BISPECIFIC ANTIBODIES SPECIFIC FOR A COSTIMULATORY TNF RECEPTOR

• US 20170114141A1
• Filed: 09/29/2016
• Published: 04/27/2017
• Est. Priority Date: 10/02/2015
• Status: Active Grant

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: AMANN, MARIA, BRUENKER, PETER, CLAUS, CHRISTINA, FERRARA KOLLER, CLAUDIA, GRAU-RICHARDS, SANDRA, HOSSE, RALF, KLEIN, CHRISTIAN, LEVITSKI, VIKTOR, UMANA, PABLO
• Granted Patent: US 10,526,413 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 16/2878 against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/3053 Skin, nerves, brain
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C12Y 304/14005 Dipeptidyl-peptidase IV (3.4.14.5)

Abstract

The invention relates to novel bispecific antigen binding molecules, comprising (a) at least one moiety capable of specific binding to a target cell antigen, (b) at least one moiety capable of specific binding to a costimulatory TNF receptor family member, and (c) a Fc domain composed of a first and a second subunit capable of stable association, and to methods of producing these molecules and to methods of using the same.

Claims

1. A bispecific antigen binding molecule, comprising

(a) at least one moiety capable of specific binding to a costimulatory TNF receptor family member,

(b) at least one moiety capable of specific binding to a target cell antigen, and

(c) a Fc domain composed of a first and a second subunit capable of stable association.

2. The bispecific antigen binding molecule of claim 1, wherein the costimulatory TNF receptor family member is selected from the group consisting of OX40 and 4-1BB.

3. The bispecific antigen binding molecule of claim 1, wherein the costimulatory TNF receptor family member is OX40.

4. The bispecific antigen binding molecule of claim 1, wherein the moiety capable of specific binding to a costimulatory TNF receptor family member binds to a polypeptide comprising the amino acid sequence of SEQ ID NO:

1.

5. The bispecific antigen binding molecule of claim 1, comprising at least one moiety capable of specific binding to OX40, wherein said moiety comprises a VH domain comprising

(i) a CDR-H1 comprising the amino acid sequence selected from the group consisting of SEQ ID NO:

2 and SEQ ID NO;

3,

(ii) a CDR-H2 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

4 and SEQ ID NO;

5, and

(iii) a CDR-H3 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11 and SEQ ID NO;

12,

and a VL domain comprising

(iv) a CDR-L1 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

13, SEQ ID NO;

14 and SEQ ID NO;

15,

(v) a CDR-L2 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

16, SEQ ID NO;

17 and SEQ ID NO;

18, and

(vi) a CDR-L3 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

19, SEQ ID NO;

20, SEQ ID NO;

21, SEQ ID NO;

22, SEQ ID NO;

23 and SEQ ID NO;

24.

6. The bispecific antigen binding molecule of claim 3, wherein the moiety capable of specific binding to OX40 comprises a heavy chain variable region VH comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:

25, SEQ ID NO;

27, SEQ ID NO;

29, SEQ ID NO;

31, SEQ ID NO;

33, SEQ ID NO;

35 and SEQ ID NO;

37 and a light chain variable region VL comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence of SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

30, SEQ ID NO;

32, SEQ ID NO;

34, SEQ ID NO;

36 and SEQ ID NO;

38.

7. The bispecific antigen binding molecule of claim 3, wherein the moiety capable of specific binding to OX40 comprises

(i) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO:

25 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

26,

(ii) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

27 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

28,

(iii) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

29 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

30,

(iv) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

31 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

32,

(v) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

33 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

34,

(vi) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

35 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

36, or

(vii) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

37 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

38.

8. The bispecific antigen binding molecule of claim 1, wherein the costimulatory TNF receptor family member is 4-1BB.

9. The bispecific antigen binding molecule of claim 8, wherein the moiety capable of specific binding to a costimulatory TNF receptor family member binds to a polypeptide comprising the amino acid sequence of SEQ ID NO:

39.

10. The bispecific antigen binding molecule of claim 8, comprising at least one moiety capable of specific binding to 4-1BB, wherein said moiety comprises a VH domain comprising

(i) a CDR-H1 comprising the amino acid sequence selected from the group consisting of SEQ ID NO:

40 and SEQ ID NO;

41,

(ii) a CDR-H2 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

42 and SEQ ID NO;

43, and

(iii) a CDR-H3 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

44, SEQ ID NO;

45, SEQ ID NO;

46, SEQ ID NO;

47 and SEQ ID NO;

48

and a VL domain comprising

(iv) a CDR-L1 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

49 and SEQ ID NO;

50,

(v) a CDR-L2 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

51 and SEQ ID NO;

52, and

(vi) a CDR-L3 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

53, SEQ ID NO;

54, SEQ ID NO;

55, SEQ ID NO;

56 and SEQ ID NO;

57.

