Humanized antibodies that recognize beta amyloid peptide

• US 7,179,892 B2
• Filed: 03/12/2003
• Issued: 02/20/2007
• Est. Priority Date: 12/06/2000
• Status: Expired due to Term

• Current Assignee: Janssen Alzheimer Immunotherapy (Johnson & Johnson), Wyeth LLC (Pfizer Inc.)
• Original Assignee: Neuralab Ltd. (Perrigo Company PLC), Wyeth (Pfizer Inc.)
• Inventors: Saldanha, Jose William, Basi, Guriq
• Primary Examiner(s): Andres; Janet L.
• Assistant Examiner(s): BALLARD, KIMBERLY
• Application Number:
  US10/388,389
• Publication Number:
  US 20040087777A1
• Time in Patent Office: 1,441 Days
• Field of Search: None
• US Class Current: 530/387.3
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• 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 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/77 Internalization into the cell
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C12N 2510/02 Cells for production

Abstract

The invention provides improved agents and methods for treatment of diseases associated with amyloid deposits of Aβ in the brain of a patient. Preferred agents include humanized antibodies.

Claims

1. A humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising;

(a) a light chain comprising a variable light chain region comprising the complementarity determining regions (CDRs) and at least one variable light chain framework residue selected from the group consisting of L1, L2, L4, L15, L35, L36, L38, L44, L46, L47, L48, L49, L64, L66, L68, L69, L71, L83, L85, L87, and L98 (Kabat numbering convention) from the 3D6 immunoglobulin light chain variable region sequence set forth as SEQ ID NO;

2, wherein the remainder of the variable light chain region is from a human acceptor immunoglobulin light chain; and

(b) a heavy chain comprising a variable heavy chain region having the sequence as set forth in residues 1–

119 of SEQ ID NO;

8 or SEQ ID NO;

12.

3. The humanized immunoglobulin or antigen binding fragment of claim 1, wherein at least three variable light chain framework residues are selected from the group consisting of L1, L2, L4, L15, L35, L36, L38, L44, L46, L47, L48, L49, L64, L66, L68, L69, L71, L83, L85, L87, and L98 (Kabat numbering convention).

15. The humanized immunoglobulin or antigen binding fragment of any one of claims 3 and 8, wherein the human acceptor light chain is of the subtype kappa II (Kabat convention).

17. The humanized immunoglobulin or antigen binding fragment of claim 15, wherein the human acceptor light chain is selected from the group consisting of Kabat ID 019230, Kabat ID 005131, Kabat ID 005058, Kabat ID 005057, Kabat ID 005059, Kabat ID U21040 and Kabat ID U41645.

18. The humanized immunoglobulin or antigen binding fragment of claim 15, wherein the human acceptor light chain is Kabat ID 019230.

22. The humanized immunoglobulin or antigen binding fragment of any one of claims 1, 3, 5, 7, 8, 11, 13, and 21, wherein the light chain variable region comprises at least one rare human framework residue and wherein the rare human framework residue is substituted with an amino acid residue which is common for human variable light chain sequences at that position.

23. The humanized immunoglobulin or antigen binding fragment of any one of claims 1, 3, 5, 7, 8, 11, 13, and 21, wherein the light chain variable region comprises at least one rare human framework residue and wherein the rare human framework residue is substituted with a corresponding amino acid residue from a germline variable light chain sequence.

24. The humanized immunoglobulin or antigen binding fragment of claim 23, wherein the germline variable light chain sequence is selected from the group consisting of A1, A17, A18, A2, and A19.

29. The humanized immunoglobulin or antigen binding fragment of claim 22, wherein the rare framework residue is selected based on occurrence at that position in less than 10% of human light chain variable region sequences in the light chain variable region subgroup, and the common residue is selected based on an occurrence at that position in greater than 50% of sequences in the light chain variable region subgroup.

31. The humanized immunoglobulin or antigen binding fragment of any one of claims 1, 3, 5, 7, 8, 11, 13, and 21, wherein the human light chain variable region framework is a human kappa light chain variable region.

37. The humanized immunoglobulin or antigen binding fragment of any one of claims 1–

14 and 21, wherein the heavy chain isotype is γ

1.

38. The humanized immunoglobulin or antigen binding fragment of any one of claims 1–

14 and 21, which binds to both soluble beta amyloid peptide (Aβ

) and aggregated Aβ

.

