BISPECIFIC ANTIBODIES SPECIFIC FOR FAP AND DR5, ANTIBODIES SPECIFIC FOR DR5 AND METHODS OF USE

• US 20140370019A1
• Filed: 04/01/2014
• Published: 12/18/2014
• Est. Priority Date: 04/03/2013
• Status: Active Grant

• Current Assignee: Roche Glycart AG (Roche Holding AG)
• Original Assignee: Roche Glycart AG (Roche Holding AG)
• Inventors: Bruenker, Peter, Ferrara Koller, Claudia, Georges, Guy, Hosse, Ralf, Klein, Christian, Moelleken, Joerg, Moessner, Ekkehard, Niu, Huifeng, Seeber, Stefan, Umana, Pablo, Waldhauer, Inja, Daouti, Sherif, Feng, Ningping, Grau-Richards, Sandra, Koenig, Maximiliane, Packman, Kathryn E., Runza, Valeria, Wang, Huisheng, Weiser, Barbara
• Granted Patent: US 9,926,379 B2
• Time in Patent Office: Days
• US Class Current: 424/136.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 16/2875 against the NGF/TNF superfamily, e.g. CD70, CD95L, CD153, CD154 against NGF C07K16/22, against TNF C07K16/241
• 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 16/40 against enzymes
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/35 Valency
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/624 Disulfide-stabilized antibody (dsFv)
• CPC Class Codes: C07K 2317/64 comprising a combination of variable region and constant region components
• CPC Class Codes: C07K 2317/66 comprising a swap of domains, e.g. CH3-CH2, VH-CL or VL-CH1
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/75 : Agonist effect on antigen
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2317/94 : Stability, e.g. half-life, pH, temperature or enzyme-resistance
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide

Abstract

The present invention relates to bispecific antibodies comprising at least one antigen binding site specific for DR5 and at least one antigen binding site specific for FAP, antibodies specific for DR5, methods for their production, pharmaceutical compositions containing said antibodies, and uses thereof.

Claims

1. A bispecific antibody that binds to death receptor 5 (DR5) and Fibroblast Activation Protein (FAP), comprising at least one antigen binding site specific for DR5, comprising

(a) a heavy chain complementarity determining region 1 (CDR1) selected from the group consisting of SEQ ID NO.:

1, SEQ ID NO.;

17 and SEQ ID NO.;

75;

(b) a heavy chain complementarity determining region 2 (CDR2) selected from the group of SEQ ID NO.;

2, SEQ ID NO.;

18, SEQ ID NO.;

25 and SEQ ID NO.;

83;

(c) a heavy chain complementarity determining region 3 (CDR3) selected from the group of SEQ ID NO.;

3, SEQ ID NO.;

19, SEQ ID NO.;

84, SEQ ID NO.;

96, SEQ ID NO.;

98;

SEQ ID NO.;

104 and SEQ ID NO.;

108;

(d) a light chain CDR1 selected from the group of SEQ ID NO.;

4, SEQ ID NO.;

20, SEQ ID NO.;

27 and SEQ ID NO.;

86;

(e) a light chain CDR2 selected from the group of SEQ ID NO.;

5, SEQ ID NO.;

21 and SEQ ID NO.;

28; and

(f) a light chain CDR3 selected from the group of SEQ ID NO.;

6, SEQ ID NO.;

22, SEQ ID NO.;

87, SEQ ID NO.;

99, SEQ ID NO.;

105, SEQ ID NO.;

109 and SEQ ID NO.;

97;

and at least one antigen binding site specific for FAP, comprising

(a) a heavy chain CDR1 selected from the group of SEQ ID NO.;

9 and SEQ ID NO.;

33;

(b) a heavy chain CDR2 selected from the group of SEQ ID NO.;

10 and SEQ ID NO.;

34;

(c) a heavy chain CDR3 selected from the group of SEQ ID NO.;

11 and SEQ ID NO.;

35;

(d) a light chain CDR1 selected from the group of SEQ ID NO.;

12 and SEQ ID NO.;

36;

(e) a light chain CDR2 selected from the group of SEQ ID NO.;

13 and SEQ ID NO.;

37;

(f) a light chain CDR3 selected from the group of SEQ ID NO.;

14 and SEQ ID NO.;

38.

