HUMANIZED ANTIBODY

• US 20090093002A1
• Filed: 07/13/2007
• Published: 04/09/2009
• Est. Priority Date: 07/14/2006
• Status: Active Grant

• Current Assignee: AC Immune SA, Genentech, Inc. (Roche Holding AG)
• Original Assignee: AC Immune SA, Genentech, Inc. (Roche Holding AG)
• Inventors: Pihlgren, Maria, Muhs, Andreas, Watts, Ryan, Pfeifer, Andrea
• Granted Patent: US 7,892,544 B2
• Time in Patent Office: Days
• US Class Current: 435/7.21
• 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 25/16 Anti-Parkinson drugs
• 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 16/28 against receptors, cell surface antigens or cell surface determinants
• 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 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: G01N 33/6896 Neurological disorders, e.g. Alzheimer's disease

Abstract

The present invention is related to chimeric and humanized antibody and to methods and compositions for the therapeutic and diagnostic use in the treatment of amyloidosis, a group of disorders and abnormalities associated with amyloid protein such as Alzheimer'"'"'s disease.

Claims

1. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof, which specifically binds to at least one epitope on the β

-amyloid protein wherein the epitope comprises at least two consecutive amino acid residues predominantly involved in binding to the antibody, wherein the at least two consecutive amino, acid residues are -Lys-Leu- embedded within the following core sequence (SEQ ID NO;

     10);

Xaa₁-Xaa₂-Lys-Leu-Xaa₃ wherein

Xaa₁ is an amino acid selected from the group consisting of His, Asn, Gln Lys, and Arg,

Xaa₂ is an amino acid selected from the group consisting of Asn and Gln; and

Xaa₃ is an amino acid selected from the group consisting of Ala, Val, Leu, norleucine, Met, Phe, and Ile.

3. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to claims 1 or 2, which specifically binds to at least two epitopes on the β

-amyloid protein wherein said at least two epitopes each comprise at least two consecutive amino acid residues predominantly involved in the binding of the antibody, wherein the at least two consecutive amino acids are separated by at least one amino acid residue not involved in antibody binding or to a significantly smaller extent than said consecutive amino acid residues, which are -Phe-Phe- and -Lys-Leu-, respectively, embedded within the following core sequence;

Xaa₁-Xaa₂-Lys-Leu-Xaa₃-Phe-Phe-Xaa₄-Xaa₅-Xaa₆ (SEQ ID NO;

     80), wherein

Xaa₁ is an amino acid selected from the group consisting of His, Asn, Gln Lys, and Arg;

Xaa₂ is an amino acid selected from the group consisting of Asn and Gln;

Xaa₃ is an amino acid selected from the group consisting of Ala, Val, Leu, norleucine, Met, Phe, and Ile;

Xaa₄ is an amino acid selected from the group consisting of Ala, Val, Leu, Ser and Ile;

Xaa₅ is an amino acid selected from the group consisting of Glu and Asp, and

Xaa₆ is an amino acid selected from the group consisting of Glu and Asp.

4. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of claims 1-3, wherein

Xaa₁ is an amino acid selected from the group consisting of His and Arg;

Xaa₂ is an amino acid selected from the group consisting of Asn and Gln;

Xaa₃ is an amino acid selected from the group consisting of Val and Leu,

Xaa₄ is an amino acid selected from the group consisting of Ala and Val;

Xaa₅ is an amino acid selected from the group consisting of Glu and Asp, and

Xaa₆ is an amino acid selected from the group consisting of Glu and Asp.

5. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to claim 4, wherein

Xaa₁ is His

Xaa₂ is Gln;

Xaa₃ is Val;

Xaa₄ is Ala;

Xaa₅ is Glu; and

Xaa₆ is Asp.

6. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of claims 1-5, which specifically binds to at least two epitopes on the β

-amyloid protein wherein said at least two epitopes each comprise at least two consecutive amino acid residues predominantly involved in the binding of the antibody,

which are -Phe-Phe- and -Lys-Leu-, respectively, and wherein said at least two distinct binding sites exhibit amino acid sequence -Val-Phe-Phe-Ala-Glu-Asp- shown in SEQ ID NO;

7 and amino acid sequence His-Gln-Lys-Leu-Val- shown in SEQ ID NO;

8, respectively.

8. A humanized antibody or a fragment thereof according to according to any one of claims 1-7, wherein the CDR of non-human origin is obtained from a donor antibody raised against an antigen fragment which does not contain said epitope.

34. A humanized antibody or a fragment thereof according to any one of claims 1-33, wherein at least one of the amino acids of the human- or primate-derived framework region is changed through a substitution to an amino acid from the corresponding region of murine humanized antibody ACI-01-Ab7C2 or a substitution conservative thereto.

35. A humanized antibody or a fragment thereof according to claim 34, wherein the Trp in Kabat position 47 in the human- or primate-derived framework region of the Heavy Chain Variable Region is replaced by an amino acid selected from the group consisting of Leu, norleucine, Ile, Val, Met, Ala, and Phe.

36. A humanized antibody or a fragment thereof according to claim 34, wherein the Arg in Kabat position 94 in the human- or primate-derived framework region of the Heavy Chain Variable Region as shown in SEQ ID NO:

15 is replaced by an amino acid selected from the group consisting of Ser and Thr, but especially by Ser.

37. A humanized antibody or a fragment thereof according to claim 34, wherein the Trp in Kabat position 47 in the human- or primate-derived framework region of the Heavy Chain Variable Region as shown in SEQ ID NO:

15 is replaced by an amino acid selected from the group consisting of Leu, norleucine, Ile, Val, Met, Ala, and Phe, particularly Leu and Ile, but especially Leu and Arg in Kabat position 94 in the human- or primate-derived framework region of the Heavy Chain Variable Region as shown in SEQ ID NO;

15 is replaced by an amino acid selected from the group consisting of Ser and Thr, but especially by Ser.