39. The humanized immunoglobulin or antigen binding fragment of claim 38, wherein the soluble beta amyloid peptide (Aβ

) is disaggregated Aβ

.

40. The humanized immunoglobulin or antigen binding fragment of any one of claims 1–

14 and 21, which mediates phagocytosis of beta amyloid peptide (Aβ

).

41. The humanized immunoglobulin or antigen binding fragment of any one of claims 1–

14 and 21, which crosses the blood-brain barrier in a subject.

42. The humanized immunoglobulin or antigen binding fragment of any one of claims 1–

14 and 21, which reduces beta amyloid peptide (Aβ

) burden in a subject.

43. The humanized immunoglobulin or antigen binding fragment of claim 1, wherein the variable heavy chain region is as set forth in residues 1–

119 of SEQ ID NO;

8.

44. The humanized immunoglobulin or antigen binding fragment of claim 1, wherein the variable heavy chain region is as set forth in residues 1–

119 of SEQ ID NO;

12.

47. A kit comprising the humanized immunoglobulin or antigen-binding fragment of any one of claims 1–

14, and 21, together with instructions for use.

48. The humanized immunoglobulin or antigen binding fragment of claim 1, wherein the human acceptor light chain is of the subtype kappa II (Kabat convention).

61. The humanized immunoglobulin or antigen binding fragment of claim 1, wherein the constant regions are derived from human IgG1.

70. The humanized immunoglobulin or antigen binding fragment of claim 1, wherein the constant regions are derived from human IgG4.

79. The humanized immunoglobulin or antigen binding fragment of claim 1, which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁸ M<sup>−

1</sup>.

2. A humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising;

(a) heavy chain comprising a variable heavy chain region comprising the complementarity determining regions (CDRs) and at least one variable heavy chain framework residue selected from the group consisting of H2, H24, H26, H27, H28, H29, H30, H37, H39, H40, H42, H45, H47, H48, H49, H67, H69, H71, H80, H91, H93, H94, and H103 (Kabat numbering convention) from the 3D6 immunoglobulin heavy chain variable region sequence set forth as SEQ ID NO;

4, wherein the remainder of the variable heavy chain region is from a human acceptor immunoglobulin heavy chain; and

(b) a light chain comprising a variable light chain region having the sequence as set forth in residues 1–

112 of SEQ ID NO;

5 or SEQ ID NO;

11.

4. The humanized immunoglobulin or antigen binding fragment of claim 2, wherein at least three variable heavy chain framework residues are selected from the group consisting of H2, H24, H26, H27, H28, H29, H30, H37, H39, H40, H42, H45, H47, H48, H49, H67, H69, H71, H80, H91, H93, H94, and H103 (Kabat numbering convention).

45. The humanized immunoglobulin or antigen binding fragment of claim 2, wherein the variable light chain region is as set forth in residues 1–

112 of SEQ ID NO;

5.

46. The humanized immunoglobulin or antigen binding fragment of claim 2, wherein the variable light chain region is as set forth in residues 1–

112 of SEQ ID NO;

11.

54. The humanized immunoglobulin or antigen binding fragment of claim 2, wherein the human heavy chain variable region framework is a human immunoglobulin IgG1 heavy chain variable region.

62. The humanized immunoglobulin or antigen binding fragment of claim 2, wherein the constant regions are derived from human IgG1.

71. The humanized immunoglobulin or antigen binding fragment of claim 2, wherein the constant regions are derived from human IgG4.

80. The humanized immunoglobulin or antigen binding fragment of claim 2, which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁸ M<sup>−

1</sup>.

5. A humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising;

(i) a light chain comprising a variable light chain region comprising

(a) the complementarity determining regions (CDRs) from the 3D6 immunoglobulin light chain variable region sequence set forth as SEQ ID NO;

2;

(b) at least one variable light chain canonical framework residue selected from the group consisting of L2, L48, L64, and L71 (Kabat numbering convention) from SEQ ID NO;

2; and

(c) at least one variable light chain interchain packing framework residue selected from the group consisting of L36, L38, L44, L46, L87, and L98 (Kabat numbering convention) from SEQ ID NO;

2; and

, optionally,

(d) at least one variable light chain vernier zone framework residue selected from the group consisting of L4, L35, L47, L49, L66, L68, and L69 (Kabat numbering convention) from SEQ ID NO;

2, or at least one variable light chain rare framework residue selected from the group consisting of L1, L15, L83, and L85 (Kabat numbering convention) from SEQ ID NO;

2,

wherein the remainder of the variable light chain region is from a human acceptor immunoglobulin light chain; and

(ii) a heavy chain comprising a variable heavy chain region having the sequence as set forth in residues 1–

119 of SEQ ID NO;

8 or SEQ ID NO;

12.