11. The bispecific antigen binding molecule of claim 8, wherein the moiety capable of specific binding to 4-1BB comprises a heavy chain variable region VH comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:

58, SEQ ID NO;

60, SEQ ID NO;

62, SEQ ID NO;

64 and SEQ ID NO;

66 and a light chain variable region comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NO;

59, SEQ ID NO;

61, SEQ ID NO;

63, SEQ ID NO;

65 and SEQ ID NO;

67.

12. The bispecific antigen binding molecule of claim 8, wherein the moiety capable of specific binding to 4-1BB comprises

(i) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO:

58 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

59,

(ii) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

60 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

61,

(iii) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

62 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

63,

(iv) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

64 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

65, or

(v) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

66 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

67.

13. The bispecific antigen binding molecule of claim 12, wherein the moiety capable of specific binding to a costimulatory TNF receptor family member is a Fab fragment.

14. The bispecific antigen binding molecule of claim 13, wherein the target cell antigen is selected from the group consisting of Fibroblast Activation Protein (FAP), Melanoma-associated Chondroitin Sulfate Proteoglycan (MCSP), Epidermal Growth Factor Receptor (EGFR), Carcinoembryonic Antigen (CEA), CD19, CD20 and CD33.

15. The bispecific antigen binding molecule of claim 3, wherein the target cell antigen is Fibroblast Activation Protein (FAP).

16. The bispecific antigen binding molecule of claim 15, wherein the moiety capable of specific binding to FAP comprises a VH domain comprising

(i) a CDR-H1 comprising the amino acid sequence selected from the group consisting of SEQ ID NO:

68 and SEQ ID NO;

69,

(ii) a CDR-H2 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

70 and SEQ ID NO;

71, and

(iii) a CDR-H3 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

72 and SEQ ID NO;

73,

and a VL domain comprising

(iv) a CDR-L1 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

74 and SEQ ID NO;

75,

(v) a CDR-L2 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

76 and SEQ ID NO;

77, and

(vi) a CDR-L3 comprising the amino acid sequence selected from the group consisting of SEQ ID NO;

78 and SEQ ID NO;

79.

17. The bispecific antigen binding molecule of claim 15, wherein

(i) the moiety capable of specific binding to OX40 comprises a heavy chain variable region VH comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:

25, SEQ ID NO;

27, SEQ ID NO;

29, SEQ ID NO;

31, SEQ ID NO;

33, SEQ ID NO;

35 or SEQ ID NO;

37 and a light chain variable region VL comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

32, SEQ ID NO;

34, SEQ ID NO;

36 or SEQ ID NO;

38 and

(ii) the moiety capable of specific binding to FAP comprises a heavy chain variable region VH comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO;

80 or SEQ ID NO;

82 and a light chain variable region VL comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO;

81 or SEQ ID NO;

83.

18. The bispecific antigen binding molecule of claim 15, wherein

(i) the moiety capable of specific binding to OX40 comprises a heavy chain variable region VH comprising the amino acid sequence of SEQ ID NO:

27 and a light chain variable region comprising an amino acid sequence of SEQ ID NO;

28 and

(ii) the moiety capable of specific binding to FAP comprises a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

82 and a light chain variable region comprising an amino acid sequence of SEQ ID NO;

83.

19. The bispecific antigen binding molecule of claim 8, wherein

(i) the moiety capable of specific binding to 4-1BB comprises a heavy chain variable region VH comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:

58, SEQ ID NO;

60, SEQ ID NO;

62, SEQ ID NO;

64 or SEQ ID NO;

66 and a light chain variable region comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO;

59, SEQ ID NO;

61, SEQ ID NO;

63, SEQ ID NO;

65 or SEQ ID NO;

67 and

(ii) the moiety capable of specific binding to FAP comprises a heavy chain variable region VH comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO;

80 or SEQ ID NO;

82 and a light chain variable region comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO;

81 or SEQ ID NO;

83.

20. The bispecific antigen binding molecule of claim 1, wherein said molecule comprises

(a) a first Fab fragment capable of specific binding to a costimulatory TNF receptor family member,

(b) a second Fab fragment capable of specific binding to a target cell antigen, and

(c) a Fc domain composed of a first and a second subunit capable of stable association.

21. The bispecific antigen binding molecule of claim 1, wherein the Fc domain is an IgG1 Fc domain or an IgG4 Fc domain.

22. The bispecific antigen binding molecule of claim 1, wherein the Fc domain comprises one or more amino acid substitution that reduces the binding affinity of the antibody to an Fc receptor and/or effector function.

