Mesothelin-targeted CD40 agonistic multispecific antibody constructs for the treatment of solid tumors

1. An antigen binding protein that specifically binds and agonizes human CD40 (SEQ ID NO: 1), the antigen binding protein comprising a light chain variable region (VL) and a heavy chain variable region (VH), wherein

the VL and VH are selected from the group consisting of:

a) a VL comprising a CDRL1, a CDRL2, and a CDRL3 selected from the group consisting of SEQ ID NOs: 58, 59, and 60, respectively; and a VH comprising a CDRH1, a CDRH2, and a CDRH3 selected from the group consisting of SEQ ID NOs: 136, 137, and 138, respectively;

b) a VL comprising a CDRL1, a CDRL2, and a CDRL3 selected from the group consisting of SEQ ID NOs: 64, 65, and 66, respectively; and a VH comprising a CDRH1, a CDRH2, and a CDRH3 selected from the group consisting of SEQ ID NOs: 142, 143, and 144, respectively;

c) a VL comprising a CDRL1, a CDRL2, and a CDRL3 selected from the group consisting of SEQ ID NOs: 70, 71, and 72, respectively; and a VH comprising a CDRH1, a CDRH2, and a CDRH3 selected from the group consisting of SEQ ID NOs: 148, 149, and 150, respectively;

d) a VL comprising a CDRL1, a CDRL2, and a CDRL3 selected from the group consisting of SEQ ID NOs: 76, 77, and 78, respectively; and a VH comprising a CDRH1, a CDRH2, and a CDRH3 selected from the group consisting of SEQ ID NOs: 154, 155, and 156, respectively;

e) a VL comprising a CDRL1, a CDRL2, and a CDRL3 selected from the group consisting of SEQ ID NOs: 82, 83, and 84, respectively; and a VH comprising a CDRH1, a CDRH2, and a CDRH3 selected from the group consisting of SEQ ID NOs: 160, 161, and 162, respectively;

f) a VL comprising a CDRL1, a CDRL2, and a CDRL3 selected from the group consisting of SEQ ID NOs: 88, 89, and 90, respectively; and a VH comprising a CDRH1, a CDRH2, and a CDRH3 selected from the group consisting of SEQ ID NOs: 166, 167, and 168, respectively;

g) a VL comprising a CDRL1, a CDRL2, and a CDRL3 selected from the group consisting of SEQ ID NOs: 94, 95, and 96, respectively; and a VH comprising a CDRH1, a CDRH2, and a CDRH3 selected from the group consisting of SEQ ID NOs: 172, 173, and 174, respectively;

h) a VL comprising a CDRL1, a CDRL2, and a CDRL3 selected from the group consisting of SEQ ID NOs: 100, 101, and 102, respectively; and a VH comprising a CDRH1, a CDRH2, and a CDRH3 selected from the group consisting of SEQ ID NOs: 178, 179, and 180, respectively;

i) a VL comprising a CDRL1, a CDRL2, and a CDRL3 selected from the group consisting of SEQ ID NOs: 106, 107, and 108, respectively; and a VH comprising a CDRH1, a CDRH2, and a CDRH3 selected from the group consisting of SEQ ID NOs: 184, 185, and 186, respectively;

j) a VL comprising a CDRL1, a CDRL2, and a CDRL3 selected from the group consisting of SEQ ID NOs: 112, 113, and 114, respectively; and a VH comprising a CDRH1, a CDRH2, and a CDRH3 selected from the group consisting of SEQ ID NOs: 190, 191, and 192, respectively;

k) a VL comprising a CDRL1, a CDRL2, and a CDRL3 selected from the group consisting of SEQ ID NOs: 118, 119, and 120, respectively; and a VH comprising a CDRH1, a CDRH2, and a CDRH3 selected from the group consisting of SEQ ID NOs: 196, 197, and 198, respectively;