2. The bispecific antibody of claim 1, wherein the antigen binding site specific for DR5 comprises a variable heavy chain and a variable light chain comprising an amino acid sequence selected from the group of:

SEQ ID NO.;

7 and SEQ ID NO.;

8;

SEQ ID NO.;

23 and SEQ ID NO.;

24;

SEQ ID NO.;

26 and SEQ ID NO.;

24;

SEQ ID NO.;

23 and SEQ ID NO.;

29;

SEQ ID NO.;

23 and SEQ ID NO.;

30;

SEQ ID NO.;

26 and SEQ ID NO.;

31;

SEQ ID NO.;

26 and SEQ ID NO.;

32;

SEQ ID NO.;

26 and SEQ ID NO.;

30;

SEQ ID NO.;

23 and SEQ ID NO.;

31;

SEQ ID NO.;

82 and SEQ ID NO.;

85;

SEQ ID NO.;

100 and SEQ ID NO.;

101;

SEQ ID NO.;

102 and SEQ ID NO.;

103;

SEQ ID NO.;

106 and SEQ ID NO.;

107;

SEQ ID NO.;

94 and SEQ ID NO.;

95;

and wherein the antigen binding site specific for FAP comprises a variable heavy chain comprising an amino acid sequence selected from the group of;

SEQ ID NO.;

15 and SEQ ID NO.;

39; and

a variable light chain comprising an amino acid sequence selected from the group of SEQ ID NO.;

16 and SEQ ID NO.;

40.

3. The bispecific antibody of claim 1, wherein the antigen binding site specific for DR5 comprises

(a) a heavy chain CDR1 of SEQ ID NO.:

1;

(b) a heavy chain CDR2 of SEQ ID NO.;

2;

(c) a heavy chain CDR3 of SEQ ID NO.;

3;

(d) a light chain CDR1 of SEQ ID NO.;

4;

(e) a light chain CDR2 of SEQ ID NO.;

5;

(f) a light chain CDR3 of SEQ ID NO.;

6

and the antigen binding site specific for FAP comprises

(a) a heavy chain CDR1 of SEQ ID NO.;

9;

(b) a heavy chain CDR2 of SEQ ID NO.;

10;

(c) a heavy chain CDR3 of SEQ ID NO.;

11;

(d) a light chain CDR1 of SEQ ID NO.;

12;

(e) a light chain CDR2 of SEQ ID NO.;

13;

(f) a light chain CDR3 of SEQ ID NO.;

14.

4. The bispecific antibody of claim 1, wherein the antigen binding site specific for DR5

comprises a variable heavy chain comprising an amino acid sequence of SEQ ID NO.:

7 and a variable light chain comprising an amino acid sequence of SEQ ID NO.;

8; and

the antigen binding site specific for FAP comprises a heavy chain variable region comprising an amino acid sequence of SEQ ID NO.;

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

16.

5. The bispecific antibody of claim 1, wherein the antibody is human.

6. The bispecific antibody of claim 1, wherein the antibody is humanized.

7. The bispecific antibody of claim 1, comprising an Fc domain, at least one Fab fragment comprising the antigen binding site specific for DR5, and at least one Fab fragment comprising the antigen binding site specific for FAP.

8. The bispecific antibody of claim 7, wherein at least one of the Fab fragments is connected to the first or second subunit of the Fc domain via the light chain (VLCL) and at least one Fab fragment is connected to the first or second subunit of the Fc domain via the heavy chain (VHCH1).

