HUMANIZED ANTIBODIES SPECIFIC FOR VON WILLEBRAND FACTOR

• US 20090232804A1
• Filed: 01/23/2009
• Published: 09/17/2009
• Est. Priority Date: 01/23/2008
• Status: Active Grant

• Current Assignee: Ichnos Sciences SA
• Original Assignee: Glenmark Pharmaceuticals SA (Glenmark Pharmaceuticals Limited)
• Inventors: Blein, Stanislas, Fan, Xiaomin, Woods, Catherine, Bertschinger, Martin, Hou, Samuel, Lazarides, Elias, Mottl, Harald
• Granted Patent: US 8,236,315 B2
• Time in Patent Office: Days
• US Class Current: 424/133.100
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• 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 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/08 Vasodilators for multiple indications
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 16/36 against blood coagulation factors
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/52 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

Abstract

The present disclosure relates to humanized antibodies or binding fragments thereof specific for human von Willebrand factor (vWF), methods for their preparation and use, including methods for treating vWF mediated diseases or disorders.

Claims

1. A humanized antibody or binding fragment thereof specific for von Willebrand factor (vWF), the humanized antibody comprising:

(a) a heavy chain variable region sequence as set forth in SEQ ID NO;

19; and

(b) a light chain variable region sequence as set forth in SEQ ID NO;

28.

18. The humanized antibody or binding fragment thereof of claim 1, wherein the humanized antibody binds to vWF with an affinity (Kd) of 10 nM or less.

19. The humanized antibody or binding fragment thereof of claim 1, wherein the humanized antibody competes for binding to vWF with an affinity (Ki) of 100 nM or less.

20. The humanized antibody or binding fragment thereof of claim 1, wherein the humanized antibody binds to the A1 domain of vWF with an affinity (Kd) of 10 nM or less.

21. The humanized antibody or binding fragment thereof of claim 1, wherein the humanized antibody competes for binding to the A1 domain of vWF with an affinity (Ki) of 100 nM or less.

22. The humanized antibody or binding fragment thereof of claim 1, wherein the humanized antibody has a FAB fragment thermostability temperature greater than 65°

C.

23. The humanized antibody or binding fragment thereof of claim 1, wherein the humanized antibody has a FAB fragment thermostability temperature greater than the parent non-humanized antibody.

42. The humanized antibody or binding fragment thereof of claim 1, wherein HCDR1 comprises one or more amino acid substitutions selected from the group consisting of F27G, L291, T30S and V34W.

43. The humanized antibody or binding fragment thereof of claim 1, wherein HCDR2 comprises one or more amino acid substitutions selected from the group consisting of S61 P and A62S.

44. The humanized antibody or binding fragment thereof of claim 1, wherein LCDR1 comprises one or more amino acid substitutions selected from the group consisting of S24Q, N30S and K31N.

45. The humanized antibody or binding fragment thereof of claim 1, wherein HCDR1 comprises one or more amino acid substitutions selected from the group consisting of F27G, L291, T30S and V34W, HCDR2 comprises one or more amino acid substitutions selected from the group consisting of S61 P and A62S, and LCDR1 comprises one or more amino acid substitutions selected from the group consisting of S24Q, N30S and K31N.

54. The humanized antibody or binding fragment thereof of claim 1, wherein the humanized antibody retains the same activity as the parent non-humanized antibody or as a chimeric antibody comprising variable regions from the parent non-humanized antibody and a human Fc region.

55. The humanized antibody or binding fragment thereof of claim 54, wherein the activity is measured as ristocetin-induced platelet agglutination activity.

56. The humanized antibody or binding fragment thereof of claim 1, wherein the humanized antibody lacks effector function.

57. The humanized antibody or binding fragment thereof of claim 1, wherein the humanized antibody comprises an Fc region derived from IgG4.

58. The humanized antibody or binding fragment thereof of claim 1, wherein the humanized antibody is specific for the A1 domain of human vWF.

59. The humanized antibody of claim 1, wherein the humanized antibody is a full length antibody.

60. The binding fragment of claim 1, wherein the binding fragment is an antibody fragment selected from the group consisting of Fab, Fab′

, Fab′

-SH, Fv, scFv, F(ab′

)2, and a diabody.