7. The humanized immunoglobulin or antigen binding fragment of claim 5, wherein the light chain comprises at least one substitution of a vernier zone framework residue selected from the group consisting of L4, L35, L47, L49, L66, L68, and L69 (Kabat numbering convention).

8. The humanized immunoglobulin or antigen-binding fragment of claim 5, wherein the light chain comprises at least one rare framework residue selected from the group consisting of L1, L15, L83, and L85 (Kabat numbering convention).

49. The humanized immunoglobulin or antigen binding fragment of claim 5, wherein the human acceptor light chain is of the subtype kappa II (Kabat convention).

50. The humanized immunoglobulin or antigen binding fragment of claim 7, wherein the human acceptor light chain is of the subtype kappa II (Kabat convention).

63. The humanized immunoglobulin or antigen binding fragment of claim 5, wherein the constant regions are derived from human IgG1.

72. The humanized immunoglobulin or antigen binding fragment of claim 5, wherein the constant regions are derived from human IgG4.

81. The humanized immunoglobulin or antigen binding fragment of claim 5, which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁸ M<sup>−

1</sup>.

88. The humanized immunoglobulin or antigen binding fragment of claim 5, wherein the variable heavy chain region is as set forth in residues 1–

119 of SEQ ID NO;

8.

92. The humanized immunoglobulin or antigen binding fragment of claim 5, wherein the variable heavy chain region is as set forth in residues 1–

119 of SEQ ID NO;

12.

6. A humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising;

(i) a heavy chain comprising a variable heavy chain region comprising

(a) the complementarity determining regions (CDRs) from the 3D6 immunoglobulin heavy chain variable region sequence set forth as SEQ ID NO;

4;

(b) at least one variable heavy chain canonical framework residue selected from the group consisting of H24, H26, H27, H29, H71, and H94 (Kabat numbering convention) from SEQ ID NO;

4;

(c) at least one variable heavy chain interchain packing framework residue selected from the group consisting of H37, H39, H45, H47, H91, H93, and H103 (Kabat numbering convention) from SEQ ID NO;

4; and

,

(d) at least one variable heavy chain vernier zone framework residue selected from the group consisting of H2, H28, H30, H48, H49, H67, H69, and H80 (Kabat numbering convention) from SEQ ID NO;

4, or at least one variable heavy chain rare framework residue selected from the group consisting of H40 and H42 (Kabat numbering convention) from SEQ ID NO;

4, wherein the remainder of the variable heavy chain region is from a human acceptor immunoglobulin heavy chain; and

(ii) a light chain comprising a variable light chain region having the sequence as set forth in residues 1–

112 of SEQ ID NO;

5 or SEQ ID NO;

11.

9. The humanized immunoglobulin or antigen-binding fragment of claim 6, wherein the heavy chain comprises at least one vernier zone framework residue selected from the group consisting of H2, H28, H30, H48, H49, H67, H69, and H80 (Kabat numbering convention).

10. The humanized immunoglobulin or antigen-binding fragment of claim 6, wherein the heavy chain comprises at least one rare framework residue selected from the group consisting of H40 and H42 (Kabat numbering convention).

16. The humanized immunoglobulin or antigen binding fragment of any one of claims 2, 4, 6, 9, 10, 12, and 14, wherein the human acceptor heavy chain is of the subtype III (Kabat convention).

19. The humanized immunoglobulin or antigen binding fragment of claim 16, wherein the human acceptor heavy chain is selected from the group consisting of Kabat ID 045919, Kabat ID 000459, Kabat ID 000553, Kabat ID 000386 and Kabat ID M23691.

20. The humanized immunoglobulin or antigen binding fragment of claim 16, wherein the human acceptor heavy chain is Kabat ID 045919.

25. The humanized immunoglobulin or antigen binding fragment of any one of claims 2, 4, 6, 9, 10, 12, and 14, wherein the heavy chain variable region comprises at least one rare human framework residue and wherein the rare human framework residue is substituted with an amino acid residue which is common for human variable heavy chain sequences at that position.