23. The bispecific antigen binding molecule of claim 21, wherein the Fc domain is of human IgG1 subclass with the amino acid mutations L234A, L235A and P329G (numbering according to Kabat EU index).

24. The bispecific antigen binding molecule of claim 21, wherein the Fc domain comprises a modification promoting the association of the first and second subunit of the Fc domain.

25. The bispecific antigen binding molecule of claim 24, wherein the first subunit of the Fc domain comprises knobs and the second subunit of the Fc domain comprises holes according to the knobs into holes method.

26. The bispecific antibody of claim 25, wherein the first subunit of the Fc domain comprises the amino acid substitutions S354C and T366W (numbering according to Kabat EU index) and the second subunit of the Fc domain comprises the amino acid substitutions Y349C, T366S and Y407V (numbering according to Kabat EU index).

27. The bispecific antigen binding molecule of claim 1, comprising

(a) two moieties capable of specific binding to a costimulatory TNF receptor family member,

(b) two moieties capable of specific binding to a target cell antigen,

and

(c) a Fc domain composed of a first and a second subunit capable of stable association.

28. The bispecific antigen binding molecule of claim 27, wherein the bispecific antigen binding molecule is bivalent both for the costimulatory TNF receptor family member and for the target cell antigen.

29. The bispecific antigen binding molecule of claim 1, comprising

(a) two light chains and two heavy chains of an antibody comprising two Fab fragments capable of specific binding to a costimulatory TNF receptor family member and the Fc domain, and

(b) two additional Fab fragments capable of specific binding to a target cell antigen, wherein said additional Fab fragments are each connected via a peptide linker to the C-terminus of the heavy chains of (a).

30. The bispecific antigen binding molecule of claim 29, wherein the two additional Fab fragments capable of specific binding to a target cell antigen are crossover Fab fragments wherein the variable domains VL and VH are replaced by each other and the VL-CH chains are each connected via a peptide linker to the C-terminus of the heavy chains of (a).

31. The bispecific antigen binding molecule of claim 30, wherein the two Fab fragments capable of specific binding to a costimulatory TNF receptor family member are two Fab fragments capable of specific binding to OX40 or 4-1BB and the two additional Fab fragments capable of specific binding to a target cell antigen are crossover Fab fragments capable of specific binding to FAP.

32. The bispecific antigen binding molecule of claim 1, comprising

(a) two moieties capable of specific binding to a costimulatory TNF receptor family member,

(b) one moiety capable of specific binding to a target cell antigen,

and

(c) a Fc domain composed of a first and a second subunit capable of stable association.

33. The bispecific antigen binding molecule of claim 32, wherein the bispecific antigen binding molecule is bivalent for the costimulatory TNF receptor family member and monovalent for the target cell antigen.

34. The bispecific antigen binding molecule of claim 1, comprising

(a) two light chains and two heavy chains of an antibody comprising two Fab fragments capable of specific binding to a costimulatory TNF receptor family member and the Fc domain, and

(b) a VH and VL domain capable of specific binding to a target cell antigen, wherein the VH domain is connected via a peptide linker to the C-terminus of one of the heavy chains and wherein the VL domain is connected via a peptide linker to the C-terminus of the second heavy chain.

35. A polynucleotide encoding the bispecific antigen binding molecule of claim 1.

39. A pharmaceutical composition comprising a bispecific antigen binding molecule of claim 1 and at least one pharmaceutically acceptable excipient.

44. A method of inhibiting the growth of tumor cells in an individual comprising administering to the individual an effective amount of the bispecific antigen binding molecule of claim 1 to inhibit the growth of the tumor cells.

36. An antibody that specifically binds to OX40, wherein said antibody comprises

(i) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO:

25 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

26,

(ii) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

27 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

28,

(iii) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

29 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

30,

(iv) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

31 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

32,

(v) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

33 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

34,

(vi) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

35 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

36, or

(vii) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

37 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

38.

38. A polynucleotide encoding the antibody of claim 36.

46. A method of inhibiting the growth of tumor cells in an individual comprising administering to the individual an effective amount of the antibody of claim 36 to inhibit the growth of the tumor cells.

37. An antibody that specifically binds to 4-1BB, wherein said antibody comprises

(i) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO:

58 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

59,

(ii) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

60 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

61,

(iii) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

62 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

63,

(iv) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

64 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

65, or

(v) a heavy chain variable region VH comprising an amino acid sequence of SEQ ID NO;

66 and and a light chain variable region VL comprising an amino acid sequence of SEQ ID NO;

67.

45. A polynucleotide encoding the antibody of claim 37.

47. A method of inhibiting the growth of tumor cells in an individual comprising administering to the individual an effective amount of the antibody of claim 37 to inhibit the growth of the tumor cells.

40. (canceled)

41. (canceled)

42. (canceled)

43. (canceled)