Steap2 directed t-cell engagers and compositions thereof

Abstract

Provided are T cell engaging molecules that bind an antigen on a non-immune cell and an antigen on an immune cell. The T cell engaging molecules can bind STEAP2 on a cancer cell and, e.g., a CD3 on a T cell. The T cell engaging molecules can also bind STEAP2 on a cancer cell and, e.g., a CD8 on a T cell. Alternatively, the T cell engaging molecules can bind STEAP2 on a cancer cell and both CD3 and CD8 on a T cell.

Description

This abstract provides T-cell adaptor molecules that bind antigens on both non-immune cells and immune cells. These T-cell adaptor molecules can bind STEAP2 on cancer cells and, for example, CD3 on T cells. They can also bind STEAP2 on cancer cells and, for example, CD8 on T cells. Alternatively, they can bind STEAP2 on cancer cells and both CD3 and CD8 on T cells.

Claims

WHAT IS CLAIMED IS:

1. A T cell engaging molecule comprising:(a) an antigen binding arm that binds an epitope on human six transmembrane epithelial antigen of prostate-2 (STEAP2) and comprises a heavy chain comprising a heavy chain variable domain (VH) comprising a variable heavy chain complementarity determining region 1 (VH- CDR1) selected from SEQ ID NOs: 1, 9, 17, 103, 111, 127, 135, and 143; a VH-CDR2 selected from SEQ ID NOs: 2, 10, 18, 104, 112, 128, 136, and 144; a VH-CDR3 selected from SEQ ID NOs: 3, 11, 19, 94, 96, 98, 105, 113, 129, 137, and 145; a heavy chain CHI domain; and a light chain comprising a light chain variable domain (VL) comprising a variable light chain complementarity determining region 1 (VL-CDR1) selected from SEQ ID NOs: 4, 12, 20, 100, 108, 130, 138, and 146; a VL-CDR2 selected from SEQ ID NOs: 5, 13, 21, 101, 109, 131, 139, and 147; a VL-CDR3 selected from SEQ ID NOs: 6, 14, 22, 102, 110, 132, 140, and 148; and a light chain constant domain;(b) a first T cell binding arm that binds to cluster of differentiation 3 (CD3) comprising a heavy chain comprising a heavy chain variable domain (VH) comprising a VH-CDR1 selected from SEQ ID NOs: 36, 40, and 44; a VH-CDR2 selected from SEQ ID NOs: 37, 41, and 45; a VH-CDR3 selected from SEQ ID NOs: 38, 42, and 46; and a heavy chain CHI domain; and a light chain comprising a light chain variable domain (VL) comprising a VL-CDR1 selected from SEQ ID NOs: 27 and 31; a VL-CDR2 selected from SEQ ID NOs: 28 and 32; a VL-CDR3 selected from SEQ ID NOs: 29 and 33; and a light chain constant domain; and(c) a Fc domain, comprising a first Fc region and a second Fc region, each Fc region comprising a CH2 domain and a CH3 domain; said Fc domain further comprising at least one modification to promote heterodimerization.

2. A trivalent T cell engaging molecule comprising the T cell engaging molecule according to claim 1, further comprising (d) a second T cell binding arm that binds to cluster of differentiation 8 (CD8) and comprises a heavy chain comprising a heavy chain variable domain (VH) comprising a VH-CDR1 set forth in SEQ ID NO: 48, a VH-CDR2 set forth in SEQ ID NO: 49, and a VH-CDR3 set forth in SEQ ID NO: 50, and a heavy chain CHIdomain; and a light chain comprising a light chain variable domain (VL) comprising a VL- CDR1 set forth in SEQ ID NO: 51, a VL-CDR2 set forth in SEQ ID NO: 52, and a VL- CDR3 set forth in SEQ ID NO: 53, and a light chain constant domain.

3. The trivalent T cell engaging molecule according to claim 2, wherein the heavy chain of the second T cell binding arm is attached to the heavy chain of the first T cell binding arm through a linker.

4. The trivalent T cell engaging molecule according to claim 2, wherein the heavy chain of the second T cell binding arm is attached to the heavy chain of the antigen binding arm through a linker.

5. The trivalent T cell engaging molecule according to claim 3 or 4, wherein the linker comprises the amino acid sequence of SEQ ID NO: 89.

6. The trivalent T cell engaging molecule according to any one of claims 3 to 5, wherein the linker comprises from 1 to about 10 copies of SEQ ID NO: 89.

7. The trivalent T cell engaging molecule according to any one of claims 3 to 6, wherein the linker comprises 2 copies of SEQ ID NO: 89.

8. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 7, wherein one of the two CH3 domains comprises a knob mutation, and the other of the two CH3 domains comprises a hole mutation.

9. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 8, wherein the CHI domain and the light chain constant domain of one or more of each of the (a) antigen binding arm, (b) the first T cell binding arm, and (d) the second T cell binding arm further comprises a charge pair substitution comprising a first charged amino acid substitution in the CHI domain and a second charged amino acid substitution in the light chain constant domain, wherein the first charged amino acid substitution and the second charged amino acid substitution have an opposite charge.

