Bispecific anti-VEGF/anti-ANG-2 antibodies

• US 8,268,314 B2
• Filed: 10/02/2009
• Issued: 09/18/2012
• Est. Priority Date: 10/08/2008
• Status: Active Grant

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: Baehner, Monika, Brinkmann, Ulrich, Georges, Guy, Griep, Remko Albert, Imhof-Jung, Sabine, Kavlie, Anita, Klein, Christian, Regula, Joerg Thomas, Schaefer, Wolfgang, Schanzer, Juergen Michael, Scheuer, Werner, Seeber, Stefan, Thomas, Markus, Kettenberger, Hubert
• Primary Examiner(s): Goddard, Laura B
• Assistant Examiner(s): Natarajan, Meera
• Application Number:
  US12/572,289
• Publication Number:
  US 20100111967A1
• Time in Patent Office: 1,082 Days
• Field of Search: None
• US Class Current: 424/136.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/507 Comprising a combination of two or more separate antibodies
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 27/00 Drugs for disorders of the senses
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/14 Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 2317/21 : from primates, e.g. man
• CPC Class Codes: C07K 2317/31 : multispecific
• 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 2317/622 : Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/73 : Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/76 : Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/30 : Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Abstract

The present invention relates to bispecific antibodies against human VEGF and against human ANG-2, methods for their production, pharmaceutical compositions containing said antibodies, and uses thereof.

Claims

1. A bispecific antibody that binds specifically to human vascular endothelial growth factor (VEGF) and human angiopoietin-2 (ANG-2), said antibody comprising a first antigen-binding site that specifically binds to human VEGF and a second antigen-binding site that specifically binds to human ANG-2, wherein:

i) said antigen-binding sites each comprise an antibody heavy chain variable domain and an antibody light chain variable domain;

ii) said first antigen-binding site comprises in the heavy chain variable domain;

a CDR3 region having an amino acid sequence selected from the group consisting of;

SEQ ID NO;

1, SEQ ID NO;

9, SEQ ID NO;

17, and SEQ ID NO;

94;

a CDR2 region having an amino acid sequence selected from the group consisting of;

SEQ ID NO;

2, SEQ ID NO;

10, SEQ ID NO;

18, and SEQ ID NO;

95;

and a CDR1 region having an amino acid sequence selected from the group consisting of;

SEQ ID NO;

3, SEQ ID NO;

11, SEQ ID NO;

19, and SEQ ID NO;

96, and

in the light chain variable domain;

a CDR3 region having an amino acid sequence selected from the group consisting of;

SEQ ID NO;

4, SEQ ID NO;

12, SEQ ID NO;

20, and SEQ ID NO;

97,

a CDR2 region having an amino acid sequence selected from the group consisting of;

SEQ ID NO;

5, SEQ ID NO;

13, SEQ ID NO;

21, and SEQ ID NO;

98; and

a CDR1 region having an amino acid sequence selected from the group consisting of;

SEQ ID NO;

6, SEQ ID NO;

14, SEQ ID NO;

22, and SEQ ID NO;

99; and

iii) said second antigen-binding site comprises in the heavy chain variable domain;

a CDR3 region having an amino acid sequence selected from the group consisting of;

SEQ ID NO;

25, SEQ ID NO;

38, SEQ ID NO;

46, SEQ ID NO;

54, SEQ ID NO;

62, SEQ ID NO;

70, SEQ ID NO;

78, and SEQ ID NO;

86;

a CDR2 region having an amino acid sequence selected from the group consisting of;

SEQ ID NO;

26, SEQ ID NO;

39, SEQ ID NO;

47, SEQ ID NO;

55, SEQ ID NO;

63, SEQ ID NO;

71, SEQ ID NO;

79, and SEQ ID NO;

87; and

a CDR1 region having an amino acid sequence selected from the group consisting of;

SEQ ID NO;

27, SEQ ID NO;

40, SEQ ID NO;

48, SEQ ID NO;

56, SEQ ID NO;

64, SEQ ID NO;

72, SEQ ID NO;

80, and SEQ ID NO;

88; and

in the light chain variable domain;

a CDR3 region having an amino acid sequence selected from the group consisting of;

SEQ ID NO;

28, SEQ ID NO;

28 with the mutations T92L, H93Q and W94T, SEQ ID NO;

41, SEQ ID NO;

49, SEQ ID NO;

57, SEQ ID NO;

65, SEQ ID NO;

73, SEQ ID NO;

81, and SEQ ID NO;

89;

a CDR2 region having an amino acid sequence selected from the group consisting of;

SEQ ID NO;

29, SEQ ID NO;

42, SEQ ID NO;

50, SEQ ID NO;

58, SEQ ID NO;

66, SEQ ID NO;

74, SEQ ID NO;

82 and SEQ ID NO;

90; and

a CDR1 region having an amino acid sequence selected from the group consisting of;

SEQ ID NO;

30, SEQ ID NO;

43, SEQ ID NO;

51, SEQ ID NO;

59, SEQ ID NO;

67, SEQ ID NO;

75, SEQ ID NO;

83, and SEQ ID NO;

91.

2. A bispecific antibody according to claim 1, characterized in that

said first antigen-binding site comprises, in the heavy chain variable domain, a CDR3 region of SEQ ID NO:

1, a CDR2 region of SEQ ID NO;

2, and a CDR1 region of SEQ ID NO;

3, and, in the light chain variable domain, a CDR3 region of SEQ ID NO;

4, a CDR2 region of SEQ ID NO;

5, and a CDR1 region of SEQ ID NO;

6; and

said second antigen-binding site comprises, in the heavy chain variable domain, a CDR3 region of SEQ ID NO;

46, a CDR2 region of SEQ ID NO;

47, and a CDR1 region of SEQ ID NO;

48, and, in the light chain variable domain, a CDR3 region of SEQ ID NO;

49, a CDR2 region of SEQ ID NO;

50, and a CDR1 region of SEQ ID NO;

51.