38. A humanized antibody or a fragment thereof according to claim 34, wherein the Tyr in Kabat position 87 in the human- or primate-derived framework region of the Light Chain Variable Region as shown in SEQ ID NO:

12 is replaced by an amino acid selected from the group consisting of Phe, Leu, Val, Ile, and Ala, particularly by Leu and Phe, but especially by Phe.

39. A humanized antibody or a fragment thereof according to claim 34, wherein the Trp in Kabat position 47 in the human- or primate-derived framework region of the Heavy Chain Variable Region as shown in SEQ ID NO:

15 is replaced by an amino acid selected from the group consisting of Leu, norleucine, Ile, Val, Met, Ala, and Phe, particularly Leu and Ile, but especially Leu and the Arg in Kabat position 94 in the human- or primatederived framework region of the Heavy Chain Variable Region as shown in SEQ ID NO;

15 is replaced by an amino acid selected from the group consisting of Ser and Thr, but especially by Ser, and the Tyr in Kabat position 87 in the human- or primate-derived framework region of the Light Chain Variable Region as shown in SEQ ID NO;

12 is replaced by an amino acid selected from the group consisting of Phe, Leu, Val, Ile, and Ala, particularly by Leu and Phe, but especially by Phe.

40. A humanized antibody or a fragment thereof according to claim 35, wherein the Trp in Kabat position 47 in the human- or primate-derived framework region of the Heavy Chain Variable Region as shown in SEQ ID NO:

15 is replaced by an amino acid selected from the group consisting of Leu, norleucine, Ile, Val, Met, Ala, and Phe, particularly Leu and Ile, but especially Leu.

41. A humanized antibody or a fragment thereof according to claim 36, wherein the Arg in Kabat position 94 in the human- or primate-derived framework region of the Heavy Chain Variable Region as shown in SEQ ID NO:

15 is replaced by an amino acid selected from the group consisting of Ser and Thr, but especially by Ser.

42. A humanized antibody or a fragment thereof according to claim 37, wherein the Trp in Kabat position 47 in the human- or primate-derived framework region of the Heavy Chain Variable Region as shown in SEQ ID NO:

15 is replaced by an amino acid selected from the group consisting of Leu and Ile, but especially Leu.

43. A humanized antibody or a fragment thereof according to claim 38, wherein the Tyr in Kabat position 87 in the human- or primate-derived framework region of the Light Chain Variable Region as shown in SEQ ID NO:

12 is replaced by Phe.

44. A humanized antibody or a fragment thereof according to claim 39, wherein the Trp in Kabat position 47 in the human- or primate-derived framework region of the Heavy Chain Variable Region as shown in SEQ ID NO:

15 is replaced by an amino acid selected from the group consisting of Leu and Ile, but especially Leu and the Arg in Kabat position 94 in the human- or primate-derived framework region of the Heavy Chain Variable Region as shown in SEQ ID NO;

15 is replaced by Ser and the Tyr in Kabat position 87 in the human- or primate-derived framework region of the Light Chain Variable Region as shown in SEQ ID NO;

12 is replaced by Phe.

45. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof antibody according to any one of claims 1-44, wherein the Heavy Chain Variable Region (HCVR) has an amino acid sequence that is 90%, particularly 95%, more particularly 98% identical to the sequence given in SEQ ID NO:

15 and 16, respectively.

46. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of claims 1-44, wherein the Light Chain Variable Region (LCVR) has an amino acid sequence that is 90%, particularly 95%, more particularly 98% identical to the sequence given in SEQ ID NO:

12 and 13, respectively.

47. A humanized antibody or a fragment thereof according to any one of claims 1-44, wherein at least two, but especially three, of the CDR regions of the Heavy Chain Variable Region (HCVR) have an amino acid sequence that is 90%, particularly 95%, more particularly 98% identical to the corresponding CDR region as given in SEQ ID NO:

1-3.

48. A humanized antibody or a fragment thereof according to any one of claims 1-44, wherein at least two, but especially three, of the CDR regions of the Light Chain Variable Region (LCVR) have an amino acid sequence that is 90%, particularly 95%, more particularly 98% identical to the corresponding CDR region as given in SEQ ID NO:

4-6.

56. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody is of the IgG1, IgG2, IgG3, or IgG4 isotype including a mutated IgG4, but particularly of the IgG 4 isotype.

57. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody is bi-effective in that it exhibits both an aggregation inhibition property as well as a disaggregation property, particularly paired with a high degree of conformational sensitivity.

58. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody, upon co-incubation with amyloid monomeric and/or polymeric soluble amyloid peptides, particularly with β

-amyloid monomeric peptides such as, for example, Aβ

monomeric peptides 1-39;

1-40, 1-41, or 1-42, and/or a polymeric soluble β

-amyloid peptide comprising a plurality of said Aβ

monomeric units, but especially with an Aβ

_1-42 monomeric and/or an Aβ

polymeric soluble amyloid peptide comprising a plurality of said Aβ

_1-42 monomeric units, inhibits the aggregation of the Aβ

monomers into high molecular polymeric fibrils or filaments and, in addition, upon co-incubation with preformed high molecular polymeric amyloid fibrils or filaments formed by the aggregation of amyloid monomeric peptides, particularly β

-amyloid monomeric peptides such as, for example, Aβ

monomeric peptides 1-39;

1-40, 1-41, or 1-42, but especially Aβ

_1-42 monomeric peptides, is capable of disaggregating the preformed polymeric fibrils or filaments.

59. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody substantially binds to aggregated Aβ

in a mammalian brain, particularly a human brain.

60. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody reduces Aβ

plaque burden in a mammalian brain, particularly a human brain.

61. A nucleic acid molecule comprising a nucleotide sequence encoding a chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of claims 1-60.

70. An expression vector comprising the nucleotide sequence of any one of claims 60 to 69.

71. A cell comprising an expression vector comprising the nucleotide sequence of any one of claims 60 to 68 or an expression vector of claim 70.

72. A composition comprising the antibody according to any one of claims 1 to 60 including any functionally equivalent antibody or functional parts thereof in a therapeutically effective amount.

73. A composition according to claim 72 which is a pharmaceutical composition optionally further comprising a pharmaceutically acceptable carrier.

74. A composition according to any one of claims 72 to 73 comprising the antibody in a therapeutically effective amount.

75. A mixture comprising a chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of claims 1 to 60 including any functionally equivalent antibody or functional parts thereof in a therapeutically effective amount and, optionally, a further biologically active substance and/or a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

76. A mixture according to claim 75, wherein the further biologically active substance is a compound used in the treatment of amyloidosis, a group of diseases and disorders associated with amyloid or amyloid-like protein such as the AO protein involved in Alzheimer'"'"'s disease.

77. A mixture according to claim 76 comprising at least one compound selected from the group consisting of compounds against oxidative stress, anti-apoptotic compounds, metal chelators, inhibitors of DNA repair such as pirenzepin and metabolites, 3-amino-1-propanesulfonic acid (3APS), 1,3-propanedisulfonate (1,3PDS), α

-secretase activators, β and

γ

-secretase inhibitors, tau proteins, neurotransmitter, β

-sheet breakers, attractants for amyloid beta clearing/depleting cellular components, inhibitors of N-terminal truncated amyloid beta including pyroglutamated amyloid beta 3-42, anti-inflammatory molecules, or cholinesterase inhibitors (ChEIs) such as tacrine, rivastigmine, donepezil, and/or galantamine, M1 agonists and other drugs including any amyloid or tau modifying drug and nutritive supplements, together with an antibody according to the present invention and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

78. A mixture according to claim 77, wherein the compound is a cholinesterase inhibitor (ChEIs).

79. A mixture according to any one of claims 75 to 77, wherein the compound is one selected from the group consisting of tacrine, rivastigmine, donepezil, galantamine, niacin and memantine.

80. A mixture according to any one of claims 75 to 79 comprising the antibody and/or the biologically active substance in a therapeutically effective amount.

81. Use of a chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof and/or a functional part thereof according to any one of claims 1 to 60 and/or a pharmaceutical composition according to any one of claims 72 to 74, or a mixture according to any one of claims 75 to 80 for the preparation of a medicament for treating or alleviating the effects of amyloidosis, a group of diseases and disorders associated with amyloid plaque formation including secondary amyloidosis and age-related amyloidosis such as diseases including, but not limited to, neurological disorders such as Alzheimer'"'"'s Disease (AD), Lewy body dementia, Down'"'"'s syndrome, hereditary cerebral hemorrhage with amyloidosis (Dutch type);

the Guam Parkinson-Dementia complex;

as well as other diseases which are based on or associated with amyloid-like proteins such as progressive supranuclear palsy, multiple sclerosis;

Creutzfeld Jacob disease, Parkinson'"'"'s disease, HIV-related dementia, ALS (amyotropic lateral sclerosis), Adult Onset Diabetes;

senile cardiac amyloidosis;

endocrine tumors, and others, including macular degeneration.

82. Use according to claim 81, wherein the amyloid associated disease or condition is Alzheimer'"'"'s disease.

83. Use according to claim 81, wherein the amyloid-associated condition is characterized by a loss of cognitive memory capacity in an animal, particularly a mammal or a human.

84. Use according to claim 81, wherein treatment of an animal, particularly a mammal or a human, suffering from an amyloid-associated condition characterized by a loss of cognitive memory capacity leads to an increase of the cognitive memory capacity.

85. Use according to claim 81, wherein treatment of an animal, particularly a mammal or a human, suffering from an amyloid-associated condition characterized by a loss of cognitive memory capacity leads to a complete restoration of the cognitive memory capacity.

86. A method for the preparation of a pharmaceutical composition according to any one of claims 72 to 74, or a mixture according to any one of claims 75 to 80 for use in a method of preventing, treating or alleviating the effects of amyloidosis, a group of diseases and disorders associated with amyloid plaque formation including secondary amyloidosis and age-related amyloidosis such as diseases including, but not limited to, neurological disorders such as Alzheimer'"'"'s Disease (AD), Lewy body dementia, Down'"'"'s syndrome, hereditary cerebral hemorrhage with amyloidosis (Dutch type);

the Guam Parkinson-Dementia complex;

as well as other diseases which are based on or associated with amyloid-like proteins such as progressive supranuclear palsy, multiple sclerosis;

Creutzfeld Jacob disease, Parkinson'"'"'s disease, HIV-related dementia, ALS (amyotropic lateral sclerosis), Adult Onset Diabetes;

senile cardiac amyloidosis;

endocrine tumors, and others, including macular degeneration comprising formulating an antibody according to the invention in a pharmaceutically acceptable form.

87. A method according to claim 86, wherein the antibody is comprised in the composition in a therapeutically effective amount.

88. A method according to claim 86, wherein the amyloid associated disease or condition is Alzheimer'"'"'s Disease.

89. A method according to claim 86, wherein the amyloid-associated condition is characterized by a loss of cognitive memory capacity in an animal, particularly a mammal or a human.