26. The humanized immunoglobulin or antigen binding fragment of any one of claims 2, 4, 6, 9, 10, 12, and 14, wherein the heavy chain variable region comprises at least one rare human framework residue and wherein the rare human framework residue is substituted with a corresponding amino acid residue from a germline variable heavy chain sequence.

27. The humanized immunoglobulin or antigen binding fragment of claim 26, wherein the germline variable heavy chain sequence is selected from the group consisting of VH3-48, VH3-23, VH3-7, VH3-21 and VH3-11.

28. The humanized immunoglobulin or antigen binding fragment of claim 26, wherein the germline variable heavy chain sequence is VH3-23.

30. The humanized immunoglobulin or antigen binding fragment of claim 25, wherein the rare framework residue is selected based on occurrence at that position in less than 10% of human heavy chain variable region sequences in the heavy chain variable region subgroup, and the common residue is selected based on an occurrence at that position in greater than 50% of sequences in the heavy chain variable region subgroup.

32. The humanized immunoglobulin or antigen binding fragment of any one of claims 4, 9, and 10, wherein the human heavy chain variable region framework is a human immunoglobulin IgG1 heavy chain variable region.

33. The humanized immunoglobulin or antigen binding fragment of any one of claims 3, 4 and 7–

10, wherein the constant regions are derived from human IgG1.

34. The humanized immunoglobulin or antigen binding fragment of any one of claims 3, 4 and 7–

10, wherein the constant regions are derived from human IgG4.

35. The humanized immunoglobulin or antigen binding fragment of any one of claims 3, 4 and 7–

10, which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁸ M<sup>−

1</sup>.

36. The humanized immunoglobulin or antigen binding fragment of claim 35, which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁹ M<sup>−

1</sup>.

55. The humanized immunoglobulin or antigen binding fragment of claim 6, wherein the human heavy chain variable region framework is a human immunoglobulin IgG1 heavy chain variable region.

58. The humanized immunoglobulin or antigen binding fragment of claim 16, wherein the human heavy chain variable region framework is a human immunoglobulin IgG1 heavy chain variable region.

59. The humanized immunoglobulin or antigen binding fragment of claim 19, wherein the human heavy chain variable region framework is a human immunoglobulin IgG1 heavy chain variable region.

60. The humanized immunoglobulin or antigen binding fragment of claim 20, wherein the human heavy chain variable region framework is a human immunoglobulin IgG1 heavy chain variable region.

64. The humanized immunoglobulin or antigen binding fragment of claim 6, wherein the constant regions are derived from human IgG1.

73. The humanized immunoglobulin or antigen binding fragment of claim 6, wherein the constant regions are derived from human IgG4.

82. The humanized immunoglobulin or antigen binding fragment of claim 6, which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁸ M<sup>−

1</sup>.

96. The humanized immunoglobulin or antigen binding fragment of claim 6, wherein the variable light chain region is as set forth in residues 1–

112 of SEQ ID NO;

5.

99. The humanized immunoglobulin or antigen binding fragment of claim 6, wherein the variable light chain region is as set forth in residues 1–

1 12 of SEQ ID NO;

11.

11. A humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising;

(a) a light chain comprising a variable light chain region comprising the complementarity determining regions (CDRs) and at least one variable light chain framework residue selected from the group consisting of L1, L2, L36, and L46 (Kabat numbering convention) from the 3D6 immunoglobulin light chain variable region sequence set forth as SEQ ID NO;

2, wherein the remainder of the variable framework region is from a human acceptor immunoglobulin light chain; and

(b) a heavy chain comprising a variable heavy chain region having the sequence as set forth in residues 1–

119 of SEQ ID NO;

8 or SEQ ID NO;

12.

51. The humanized immunoglobulin or antigen binding fragment of claim 11, wherein the human acceptor light chain is of the subtype kappa II (Kabat convention).

65. The humanized immunoglobulin or antigen binding fragment of claim 11, wherein the constant regions are derived from human IgG1.

74. The humanized immunoglobulin or antigen binding fragment of claim 11, wherein the constant regions are derived from human IgG4.

83. The humanized immunoglobulin or antigen binding fragment of claim 11, which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁸ M<sup>−

1</sup>.

89. The humanized immunoglobulin or antigen binding fragment of claim 11, wherein the variable heavy chain region is as set forth in residues 1–

119 of SEQ ID NO;

8.