10. The T cell engaging molecule or trivalent T cell engaging molecule according to claim 9, wherein the light chain constant domain of one or more of each of (a), (b), and (d) is a lambda light chain constant domain (CLX) and wherein the charge pair is a lambda charge pair comprising a positively charged amino acid residue selected from arginine, lysine or histidine, and a negatively charged amino acid residue selected from aspartic acid, glutamic acid, serine or threonine, wherein the numbering is according to the EU index and wherein the charged amino acids of the lambda charge pair are located at one or more of the following positions:(i) position 117 in the CLX and position 141 in the CHI domain;(ii) position 117 in the CLX and position 185 in the CHI domain;(iii) position 119 in the CLX and position 128 in the CHI domain;(iv) position 134 in the CLX and position 128 in the CHI domain;(v) position 134 in the CLX and position 145 in the CHI domain;(vi) position 134 in the CLX and position 183 in the CHI domain;(vii) position 136 in the CLX and position 185 in the CHI domain;(viii) position 178 in the CLX and position 173 in the CHI domain; and / or(ix) position 117 in the CLX and position 187 in the CHI domain.

11. The T cell engaging molecule or trivalent T cell engaging molecule according to claim 10, wherein (i) the charged amino acid at position 117 is arginine and the charged amino acid at position 141 is aspartic acid; the charged amino acid at position 117 is arginine and the charged amino acid at position 141 is glutamic acid; the charged amino acid at position 117 is arginine and the charged amino acid at position 141 is serine; the charged amino acid at position 117 is arginine and the charged amino acid at position 141 is threonine; the charged amino acid at position 117 is lysine and the charged amino acid at position 141 is aspartic acid; the charged amino acid at position 117 is lysine and the charged amino acid at position141 is glutamic acid; the charged amino acid at position 117 is lysine and the charged amino acid at position 141 is serine; or the charged amino acid at position 117 is lysine and the charged amino acid at position 141 is threonine;(ii) the charged amino acid at position 117 is arginine and the charged amino acid at position 185 is aspartic acid; the charged amino acid at position 117 is arginine and the charged amino acid at position 185 is glutamic acid; the charged amino acid at position 117 is arginine and the charged amino acid at position 185 is serine; the charged amino acid at position 117 is arginine and the charged amino acid at position 185 is threonine; the charged amino acid at position 117 is lysine and the charged amino acid at position 185 is aspartic acid; the charged amino acid at position 117 is lysine and the charged amino acid at position 185 is glutamic acid; the charged amino acid at position 117 is lysine and the charged amino acid at position 185 is serine; or the charged amino acid at position 117 is lysine and the charged amino acid at position 185 is threonine;(iii) the charged amino acid at position 119 is arginine and the charged amino acid at position 128 is aspartic acid; the charged amino acid at position 119 is arginine and the charged amino acid at position 128 is glutamic acid; the charged amino acid at position 119 is arginine and the charged amino acid at position 128 is serine; the charged amino acid at position 119 is arginine and the charged amino acid at position 128 is threonine; the charged amino acid at position 119 is lysine and the charged amino acid at position 128 is aspartic acid; the charged amino acid at position 119 is lysine and the charged amino acid at position 128 is glutamic acid; the charged amino acid at position 119 is lysine and the charged amino acid at position 128 is serine; or the charged amino acid at position 119 is lysine and the charged amino acid at position 128 is threonine;(iv) the charged amino acid at position 134 is arginine and the charged amino acid at position 128 is aspartic acid; the charged amino acid at position 134 is arginine and the charged amino acid at position 128 is glutamic acid; the charged amino acid at position 134 is arginine and the charged amino acid at position 128 is serine; the charged amino acid at position 134 is arginine and the charged amino acid at position 128 is threonine; the charged amino acid at position 134 is lysine and the charged amino acid at position 128 is aspartic acid; the chargedamino acid at position 134 is lysine and the charged amino acid at position 128 is glutamic acid; the charged amino acid at position 134 is lysine and the charged amino acid at position 128 is serine; or the charged amino acid at position 134 is lysine and the charged amino acid at position 128 is threonine;(v) the charged amino acid at position 134 is arginine and the charged amino acid at position 145 is aspartic acid; the charged amino acid at position 134 is arginine and the charged amino acid at position 145 is glutamic acid; the charged amino acid at position 134 is arginine and the charged amino acid at position 145 is serine; the charged amino acid at position 134 is arginine and the charged amino acid at position 145 is threonine; the charged amino acid at position 134 is lysine and the charged amino acid at position 145 is aspartic acid; the charged amino acid at position 134 is lysine and the charged amino acid at position 145 is glutamic acid; the charged amino acid at position 134 is lysine and the charged amino acid at position 145 is serine; or the charged amino acid at position 134 is lysine and the charged amino acid at position 145 is threonine;(vi) the charged amino acid at position 134 is arginine and the charged amino acid at position 183 is aspartic acid; the charged amino acid at position 134 is arginine and the charged amino acid at position 183 is glutamic acid; the charged amino acid at position 134 is arginine and the charged amino acid at position 183 is serine; the charged amino acid at position 134 is arginine and the charged amino acid at position 183 is threonine; the charged amino acid at position 134 is lysine and the charged amino acid at position 183 is aspartic acid; the charged amino acid at position 134 is lysine and the charged amino acid at position 183 is glutamic acid; the charged amino acid at position 134 is lysine and the charged amino acid at position 183 is serine; or the charged amino acid at position 134 is lysine and the charged amino acid at position 183 is threonine;(vii) the charged amino acid at position 136 is arginine and the charged amino acid at position 185 is aspartic acid; the charged amino acid at position 136 is arginine and the charged amino acid at position 185 is glutamic acid; the charged amino acid at position 136 is arginine and the charged amino acid at position 185 is serine; the charged amino acid at position 136 is arginine and the charged amino acid at position 185 is threonine; the chargedamino acid at position 136 is lysine and the charged amino acid at position 185 is aspartic acid; the charged amino acid at position 136 is lysine and the charged amino acid at position 185 is glutamic acid; the charged amino acid at position 136 is lysine and the charged amino acid at position 185 is serine; or the charged amino acid at position 136 is lysine and the charged amino acid at position 185 is threonine;(viii) the charged amino acid at position 178 is arginine and the charged amino acid at position 173 is aspartic acid; the charged amino acid at position 178 is arginine and the charged amino acid at position 173 is glutamic acid; the charged amino acid at position 178 is arginine and the charged amino acid at position 173 is serine; the charged amino acid at position 178 is arginine and the charged amino acid at position 173 is threonine; the charged amino acid at position 178 is lysine and the charged amino acid at position 173 is aspartic acid; the charged amino acid at position 178 is lysine and the charged amino acid at position 173 is glutamic acid; the charged amino acid at position 178 is lysine and the charged amino acid at position 173 is serine; or the charged amino acid at position 178 is lysine and the charged amino acid at position 173 is threonine; and / or(ix) the charged amino acid at position 117 is arginine and the charged amino acid at position 187 is aspartic acid; the charged amino acid at position 117 is arginine and the charged amino acid at position 187 is glutamic acid; the charged amino acid at position 117 is arginine and the charged amino acid at position 187 is serine; the charged amino acid at position 117 is arginine and the charged amino acid at position 187 is threonine; the charged amino acid at position 117 is lysine and the charged amino acid at position 187 is aspartic acid; the charged amino acid at position 117 is lysine and the charged amino acid at position 187 is glutamic acid; the charged amino acid at position 117 is lysine and the charged amino acid at position 187 is serine; or the charged amino acid at position 117 is lysine and the charged amino acid at position 187 is threonine.