90. A method according to claim 86, wherein treatment of an animal, particularly a mammal or a human, suffering from an amyloid-associated condition characterized by a loss of cognitive memory capacity leads to an increase of the cognitive memory capacity.

91. A method according to claim 86, wherein the amyloid-associated condition is characterized by an impairment of the cognitive memory capacity in an animal, particularly a mammal or a human.

92. A method according to claim 86, wherein treatment of an animal, particularly a mammal or a human, suffering from an amyloid-associated condition characterized by a loss of cognitive memory capacity leads to a reversal of the cognitive memory capacity and, particularly, a complete restoration of the cognitive memory capacity.

93. A method for preventing, treating or alleviating the effects of amyloidosis, a group of diseases and disorders associated with amyloid plaque formation including secondary amyloidosis and age-related amyloidosis such as diseases including, but not limited to, neurological disorders such as Alzheimer'"'"'s Disease (AD), Lewy body dementia, Dawn'"'"'s syndrome, hereditary cerebral hemorrhage with amyloidosis (Dutch type);

the Guam Parkinson-Dementia complex;

as well as other diseases which are based on or associated with amyloid-like proteins such as progressive supranuclear palsy, multiple sclerosis;

Creutzfeld Jacob disease, Parkinson'"'"'s disease, HIV-related dementia, ALS (amyotropic lateral sclerosis), Adult Onset Diabetes;

senile cardiac amyloidosis;

endocrine tumors, and others, including macular degeneration by administering a chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof and/or a functional part thereof according to any one of claims 1 to 60 and/or a pharmaceutical composition according to any one of claims 72 to 74, or a mixture according to any one of claims 75 to 80, to a animal or a human affected by such a disorder comprising administering the antibody in a therapeutically effective amount.

94. A method according to claim 93, wherein the amyloid associated disease or condition is Alzheimer'"'"'s Disease.

95. A method according to claim 93, wherein the amyloid-associated condition is characterized by a loss of cognitive memory capacity in an animal, particularly a mammal or a human.

96. A method according to claim 93, wherein treatment of an animal, particularly a mammal or a human, suffering from an amyloid-associated condition characterized by a loss of cognitive memory capacity leads to an increase in the retention or cognitive memory capacity.

97. A method according to claim 93, wherein the amyloid-associated condition is characterized by an impairment of cognitive memory capacity in an animal, particularly a mammal or a human.

98. A method according to claim 93, wherein treatment of an animal, particularly a mammal or a human, suffering from an amyloid-associated condition characterized by a loss of cognitive memory capacity leads to a complete restoration of cognitive memory capacity.

99. Method of diagnosis of an amyloid-associated disease or condition in a patient comprising

(a) bringing the sample or a specific body part or body area suspected to contain the amyloid protein into contact with an antibody according to any one of claims 1 to 60;

(b) allowing the antibody to bind to the amyloid protein;

(c) detecting the antibody bound to the protein; and

(d) correlating the presence or absence of antibody binding with the presence or absence of amyloid protein in the sample or specific body part or area.

100. Method of determining the extent of amyloidogenic plaque burden in a tissue and/or body fluids comprising

(a) obtaining a sample representative of the tissue and/or body fluids under investigation;

(b) testing said sample for the presence of amyloid protein with an antibody according to any one of claims 1-60;

(c) determining the amount of antibody bound to the protein; and

(d) calculating the plaque burden in the tissue and/or body fluids.

101. Test kits for detection and diagnosis of amyloid-associated diseases and conditions comprising an antibody according to any one of claims 1-60.

102. Test kit according to claim 101 comprising a container holding one or more antibodies according to the present invention and instructions for using the antibodies for the purpose of binding to amyloid protein to form an immunological complex and detecting the formation of the immunological complex such that presence or absence of the immunological complex correlates with presence or absence of amyloid protein.

121. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody substantially binds Aβ

monomers but, preferably, does not show any significant cross-reactivity with amyloid precursor protein (APP).

123. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody substantially binds Aβ

fiber, fibril or filament but, preferably, does not show any significant cross-reactivity with amyloid precursor protein (APP).

124. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to claim 123, wherein said antibody or fragment binds to an Aβ

fiber, fibril or filament with a binding affinity represented by a K_D in a range of at least about 1×

10<sup>−

7</sup>M to at least about 1×

10<sup>−

12</sup> M, particularly of at least about 1×

10<sup>−

8</sup>M to at least about 1×

10<sup>−

11</sup> M, more particularly of at least about 1×

10<sup>−

9</sup>M to at least about 1×

10-10M, even more particularly of at least about 2×

10<sup>−

9</sup>M to at least about 8×

10<sup>−

9</sup>M, but, preferably, does not show any significant cross-reactivity with amyloid precursor protein (APP).

125. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody substantially binds soluble polymeric amyloid beta but, preferably, does not show any significant crossreactivity with amyloid precursor protein (APP).

126. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody substantially binds Aβ

monomers and/or Aβ

fibers, fibrils or filaments and/or soluble polymeric amyloid beta but, preferably, does not show any significant cross-reactivity with amyloid precursor protein (APP).

127. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to claim 126, wherein the binding affinity increases in ascending order.

128. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to claim 127, wherein said antibody or fragment thereof exhibits an binding affinity to an Aβ

fiber, fibril or filament which is at least 10 times, particularly at least 15 times, more particularly at least 20 times, but especially at least 25 times higher than the binding affinity to an Aβ

monomer.

129. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody substantially binds to aggregated Aβ

, including Aβ

plaques, in the mammalian, particularly the human brain but, preferably, does not show any significant cross-reactivity with amyloid precursor protein (APP).

130. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody substantially binds to soluble fibers, including Aβ

monomers, in the mammalian, particularly the human brain but, preferably, does not show any significant cross-reactivity with amyloid precursor protein (APP).

131. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody or fragment thereof is capable of shifting the equilibrium between Aβ

in its soluble and aggregated state towards its soluble form by disaggregating fibers to soluble poly- and monomeric forms by inducing a shift in conformation and binding and stabilizing the disaggregated and solubilized Aβ

forms in the tissue and/or body fluids, particularly the brain.

132. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to claim 131, which antibody or fragment thereof is capable of inducing a transition of the β

-sheet conformation towards an a-helix and/or a random coil conformation, but particularly a random coil conformation, even more particularly a random coil conformation at a given location in the molecule, especially in the environment of Tyr 10 and Val12 of the Aβ

protein, which leads to an increase of the random coil conformation at the expense of the β

-sheet conformation and an improved solubilization of the preformed high molecular polymeric amyloid fibrils or filaments.

133. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to claim 132, wherein the decrease of the β

-sheet conformation amounts to at least 30%, particularly to at least 35%, and more particularly to at least 40% and more as compared to the respective preformed amyloid polymeric fibrils or filaments incubated in buffer (control).

134. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to claim 131, which antibody or fragment thereof substantially bind to polymeric and less aggregated soluble Aβ

protein species including monomeric forms thus modifying the peripheral clearing and catabolism of these Aβ

protein species and reducing their toxic effects.

135. A humanized antibody or a fragment thereof according to any of the preceding claims comprising at least one light chain or a fragment thereof or at least one heavy chain or a fragment thereof incorporating at least one, particularly two and more particularly three CDR regions obtained form a mouse donor antibody, particularly from mouse antibody ACI-01-Ab7C2, wherein said antibody or fragment thereof has an affinity to the Aβ

antigen which is at least 5 times, particularly at least 8 times, more particularly at least 10 times, but especially at least 15 times higher than that of the mouse donor antibody.

136. Use according to any one of claims 83 to 85, wherein the humanized antibody, or a fragment thereof does not necessarily bind aggregated amyloid beta in the brain.

137. A method according to any one of claims 89 to 91, wherein the humanized antibody or a fragment thereof does not necessarily bind aggregated amyloid beta in the brain.

138. Use according to claim 81, wherein treatment of an animal, particularly a mammal or a human, suffering from an amyloid-associated condition characterized by a loss of cognitive memory capacity leads to the retention of cognitive memory capacity.

139. A method according to claim 86, wherein treatment of an animal, particularly a mammal or a human, suffering from an amyloid-associated condition characterized by a loss of cognitive memory capacity leads to the retention of the cognitive memory capacity.

140. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody or fragment thereof is capable of shifting the equilibrium between Aβ

in its soluble and aggregated state towards its soluble form.

141. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody or fragment thereof is capable of shifting the equilibrium between Aβ

in its soluble and aggregated state towards its soluble form by disaggregating fibers to soluble poly- and monomeric forms.

142. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of the preceding claims, which antibody or fragment thereof does not substantially bind Aβ

in human tissue and/or body fluids, particularly the brain, but is capable of shifting the equilibrium between Aβ

in its soluble and aggregated state towards its soluble form by disaggregating fibers to soluble poly- and monomeric forms by inducing a shift in conformation and binding and stabilizing the disaggregated and solubilized Aβ

forms in the tissue and/or body fluids, particularly the brain.

143. A humanized antibody or a fragment thereof according to any of the preceding claims, wherein said antibody or fragment thereof exhibits an binding affinity which is at least 5 times, 10 times, particularly at least 15 times, more particularly at least 20 times, but especially at least 100 times higher than the binding affinity of the mouse antibody.

144. A humanized antibody or a fragment thereof according to any of the preceding claims, wherein the affinity, specificity and stability of said antibody or fragment thereof has been modified by a change of the glycosylation profile.

150. A humanized antibody or a fragment thereof according to any of the preceding claims, wherein said antibody or fragment thereof protects neurons from Abeta-induced degradation.

151. A method of preventing Abeta-induced neuron degradation comprising treating neurons with an effective amount of a humanized antibody or a fragment thereof according to any of the preceding claims.

152. Use of a humanized antibody or a fragment thereof according to any of the preceding claims for the preparation of a medicament for preventing degeneration of neurons upon exposure to Abeta oligomer.

2. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof, which specifically binds to at least one epitope on the β

-amyloid protein wherein the said epitope comprises at least two consecutive amino acid residues predominantly involved in the binding to the antibody, wherein the at least two consecutive amino acid residues are -Phe-Phe- embedded within the following core sequence (SEQ ID NO;

     9);

Xaa₃-Phe-Phe-Xaa₄-Xaa₅-Xaa₆, wherein

Xaa₃ is an amino acid residue selected from the group consisting of Ala, Val, Leu, norleucine, Met, Phe, and Ile;

Xaa₄ is an amino acid residue selected from the group consisting of Ala, Val, Leu, Ser and Ile;

Xaa₅ is an amino acid residue selected from the group consisting of Glu and Asp, and

Xaa₆ is an amino acid residue selected from the group consisting of Glu and Asp.