93. The humanized immunoglobulin or antigen binding fragment of claim 11, wherein the variable heavy chain region is as set forth in residues 1–

119 of SEQ ID NO;

12.

12. A humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising;

(a) a heavy chain comprising a variable heavy chain region comprising the complementarity determining regions (CDRs) and at least one variable heavy chain framework residue selected from the group consisting of H49, H93 and H94 (Kabat numbering convention) from the 3D6 immunoglobulin heavy chain variable region sequence set forth as SEQ ID NO;

4, wherein the remainder of the variable framework region is from a human acceptor immunoglobulin heavy chain; and

(b) a light chain comprising a variable light chain region having the sequence as set forth in residues 1–

112 of SEQ ID NO;

5 or SEQ ID NO;

11.

56. The humanized immunoglobulin or antigen binding fragment of claim 12, wherein the human heavy chain variable region framework is a human immunoglobulin IgG1 heavy chain variable region.

66. The humanized immunoglobulin or antigen binding fragment of claim 12, wherein the constant regions are derived from human IgG1.

75. The humanized immunoglobulin or antigen binding fragment of claim 12, wherein the constant regions are derived from human IgG4.

84. The humanized immunoglobulin or antigen binding fragment of claim 12, which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁸ M<sup>−

1</sup>.

97. The humanized immunoglobulin or antigen binding fragment of claim 12, wherein the variable light chain region is as set forth in residues 1–

112 of SEQ ID NO;

5.

100. The humanized immunoglobulin or antigen binding fragment of claim 12, wherein the variable light chain region is as set forth in residues 1–

112 of SEQ ID NO;

11.

13. A humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising;

(a) a light chain comprising a variable light chain region comprising the complementarity determining regions (CDRs) and variable framework residues L2, L36, and L46 (Kabat numbering convention) from the 3D6 immunoglobulin light chain variable region sequence set forth as SEQ ID NO;

2, wherein the remainder of the variable light chain region is from a human acceptor immunoglobulin light chain; and

(b) a heavy chain comprising a variable heavy chain region having the sequence as set forth in residues 1–

119 of SEQ ID NO;

8 or SEQ ID NO;

12.

52. The humanized immunoglobulin or antigen binding fragment of claim 13, wherein the human acceptor light chain is of the subtype kappa II (Kabat convention).

67. The humanized immunoglobulin or antigen binding fragment of claim 13, wherein the constant regions are derived from human IgG1.

76. The humanized immunoglobulin or antigen binding fragment of claim 13, wherein the constant regions are derived from human IgG4.

85. The humanized immunoglobulin or antigen binding fragment of claim 13, which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁸ M<sup>−

1</sup>.

90. The humanized immunoglobulin or antigen binding fragment of claim 13, wherein the variable heavy chain region is as set forth in residues 1–

119 of SEQ ID NO;

8.

94. The humanized immunoglobulin or antigen binding fragment of claim 13, wherein the variable heavy chain region is as set forth in residues 1–

119 of SEQ ID NO;

12.

14. A humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising;

(a) a heavy chain comprising a variable heavy chain region comprising the complementarity determining regions (CDRs) and variable framework residues H49, H93 and H94 (Kabat numbering convention) from the 3D6 immunoglobulin heavy chain variable region sequence set forth as SEQ ID NO;

4, wherein the remainder of the variable heavy chain region is from a human acceptor immunoglobulin heavy chain; and

(b) a light chain comprising a variable light chain region having the sequence as set forth in residues 1–

112 of SEQ ID NO;

5 or SEQ ID NO;

11.

57. The humanized immunoglobulin or antigen binding fragment of claim 14, wherein the human heavy chain variable region framework is a human immunoglobulin IgG1 heavy chain variable region.

68. The humanized immunoglobulin or antigen binding fragment of claim 14, wherein the constant regions are derived from human IgG1.

77. The humanized immunoglobulin or antigen binding fragment of claim 14, wherein the constant regions are derived from human IgG4.

86. The humanized immunoglobulin or antigen binding fragment of claim 14, which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁸ M<sup>−

1</sup>.

98. The humanized immunoglobulin or antigen binding fragment of claim 14, wherein the variable light chain region is as set forth in residues 1–

1 12 of SEQ ID NO;

5.

101. The humanized immunoglobulin or antigen binding fragment of claim 14, wherein the variable light chain region is as set forth in residues 1–

112 of SEQ ID NO;

11.

