BISPECIFIC HER2 ANTIBODIES AND METHODS OF USE

• US 20170029529A1
• Filed: 06/17/2016
• Published: 02/02/2017
• Est. Priority Date: 12/20/2013
• Status: Active Grant

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: Hosse, Ralf, Moser, Samuel, Schanzer, Juergen Michael, Sustmann, Claudio, Croasdale, Rebecca, Duerner, Lydia Jasmin, Georges, Guy, Hofer, Thomas, Klein, Christian, Moessner, Ekkehard, Schaefer, Wolfgang, Scheuer, Werner, Umana, Pablo
• Granted Patent: US 10,584,178 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• 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 16/32 against translation products of oncogenes
• 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 2317/35 Valency
• CPC Class Codes: C07K 2317/41 Glycosylation, sialylation, or fucosylation
• CPC Class Codes: C07K 2317/51 Complete heavy chain or Fd fragment, i.e. VH + CH1
• CPC Class Codes: C07K 2317/515 Complete light chain, i.e. VL + CL
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/522 CH1 domain
• 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 2317/567 Framework region [FR]
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• 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 2317/732 : Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/734 : Complement-dependent cytotoxicity [CDC]
• 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: C12N 15/63 : Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
• CPC Class Codes: C12N 5/10 : Cells modified by introduction of foreign genetic material

Abstract

The present invention relates to bispecific HER2 antibodies, novel HER2 antibody variants, methods for their production, pharmaceutical compositions containing said antibodies, and uses thereof.

Claims

1. A bispecific antibody specifically binding to HER2 comprising a first antigen binding site specific for extracellular domain II of HER2 and a second antigen binding site specific for extracellular domain IV of HER2, wherein the bispecific antibody is monovalent for both the extracellular domain II and IV of HER2.

2. The bispecific antibody of claim 1, wherein said antibody induces complement-dependent cytotoxicity to a higher degree than the combination of Pertuzumab or Trastuzumab.

3. The bispecific antibody of claim 2, wherein the complement dependent cytotoxicity of the bispecific antibody is determined by a LDH assay or a complement assay and compared to the complement dependent cytotoxicity of the combination of Pertuzumab and Trastuzumab as determined by the same assay.

4. The bispecific antibody of claim 3, wherein the complement dependent cytotoxicity is determined in vitro on cancer cells, preferably on breast cancer cells.

5. The bispecific antibody of claim 1, comprising a first Fab molecule capable of specific binding to extracellular domain II of HER2 and a second Fab molecule capable of specific binding to extracellular domain IV of HER2, wherein the sequence of the variable light chain of the first Fab molecule is identical to the sequence of the variable light chain of the second Fab molecule.

6. The bispecific antibody of claim 5, comprising

(a) a first heavy chain comprising a heavy chain CDR1 selected from the group consisting of SEQ ID NO:

55, SEQ ID NO;

58 and SEQ ID NO;

14;

a heavy chain CDR 2 selected from the group consisting of SEQ ID NO;

77;

SEQ ID NO;

15 and SEQ ID NO;

60 and

a heavy chain CDR 3 selected from the group consisting of SEQ ID NO;

56 or SEQ ID NO;

59 and SEQ ID NO;

16, and

(b) a second heavy chain comprising a heavy chain CDR1 of SEQ ID NO;

20, a heavy chain CDR2 of SEQ ID NO;

29 and a heavy chain CDR3 selected from the group consisting of SEQ ID NO;

30 and SEQ ID NO;

79

(c) a first and a second light chain, wherein the variable light chains of the first and second light chain comprise the CDRs of SEQ ID NO;

89, SEQ ID NO;

90 and SEQ ID NO;

19.

7. The bispecific antibody of claim 5, comprising two variable light chains comprising an amino acid sequence of SEQ ID NO:

54, a first heavy chain comprising a variable heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO;

64, SEQ ID NO;

70 and SEQ ID NO;

68, and a second heavy chain comprising a variable heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO;

92 and SEQ ID NO;

117.

8. The bispecific antibody of claim 1, comprising a first Fab molecule capable of specific binding to extracellular domain II of HER2 and a second Fab molecule capable of specific binding to extracellular domain IV of HER2, wherein either the variable regions or the constant regions of the heavy and light chain of at least one Fab fragment are exchanged.

9. The bispecific antibody of claim 8, wherein the first Fab molecule comprises a heavy chain CDR1 of SEQ ID NO:

14, a heavy chain CDR2 of SEQ ID NO;

15 and a heavy chain CDR3 of SEQ ID NO;

16; and

a light chain CDR1 of SEQ ID NO;

11;

a light chain CDR2 of SEQ ID NO;

12 and a light chain CDR3 of SEQ ID NO;

13, and wherein the second Fab molecule comprises

a heavy chain CDR1 of SEQ ID NO;

20;

a heavy chain CDR2 of SEQ ID NO;

108;

a heavy chain CDR3 of SEQ ID NO;

79; and

a light chain CDR1 of SEQ ID NO;

107, a light chain CDR2 of SEQ ID NO;

18 and a light chain CDR3 of SEQ ID NO;

19.

10. The bispecific antibody of claim 8, wherein the first Fab molecule comprises a heavy chain CDR1 of SEQ ID NO:

14, a heavy chain CDR2 of SEQ ID NO;

15 and a heavy chain CDR3 of SEQ ID NO;

16; and

a light chain CDR1 of SEQ ID NO;

11;

a light chain CDR2 of SEQ ID NO;

12 and a light chain CDR3 of SEQ ID NO;

13, and wherein the second Fab molecule comprises

a heavy chain CDR1 of SEQ ID NO;

20, a heavy chain CDR2 of SEQ ID NO;

29, and a heavy chain CDR3 selected from the group consisting of SEQ ID NO;

79, SEQ ID NO;

78, SEQ ID NO;

80, SEQ ID NO;

87, SEQ ID NO;

88; and

a light chain CDR1 selected from the group consisting of SEQ ID NO;

104, SEQ ID NO;

103 and SEQ ID NO;

158;

a light chain CDR2 of SEQ ID NO;

18 and a light chain CDR3 of SEQ ID NO;

19.

11. The bispecific antibody of claim 8, wherein the first Fab molecule comprises

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

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

24 and wherein the second Fab molecule comprises an amino acid sequence of SEQ ID NO;

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

106.

12. A pharmaceutical composition comprising a bispecific antibody of claim 1.

17. A nucleic acid sequence comprising a sequence encoding a heavy chain of a bispecific antibody of claim 1.

18. A nucleic acid sequence comprising a sequence encoding a light chain of a bispecific antibody of claim 1.

19. An expression vector comprising a nucleic acid sequence of claim 17.

20. A prokaryotic or eukaryotic host cell comprising a vector according to claim 19.

21. A method of producing an antibody comprising culturing the host cell of claim 20 so that the antibody is produced.

23. An expression vector comprising a nucleic acid sequence of claim 18.

24. A prokaryotic or eukaryotic host cell comprising a vector according to claim 23.

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

13. (canceled)

14. (canceled)

15. (canceled)

16. (canceled)

22. (canceled)