. A pharmaceutical composition comprising the T cell of claim 25 and a carrier.

27. A use of a T cell for treating cancer in a subject in need thereof, wherein the T cell expresses a nucleic acid comprising:

a. a first polynucleotide encoding a target-specific ligand;

b. a second polynucleotide encoding a ligand that binds a protein associated with the TCR complex; and

c. a third polynucleotide encoding a T cell receptor signaling domain polypeptide.

28. The use of claim 27, wherein the target-specific ligand binds an antigen on a cancerous cell.

29. The use of claim 28, wherein the target-specific ligand is a designed ankyrin repeat (DARPin) polypeptide or scFv.

30. The use of claim 29, wherein the protein associated with the TCR complex is CD3.

31. The use of claim 27, wherein the ligand that binds a protein associated with the TCR complex is a single chain antibody.

32. The use of claim 27, wherein the ligand that binds a protein associated with the TCR complex is UCHT1, or a variant thereof.

33. The use of claim 27, wherein the T cell receptor signaling domain polypeptide comprises a cytosolic domain and a transmembrane domain.

34. The use of claim 33, wherein the cytosolic domain is a CD4 cytosolic domain and the transmembrane domain is a CD4 transmembrane domain.

35. The use of claim 27, wherein the first polynucleotide and third polynucleotide are fused to the second polynucleotide.

36. The use of claim 27, wherein the second polynucleotide and third polynucleotide are fused to the first polynucleotide.