70. An isolated nucleic acid encoding the antibody or binding fragment thereof of claim 1.

75. A vector comprising the isolated nucleic acid of claim 70.

76. A host cell comprising the isolated nucleic acid of claim 70 or the vector of claim 75.

77. A method of producing a humanized antibody or binding fragment thereof comprising culturing the host cell of claim 76 so that the nucleic acid is expressed and the antibody produced.

78. The method of claim 77 further comprising recovering the antibody from the host cell culture.

79. The method of claim 77, wherein the antibody is recovered from the host cell medium.

80. The method of claim 77, wherein before culturing, the host cell is co-transfected with a vector comprising nucleic acid encoding a heavy chain variable region and with a vector comprising nucleic acid encoding a light chain variable region.

81. A composition comprising the humanized antibody or binding fragment thereof of claim 1 and a pharmaceutically acceptable carrier.

82. A composition comprising a first humanized antibody or binding fragment thereof of claim 1 and a second antibody that binds to the A1 domain of vWF.

83. The composition of claim 82, wherein, the second antibody is AJW-200.

84. A method for treating a vWF mediated disease or disorder in a subject, the method comprising administering to the subject a therapeutically effective amount of the humanized antibody or binding fragment thereof of claim 1.

85. The method of claim 84, wherein the subject is a human.

86. The method of claim 84, wherein the vWF mediated disorder is a thrombotic disease or disorder.

87. The method of claim 86, wherein the thrombotic disease or disorder is cardiovascular disease or cerebrovascular disease such as ischemic stroke.

88. The method of claim 87, wherein the cardiovascular disease is atherosclerosis, restenosis, angina, acute myocardial infarction, acute coronary syndrome or cardiovascular disorders associated with diabetes.

89. The method of claim 86, wherein the thrombotic disease or disorder is vascular inflammation, venous thrombosis, sickle cell disease, xenograft rejection, peripheral vascular disease, thrombotic thrombocytopenic purpura, cystic fibrosis, vascular dementia, Raynaud'"'"'s disease, rheumatoid arthritis or diabetes.

90. The method of claim 87, wherein the cerebrovascular disease is vascular dementia, ischemic stroke, or prevention of recurrent strokes.

91. The method of claim 84, wherein the therapeutically effective amount is from about 0.001 to about 100 mg/kg.

92. The method of claim 84, wherein the therapeutically effective amount is from about 0.002 to about 20 mg/kg.

93. The method of claim 84, wherein the therapeutically effective amount is from about 0.002 to about 10 mg/kg.

94. The method of claim 84, wherein a single or multiple sub-doses of the therapeutically effective amount of the humanized antibody or binding fragment thereof are administered to the subject.

95. The method of claim 84, wherein the therapeutically effective amount is sufficient to inhibit platelet aggregation but insufficient to cause significant clinical signs of bleeding.

96. The method of claim 84, wherein the therapeutically effective amount is from about 1 to about 250 times an ED₁₀₀ without causing significant clinical signs of bleeding.

97. A pharmaceutical composition comprising a therapeutically effective amount of the humanized antibody or binding fragment thereof of claim 1 and a pharmaceutically acceptable carrier or diluent.

98. A pharmaceutical composition for the treatment of a vWF mediated disease or disorder comprising a therapeutically effective amount of the humanized antibody or binding fragment thereof of claim 1 and a pharmaceutically acceptable carrier or diluent.

99. The composition of claim 98, wherein the vWF mediated disorder is a thrombotic disease or disorder.

100. The composition of claim 99, wherein the thrombotic disorder is cardiovascular disease or cerebrovascular disease such as ischemic stroke.

101. The composition of claim 100, wherein the cardiovascular disease is atherosclerosis, restenosis, angina, acute myocardial infarction, acute coronary syndrome or cardiovascular disorders associated with diabetes.

102. The composition of claim 99, wherein the thrombotic disease is vascular inflammation, venous thrombosis, sickle cell disease, xenograft rejection, peripheral vascular disease, thrombotic thrombocytopenic purpura, cystic fibrosis, vascular dementia, Raynaud'"'"'s disease, rheumatoid arthritis or diabetes.

103. The composition of claim 100, wherein the cerebrovascular disease is vascular dementia, ischemic stroke, or prevention of recurrent strokes.