l) a VL comprising a CDRL1, a CDRL2, and a CDRL3 selected from the group consisting of SEQ ID NOs: 124, 125, and 126, respectively; and a VH comprising a CDRH1, a CDRH2, and a CDRH3 selected from the group consisting of SEQ ID NOs: 202, 203, and 204, respectively;

m) a VL comprising a CDRL1, a CDRL2, and a CDRL3 selected from the group consisting of SEQ ID NOs: 130, 131, and 132, respectively; and a VH comprising a CDRH1, a CDRH2, and a CDRH3 selected from the group consisting of SEQ ID NOs: 208, 209, and 210, respectively; and

n) a VL comprising a CDRL1 comprising SEQ ID NO:100, a CDRL2 comprising SEQ ID NO:101, and a CDRL3 comprising the CDRL3 of SEQ ID NO: 370; and a VH comprising a CDRH1 comprising SEQ ID NO:178, a CDRH2 comprising SEQ ID NO: 179, and a CDRH3 comprising SEQ ID NO: 180.

2. The antigen binding protein according to claim 1 , wherein: the VL and VH are selected from the group consisting of:

a) a VL comprising SEQ ID NO: 5 and a VH comprising SEQ ID NO: 6;

b) a VL comprising SEQ ID NO: 9 and a VH comprising SEQ ID NO: 10;

c) a VL comprising SEQ ID NO: 13 and a VH comprising SEQ ID NO: 14;

d) a VL comprising SEQ ID NO: 17 and a VH comprising SEQ ID NO: 18;

e) a VL comprising SEQ ID NO: 21 and a VH comprising SEQ ID NO: 22;

f) a VL comprising SEQ ID NO: 25 and a VH comprising SEQ ID NO: 26;

g) a VL comprising SEQ ID NO: 29 and a VH comprising SEQ ID NO: 30;

h) a VL comprising SEQ ID NO: 33 and a VH comprising SEQ ID NO: 34;

i) a VL comprising SEQ ID NO: 37 and a VH comprising SEQ ID NO: 38;

j) a VL comprising SEQ ID NO: 41 and a VH comprising SEQ ID NO: 42;

k) a VL comprising SEQ ID NO: 45 and a VH comprising SEQ ID NO: 46;

l) a VL comprising SEQ ID NO: 49 and a VH comprising SEQ ID NO: 50;

m) a VL comprising SEQ ID NO: 53 and a VH comprising SEQ ID NO: 54; and

n) a VL comprising the VL sequence of SEQ ID NO: 370 and a VH comprising SEQ ID NO: 34.

3. The antigen binding protein according to claim 1 , wherein the heavy chain comprises an amino acid substitution selected from the group consisting of:

(i) N297G or N297A;

(ii) L234A and L235A; and

(iii) R292C and V302C;

wherein the amino acid numbering is EU numbering according to Kabat.

4. The antigen binding protein according to claim 3 , wherein the heavy chain comprises N297G, R292C, and V302C mutations, wherein the amino acid numbering is EU numbering according to Kabat.

5. The antigen binding protein according to claim 1 , wherein the antigen binding protein further comprises a light chain constant domain (CL) polypeptide linked to the VL, wherein the CL polypeptide is selected from the group consisting of SEQ ID NO: 883 and SEQ ID NO: 884.

6. The antigen binding protein according to claim 1 , wherein the antigen binding protein further comprises a heavy chain CH1-hinge-CH2-CH3 polypeptide wherein the CH1-hinge-CH2-CH3 polypeptide is selected from the group consisting of SEQ ID NO: 885 and SEQ ID NO: 886.

7. The antigen binding protein according to claim 1 , wherein said VH comprises a human framework (FR) sequence, and said VL comprises a human framework (FR) sequence.

8. The antigen binding protein according to claim 1 , wherein said antigen binding protein comprises a human constant domain.

9. The antigen binding protein according to claim 1 , wherein said antigen binding protein comprises a human light chain constant domain (CL).

