T-CELL RECEPTORS TO MAGE-A4 AND METHODS OF THEIR USE

Abstract

The present invention provides isolated T cell receptors (TCRs) that specifically bind to the HLA-presented cancer antigen testicular melanoma-associated antigen 4 (MAGE-A4) peptide, as well as therapeutic and diagnostic methods for using these isolated TCRs.

Claims

1. A T-cell receptor (TCR) that specifically binds to the HLA-A2-presented testicular cancer antigen melanoma-associated antigen 4 (MAGE-A4) peptide, The TCR comprises CDR1, CDR2 and CDR3 of the alpha chain variable domain and CDR1, CDR2 and CDR3 of the beta chain variable domain contained in the amino acid sequence pair of the TCR alpha chain variable domain / TCR beta chain variable domain SEQ ID NO: 748 / 756 or 716 / 724.

2. The TCR according to claim 1, comprising a pair of amino acid sequences SEQ ID NO: 748 / 756 or 716 / 724 of the variable domain of the TCR alpha chain / variable domain of the TCR beta chain.

3. The TCR of claim 1, wherein the TCR has a property selected from the group consisting of: (a) has a target binding / off-target binding ratio of greater than 5; (b) does not bind to cells expressing putative off-target peptides as determined by fluorescence assay; (c) does not bind to cells expressing putative off-target peptides as determined by flow cytometric analysis; (d) activates a T cell response approximately two-fold greater than that of a patient-derived MAGE-A4-specific TCR, as determined by a TCR-mediated luminescence T cell signaling bioassay; and (e) activates a T cell response approximately two-fold greater than that of an affinity-matured MAGE-A4-specific TCR, as determined by a TCR-mediated luminescence T cell signaling bioassay.

4. The TCR according to any one of claims 1-3, further comprising a detectable fragment.

5. A pharmaceutical composition comprising a TCR according to any one of claims 1 to 4 and a pharmaceutically acceptable carrier or diluent.

6. A cell presenting the TCR according to any one of claims 1 to 4.

7. An isolated polynucleotide molecule containing a polynucleotide sequence that encodes the variable domain of the TCR alpha chain as set forth in any one of claims 1 to 4, and a polynucleotide sequence that encodes the variable domain of the TCR beta chain as set forth in any one of claims 1 to 4.

8. The isolated polynucleotide molecule of claim 7, wherein the polynucleotide sequence that encodes the variable domain of the TCR alpha chain comprises the nucleotide sequence of SEQ ID NO: 749, and wherein the polynucleotide sequence that encodes the variable domain of the TCR beta chain comprises SEQ ID NO:

757.

9. The isolated polynucleotide molecule according to claim 7, wherein the polynucleotide sequence that encodes the variable domain of the TCR alpha chain comprises the nucleotide sequence of SEQ ID NO: 717, and wherein the polynucleotide sequence that encodes the variable domain of the TCR beta chain comprises the nucleotide sequence of SEQ ID NO:

725.

10. A vector containing a polynucleotide molecule according to any one of claims 7 to 9.

11. Isolated cell containing: vector according to item 10; a vector containing a polynucleotide molecule according to claim 7; a vector containing a polynucleotide molecule according to claim 8; a vector containing a polynucleotide molecule according to claim 9; a polynucleotide molecule according to claim 7; a polynucleotide molecule according to claim 8; or a polynucleotide molecule according to paragraph 9.

12. Isolated cell containing: a first polynucleotide molecule, wherein the first polynucleotide sequence comprises a polynucleotide sequence that encodes a variable domain of a TCR alpha chain as set forth in any one of paragraphs 7-9, and a second polynucleotide molecule, wherein the second polynucleotide molecule comprises a polynucleotide sequence that encodes a variable domain of a TCR beta chain as set forth in any one of claims 7 to 9.

13. The isolated cell according to paragraph 12, the first polynucleotide molecule is contained in a vector.

14. The isolated cell of claim 13, wherein the second polynucleotide molecule is contained in a second vector.

15. An isolated cell according to any one of claims 12-14, wherein the first polynucleotide molecule and the second polynucleotide molecule are contained in a vector.

16. A pharmaceutical composition comprising a cell according to any one of claims 6 and 11-15 and a pharmaceutically acceptable carrier or diluent.

17. A method of treating a subject who has a MAGE-A4 associated disease or disorder, comprising administering to the subject a therapeutically effective amount of a TCR as set forth in any of claims 1-4, a pharmaceutical composition according to claim 5 or 16, or a cell according to any of claims 6 and 11-15, thereby treating the subject.

18. The method of claim 17, wherein the MAGE-A4-associated disease or disorder is a MAGE-A4-associated cancer.

19. The method of claim 18, wherein the MAGE-A4-associated cancer is liposarcoma, neuroblastoma, myeloma, melanoma, metastatic melanoma, synovial sarcoma, bladder cancer, esophageal cancer, esophageal squamous cell carcinoma, hepatocellular carcinoma, head and neck cancer, non-small cell lung cancer, ovarian cancer, epithelial ovarian cancer, prostate cancer, breast cancer, astrocytic tumor, glioblastoma multiforme, anaplastic astrocytoma, brain tumor, fallopian tube cancer, primary abdominal cancer, advanced solid tumors, soft tissue sarcoma, sarcoma, myelodysplastic syndrome, acute myeloid leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, Hodgkin's disease, multiple myeloma, metastatic solid tumors, colorectal carcinoma, gastric cancer, rhabdomyosarcoma, myxoid round cell liposarcoma, or recurrent non-small cell lung cancer.

20. The method according to any one of claims 17-19, wherein the TCR, pharmaceutical composition or cell is administered to the subject in combination with a second therapeutic agent.

21. The method according to any one of claims 17-20, wherein the TCR, pharmaceutical composition or cell is administered to the subject subcutaneously, intravenously, intradermally, intraperitoneally, orally, intramuscularly or intracranially.

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