104. The composition of claim 97, wherein the therapeutically effective amount is from about 0.001 to about 100 mg/kg.

105. The composition of claim 97, wherein the therapeutically effective amount is from about 0.002 to about 20 mg/kg.

106. The composition of claim 97, wherein the therapeutically effective amount is from about 0.002 to about 10 mg/kg.

107. The composition of claim 97, wherein a single or multiple sub-doses of the therapeutically effective amount of the humanized antibody or binding fragment thereof are administered to the subject.

108. The composition of claim 97, wherein the therapeutically effective amount is sufficient to inhibit platelet aggregation but insufficient to cause significant clinical signs of bleeding.

109. The composition of claim 97, wherein the humanized antibody or binding fragment thereof is administered in a therapeutically effective amount from about 1 to about 250 times an ED₁₀₀ without causing significant clinical signs of bleeding.

112. A method for administering the humanized antibody or binding fragment thereof of claim 1 to a subject in need thereof comprising administering a therapeutically effective amount of the humanized antibody or binding fragment thereof sufficient to inhibit platelet aggregation without significant clinical signs of bleeding.

113. The method of claim 112, wherein the humanized antibody or binding fragment thereof is administered subcutaneously.

114. The method of claim 112, wherein the humanized antibody or binding fragment thereof is administered intravenously.

115. The method of claim 112, wherein the humanized antibody or binding fragment thereof is administered intravenously in combination with radiological treatments.

116. The method of claim 112, wherein the therapeutically effective amount of the humanized antibody or binding fragment thereof is from about 1 to about 250 times the ED₁₀₀.

117. The composition of claim 97, wherein the humanized antibody or binding fragment thereof is administered in an amount from about 1 to about 250 times the ED₁₀₀.

118. An article of manufacture comprising the humanized antibody or binding fragment thereof of claim 1 for the treatment of a vWF mediated disease or disorder.

119. A kit comprising the humanized antibody or binding fragment thereof of claim 1 for the treatment of a vWF mediated disease or disorder.

120. A kit comprising the humanized antibody or binding fragment thereof of claim 1 for use in non-therapeutic applications.

121. The kit of claim 120, wherein the non-therapeutic application is a diagnostic assay.

2. A humanized antibody or binding fragment thereof specific for vWF, the humanized antibody comprising:

(a) a heavy chain sequence as set forth in SEQ ID NO;

237; and

(b) a light chain sequence as set forth in SEQ ID NO;

238.

3. A humanized antibody or binding fragment thereof specific for vWF, the humanized antibody comprising:

(a) heavy and light chain complementarity determining regions (CDRs) corresponding to the CDRs present in the heavy and light chain variable regions of murine antibody NMC-4 (SEQ ID NO;

1 and 2, respectively); and

(b) heavy chain framework regions corresponding to the framework regions present in the variable region of human antibody AAC18165.1 (SEQ ID NO;

     4) and/or light chain framework regions corresponding to the framework regions present in the variable region of human antibody AAK94808 (SEQ ID NO;

     6).

4. A humanized antibody or binding fragment thereof specific for vWF, the humanized antibody comprising one or more heavy chain CDRs selected from the group consisting of:

HCDR1;

GFSLTDYGVD (SEQ ID NO;

     7), HCDR2;

MIWGDGSTDYNSALKS (SEQ ID NO;

     8) and HCDR3;

DPADYGNYDYALDY (SEQ ID NO;

     9); and

/or one or more light chain CDRs selected from the group consisting of;

LCDR1;

SASQDINKYLN (SEQ ID NO;

     10), LCDR2;

YTSSLHS (SEQ ID NO;

     11) and LCDR3;

QQYEKLPWT (SEQ ID NO;

     12).

6. The humanized antibody or binding fragment thereof of claim 4, further comprising a heavy chain framework region from human antibody AAC18165.1 (SEQ ID NO:

4) and/or a light chain framework region from human antibody AAK94808 (SEQ ID NO;

6).

7. The humanized antibody or binding fragment thereof of claim 6, wherein the heavy chain framework region further comprises one or more murine residues.

8. The humanized antibody or binding fragment thereof of claim 6, wherein the heavy chain framework region does not comprise one or more murine residues.