21. A humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising;

(a) a light chain comprising a variable light chain region comprising the complementarity determining regions (CDRs) and variable framework residues L1, L2, L36, and L46 (Kabat numbering convention) from the 3D6 immunoglobulin light chain variable region sequence set forth as SEQ ID NO;

2, wherein the remainder of the variable light chain region is from a human acceptor immunoglobulin light chain; and

(b) a heavy chain comprising a variable heavy chain region having the sequence as set forth in residues 1–

119 of SEQ ID NO;

8 or SEQ ID NO;

12.

53. The humanized immunoglobulin or antigen binding fragment of claim 21, wherein the human acceptor light chain is of the subtype kappa II (Kabat convention).

69. The humanized immunoglobulin or antigen binding fragment of claim 21, wherein the constant regions are derived from human IgG1.

78. The humanized immunoglobulin or antigen binding fragment of claim 21, wherein the constant regions are derived from human IgG4.

87. The humanized immunoglobulin or antigen binding fragment of claim 21, which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁸ M<sup>−

1</sup>.

91. The humanized immunoglobulin or antigen binding fragment of claim 21, wherein the variable heavy chain region is as set forth in residues 1–

119 of SEQ ID NO;

8.

95. The humanized immunoglobulin or antigen binding fragment of claim 21, wherein the variable heavy chain region is as set forth in residues 1–

119 of SEQ ID NO;

12.

102. An immunoglobulin which specifically binds to beta amyloid peptide (Aβ

), or antigen-binding fragment thereof, comprising a variable heavy chain region set as forth in residues 1–

119 of SEQ ID NO;

8 and a variable light chain region as set forth in residues 1–

112 of SEQ ID NO;

11.

103. An immunoglobulin which specifically binds to beta amyloid peptide (Aβ

), or antigen-binding fragment thereof, comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

12, and a variable light chain region as set forth in residues 1–

112 of SEQ ID NO;

5.

104. An immunoglobulin comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

8, a variable light chain region as set forth in residues 1–

112 of SEQ ID NO;

11, and constant regions from human IgG1.

105. An immunoglobulin comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

12, a light chain region as set forth in residues 1–

112 of SEQ ID NO;

5, and constant regions from human IgG1.

106. An immunoglobulin which specifically binds to beta amyloid peptide (Aβ

), or antigen-binding fragment thereof, comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

8 and a variable light chain region as set forth in residues 1–

112 of SEQ ID NO;

5.

107. An immunoglobulin which specifically binds to beta amyloid peptide (Aβ

), or antigen-binding fragment thereof, comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

12 and a light chain region as set forth in residues 1–

112 of SEQ ID NO;

11.

108. An immunoglobulin comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

8, a variable light chain region as set forth in residues 1–

112 of SEQ ID NO;

5, and constant regions from human IgG1.

109. An immunoglobulin comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

12, a light chain region as set forth in residues 1–

112 of SEQ ID NO;

11, and constant regions from human IgG1.

110. A pharmaceutical composition comprising:

(i) a humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen-binding fragment thereof, the humanized immunoglobulin or antigen-binding fragment comprising

(a) a light chain comprising a variable light chain region comprising the complementarity determining regions (CDRs) and at least one variable light chain framework residue selected from the group consisting of L1, L2, L4, L15, L35, L36, L38, L44, L46, L47, L48, L49, L64, L66, L68, L69, L71, L83, L85, L87, and L98 (Kabat numbering convention) from the 3D6 immunoglobulin light chain variable region sequence set forth as SEQ ID NO;

2, wherein the remainder of the variable light chain region is from a human acceptor immunoglobulin light chain, and

(b) a heavy chain comprising a variable heavy chain region having the sequence as set forth in residues 1–

119 of SEQ ID NO;

8 or SEQ ID NO;

12; and

(ii) a pharmaceutical carrier.

111. A pharmaceutical composition comprising:

(i) a humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising

(a) heavy chain comprising a variable heavy chain region comprising the complementarity determining regions (CDRs) and at least one variable heavy chain framework residue selected from the group consisting of H2, H24, H26, H27, H28, H29, H30, H37, H39, H40, H42, H45, H47, H48, H49, H67, H69, H71, H80, H91, H93, H94, and H103 (Kabat numbering convention) from the 3D6 immunoglobulin heavy chain variable region sequence set forth as SEQ ID NO;

4, wherein the remainder of the variable heavy chain region is from a human acceptor immunoglobulin heavy chain, and

(b) a light chain comprising a variable light chain region having the sequence as set forth in residues 1–

112 of SEQ ID NO;

5 or SEQ ID NO;

11; and

(ii) a pharmaceutical carrier.

