Single variable domains of immunoglobulin targeting the T-cell receptor

Claims

1. An immunoglobulin single variable domain (ISVD) that specifically binds to the constant domain of a human and / or non-human primate T cell receptor (TCR) present on a T cell, wherein said ISVD essentially consists of 4 framework regions (FR1-FR4, respectively) and 3 complementarity determining regions (CDR1-CDR3, respectively), wherein a. The amino acid sequence of CDR1 (according to Rabat) is INFYG (SEQ ID NO: 79); b. the amino acid sequence of CDR2 (according to Rabat) is HISIGDQTDYAX1SAKG (SEQ ID NO: 80); and c. the amino acid sequence of CDR3 (according to Rabat) is LSRIX2PYDY (SEQ ID NO: 81); Where - amino acid residue X1 is selected from E, D, N, P, K, R, I, T, H, V, A, Y, L, Q, F and S; and / or - amino acid residue X2 is selected from Y, A, P, D, Q, E, R, F, S, G, T, H, V, K, L and I.

2. An immunoglobulin single variable domain (ISVD) that specifically binds to the constant domain of a human and / or non-human primate T cell receptor (TCR) present on a T cell, wherein said ISVD essentially consists of 4 framework regions (FR1-FR4, respectively) and 3 complementarity determining regions (CDR1-CDR3, respectively), wherein a. The amino acid sequence of CDR1 (according to AbM) is GYVHKINFYG (SEQ ID NO: 82); b. the amino acid sequence of CDR2 (according to AbM) is HISIGDQTD (SEQ ID NO: 83); and c. the amino acid sequence of CDR3 (according to AbM) is LSRIX2PYDY (SEQ ID NO: 84); and where - the amino acid residue at position 61 (according to Kabat) is selected from E, D, N, P, K, R, I, T, H, V, A, Y, L, Q, F and S; and / or - amino acid residue X2 is selected from Y, A, P, D, Q, E, R, F, S, G, T, H, V, K, L and I.

3. ISVD according to claim 1, where X1 is E or D.

4. ISVD according to claim 1, where X1 represents E.

5. ISVD according to claim 2, wherein the amino acid residue at position 61 (according to Kabat) is E or D.

6. ISVD according to claim 2, wherein the amino acid residue at position 61 (according to Kabat) is E.

7. ISVD according to any one of paragraphs 1-6, where X2 is Y, A, Q, F, S, T or H.

8. ISVD according to any one of paragraphs 1-6, where X2 is Y, Q, S or T.

9. ISVD according to any one of paragraphs 1-6, where X2 is Y.

10. An ISVD according to any one of claims 1 to 9, wherein the amino acid residue in the ISVD at position 103 (numbering according to Kabat) is selected from the group consisting of W, R, A, E, Y, L, H, I, Q, V, K, S, G, P, F, T.

11. An ISVD according to any one of claims 1 to 9, wherein the amino acid residue in the ISVD at position 103 (numbering according to Kabat) is W.

12. An ISVD according to claim 1, wherein X1 is E, X2 is Y, and the amino acid residue at position 103 (numbering according to Kabat) is W.

13. An ISVD according to claim 2, wherein the amino acid residue at position 61 (numbering according to Kabat) is E, X2 is Y, and the amino acid residue at position 103 (numbering according to Kabat) is W.

14. ISVD according to any one of claims 1-13, wherein the amino acid residue at position 1 (numbering according to Kabat) is selected from E and D.

15. An ISVD according to any one of claims 1 to 14, wherein the ISVD is a heavy chain ISVD.

16. The ISVD of claim 15, wherein the heavy chain ISVD is selected from V HH humanized V HH camelized V H , domain antibody, single domain antibody and dAb.

17. An ISVD according to any one of the preceding claims, which is characterized by a degree of sequence identity with the sequence of any one of SEQ ID NOs: 2-57 of at least 85%, preferably at least 90%, more preferably at least 95%, wherein for the purposes of determining the degree of sequence identity, amino acid residues that form the CDR sequences are not taken into account, and wherein preferably the sequence is SEQ ID NO: 37, SEQ ID NO: 42, SEQ ID NO: 46, SEQ ID NO: 50 or SEQ ID NO:

52.

18. An ISVD according to any of the preceding claims, which is characterized by a degree of sequence identity with the sequence of SEQ ID NO: 32, 33 and / or 35-57 of at least 85%, preferably at least 90%, more preferably at least 95%, wherein for the purposes of determining the degree of sequence identity, amino acid residues that form the CDR sequences are not taken into account, and where preferably the sequence comprises SEQ ID NO: 37, SEQ ID NO: 42, SEQ ID NO: 46, SEQ ID NO: 50 or SEQ ID NO:

52.

19. The ISVD according to claim 17 or 18, wherein the ISVD comprises or consists of SEQ ID NO: 37 or SEQ ID NO:

42.

20. An ISVD according to claim 17 or 18, wherein the ISVD comprises or consists of SEQ ID NO: 46, SEQ ID NO: 50 or SEQ ID NO:

52.

