T cell recruiting polypeptides based on TCR alpha/beta reactivity
T cell recruiting polypeptides are provided that bind the constant domain of TCR on a T cell. The polypeptides can be used in methods for treatment of cancers.
1. Polypeptide comprising a first and a second immunoglobulin single variable domain (ISV), wherein
said first ISV specifically binds to the constant domain of the T cell receptor (TCR) present on a T cell;
said second ISV specifically binds to a first antigen on a target cell;
wherein said first antigen is different from said TCR;
wherein said target cell is different from said T cell, and
wherein said first ISV essentially consists of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively), in which:
(i) CDR1 is chosen from the group consisting of:
(a) SEQ ID NOs: 119-123, 125-127, 129, 132 and 133, or
(b) amino acid sequences that have 2 or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO: 123; and
(ii) CDR2 is chosen from the group consisting of:
(c) SEQ ID NOs: 134-141, 143-144, 146-156, and 159-163, or
(d) amino acid sequences that have 2 or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO:153; and
(iii) CDR3 is chosen from the group consisting of:
(e) SEQ ID NOs: 164-166, 169-171, 173-174, or
(f) amino acid sequences that have 2 or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO: 170.
2. The polypeptide according to claim 1 , wherein said first ISV binds to the constant domain of a T cell receptor α (TCR-α) (SEQ ID NO: 348 and/or SEQ ID NO: 484) and/or the constant domain of the T cell receptor β (TCR-β) (SEQ ID NO: 349 and/or SEQ ID NO: 485), or polymorphic variants or isoforms thereof.
3. The polypeptide according to claim 1 , wherein:
(i) CDR1 is chosen from the group consisting of:
(a) SEQ ID NO: 123; or
(b) amino acid sequences that have 2 or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO: 123; and
(ii) CDR2 is chosen from the group consisting of:
(c) SEQ ID NO: 153; or
(d) amino acid sequences that have 2 or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO: 153; and
(iii) CDR3 is chosen from the group consisting of:
(e) SEQ ID NOs: 170; or
(f) amino acid sequences that have 2 or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO: 170.
4. The polypeptide according to claim 1 , further comprising a third ISV, which specifically binds to a second antigen on a target cell, wherein said second antigen is different from said first antigen.
5. The polypeptide according to claim 1 , wherein said first antigen on a target cell is a tumour antigen, preferably a tumour associated antigen (TAA).
6. The polypeptide according to claim 4 , wherein said second antigen on a target cell is a tumour antigen, preferably a tumour associated antigen (TAA).
7. The polypeptide according to claim 4 , wherein said first antigen and said second antigen are present on the same target cell.
8. The polypeptide according to claim 4 , wherein said first antigen and said second antigen are present on different target cells.
9. The polypeptide according to claim 1 , further comprising a serum protein binding moiety.
10. The polypeptide according to claim 9 , wherein said serum protein binding moiety is an ISV binding serum albumin.
11. The polypeptide according to claim 1 , wherein said first ISV and said second ISV are linked via a linker.
12. The polypeptide according to claim 11 , wherein said linker is chosen from the group consisting of linkers of 5GS, 7GS, 9GS, 10GS, 15GS, 18GS, 20GS, 25GS, 30GS and 35GS (SEQ ID NOs: 376 to 385).
13. The polypeptide according to claim 1 , wherein said ISV is a Nanobody, a V HH , a humanized V HH , or a camelized V H .
14. A polypeptide that specifically binds the constant domain of the T cell receptor (TCR) and that comprises or essentially consists of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively), in which:
(i) CDR1 is chosen from the group consisting of:
(a) SEQ ID NOs: 119-123, 125-127, 129, 132 and 133; or
(b) amino acid sequences that have 2 or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO: 123; and
(ii) CDR2 is chosen from the group consisting of:
(c) SEQ ID NOs: 134-141, 143-144, 146-156, and 159-163; or
(d) amino acid sequences that have 2 or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO: 153; and
(iii) CDR3 is chosen from the group consisting of:
(e) SEQ ID NOs: 164-166, 169-171, 173-174; or
(f) amino acid sequences that have 2 or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO: 170.
15. The polypeptide according to claim 14 , which is a Nanobody, a V HH , a humanized V HH , or a camelized V H .
16. The polypeptide according to claim 14 , further comprising a serum protein binding moiety.
17. The polypeptide according to claim 16 , wherein said serum protein binding moiety is an ISV that binds serum albumin.
18. A nucleic acid or nucleic acid sequence encoding the polypeptide as defined in claim 1 .
19. A pharmaceutical composition comprising the polypeptide according to claim 1 .
20. The polypeptide according to claim 14 , wherein:
(i) CDR1 is chosen from the group consisting of:
(a) SEQ ID NO: 123; or
(b) amino acid sequences that have 2 or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO: 123; and
(ii) CDR2 is chosen from the group consisting of:
(c) SEQ ID NO: 153; or
(d) amino acid sequences that have 2 or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO: 153; and
(iii) CDR3 is chosen from the group consisting of:
(e) SEQ ID NOs: 170; or
(f) amino acid sequences that have 2 or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO: 170.