9. The humanized antibody or binding fragment thereof of claim 6, wherein the light chain framework region further comprises one or more murine residues.

10. The humanized antibody or binding fragment thereof of claim 6, wherein the light chain framework region does not comprise one or more murine residues.

11. The humanized antibody or binding fragment thereof of claim 4, wherein the humanized antibody comprises a heavy chain variable region selected from the group consisting of:

H2 (SEQ ID NO;

     13), H4 (SEQ ID NO;

     14), H5 (SEQ ID NO;

     15), H6 (SEQ ID NO;

     16), H7 (SEQ ID NO;

     17), H8 (SEQ ID NO;

     18), H9 (SEQ ID NO;

     19), H12 (SEQ ID NO;

     20), H13 (SEQ ID NO;

     21), H14 (SEQ ID NO;

     22), H15 (SEQ ID NO;

     145) or H16 (SEQ ID NO;

     146).

12. The humanized antibody or binding fragment thereof of claim 4, wherein the humanized antibody comprises a light chain variable region selected from the group consisting of:

L5 (SEQ ID NO;

     23), L4 (SEQ ID NO;

     24), L6 (SEQ ID NO;

     25), L7 (SEQ ID NO;

     26), L8 (SEQ ID NO;

     27), L9 (SEQ ID NO;

     28), L10 (SEQ ID NO;

     29) or L11 (SEQ ID NO;

     30).

13. The humanized antibody or binding fragment thereof of claim 4, wherein the humanized antibody comprises a heavy chain variable region selected from the group consisting of:

H2 (SEQ ID NO;

     13), H4 (SEQ ID NO;

     14), H5 (SEQ ID NO;

     15), H6 (SEQ ID NO;

     16), H7 (SEQ ID NO;

     17), H8 (SEQ ID NO;

     18), H9 (SEQ ID NO;

     19), H12 (SEQ ID NO;

     20), H13 (SEQ ID NO;

     21), H14 (SEQ ID NO;

     22), H15 (SEQ ID NO;

     145) or H16 (SEQ ID NO;

     146); and

a light chain variable region selected from the group consisting of;

L5 (SEQ ID NO;

     23), L4 (SEQ ID NO;

     24), L6 (SEQ ID NO;

     25), L7 (SEQ ID NO;

     26), L8 (SEQ ID NO;

     27), L9 (SEQ ID NO;

     28), L10 (SEQ ID NO;

     29) or L11 (SEQ ID NO;

     30).

24. The humanized antibody or binding fragment thereof of claim 4, wherein the humanized antibody binds to vWF with an affinity (Kd) of 10 nM or less.

25. The humanized antibody or binding fragment thereof of claim 4, wherein the humanized antibody competes for binding to vWF with an affinity (Ki) of 100 nM or less.

26. The humanized antibody or binding fragment thereof of claim 4, wherein the humanized antibody binds to the A1 domain of vWF with an affinity (Kd) of 10 nM or less.

27. The humanized antibody or binding fragment thereof of claim 4, wherein the humanized antibody competes for binding to the A1 domain of vWF with an affinity (Ki) of 100 nM or less.

28. The humanized antibody or binding fragment thereof of claim 4, wherein the humanized antibody has a FAB fragment thermostability temperature greater than 65°

C.

29. The humanized antibody or binding fragment thereof of claim 4, wherein the humanized antibody has a FAB fragment thermostability temperature greater than the parent non-humanized antibody.

46. The humanized antibody or binding fragment thereof of claim 4, wherein HCDR1 comprises one or more amino acid substitutions selected from the group consisting of F27G, L291, T30S and V34W.

47. The humanized antibody or binding fragment thereof of claim 4, wherein HCDR2 comprises one or more amino acid substitutions selected from the group consisting of S61 P and A62S.

48. The humanized antibody or binding fragment thereof of claim 4, wherein LCDR1 comprises one or more amino acid substitutions selected from the group consisting of S24Q, N30S and K31N.

49. The humanized antibody or binding fragment thereof of claim 4, wherein HCDR1 comprises one or more amino acid substitutions selected from the group consisting of F27G, L291, T30S and V34W, HCDR2 comprises one or more amino acid substitutions selected from the group consisting of S61 P and A62S, and LCDR1 comprises one or more amino acid substitutions selected from the group consisting of S24Q, N30S and K31N.