7. A humanized antibody or a fragment thereof which comprises in the variable region at least one CDR of non-human origin and one or more human- or primate-derived framework regions and, optionally, a constant region derived from a human or primate source antibody, which humanized antibody or fragment thereof is capable of specifically binding β

-amyloid protein, β

-amyloid monomeric peptide, polymeric soluble amyloid peptides comprising a plurality of β

-amyloid monomeric units, β

-amyloid fibers, fibrils or filaments, and a β

-amyloid polymeric peptide in isolation or as part of a β

-amyloid plaque, at an epitope comprising the following amino acid sequence (SEQ ID NO;

     11);

Xaa₁-Xaa₂-Lys-Leu-Xaa₃-Phe-Phe-Xaa₄-Xaa₅-Xaa₆, wherein

Xaa₁ is an amino acid residue selected from the group consisting of His, Asn, Gln, but particularly His;

Xaa₂ is an amino acid residue selected from the group consisting of Asn and Gln, but particularly Gln; and

Xaa₃ is an amino acid residue selected from the group consisting of Val, Leu, and Ile, but particularly Val;

Xaa₄ is an amino acid residue selected from the group consisting of Ala and Val, but particularly Ala;

Xaa₅ is an amino acid residue selected from the group consisting of Glu and Asp, but particularly Glu; and

Xaa₆ is an amino acid residue selected from the group consisting of Glu and Asp, but particularly Asp.

9. A humanized antibody or a fragment thereof comprising variable regions with human- or primate-derived framework regions and at least one CDR with an amino acid sequence selected from the group of sequences consisting of SEQ ID NO:

2 representing CDR2 and SEQ ID NO;

3 representing CDR3 of the Heavy Chain Variable Region (HCVR) and SEQ ID NO;

4 representing CDR1 of the Light Chain Variable Region (LCVR)

10. A humanized antibody or a fragment thereof according to claim 9, wherein said humanized antibody comprises a heavy chain variable region with human- or primate derived heavy chain framework regions and at least one CDR with an amino acid sequence selected from the group of sequences consisting of SEQ ID NO:

2 representing CDR2 and SEQ ID NO;

3 representing CDR3 of the Heavy Chain Variable Region (HCVR).

11. A humanized antibody or a fragment thereof according to claim 9, wherein said humanized antibody comprises a light chain variable region with human- or primate-derived light chain framework regions and a CDR with an amino acid sequence of, SEQ ID NO:

4 representing CDR1 of the Light Chain Variable Region (LCVR).

12. A humanized antibody or a fragment thereof according to claim 9, wherein the humanized antibody comprises variable regions with human- or primate-derived framework regions and at least two CDRs with an amino acid sequence selected from the group of sequences consisting of SEQ ID NO:

1 representing CDR1, SEQ ID NO;

2 representing CDR2 and SEQ ID NO;

3 representing CDR3 of the Heavy Chain Variable Region (HCVR) and SEQ ID NO;

4 representing CDR1, SEQ ID NO;

5 representing CDR2 and SEQ ID NO;

6 representing CDR3 of the Light Chain Variable Region (LCVR).

13. A humanized antibody or a fragment thereof according to claim 12, wherein the humanized antibody comprises heavy chain variable region with human- or primatederived heavy chain framework regions and at least two CDRs with an amino acid sequence selected from the group of sequences consisting of SEQ ID NO:

1 representing CDR1, SEQ ID NO;

2 representing CDR2 and SEQ ID NO;

3 representing CDR3 of the Heavy Chain Variable Region (HCVR), wherein the same CDR cannot be present twice in the antibody.

14. A humanized antibody or a fragment thereof according to claim 12, wherein the humanized antibody comprises light chain variable regions with human- or primate derived light chain framework regions and at least two CDRs with an amino acid sequence selected from the group of sequences consisting of SEQ ID NO:

4 representing CDR1, SEQ ID NO;

5 representing CDR2 and SEQ ID NO;

6 representing CDR3 of the Light Chain Variable Region (LCVR), wherein the same CDR cannot be present twice in the antibody.

15. A humanized antibody or a fragment thereof according to claim 9, wherein the humanized antibody comprises heavy chain variable regions with human- or primate-derived heavy chain framework regions and CDRs with an amino acid sequence of SEQ ID NO:

1 representing CDR1, SEQ ID NO;

2 representing CDR2 and SEQ ID NO;

3 representing CDR3 of the Heavy Chain Variable Region (HCVR), particularly in the order indicated.

16. A humanized antibody or a fragment thereof according to claim 9, wherein the humanized antibody comprises light chain variable regions with human- or primate-derived light chain framework regions and CDRs with an amino acid sequence of SEQ ID NO:

4 representing CDR1, SEQ ID NO;

5 representing CDR2 and SEQ ID NO;

6 representing CDR3 of the Light Chain Variable Region (LCVR), particularly in the order indicated.

17. A humanized antibody or a fragment thereof according to claim 9, wherein the humanized antibody comprises variable regions with human- or primate-derived framework regions and at least three CDRs with an amino acid sequence selected from the group of sequences consisting of SEQ ID NO:

1 representing CDR1, SEQ ID NO;

2 representing CDR2 and SEQ ID NO;

3 representing CDR3 of the Heavy Chain Variable Region (HCVR) and SEQ ID NO;

4 representing CDR1, SEQ ID NO;

5 representing CDR2 and SEQ ID NO;

6 representing CDR3 of the Light Chain Variable Region (LCVR), wherein the same CDR cannot be present twice in the antibody.

18. A humanized antibody or a fragment thereof according to claim 9, wherein the humanized antibody comprises variable regions with human- or primate-derived framework regions and at least four CDRs with an amino acid sequence selected from the group of sequences consisting of SEQ ID NO:

1 representing CDR1, SEQ ID NO;

2 representing CDR2 and SEQ ID NO;

3 representing CDR3 of the Heavy Chain Variable Region (HCVR) and SEQ ID NO;

4 representing CDR1, SEQ ID NO;

5 representing CDR2 and SEQ ID NO;

6 representing CDR3 of the Light Chain Variable Region (LCVR), wherein the same CDR cannot be present twice in the antibody.