9. The bispecific antibody of claim 7, comprising

a) an Fc domain,

b) two Fab fragments comprising an antigen binding site specific for DR5,

wherein said Fab fragments are connected at the C-terminus of the constant light chain (CL) to the first or second subunit of the Fc domain,

c) two Fab fragments comprising the antigen binding site specific for FAP, wherein the two Fab fragments are connected at the C-terminus of the constant heavy chain (CH1) to the first or second subunit of the Fc domain.

10. The bispecific antibody of claim 7, comprising

a) an Fc domain,

b) two Fab fragments comprising an antigen binding site specific for DR5,

wherein said Fab fragments are connected at the C-terminus of the constant heavy chain (CH1) to the first or second subunit of the Fc domain,

c) two Fab fragments comprising the antigen binding site specific for FAP, wherein the two Fab fragments are connected at the C-terminus of the constant light chain (CL) to the first or second subunit of the Fc domain.

11. The bispecific antibody of claim 1, comprising an Fc domain, at least one Fab fragment comprising the antigen binding site specific for DR5, and at least one Fab fragment comprising the antigen binding site specific for FAP wherein either the variable regions or the constant regions of the heavy and light chain of at least one Fab fragment are exchanged.

12. The bispecific antibody of claim 11, comprising two Fab fragments comprising each an

antigen binding site specific for DR5, and two Fab fragments comprising each an antigen binding site specific for FAP.

13. The bispecific antibody of claim 12, wherein the bispecific antibody is bivalent both for DR5 and FAP.

14. The bispecific antibody of claim 11, comprising two Fab fragments comprising each an

antigen binding site specific for DR5, and one Fab fragment comprising an antigen binding site specific for FAP.

15. The bispecific antibody of claim 14, wherein the bispecific antibody is bivalent for DR5 and monovalent for FAP.

16. The bispecific antibody of claim 14, additionally comprising one Fab fragment comprising an antigen binding site specific for DR5.

17. The bispecific antibody of claim 16, wherein the bispecific antibody is trivalent for DR5 and monovalent for FAP.

18. The bispecific antibody of claim 11, comprising one Fab fragment comprising an antigen binding site specific for DR5, and one Fab fragment comprising an antigen binding site specific for FAP.

19. The bispecific antibody of claim 18, wherein the bispecific antibody is monovalent for DR5 and monovalent for FAP.

20. The bispecific antibody of claim 11, wherein either the variable regions or the constant regions of the heavy and light chain of the Fab fragment(s) comprising an antigen binding site specific for FAP are exchanged.

21. The bispecific antibody of claim 7, wherein at least one of the Fab fragments are connected to the Fc domain via a peptide linker.

22. The bispecific antibody of claim 7, wherein the Fc domain comprises one or more amino acid substitution that reduces binding to an Fc receptor and/or effector function.

23. The bispecific antibody of claim 22, wherein said one or more amino acid substitution is at one or more position selected from the group of L234, L235, and P329.

24. The bispecific antibody of claim 23, wherein each subunit of the Fc domain comprises three amino acid substitutions that reduce binding to an activating or inhibitory Fc receptor and/or effector function wherein said amino acid substitutions are L234A, L235A and P329G.

25. The bispecific antibody of claim 7, wherein the Fc part of the first heavy chain comprises a first dimerization module and the Fc part of the second heavy chain comprises a second dimerization module allowing a heterodimerization of the two heavy chains of the first antibody.

26. The bispecific antibody of claim 25, wherein the first dimerization module comprises knobs and the second dimerization module comprises holes according to the knobs into holes strategy.

31. A pharmaceutical composition comprising the bispecific antibody of claim 1.

33. A method of treating an individual having cancer comprising administering to the individual an effective amount of the bispecific antibody of claim 1.