12. The T cell engaging molecule or trivalent T cell engaging molecule according to claim 9, wherein the light chain constant domain of one or more of each of (a), (b), and (d) is a kappa light chain constant domain (CLK) and wherein the charge pair is a kappa charge pair comprising a positively charged amino acid residue selected from arginine, lysine orhistidine, and a negatively charged amino acid residue selected from aspartic acid, glutamic acid, serine or threonine, wherein the numbering is according to the EU index and wherein the kappa charge pair is located at position 133 in the CLK and position 183 in the CHI domain.

13. The T cell engaging molecule or trivalent T cell engaging molecule according to claim 12, wherein the charged amino acid at position 133 is glutamic acid and the charged amino acid at position 183 is lysine; or the charged amino acid at position 133 is lysine and the charged amino acid at position 183 is glutamic acid.

14. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 13, wherein the light chain constant domain of one of (a), (b), and (d) is a CLX and wherein the charge pair is a lambda charge pair and the light chain constant domain of a second and a third of (a), (b), and (d) is a CLX or a CLK and wherein the charge pair is a lambda or a kappa charge pair and the second of (a), (b), and (d) comprises a charged amino acid in the CHI domain of a same charge as a charged amino acid in the CHI domain of the third of (a), (b), and (d).

15. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 14, wherein the light chain constant domain of one of (a), (b), and (d) is a CLX and wherein the charge pair is a lambda charge pair and the light chain constant domain of a second and a third of (a), (b), and (d) is a kappa charge pair and the second of (a), (b), and (d) comprises a charged amino acid in the CHI domain of a same charge as a charged amino acid in the CHI domain of the third of (a), (b), and (d).

16. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 15, wherein the CHI domain of (a), (b), and / or (d) is capable of being linked to the light chain constant domain through an engineered disulfide link.

17. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 16, wherein the CHI domain of (b) and (d) is capable of being linked to the light chain constant domain through an engineered disulfide link and the CHI domain of (a) is linked to the light chain constant domain through a native disulfide link.

18. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claim 16 or 17, wherein the CHI domain that is capable of being linked to the light chain constant domain through an engineered disulfide link comprises(i) a substitution of a native cysteine to a non-cysteine amino acid,(ii) a substitution of a native non-cysteine amino acid to a cysteine; and the light chain constant domain comprises(i) a substitution of a native cysteine to a non-cysteine amino acid,(ii) a substitution of a native non-cysteine amino acid to a cysteine; and wherein the substituted cysteine of the light chain constant domain and the substituted cysteine of the CHI domain can form a disulfide bond.

19. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 16 to 18, wherein the CHI domain that is capable of being linked to the light chain constant domain through an engineered disulfide link comprises:(i) a substitution of a native cysteine to a non-cysteine amino acid at position 220,(ii) a substitution of a native non-cysteine amino acid to a cysteine at position 126; and the light chain constant domain comprises:(i) a substitution of a native cysteine to a non-cysteine amino acid at position 212,(ii) a substitution of a native non-cysteine amino acid to a cysteine at position 122, wherein the cysteine at position 126 of the CHI domain and the cysteine at position 122 of the light chain constant domain can form a disulfide bond; wherein the numbering is according to the EU index.

20. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 16 to 18 wherein the CHI domain that is capable of being linked to the light chain constant domain through an engineered disulfide link comprises:(i) a substitution of a native cysteine to a non-cysteine amino acid at position 220,(ii) a substitution of a native non-cysteine amino acid to a cysteine at position 126; and the light chain constant domain comprises:(i) a substitution of a native cysteine to a non-cysteine amino acid at position 214,(ii) a substitution of a native non-cysteine amino acid to a cysteine at position 121, wherein the cysteine at position 126 of the CHldomain and the cysteine at position 121 of the light chain constant domain can form a disulfide bond; wherein the numbering is according to the EU index.

21. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 20, wherein the antigen binding arm comprises a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 7, 15, 23, 95, 97, 99, 106, 114, 133, 141, and 149.

22. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 21, wherein the antigen binding arm is attached to the Fc domain and comprises a heavy chain constant region comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 66, 67, 69, 72, 73, 76, 80, 82, 88, and 91.

23. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 22, wherein the antigen binding arm comprises a VL comprising an amino acidsequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 8, 16, 24, 107, 115, 134, 142, and 150.

24. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 23, wherein the antigen binding arm comprises a light chain constant domain comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence set forth in SEQ ID NOs: 63.

25. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 24, wherein the VL of the T cell binding arm of the T cell engaging molecule or the VL of the first or second T cell binding arm of the trivalent T cell engaging molecule comprises a VL-CDR1 selected from SEQ ID NOs: 27 and 31; a VL-CDR2 selected from SEQ ID NOs: 28 and 32; and a VL-CDR3 selected from SEQ ID NOs: 29 and 33; and the VH comprises a VH-CDR1 selected from SEQ ID NOs: 36, 40, and 44; a VH-CDR2 selected from SEQ ID NOs: 37, 41 and 45; and a VH-CDR3 selected from SEQ ID NOs: 38, 42, and 46.

26. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 25, wherein the VL of the T cell binding arm of the T cell engaging molecule or the VL of the first or second T cell binding arm of the trivalent T cell engaging molecule comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 30 or 34.

27. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 26, wherein the light chain constant domain of the T cell engaging molecule or the light chain constant domain of the first or second T cell binding arm of the trivalent T cell engaging molecule comprises an amino acid sequence having at least about 80%, at leastabout 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 60 or 61.

28. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 27, wherein the VH of the T cell binding arm of the T cell engaging molecule or the VH of the first or second T cell binding arm of the trivalent T cell engaging molecule comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence set forth in SEQ ID NOs: 39, 43, or 47.

29. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 28, wherein the heavy chain of the T cell binding arm of the T cell engaging molecule or the first or second T cell binding arm of the trivalent T cell engaging molecule is attached to the Fc domain and comprises a heavy chain constant region comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID Nos: 66, 67, 69, 74, 75, 77, 78, and 79.

30. The trivalent T cell engaging molecule according to any one of claims 1 to 29, wherein the VH of the first or second T cell binding arm comprises a VH-CDR1 set forth in SEQ ID NO: 48; a VH-CDR2 set forth in SEQ ID NO: 49; a VH-CDR3 set forth in SEQ ID NO: 50; and the VL comprises a VL-CDR1 set forth in SEQ ID NO: 51; a VL-CDR2 set forth in SEQ ID NO: 52; and a VL-CDR3 set forth in SEQ ID NO: 53.

31. The trivalent T cell engaging molecule according to any one of claims 1 to 30, wherein the VL of the first or second T cell binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 54.

32. The trivalent T cell engaging molecule according to any one of claims 1 to 31, wherein the constant light chain domain of the first or second T cell binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 64.

33. The trivalent T cell engaging molecule according to any one of claims 1 to 32, wherein the VH of the first or second T cell binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 56.

34. The trivalent T cell engaging molecule according to any one of claims 1 to 33, wherein the heavy chain constant domain of the first or second T cell binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 57, 66, 67, 69, 74, 75, 77, 78, and 79.

35. The T cell engaging molecule or trivalent T cell engaging molecule according to any one of claims 1 to 34, wherein the antigen binding arm binds an epitope on an extracellular loop of STEAP2.

36. The T cell engaging molecule according to any one of claims 1 to 35, wherein the T cell engaging molecule comprises SEQ ID NOs: 25, 26, 35, 81 and 92.

37. The trivalent T cell engaging molecule according to any one of claims 1 to 35, wherein the trivalent T cell engaging molecule comprises SEQ ID NOs: 25, 26, 35, 55 and 58.