10. The antigen binding protein according to claim 9 , wherein said CL is a kappa CL or lambda CL.

11. The antigen binding protein according to claim 1 , wherein said antigen binding protein comprises a human heavy chain constant domain CH1.

12. The antigen binding protein according to claim 11 , wherein said CH1 is an IgG CH1.

13. The antigen binding protein according to claim 11 , wherein said CH1 is an IgG1 CH1.

14. The antigen binding protein according to claim 1 , wherein said antigen binding protein comprises a human heavy chain Fc domain.

15. The antigen binding protein according to claim 14 , wherein said Fc is an IgG Fc.

16. The antigen binding protein according to claim 14 , wherein said Fc is an IgG1 Fc.

17. The antigen binding protein according to claim 1 , comprising a VH and a VL, wherein said VL comprises a CDRL1 comprising SEQ ID NO:100, a CDRL2 comprising SEQ ID NO:101, and a CDRL3 comprising the CDRL3 of SEQ ID NO: 370; and said VH comprises a CDRH1 comprising SEQ ID NO:178, a CDRH2 comprising SEQ ID NO: 179, and a CDRH3 comprising SEQ ID NO: 180.

18. The antigen binding protein according to claim 1 , comprising a VH that comprises SEQ ID NO: 34, and a VL that comprises the VL sequence of SEQ ID NO: 370.

19. A fusion protein comprising the VL and VH of claim 1 .

20. An antigen binding protein that specifically binds human CD40 (SEQ ID NO: 1), wherein said antigen binding protein comprises:

(i) a light chain variable region (VL) that comprises a CDRL1 comprising SEQ ID NO:100, a CDRL2 comprising SEQ ID NO:101, and a CDRL3 comprising amino acid residues 91 to 101 of SEQ ID NO: 370; and

(ii) a heavy chain variable region (VH) that comprises a CDRH1 comprising SEQ ID NO:178, a CDRH2 comprising SEQ ID NO: 179, and a CDRH3 comprising SEQ ID NO: 180.

21. The antigen binding protein of claim 20 , wherein said VL comprises amino acid residues 1 to 112 of SEQ IDNO:370, and said VH comprises SEQ ID NO:34.

22. The antigen binding protein according to claim 20 , wherein the heavy chain comprises an amino acid substitution selected from the group consisting of:

(i) N297G or N297A;

(ii) L234A and L235A; and

(iii) R292C and V302C;

wherein the amino acid numbering is EU numbering according to Kabat.

23. The antigen binding protein according to claim 20 , wherein the heavy chain comprises N297G, R292C, and V302C mutations, wherein the amino acid numbering is EU numbering according to Kabat.

24. The antigen binding protein according to claim 20 , wherein said VH comprises a human framework (FR) sequence, and said VL comprises a human framework (FR) sequence.

25. The antigen binding protein according to claim 20 , wherein said antigen binding protein comprises a human constant domain.

26. The antigen binding protein according to claim 20 , wherein said antigen binding protein comprises a human light chain constant domain (CL).

27. The antigen binding protein according to claim 26 , wherein said CL is a kappa CL or lambda CL.

28. The antigen binding protein according to claim 20 , wherein said antigen binding protein comprises a human heavy chain constant domain CH1.

29. The antigen binding protein according to claim 28 , wherein said CH1 is an IgG CH1.

30. The antigen binding protein according to claim 28 , wherein said CH1 is an IgG1 CH1.

31. The antigen binding protein according to claim 20 , wherein said antigen binding protein comprises a human heavy chain Fc domain.

32. The antigen binding protein according to claim 31 , wherein said Fc is an IgG Fc.

33. The antigen binding protein according to claim 31 , wherein said Fc is an IgG1 Fc.

34. A fusion protein comprising the VL and VH of claim 20 .

35. The fusion protein of claim 34 , comprising a heavy chain fusion protein that comprises a Fab fragment, a single-chain FIT (scFv), or both.