19. A humanized antibody or a fragment thereof according to claim 9, wherein the humanized antibody comprises variable regions with human- or primate-derived framework regions and at least five CDRs with an amino acid sequence selected from the group of sequences consisting of SEQ ID NO:

1 representing CDR1, SEQ HD NO;

2 representing CDR2 and SEQ ID NO;

3 representing CDR3 of the Heavy Chain Variable Region (HCVR) and SEQ ID NO;

4 representing CDR1, SEQ ID NO;

5 representing CDR2 and SEQ ID NO;

6 representing CDR3 of the Light Chain Variable Region (LCVR), wherein the same CDR cannot be present twice in the antibody.

20. A humanized antibody or a fragment thereof according to claim 9, wherein the humanized antibody comprises variable regions with human- or primate-derived framework regions and CDRs with an amino acid sequence of SEQ ID NO:

1 representing CDR1, SEQ ID NO;

2 representing CDR2 and SEQ ID NO;

3 representing CDR3 of the Heavy Chain Variable Region (HCVR) and SEQ ID NO;

4 representing CDR1, SEQ ID NO;

5 representing CDR2 and SEQ ID NO;

6 representing CDR3 of the Light Chain Variable Region (LCVR).

21. A humanized antibody or a fragment thereof according to claim 10, wherein the at least one CDR is a CDR with an amino acid sequence of SEQ ID NO:

2 representing CDR2 of the Heavy Chain Variable Region (HCVR).

22. A humanized antibody or a fragment thereof according to claim 10, wherein the at least one CDR is a CDR with an amino acid sequence of SEQ ID NO:

3 representing CDR3 of the Heavy Chain Variable Region (HCVR).

23. A humanized antibody or a fragment thereof according to claim 13, wherein the at least two CDRs are CDRs with an amino acid sequence of SEQ ID NO:

1 representing CDR1 and SEQ ID NO;

2 representing CDR2 of the Heavy Chain Variable Region (HCVR).

24. A humanized antibody or a fragment thereof according to claim 13, wherein the at least two CDRs are CDRs with an amino acid sequence of SEQ ID NO:

1 representing CDR1 and SEQ ID NO;

3 representing CDR3 of the Heavy Chain Variable Region (HCVR).

25. A humanized antibody or a fragment thereof according to claim 13, wherein the at least two CDRs are CDRs with an amino acid sequence of SEQ ID NO:

2 representing CDR2 and SEQ ID NO;

3 representing CDR3 of the Heavy Chain Variable Region (HCVR).

26. A humanized antibody or a fragment thereof according to claim 14, wherein the at least two CDRs are CDRs with an amino acid sequence of SEQ ID NO:

4 representing CDR1 and SEQ ID NO;

5 representing CDR2 of the Light Chain Variable Region (LCVR).

27. A humanized antibody or a fragment thereof according to claim 14, wherein the at least two CDRs are CDRs with an amino acid sequence of SEQ ID NO:

4 representing CDR1 and SEQ ID NO;

6 representing CDR3 of the Light Chain Variable Region (LCVR).

28. A humanized antibody or a fragment thereof according to claim 12, wherein the at least two CDRs are selected from the group of CDRs according to any one of claims 23 to 27.

29. A humanized antibody or a fragment thereof according to any one of claims 9 to 28 comprising a light chain and/or a heavy chain constant region of human or primate origin.

32. A humanized antibody or a fragment thereof according to any one of claims 9 to 31, wherein the framework sequences are human germline V_K sequences of KABAT subgroup VKII.

33. A humanized antibody or a fragment thereof according to any one of claims 9 to 31, wherein the framework sequences are human germline V_H sequences of KABAT subgroup VHIII.

30. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof, wherein at least one of the amino acids of a light chain and a heavy chain CDR region as given in SEQ ID NOs:

1-6 is changed through a conservative substitution such that the antibody maintains its full functionality.

31. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof, according to claim 30, wherein within the amino acid sequence of CDR2 of the light chain variable region (LCVR) as given in SEQ ID NO:

5, the Lys at Kabat position 50 is replaced by Arg, Gln or Glu, but particularly by Arg, and/or the Asn at Kabat position 53 is replaced by Ala, Val, Leu, Ser and Ile;

but especially Ser.

49. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof which comprises the light chain variable region of SEQ ID NO:

13.

50. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to claim 49 which, in addition, comprises the light chain constant region of SEQ ID NO:

14.

55. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to any one of claims 50, 51, 53 and 54, wherein the N-terminal Lys of the heavy chain constant region has been removed.

51. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof which comprises the light chain variable region of SEQ ID NO:

13 and the light chain constant region of SEQ ID NO;

14.

52. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof which comprises the heavy chain variable region of SEQ ID NO:

16.

53. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to claim 52 which, in addition, comprises the heavy chain constant region of SEQ ID NO:

17.

54. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof which comprises the heavy chain variable region of SEQ ID NO:

16 and the heavy chain constant region of SEQ ID NO;

17.

62. A nucleic acid molecule comprising a nucleotide sequence encoding an antibody variable region comprising SEQ ID NO:

2 and 3 representing the Complementarity Determining Regions (CDRs) 2 and 3 of the Heavy Chain Variable Region (HCVR).

64. A nucleic acid molecule according to claim 62 comprising a nucleotide sequence as given in SEQ ID NO:

18 and SEQ ID NO;

19.

63. A nucleic acid molecule comprising a nucleotide sequence encoding an antibody variable region comprising SEQ ID NO:

4 representing the Complementarity Determining Regions (CDRs) 1 of the Light Chain Variable Region (LCVR).