34. The method of claim 33, wherein the cancer is pancreatic cancer or colorectal carcinoma.

37. A nucleic acid sequence encoding the bispecific antibody of claim 1.

38. An expression vector comprising the nucleic acid sequence of claim 37.

39. A prokaryotic or eukaryotic host cell comprising a vector according to claim 38.

40. A method of producing an antibody comprising culturing the host cell of claim 39 so that the antibody is produced.

27. An antibody that specifically binds to DR5, comprising

(a) a heavy chain complementarity determining region 1 (CDR1) selected from the group consisting of SEQ ID NO.:

1, SEQ ID NO.;

17 and SEQ ID NO.;

75;

(b) a heavy chain complementarity determining region 2 (CDR2) selected from the group of SEQ ID NO.;

2, SEQ ID NO.;

18, SEQ ID NO.;

25 and SEQ ID NO.;

83;

(c) a heavy chain complementarity determining region 3 (CDR3) selected from the group of SEQ ID NO.;

3, SEQ ID NO.;

19, SEQ ID NO.;

84, SEQ ID NO.;

96, SEQ ID NO.;

98, SEQ ID NO.;

104 and SEQ ID NO.;

108;

(d) a light chain CDR1 selected from the group of SEQ ID NO.;

4, SEQ ID NO.;

20, SEQ ID NO.;

27 and SEQ ID NO.;

86;

(e) a light chain CDR2 selected from the group of SEQ ID NO.;

5, SEQ ID NO.;

21 and SEQ ID NO.;

28; and

(f) a light chain CDR3 selected from the group of SEQ ID NO.;

6, SEQ ID NO.;

22, SEQ ID NO.;

87, SEQ ID NO.;

99, SEQ ID NO.;

105, SEQ ID NO.;

109 and SEQ ID NO.;

97;

28. The antibody of claim 27, comprising

(a) a heavy chain CDR1 of SEQ ID NO.:

1;

(b) a heavy chain CDR2 of SEQ ID NO.;

2;

(c) a heavy chain CDR3 of SEQ ID NO.;

3;

(d) a light chain CDR1 of SEQ ID NO.;

4;

(e) a light chain CDR2 of SEQ ID NO.;

5;

(f) a light chain CDR3 of SEQ ID NO.;

6.

29. The antibody of claim 27, comprising a variable heavy chain and a variable light chain

comprising an amino acid sequence selected from the group of:

SEQ ID NO.;

7 and SEQ ID NO.;

8;

SEQ ID NO.;

23 and SEQ ID NO.;

24;

SEQ ID NO.;

26 and SEQ ID NO.;

24;

SEQ ID NO.;

23 and SEQ ID NO.;

29;

SEQ ID NO.;

23 and SEQ ID NO.;

30;

SEQ ID NO.;

26 and SEQ ID NO.;

31;

SEQ ID NO.;

26 and SEQ ID NO.;

32;

SEQ ID NO.;

26 and SEQ ID NO.;

30;

SEQ ID NO.;

23 and SEQ ID NO.;

31;

SEQ ID NO.;

82 and SEQ ID NO.;

85;

SEQ ID NO.;

100 and SEQ ID NO.;

101;

SEQ ID NO.;

102 and SEQ ID NO.;

103;

SEQ ID NO.;

106 and SEQ ID NO.;

107;

SEQ ID NO.;

94 and SEQ ID NO.;

95.

30. The antibody of claim 27, comprising a variable heavy chain comprising an amino acid sequence of SEQ ID NO.:

7 and a variable light chain comprising an amino acid sequence of SEQ ID NO.;

8.

32. A pharmaceutical composition comprising the antibody of claim 27.

35. A method of treating an individual having cancer comprising administering to the individual an effective amount of the antibody of claim 27.

36. The method of claim 35, wherein the cancer is pancreatic cancer or colorectal carcinoma.

41. A nucleic acid sequence encoding the antibody of claim 27.

42. An expression vector comprising a nucleic acid sequence of claim 41.

43. A prokaryotic or eukaryotic host cell comprising a vector according to claim 42.

44. A method of producing an antibody comprising culturing the host cell of claim 43 so that the antibody is produced.