38. A trivalent T cell engaging molecule comprising:(a) a first antigen binding arm and a second antigen binding arm that each bind an epitope on human six transmembrane epithelial antigen of prostate-2 (STEAP2) and each comprise a heavy chain comprising a heavy chain variable domain (VH) comprising a variable heavy chaincomplementarity determining region 1 (VH-CDR1) selected from SEQ ID NOs: 1, 9, 17, 103, 111, 127, 135, and 143; a VH-CDR2 selected from SEQ ID NOs: 2, 10, 18, 104, 112, 128, 136, and 144; a VH-CDR3 selected from SEQ ID NOs: 3, 11, 19, 94, 96, 98, 105, 113, 129, 137, and 145; a heavy chain CHI domain; and a light chain comprising a light chain variable domain (VL) comprising a variable light chain complementarity determining region 1 (VL-CDR1) selected from SEQ ID NOs: 4, 12, 20, 100, 108, 130, 138, and 146; a VL-CDR2 selected from SEQ ID NOs: 5, 13, 21, 101, 109, 131, 139, and 147; a VL-CDR3 selected from SEQ ID NOs: 6, 14, 22, 102, 110, 132, 140, and 148; and a light chain constant domain;(b) a first T cell binding arm that binds to cluster of differentiation 3 (CD3) and comprises a heavy chain comprising a VH comprising a VH-CDR1 selected from SEQ ID NOs: 36, 40, and 44; a VH-CDR2 selected from SEQ ID NOs: 37, 41, and 45; a VH-CDR3 selected from SEQ ID NOs: 38, 42, and 46; and a heavy chain CHI domain; and a light chain comprising a VL comprising a VL-CDR1 selected from SEQ ID NOs: 27 and 31; a VL-CDR2 selected from SEQ ID NOs: 28 and 32; a VL-CDR3 selected from SEQ ID NOs: 29 and 33; and a light chain constant domain; and(c) an Fc domain, comprising a first Fc region and a second Fc region, each Fc region comprising a CH2 domain and a CH3 domain; said Fc domain further comprising at least one modification to promote heterodimerization; wherein the heavy chain of the first antigen binding arm and the heavy chain of the first T cell binding arm are attached to the Fc domain, and the heavy chain of the second antigen binding arm is attached to the heavy chain of the first T cell binding arm.

39. A tetraval ent T cell engaging molecule comprising the trivalent T cell engaging molecule according to claim 38, further comprising (d) a second T cell binding arm that binds to cluster of differentiation 8 (CD8) and comprises a heavy chain comprising a variable heavy domain (VHH) comprising a VH-CDR1 set forth in SEQ ID NO: 84; a VH-CDR2 set forth in SEQ ID NO: 85; and a VH-CDR3 set forth in SEQ ID NO: 86.

40. The tetravalent T cell engaging molecule according to claim 39, wherein the heavy chain of the second T cell binding arm is attached to the heavy chain of the first antigen binding arm.

41. The tetravalent T cell engaging molecule according to claim 39 or 40, wherein the heavy chain of the second antigen binding arm is attached to the heavy chain of the first T cell binding arm through a linker.

42. The tetravalent T cell engaging molecule according to any one of claims 39 or 41, wherein the heavy chain of the second T cell binding arm is attached to the heavy chain of the first antigen binding arm through a linker.

43. The trivalent or tetraval ent T cell engaging molecule according to claim 41 or 42, wherein the linker comprises the amino acid sequence of SEQ ID NO: 89.

44. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 41 to 43, wherein the linker comprises from 1 to about 10 copies of SEQ ID NO: 89.

45. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 41 to 44, wherein the linker comprises 2 copies of SEQ ID NO: 89.

46. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 45, wherein one of the two CH3 domains comprises a knob mutation, and the other of the two CH3 domains comprises a hole mutation.

47. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 46, wherein the CHI domain and the light chain constant domain of one or more of each of the (a) first antigen binding arm or second antigen binding arm, and (b) the first T cell binding arm, comprises a charge pair substitution comprising a first charged amino acid substitution in the CHI domain and a second charged amino acid substitution in the light chain constant domain, wherein the first charged amino acid substitution and the second charged amino acid substitution have an opposite charge.

48. The trivalent or tetravalent T cell engaging molecule according to claim 47, wherein the light chain constant domain of one or more of each of (a) the first antigen binding arm or the second antigen binding arm, and (b) the first T cell binding arm is a lambda light chain constant domain (CLX) and wherein the charge pair is a lambda charge pair comprising a positively charged amino acid residue selected from arginine, lysine or histidine, and anegatively charged amino acid residue selected from aspartic acid, glutamic acid, serine or threonine, wherein the numbering is according to the EU index and wherein the lambda charge pair is located at one or more of the following positions:(i) position 117 in the CLX. and position 141 in the CHI domain;(ii) position 117 in the CLX and position 185 in the CHI domain;(iii) position 119 in the CLX and position 128 in the CHI domain;(iv) position 134 in the CLX and position 128 in the CHI domain;(v) position 134 in the CLX and position 145 in the CHI domain;(vi) position 134 in the CLX and position 183 in the CHI domain;(vii) position 136 in the CLX and position 185 in the CHI domain;(viii) position 178 in the CLX and position 173 in the CHI domain; and(ix) position 117 in the CLX and position 187 in the CHI domain.