50. The humanized antibody or binding fragment thereof of claim 4, further comprising a heavy chain framework region from a human antibody, wherein the human heavy chain framework does not comprise one or more murine residues.

51. The humanized antibody or binding fragment thereof of claim 4, further comprising a light chain framework region from a human antibody, wherein the human light chain framework does not comprise one or more murine residues.

61. The binding fragment of claim 50, wherein the antibody fragment is not a Fab.

62. The humanized antibody or binding fragment thereof of claim 4, wherein the humanized antibody retains the same activity as the parent non-humanized antibody or as a chimeric antibody comprising variable regions from the parent non-humanized antibody and a human Fc region.

63. The humanized antibody or binding fragment thereof of claim 62, wherein the activity is measured as ristocetin-induced platelet agglutination activity.

64. The humanized antibody or binding fragment thereof of claim 4, wherein the humanized antibody lacks effector function.

65. The humanized antibody or binding fragment thereof of claim 4, wherein the humanized antibody comprises an Fc region derived from IgG4.

66. The humanized antibody or binding fragment thereof of claim 4, wherein the humanized antibody is specific for the A1 domain of human vWF.

67. The humanized antibody of claim 4, wherein the humanized antibody is a full length antibody.

68. The binding fragment of claim 4, wherein the binding fragment is an antibody fragment selected from the group consisting of Fab, Fab′

, Fab′

-SH, Fv, scFv, F(ab′

)2, and a diabody.

69. The binding fragment of claim 68, wherein the antibody fragment is not a Fab.

5. A humanized antibody or binding fragment thereof specific for vWF, the humanized antibody comprising heavy chain CDRs:

HCDR1;

GFSLTDYGVD (SEQ ID NO;

     7), HCDR2;

MIWGDGSTDYNSALKS (SEQ ID NO;

     8) and HCDR3;

DPADYGNYDYALDY (SEQ ID NO;

     9); and

/or light chain CDRs;

LCDR1;

SASQDINKYLN (SEQ ID NO;

     10), LCDR2;

YTSSLHS (SEQ ID NO;

     11) and LCDR3;

QQYEKLPWT (SEQ ID NO;

     12).

14. The humanized antibody or binding fragment thereof of claim 5, wherein the humanized antibody or binding fragment thereof comprises a heavy chain variable region sequence which comprises a framework region which is at least 80% identical to the framework region of SEQ ID NO:

19 and/or a light chain variable region sequence which comprises a framework region which is at least 80% identical to the framework region of SEQ ID NO;

28.

15. The humanized antibody or binding fragment thereof of claim 5 further comprising

heavy chain framework regions corresponding to framework regions in human antibody family VH4;

and

light chain framework regions corresponding to framework regions in human antibody family VK1.

16. The humanized antibody or binding fragment thereof of claim 5 further comprising

heavy chain framework regions 1, 2 and 3 corresponding to framework regions present in human antibody heavy chain germlne sequence 4-59, wherein heavy chain framework region 1 is QVQLQESGPGLVKPSETLSLTCTVS (SEQ ID NO:

171);

heavy chain framework region 2 is WIRQPPGKGLEWIG (SEQ ID NO;

     172); and

heavy chain framework region 3 is

RVTISVDTSKNQFSLKLSSVTAADTAVYYCAR (SEQ ID NO;

     173); and

light chain framework region 1, 2 and 3 corresponding to framework regions present in human antibody light chain germlne sequence 018, wherein light chain framework region 1 is DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO;

     186);

light chain framework region 2 is WYQQKPGKAPKLLIY (SEQ ID NO;

     187); and

light chain framework region 3 is

GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC (SEQ ID NO;

     188).