112. A pharmaceutical composition comprising:

(i) a humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising;

(1) a light chain comprising a variable light chain region comprising

(a) the complementarity determining regions (CDRs) from the 3D6 immunoglobulin light chain variable region sequence set forth as SEQ ID NO;

2;

(b) at least one variable light chain canonical framework residue selected from the group consisting of L2, L48, L64, and L71 (Kabat numbering convention) from SEQ ID NO;

2; and

(c) at least one variable light chain interchain packing framework residue selected from the group consisting of L36, L38, L44, L46, L87, and L98 (Kabat numbering convention) from SEQ ID NO;

2; and

, optionally,

(d) at least one variable light chain vernier zone framework residue selected from the group consisting of L4, L35, L47, L49, L66, L68, and L69 (Kabat numbering convention) from SEQ ID NO;

2, or at least one variable light chain rare framework residue selected from the group consisting of L1, L15, L83, and L85 (Kabat numbering convention) from SEQ ID NO;

2, wherein the remainder of the variable light chain region is from a human acceptor immunoglobulin light chain; and

(2) a heavy chain comprising a variable heavy chain region having the sequence as setforth in residues 1–

119 of SEQ ID NO;

8 or SEQ ID NO;

12; and

(ii) a pharmaceutical carrier.

113. A pharmaceutical composition comprising:

(i) a humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising

(1) a heavy chain comprising a variable heavy chain region comprising

(a) the complementarity determining regions (CDRs) from the 3D6 immunoglobulin heavy chain variable region sequence set forth as SEQ ID NO;

4;

(b) at least one variable heavy chain canonical framework residue selected from the group consisting of H24, H26, H27, H29, H71, and H94 (Kabat numbering convention) from SEQ ID NO;

4;

(c) at least one variable heavy chain interchain packing framework residue selected from the group consisting of H37, H39, H45, H47, H91, H93, and H103 (Kabat numbering convention) from SEQ ID NO;

4; and

(d) at least one variable heavy chain vernier zone framework residue selected from the group consisting of H2, H28, H30, H48, H49, H67, H69, and H80 (Kabat numbering convention) from SEQ ID NO;

4, or at least one variable heavy chain rare framework residue selected from the group consisting of H40 and H42 (Kabat numbering convention) from SEQ ID NO;

4, wherein the remainder of the variable heavy chain region is from a human acceptor immunoglobulin heavy chain, and

(2) a light chain comprising a variable light chain region having the sequence as set forth in residues 1–

112 of SEQ ID NO;

5 or SEQ ID NO;

11; and

(ii) a pharmaceutical carrier.

114. A pharmaceutical composition comprising:

(i) a humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising;

(a) a light chain comprising a variable light chain region comprising the complementarity determining regions (CDRs) and at least one variable light chain framework residue selected from the group consisting of L1, L2, L36, and L46 (Kabat numbering convention) from the 3D6 immunoglobulin light chain variable region sequence set forth as SEQ ID NO;

2, wherein the remainder of the variable light chain region is from a human acceptor immunoglobulin light chain, and

(b) a heavy chain comprising a variable heavy chain region having the sequence as setforth in residues 1–

119 of SEQ ID NO;

8 or SEQ ID NO;

12; and

(ii) a pharmaceutical carrier.

115. A pharmaceutical composition comprising:

(i) a humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising

(a) a heavy chain comprising a variable heavy chain region comprising the complementarity determining regions (CDRs) and at least one variable heavy chain framework residue selected from the group consisting of H49, H93 and H94 (Kabat numbering convention) from the 3D6 immunoglobulin heavy chain variable region sequence set forth as SEQ ID NO;

4, wherein the remainder of the variable heavy chain region is from a human acceptor immunoglobulin heavy chain, and

(b) a light chain comprising a variable light chain region having the sequence as set forth in residues 1–

112 of SEQ ID NO;

5 or SEQ ID NO;

11; and

(ii) a pharmaceutical carrier.