49. The trivalent or tetravalent T cell engaging molecule according to claim 48, wherein (i) the charged amino acid at position 117 is arginine and the charged amino acid at position 141 is aspartic acid; the charged amino acid at position 117 is arginine and the charged amino acid at position 141 is glutamic acid; the charged amino acid at position 117 is arginine and the charged amino acid at position 141 is serine; the charged amino acid at position 117 is arginine and the charged amino acid at position 141 is threonine; the charged amino acid at position 117 is lysine and the charged amino acid at position 141 is aspartic acid; the charged amino acid at position 117 is lysine and the charged amino acid at position 141 is glutamic acid; the charged amino acid at position 117 is lysine and the charged amino acid at position 141 is serine; or the charged amino acid at position 117 is lysine and the charged amino acid at position 141 is threonine;(ii) the charged amino acid at position 117 is arginine and the charged amino acid at position 185 is aspartic acid; the charged amino acid at position 117 is arginine and the charged amino acid at position 185 is glutamic acid; the charged amino acid at position 117 is arginine and the charged amino acid at position 185 is serine; the charged amino acid at position 117 is arginine and the charged amino acid at position 185 is threonine; the charged amino acid at position 117 is lysine and the charged amino acid at position 185 is aspartic acid; the charged amino acid at position 117 is lysine and the charged amino acid at position 185 is glutamic acid; the charged amino acid at position 117 is lysine and the charged amino acid at position 185 is serine; or the charged amino acid at position 117 is lysine and the charged amino acid at position 185 is threonine;(iii) the charged amino acid at position 119 is arginine and the charged amino acid at position 128 is aspartic acid; the charged amino acid at position 119 is arginine and the charged amino acid at position 128 is glutamic acid; the charged amino acid at position 119 is arginine and the charged amino acid at position 128 is serine; the charged amino acid at position 119 is arginine and the charged amino acid at position 128 is threonine; the charged amino acid at position 119 is lysine and the charged amino acid at position 128 is aspartic acid; the charged amino acid at position 119 is lysine and the charged amino acid at position 128 is glutamic acid; the charged amino acid at position 119 is lysine and the charged amino acid at position 128 is serine; or the charged amino acid at position 119 is lysine and the charged amino acid at position 128 is threonine;(iv) the charged amino acid at position 134 is arginine and the charged amino acid at position 128 is aspartic acid; the charged amino acid at position 134 is arginine and the charged amino acid at position 128 is glutamic acid; the charged amino acid at position 134 is arginine and the charged amino acid at position 128 is serine; the charged amino acid at position 134 is arginine and the charged amino acid at position 128 is threonine; the charged amino acid at position 134 is lysine and the charged amino acid at position 128 is aspartic acid; the charged amino acid at position 134 is lysine and the charged amino acid at position 128 is glutamic acid; the charged amino acid at position 134 is lysine and the charged amino acid at position 128 is serine; or the charged amino acid at position 134 is lysine and the charged amino acid at position 128 is threonine;(v) the charged amino acid at position 134 is arginine and the charged amino acid at position 145 is aspartic acid; the charged amino acid at position 134 is arginine and the charged amino acid at position 145 is glutamic acid; the charged amino acid at position 134 is arginine and the charged amino acid at position 145 is serine; the charged amino acid at position 134 is arginine and the charged amino acid at position 145 is threonine; the charged amino acid at position 134 is lysine and the charged amino acid at position 145 is aspartic acid; the charged amino acid at position 134 is lysine and the charged amino acid at position 145 is glutamic acid; the charged amino acid at position 134 is lysine and the charged amino acid at position 145 is serine; or the charged amino acid at position 134 is lysine and the charged amino acid at position 145 is threonine;(vi) the charged amino acid at position 134 is arginine and the charged amino acid at position 183 is aspartic acid; the charged amino acid at position 134 is arginine and the charged amino acid at position 183 is glutamic acid; the charged amino acid at position 134 is arginine and the charged amino acid at position 183 is serine; the charged amino acid at position 134 is arginine and the charged amino acid at position 183 is threonine; the charged amino acid at position 134 is lysine and the charged amino acid at position 183 is aspartic acid; the charged amino acid at position 134 is lysine and the charged amino acid at position 183 is glutamic acid; the charged amino acid at position 134 is lysine and the charged amino acid at position 183 is serine; or the charged amino acid at position 134 is lysine and the charged amino acid at position 183 is threonine;(vii) the charged amino acid at position 136 is arginine and the charged amino acid at position 185 is aspartic acid; the charged amino acid at position 136 is arginine and the charged amino acid at position 185 is glutamic acid; the charged amino acid at position 136 is arginine and the charged amino acid at position 185 is serine; the charged amino acid at position 136 is arginine and the charged amino acid at position 185 is threonine; the charged amino acid at position 136 is lysine and the charged amino acid at position 185 is aspartic acid; the charged amino acid at position 136 is lysine and the charged amino acid at position 185 is glutamic acid; the charged amino acid at position 136 is lysine and the charged amino acid at position 185 is serine; or the charged amino acid at position 136 is lysine and the charged amino acid at position 185 is threonine;(viii) the charged amino acid at position 178 is arginine and the charged amino acid at position 173 is aspartic acid; the charged amino acid at position 178 is arginine and the charged amino acid at position 173 is glutamic acid; the charged amino acid at position 178 is arginine and the charged amino acid at position 173 is serine; the charged amino acid at position 178 is arginine and the charged amino acid at position 173 is threonine; the charged amino acid at position 178 is lysine and the charged amino acid at position 173 is aspartic acid; the charged amino acid at position 178 is lysine and the charged amino acid at position 173 is glutamic acid; the charged amino acid at position 178 is lysine and the charged amino acid at position 173 is serine; or the charged amino acid at position 178 is lysine and the charged amino acid at position 173 is threonine; and / or(ix) the charged amino acid at position 117 is arginine and the charged amino acid at position 187 is aspartic acid; the charged amino acid at position 117 is arginine and the charged amino acid at position 187 is glutamic acid; the charged amino acid at position 117 is arginine and the charged amino acid at position 187 is serine; the charged amino acid at position 117 is arginine and the charged amino acid at position 187 is threonine; the charged amino acid at position 117 is lysine and the charged amino acid at position 187 is aspartic acid; the charged amino acid at position 117 is lysine and the charged amino acid at position 187 is glutamic acid; the charged amino acid at position 117 is lysine and the charged amino acid at position 187 is serine; or the charged amino acid at position 117 is lysine and the charged amino acid at position 187 is threonine.