65. A nucleic acid molecule according to claim 63 comprising a nucleotide sequence as given in SEQ ID NO:

20.

66. A nucleic acid molecule comprising a nucleotide sequence encoding the light chain variable region of SEQ ID NO:

22.

67. A nucleic acid molecule comprising a nucleotide sequence encoding the light chain variable region of SEQ ID NO:

22 and the light chain constant region of SEQ ID NO;

23.

68. A nucleic acid molecule comprising a nucleotide sequence encoding the heavy chain variable region of SEQ ID NO:

25

69. A nucleic acid molecule comprising a nucleotide sequence encoding the heavy chain variable region of SEQ ID NO:

25 and the heavy chain constant region of SEQ ID NO;

26.

103. Light Chain Variable Region (LCVR) comprising human- or primate-derived framework regions and at least one CDR with an amino acid sequence of SEQ ID NO:

4 representing CDR1 of the Light Chain Variable Region (LCVR).

104. Heavy Chain Variable Region (HCVR) comprising human- or primate-derived framework regions and at least one CDR with an amino acid sequence selected from the group of sequences consisting of SEQ ID NO:

2 representing CDR2 and SEQ ID NO;

3 representing CDR3 of the Heavy Chain Variable Region (HCVR).

105. Heavy Chain Variable Region (HCVR) comprising human- or primate-derived heavy chain framework regions and at least two CDRs with an amino acid sequence selected from the group of sequences consisting of SEQ ID NO:

1 representing CDR1, SEQ ID NO;

2 representing CDR2 and SEQ ID NO;

3 representing CDR3 of the Heavy Chain Variable Region (HCVR).

106. Light Chain Variable Region (LCVR), comprising human- or primate-derived light chain framework regions and at least two CDRs with an amino acid sequence selected from the group of sequences consisting of SEQ ID NO:

4 representing CDR1, SEQ ID NO;

5 representing CDR2 and SEQ ID NO;

6 representing CDR3 of the Light Chain Variable Region (LCVR).

107. Heavy Chain Variable Region (HCVR) comprising human- or primate-derived heavy chain framework regions and CDRs with an amino acid sequence of SEQ ID NO:

1 representing CDR1, SEQ ID NO;

2 representing CDR2 and SEQ ID NO;

3 representing CDR3 of the Heavy Chain Variable Region (HCVR), particularly in the order indicated above.

108. The Light Chain Variable Region (LCVR) comprising integrated into human- or primate-derived light chain framework regions and CDRs with an amino acid sequence of SEQ ID NO:

4 representing CDR1, SEQ ID NO;

5 representing CDR2 and SEQ ID NO;

6 representing CDR3 of the Light Chain Variable Region (LCVR), particularly if the order indicated above.

109. The Light Chain Variable Region (LCVR) of SEQ ID NO:

12.

110. A humanized antibody or a fragment thereof is provided, which comprises the Light Chain Variable Region (LCVR) of SEQ ID NO:

12.

111. The Light Chain Variable Region (LCVR) including signal sequences as shown in SEQ ID NO:

13.

112. A humanized antibody or a fragment thereof, which comprises the complete Light Chain Variable Region (LCVR) including signal sequences as shown in SEQ ID NO:

13.

113. A humanized antibody or a fragment thereof, which comprises the Light Chain Variable Region (LCVR) of SEQ ID NO:

12 and the light chain constant region of SEQ ID NO;

14.

114. A humanized antibody or a fragment thereof, which comprises the complete Light Chain Variable Region (LCVR) of SEQ ID NO:

13 and the light chain constant region of SEQ ID NO;

14.

115. The Heavy Chain Variable Region (HCVR) of SEQ ID NO:

15.

116. A humanized antibody or a fragment thereof, which comprises the Heavy Chain Variable Region (HCVR) of SEQ ID NO:

15.

117. The Heavy Chain Variable Region (HCVR) including signal sequences as shown in SEQ ID NO:

16.

118. A humanized antibody or a fragment thereof, which comprises the complete Heavy Chain Variable Region (HCVR) including signal sequences as shown in SEQ ID NO;

16.

119. A humanized antibody or a fragment thereof, which comprises the Heavy Chain Variable Region (HCVR) of SEQ ID NO:

15 and the heavy chain constant region of SEQ ID NO;

17.

120. A humanized antibody or a fragment thereof, which comprises the Heavy Chain Variable Region (HCVR) of SEQ ID NO:

16 and the heavy chain constant region of SEQ ID NO;

17.

122. A chimeric antibody or a fragment thereof, or a humanized antibody or a fragment thereof according to claim 122, wherein said antibody or fragment binds to an Aβ

monomer with a binding affinity represented by a K_D in a range of at least about 1×

10<sup>−

7</sup> M to at least about 1×

10<sup>−

12</sup> M, particularly of at least about 1×

10<sup>−

8</sup> M to at least about 1×

10<sup>−

11</sup> M, more particularly of at least about 1×

10<sup>−

8</sup> M to at least about 1×

10<sup>−

10</sup> M, even more particularly of at least about 1×

10<sup>−

8</sup> M to at least about 5×

10<sup>−

8</sup> M but, preferably, does not show any significant cross-reactivity with amyloid precursor protein (APP).

145. An antibody comprising a variable region as recited in SEQ ID NO:

27, or a variant thereof

146. A cell line expressing the antibody of claim 145.

147. An antibody gene comprising a variable region as recited in SEQ ID NO:

29, or a variant thereof

148. A cell line expressing the antibody of claim 147.

149. A method for disaggregating preformed beta-amyloid fibers, comprising interacting an hC2 antibody with preformed beta-amyloid fibers.