17. The humanized antibody or binding fragment thereof of claim 5 further comprising

heavy chain framework region 1, 2 and 3 corresponding to framework regions present in human antibody heavy chain germlne sequence 4-34, wherein heavy chain framework region 1 is QVQLQQWGAGLLKPSETLSLTCAVY (SEQ ID NO:

165);

heavy chain framework region 2 is WIRQPPGKGLEWIG (SEQ ID NO;

     166) and heavy chain framework region 3 is

RVTISVDTSKNQFSLKLSSVTAADTAVYYCAR (SEQ ID NO;

     167); and

light chain framework region 1, 2 and 3 corresponding to framework regions present in human antibody light chain germlne sequence 018, wherein light chain framework region 1 is DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO;

     186);

light chain framework region 2 is WYQQKPGKAPKLLIY (SEQ ID NO;

     187); and

light chain framework region 3 is

GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC (SEQ ID NO;

     188).

52. The humanized antibody or binding fragment thereof of claim 15, wherein the heavy chain framework region does not comprise one or more murine residues.

53. The humanized antibody or binding fragment thereof of claim 15, wherein the light chain framework region does not comprise one or more murine residues.

30. A humanized antibody or binding fragment thereof specific for vWF, the humanized antibody comprising a heavy chain CDR1 that is GFSLTDYGVD (SEQ ID NO:

7), a heavy chain CDR2 that is MIWGDGSTDYNSALKS (SEQ ID NO;

     8), a heavy chain CDR3 that is DPADYGNYDYALDY (SEQ ID NO;

     9) and a light chain CDR1, a light chain CDR2 and a light chain CDR3 that is QQYEKLPWT (SEQ ID NO;

     12), with the proviso that the light chain CDR1 is not SASQDINKYLN (SEQ ID NO;

     10) and/or the light chain CDR2 is not YTSSLHS (SEQ ID NO;

     11).

31. The humanized antibody or binding fragment thereof of claim 30 further comprising human antibody heavy chain framework regions and/or human antibody light chain framework regions.

32. The humanized antibody or binding fragment thereof of claim 31 wherein the heavy chain framework regions correspond to heavy chain framework regions present in a 4-59 derived human antibody.

33. The humanized antibody or binding fragment thereof of claim 32 wherein the heavy chain framework regions present in a 4-59 derived human antibody further comprises one or more murine residues.

34. The humanized antibody or binding fragment thereof of claim 32 wherein the heavy chain framework regions present in a 4-59 derived human antibody does not comprise one or more murine residues.

35. The humanized antibody or binding fragment thereof of claim 31 wherein the light chain framework regions correspond to the light chain framework regions present in a 018 derived human antibody.

36. The humanized antibody or binding fragment thereof of claim 35 wherein the light chain framework regions present in a 018 derived human antibody further comprises one or more murine residues.

37. The humanized antibody or binding fragment thereof of claim 35 wherein the light chain framework regions present in a 018 derived human antibody does not comprise one or more murine residues.

38. The humanized antibody or binding fragment thereof of claim 32, wherein the 4-59 derived human antibody is AAC18165.1.

39. The humanized antibody or binding fragment thereof of claim 35, wherein the 018 derived human antibody is AAK94808.

40. The humanized antibody or binding fragment thereof of claim 30, wherein LCDR1 and/or LCDR2 are from a human antibody.

41. The humanized antibody or binding fragment thereof of claim 40, wherein LCDR2 is DASNLET (SEQ ID NO:

118).

71. An isolated nucleic acid comprising the light chain encoding nucleic acid sequence of the vector GS264 as deposited in a microorganism with DSMZ having accession No. DSM 21059.

72. An isolated nucleic acid comprising the heavy chain encoding nucleic acid sequence of the vector GS265 as deposited in a microorganism with DSMZ having accession No. DSM 21060.

73. An isolated nucleic acid encoding a humanized antibody or binding fragment thereof specific for vWF that comprises a heavy chain variable region sequence as set forth in SEQ ID NO:

19 and a light chain variable region sequence as set forth in SEQ ID NO;

28.

74. An isolated nucleic acid encoding a humanized antibody or binding fragment thereof specific for vWF that comprises a heavy chain sequence as set forth in SEQ ID NO:

237 and a light chain sequence as set forth in SEQ ID NO;

238.

110. A human antibody or binding fragment thereof specific for von Willebrand factor (vWF), which can be administered in a therapeutically effective amount ranging from between 1 to around 250 times the ED₁₀₀ without causing clinical signs of bleeding.

111. The human antibody or binding fragment thereof of claim 110, wherein the antibody or binding fragment thereof is specific for the A1 domain of human vWF.