3. A bispecific antibody according to claim 2, characterized in that said first antigen-binding site comprises, as the heavy chain variable domain, SEQ ID NO:

7, and, as the light chain variable domain, SEQ ID NO;

8, and

said second antigen-binding site comprises, as the heavy chain variable domain, SEQ ID NO;

52, and, as the light chain variable domain, SEQ ID NO;

53.

4. A bispecific antibody according to claim 1, characterized in that said first antigen-binding site comprises, in the heavy chain variable domain, a CDR3 region of SEQ ID NO:

1, a CDR2 region of SEQ ID NO;

2, and a CDR1 region of SEQ ID NO;

3, and, in the light chain variable domain, a CDR3 region of SEQ ID NO;

4, a CDR2 region of SEQ ID NO;

5, and a CDR1 region of SEQ ID NO;

6;

said second antigen-binding site comprises, in the heavy chain variable domain, a CDR3 region of SEQ ID NO;

62, a CDR2 region of SEQ ID NO;

63, and a CDR1 region of SEQ ID NO;

64, and, in the light chain variable domain, a CDR3 region of SEQ ID NO;

65, a CDR2 region of SEQ ID NO;

66, and a CDR1 region of SEQ ID NO;

67.

5. A bispecific antibody according to claim 4, characterized in that said first antigen-binding site comprises, as the heavy chain variable domain, SEQ ID NO:

7, and, as the light chain variable domain, SEQ ID NO;

8; and

said second antigen-binding site comprises, as the heavy chain variable domain, SEQ ID NO;

68, and, as the light chain variable domain, SEQ ID NO;

69.

6. A bispecific antibody according to claim 1, characterized in that said first antigen-binding site s comprises, in the heavy chain variable domain, a CDR3 region of SEQ ID NO:

1, a CDR2 region of SEQ ID NO;

2, and a CDR1 region of SEQ ID NO;

3, and, in the light chain variable domain, a CDR3 region of SEQ ID NO;

4, a CDR2 region of SEQ ID NO;

5, and a CDR1 region of SEQ ID NO;

6;

said second antigen-binding site comprises, in the heavy chain variable domain, a CDR3 region of SEQ ID NO;

78, a CDR2 region of SEQ ID NO;

79, and a CDR1 region of SEQ ID NO;

80, and, in the light chain variable domain, a CDR3 region of SEQ ID NO;

81, a CDR2 region of SEQ ID NO;

82, and a CDR1 region of SEQ ID NO;

83.

7. A bispecific antibody according to claim 6, characterized in that said first antigen-binding site comprises, as the heavy chain variable domain, SEQ ID NO:

7, and, as the light chain variable domain, SEQ ID NO;

8; and

said second antigen-binding site comprises, as the heavy chain variable domain SEQ ID NO;

84, and, as the light chain variable domain, SEQ ID NO;

85.

8. A bispecific antibody according to claim 1, characterized in that the ratio of the binding affinities K_D (antigen-binding site specific for VEGF)/ K_D (antigen-binding site specific for ANG-2) is 1.0-10.0.

9. A bispecific antibody according to claim 1, characterized in that the second antigen-binding site that specifically binds to human ANG-2does not specifically bind to human Angiopoetin 1 (ANG-1).

10. A bispecific antibody according to claim claims 1, characterized in that said antibody is bivalent, trivalent or tetravalent.

11. A pharmaceutical composition comprising an antibody according to claim 1.

12. A nucleic acid encoding a bispecific antibody according to claim 1.

13. An expression vector which comprises the nucleic acid according claim 12 and is capable of expressing said nucleic acid in a prokaryotic or eukaryotic host cell.

14. A prokaryotic or eukaryotic host cell comprising a vector according to claim 13.

15. A method for the production of a bispecific antibody according to claim 1, comprising expressing a nucleic acid encoding such a bispecific antibody in a prokaryotic or eukaryotic host cell and recovering said bispecific antibody from said cell or the cell culture supernatant.

16. An antibody obtained by the recombinant method of claim 15.

17. A bispecific antibody according to claim 1 wherein:

(A) said first antigen-binding site comprises, in the heavy chain variable domain, a CDR3 region of SEQ ID NO;

9, a CDR2 region of SEQ ID NO;

10, and a CDR1 region of SEQ ID NO;

11, and, in the light chain variable domain, a CDR3 region of SEQ ID NO;

12, a CDR2 region of SEQ ID NO;

13, and a CDR1 region of SEQ ID NO;

14; and

(B) said second antigen-binding site comprises, in the heavy chain variable domain, a CDR3 region of SEQ ID NO;

78, a CDR2 region of SEQ ID NO;

79, and a CDR1 region of SEQ ID NO;

80, and, in the light chain variable domain, a CDR3 region of SEQ ID NO;

81, a CDR2 region of SEQ ID NO;

82, and a CDR1 region of SEQ ID NO;

83.

18. A bispecific antibody according to claim 17 wherein:

(A) said first antigen-binding site comprises, as the heavy chain variable domain, SEQ ID NO;

15, and, as the light chain variable domain, SEQ ID NO;

16; and

(B) said second antigen-binding site comprises, as the heavy chain variable domain, SEQ ID NO;

84, and, as the light chain variable domain, SEQ ID NO;

85.