50. The trivalent or tetravalent T cell engaging molecule according to any one of claims 47 to 49, wherein the light chain constant domain of one or more of each of (a) the first antigen binding arm or the second antigen binding arm, and (b) the first T cell binding arm are kappa light chain constant domains (CLK) and wherein the charge pair is a kappa charge pair comprising a positively charged amino acid residue selected from arginine, lysine or histidine, and a negatively charged amino acid residue selected from aspartic acid, glutamic acid, serine or threonine, wherein the numbering is according to the EU index and wherein the kappa charge pair is located at position 133 in the CLK and position 183 in the CHI.

51. The trivalent or tetraval ent T cell engaging molecule according to claim 50, wherein the charged amino acid at position 133 is glutamic acid and the charged amino acid at position 183 is lysine; or the charged amino acid at position 133 is lysine and the charge amino acid at position 183 is glutamic acid.

52. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 51 wherein the light chain constant domain of one of (a), (b), and (d) is a CLX and wherein the charge pair is a lambda charge pair and the light chain constant domain of a second and a third of (a), (b), and (d) is a CLX or a CLK and wherein the charge pair is a lambda or a kappa charge pair and the second of (a), (b), and (d) comprises a charged amino acid in the CHI domain of a same charge as a charged amino acid in the CHI domain of the third of (a), (b), and (d).

53. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 51, wherein the light chain constant domain of one of (a), (b), and (d) is a CLX and wherein the charge pair is a lambda charge pair and the light chain constant domain of a second and a third of (a), (b), and (d) is a CLK and wherein the charge pair is a kappa charge pair and the second of (a), (b), and (d) comprises a charged amino acid in the CHI domain of a same charge as a charged amino acid in the CHI domain of the third of (a), (b), and (d).

54. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 53, wherein the CHI domain of (a) and / or (b) of the trivalent T cell engaging molecule or the CHI domain of (a), (b), and / or (d) of the tetravalent T cell engaging molecule is capable of forming a disulfide link to the light chain constant domain of (a) and / or (b) of the trivalent T cell engaging molecule or the light chain of (a), (b), and / or (d) of the tetravalent T cell engaging molecule through an engineered disulfide link.

55. The trivalent T cell engaging molecule according to any one of claims 38 to 54, wherein the CHI domain of (b) is capable of forming a disulfide link to the light chain constant domain through an engineered disulfide link and the CHI domain of (a) is linked to the light chain constant domain through a native disulfide link.

56. The tetravalent T cell engaging molecule according to any one of claims 38 to 54, wherein the CHI domain of (b) and (d) are capable of forming a disulfide link to the light chain constant domain of (b) and (d) through an engineered disulfide link and the CHI domain of (a) is linked to the light chain constant domain of (a) through a native disulfide link.

57. The trivalent or tetravalent T cell engaging molecule according to any one of claims 38 to 56, wherein CHI domain of the first T cell binding arm comprises(i) a substitution of a native cysteine to a non-cysteine amino acid,(ii) a substitution of a native non-cysteine amino acid to a cysteine; and the light chain constant domain of the second T cell binding arm comprises:(i) a substitution of a native cysteine to a non-cysteine amino acid,(ii) a substitution of a native non-cysteine amino acid to a cysteine; wherein the substituted cysteine of the CHI domain of the first T cell binding arm and the substituted cysteine of the light chain constant domain of the first T cell binding arm can form a disulfide bond.

58. The trivalent or tetravalent T cell engaging molecule according to claim 57, wherein the CHI domain comprises:(i) a substitution of a native cysteine to a non-cysteine amino acid at position 220,(ii) a substitution of a native non-cysteine amino acid to a cysteine at position 126; and the light chain constant domain comprises:(i) a substitution of a native cysteine to a non-cysteine amino acid at position 212,(ii) a substitution of a native non-cysteine amino acid to a cysteine at position 122,wherein the cysteine at position 126 of the CHI domain and the cysteine at position 122 of the light chain constant domain can form a disulfide bond; wherein the numbering is according to the EU index.

59. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 58, wherein the light chains of the first and second antigen binding arms comprise a native cysteine and the CHI domain of the first and second antigen binding arms comprise a native cysteine, wherein the native cysteine of the CHI domain of the first antigen binding arm and the native cysteine of the light chain constant domain of the first antigen binding arm can form a disulfide bond and the native cysteine of the CHI of the second antigen binding arm and the native cysteine of the light chain constant domain of the second antigen binding arm can form a disulfide bond.

60. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 59, wherein the VH of the first and / or second antigen binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 7, 15, 23, 95, 97, 99, 106, 114, 133, 141, and 149.

61. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 59 wherein the heavy chain of the first and / or second antigen binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 67 and 93.

62. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 60, wherein the VL of the first and / or second antigen binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 8, 16, 24, 107, 115, 134, 142, and 150.

63. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 61, wherein the light chain constant domain of the first and / or second antigen binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 63.

64. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 62, wherein the VL of the first T cell binding arm comprises a VL-CDR1 set forth in SEQ ID NO: 27 or 31; a VL-CDR2 set forth in SEQ ID NO: 28 or 32; a VL-CDR3 set forth in SEQ ID NO: 29 or 33; and the VH of the first T cell binding arm comprises a VH-CDR1 set forth in SEQ ID NO: 36, 40, or 44; a VH-CDR2 set forth in SEQ ID NO: 37, 41 or 45; and a VH- CDR3 set forth in SEQ ID NO: 38, 42, or 46.

65. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 63, wherein the VL of the first T cell binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 30 or 34.

66. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 64, wherein the VH of the first T cell binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 39, 43, and 47.

67. The tetravalent T cell engaging molecule according to any one of claims 38 to 65, wherein the VH of the second T cell binding arm comprises a VH-CDR1 set forth in SEQ ID NO: 84; a VH-CDR2 set forth in SEQ ID NO: 85; and a VH-CDR3 set forth in SEQ ID NO: 86.

68. The tetravalent T cell engaging molecule according to any one of claims 38 to 66, wherein the VH of the second T cell binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, atleast about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 56 or 83.

69. A tetravalent T cell engaging molecule comprising the trivalent T cell engaging molecule according to claim 38, further comprising (d) a second T cell binding arm.

70. The tetravalent T cell engaging molecule according to claim 69, wherein the second T cell binding arm comprises a VL comprising a VL-CDR1 set forth in SEQ ID NO: 51; a VL- CDR2 set forth in SEQ ID NO: 52; and a VL-CDR3 set forth in SEQ ID NO: 53, and a VH comprising a VH-CDR1 set forth in SEQ ID NO: 48; a VH-CDR2 set forth in SEQ ID NO: 49; and a VH-CDR3 set forth in SEQ ID NO: 50.

71. The tetravalent T cell engaging molecule according to claim 70, wherein the VL of the second T cell binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 54, and wherein the VH of the second T cell binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 56.

72. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 70, wherein the first and the second antigen binding arm bind an epitope on an extracellular loop of STEAP2.

73. The trivalent or tetraval ent T cell engaging molecule according to any one of claims 38 to 71, wherein the heavy chain comprising the heavy chain constant region of the second antigen binding arm and the first T cell binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 81.

74. The tetravalent T cell engaging molecule according to any one of claims 38 to 67, 71 and 72, wherein the heavy chain comprising the heavy chain constant region of the first antigen binding arm and the second T cell binding arm comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 87.

75. The trivalent T cell engaging molecule according to any one of claims 38 to 65, 71, and 72, wherein the trivalent T cell engaging molecule comprises SEQ ID NOs: 25, 35, 81, 25, and 93.

76. The tetravalent T cell engaging molecule according to any one of claims 39 to 67, 71, and 73, wherein the tetravalent T cell engaging molecule comprises SEQ ID NOs: 25, 35, 81, 25, and 87, or wherein the tetravalent T cell engaging molecule comprises SEQ ID NOs: 151, 152, 153, 154, and 152.

77. One or more nucleic acid(s) encoding the T cell engaging molecule of any one of claims 1- 76.

78. A vector comprising the nucleic acid(s) of claim 77.

79. An isolated host cell comprising the nucleic acid(s) of claim 77, or the vector of claim 78.

80. A pharmaceutical composition comprising the T cell engaging molecule according to any one of claims 1 to 76 and a pharmaceutically acceptable carrier.

81. A method of treating a disease in a patient in need thereof, the method comprising administering to the patient an effective amount of the T cell engaging molecule according to any one of claims 1 to 76, or the pharmaceutical composition according to claim 80.

82. The method of claim 81, wherein the disease is cancer or prostate cancer.

83. The T cell engaging molecule according to any one of claims 1 to 76, or pharmaceutical composition according to claim 80, for use as a medicament.

84. The T cell engaging molecule according to any one of claims 1 to 76, or pharmaceutical composition according to claim 80, for use in the treatment of cancer.

85. Use of the T cell engaging molecule according to any one of claims 1 to 76, or pharmaceutical composition according to claim 80, for the manufacture of a medicament for the treatment of cancer.

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