21. An immunoglobulin single variable domain (ISVD) comprising the following sequence: X0VQLVESGGGVVQPGGSLRLSCVASGYVHKINFYGWYRQAPGKEREKVAHISIGDQTDYA X1SAKGRFTISRDESKNTVYLQMNSLRPEDTAAYYCRALSRIX2PYDYX3GQGTLVTVSS (SEQ ID NO: 85), or consisting of it, where a. X0 is chosen from E and D; b. X1 is selected from the group consisting of E, D, N, P, K, R, I, T, H, V, A, Y, L, Q, F, and S; c. X2 is selected from the group consisting of Y, A, P, D, Q, E, R, F, S, G, T, H, V, K, L, and I; and d. X3 is selected from the group consisting of W, R, A, E, Y, L, H, I, Q, V, K, S, G, P, F, and T.

22. ISVD according to claim 21, where X0 is D.

23. ISVD according to claim 21 or 22, where X1 is selected from the group consisting of D or E.

24. ISVD according to item 21 or 22, where X1 is E.

25. ISVD according to any one of paragraphs 21-24, where X2 is selected from the group consisting of Y, T, S and Q.

26. ISVD according to any one of paragraphs 21-24, where X2 is Y.

27. ISVD according to any one of paragraphs 21-26, where X3 is W.

28. A polypeptide comprising a first ISVD capable of specifically binding to a constant domain of a human and / or non-human primate T cell receptor (TCR) present on a T cell, and a second ISVD capable of specifically binding to a first antigen on a target cell, wherein said first antigen is different from said TCR, and wherein said target cell is different from said T cell, wherein said first and second ISVDs essentially consist of 4 framework regions (FR1-FR4, respectively) and 3 complementarity determining regions (CDR1-CDR3, respectively), and wherein the first ISVD is an ISVD according to any one of claims 1-27.

29. The polypeptide of claim 28, wherein the amino acid sequence of the first ISVD is characterized by at least 80% sequence identity with at least one of the amino acid sequences of any of SEQ ID NOs: 2-57, preferably at least 85%, more preferably at least 90%, more preferably at least 95%, wherein for the purposes of determining the degree of sequence identity, amino acid residues that form CDR sequences are not taken into account, wherein preferably the sequence of the first ISVD comprises SEQ ID NO: 37, SEQ ID NO: 42, SEQ ID NO: 46, SEQ ID NO: 50 or SEQ ID NO:

52.

30. The polypeptide of claim 28 or 29, wherein the first ISVD has at least 80% sequence identity to the amino acid sequence of any of SEQ ID NOs: 32, 33 and / or 35-57, preferably at least 85%, more preferably at least 90%, more preferably at least 95%, wherein for the purposes of determining the degree of sequence identity, amino acid residues that form CDR sequences are not taken into account, and wherein preferably the sequence comprises SEQ ID NO: 37, SEQ ID NO: 42, SEQ ID NO: 46, SEQ ID NO: 50 or SEQ ID NO:

52.

31. The polypeptide of claim 28 or 29, wherein the first ISVD comprises or consists of SEQ ID NO: 37 or SEQ ID NO:

42.

32. The polypeptide of claim 28 or 29, wherein the first ISVD comprises or consists of SEQ ID NO: 46, SEQ ID NO: 50 or SEQ ID NO:

52.

33. The polypeptide of any one of claims 28-32, further comprising a third ISVD that specifically binds to a second antigen on a target cell.

34. The polypeptide of any one of claims 28-33, wherein said polypeptide further comprises one or more other groups, residues, fragments or linking units, optionally linked via one or more peptide linkers, wherein said one or more other groups, residues, fragments or linking units provide an increased half-life of the polypeptide compared to the corresponding polypeptide without said one or more other groups, residues, fragments or linking units.

35. The polypeptide of claim 34, wherein said one or more other groups, residues, fragments or binding units are an ISVD that is capable of binding to human serum albumin.

36. A method for producing an ISVD according to any one of claims 1-27 or a polypeptide according to any one of claims 28-35, wherein the method comprises a. providing for expression in a suitable host cell or host organism or other suitable expression system of a nucleic acid sequence encoding an ISVD or polypeptide; optionally followed by b. isolating and / or purifying the ISVD or polypeptide.

37. A nucleic acid encoding an ISVD according to any one of claims 1-27 or a polypeptide according to any one of claims 28-35.

38. A vector containing a nucleic acid according to claim 37.

39. A host or host cell other than a human being transformed or transfected with the nucleic acid of claim 37 or the vector of claim 38.

40. A composition comprising an ISVD according to any one of claims 1-27 or a polypeptide according to any one of claims 28-35.

41. The composition according to claim 40, wherein the composition is a pharmaceutical composition.

42. The polypeptide according to any one of claims 28-35 or the composition according to claim 40 or 41 for use as a medicinal product.

43. The polypeptide of any one of claims 28-35 or the composition of claim 40 or 41 for use in the prevention, treatment or amelioration of a disease selected from the group consisting of a proliferative disease, an inflammatory disease, an infectious disease and an autoimmune disease.

44. The polypeptide or composition for use according to claim 43, wherein the disease is cancer.