116. A pharmaceutical composition comprising:

(i) a humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising

(a) a light chain comprising a variable light chain region comprising the complementarity determining regions (CDRs) and variable framework residues L1, L2, L36, and L46 (Kabat numbering convention) from the 3D6 immunoglobulin light chain variable region sequence set forth as SEQ ID NO;

2, wherein the remainder of the variable light chain region is from a human acceptor immunoglobulin light chain, and

(b) a heavy chain comprising a variable heavy chain region having the sequence as set forth in residues 1–

119 of SEQ ID NO;

8 or SEQ ID NO;

12; and

(ii) a pharmaceutical carrier.

117. A pharmaceutical composition comprising:

(i) a humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising

(a) a light chain comprising a variable light chain region comprising the complementarity determining regions (CDRs) and variable framework residues L2, L36, and L46 (Kabat numbering convention) from the 3D6 immunoglobulin light chain variable region sequence set forth as SEQ ID NO;

2, wherein the remainder of the variable light chain region is from a human acceptor immunoglobulin light chain, and

(b) a heavy chain comprising a variable heavy chain region having the sequence as set forth in residues 1–

119 of SEQ ID NO;

8 or SEQ ID NO;

12; and

(ii) a pharmaceutical carrier.

118. A pharmaceutical composition comprising:

(i) a humanized immunoglobulin which specifically binds to beta amyloid peptide (Aβ

) with a binding affinity of at least 10⁷ M<sup>−

1</sup>, or antigen binding fragment thereof, the humanized immunoglobulin or antigen binding fragment comprising

(a) a heavy chain comprising a variable heavy chain region comprising the complementarity determining regions (CDRs) and variable framework residues H49, H93 and H94 (Kabat numbering convention) from the 3D6 immunoglobulin heavy chain variable region sequence set forth as SEQ ID NO;

4, wherein the remainder of the variable heavy chain region is from a human acceptor immunoglobulin heavy chain, and

(b) a light chain comprising a variable light chain region having the sequence as set forth in residues 1–

112 of SEQ ID NO;

5 or SEQ ID NO;

11; and

(ii) a pharmaceutical carrier.

119. A pharmaceutical composition comprising:

(i) an immunoglobulin which specifically binds to beta amyloid peptide (Aβ

), or antigen-binding fragment thereof, comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

8 and a variable light chain region as set forth in residues 1–

112 of SEQ ID NO;

11; and

(ii) a pharmaceutical carrier.

120. A pharmaceutical composition comprising:

(i) an immunoglobulin which specifically binds to beta amyloid peptide (Aβ

), or antigen-binding fragment thereof, comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

12, and a variable light chain region as set forth in residues 1–

112 of SEQ ID NO;

5; and

(ii) a pharmaceutical carrier.

121. A pharmaceutical composition comprising:

(i) an immunoglobulin comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

8, a variable light chain region as set forth in residues 1–

112 of SEQ ID NO;

1 1, and constant regions from human IgG1; and

(ii) a pharmaceutical carrier.

122. A pharmaceutical composition comprising:

(i) an immunoglobulin comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

12, a light chain region as set forth in residues 1–

112 of SEQ ID NO;

5, and constant regions from human IgG1; and

(ii) a pharmaceutical carrier.

123. A pharmaceutical composition comprising:

(i) an immunoglobulin which specifically binds to beta amyloid peptide (Aβ

), or antigen-binding fragment thereof, comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

8 and a variable light chain region as set forth in residues 1–

112 of SEQ ID NO;

5; and

(ii) a pharmaceutical carrier.

124. A pharmaceutical composition comprising:

(i) an immunoglobulin which specifically binds to beta amyloid peptide (Aβ

), or antigen-binding fragment thereof, comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

12 and a light chain region as set forth in residues 1–

112 of SEQ ID NO;

11; and

(ii) a pharmaceutical carrier.

125. A pharmaceutical composition comprising:

(i) an immunoglobulin comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

8, a variable light chain region as set forth in residues 1–

112 of SEQ ID NO;

5, and constant regions from human IgG1; and

(ii) a pharmaceutical carrier.

126. A pharmaceutical composition comprising:

(i) an immunoglobulin comprising a variable heavy chain region as set forth in residues 1–

119 of SEQ ID NO;

12, a light chain region as set forth in residues 1–

112 of SEQ ID NO;

11, and constant regions from human IgG1; and

(ii) a pharmaceutical carrier.