The present invention is related to methods and compositions for the therapeutic and diagnostic use in the treatment of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including amyloidosis, a group of disorders and abnormalities associated with amyloid protein such as Alzheimer's disease. The present invention provides novel methods and compositions comprising highly specific and highly effective antibodies having the ability to specifically recognize and bind to specific epitopes from a range of .beta.-amyloid proteins. The antibodies enabled by the teaching of the present invention are particularly useful for the treatment of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including amyloidosis, a group of diseases and disorders associated with amyloid plaque formation including secondary amyloidosis and age-related amyloidosis including, but not limited to, neurological disorders such as Alzheimer's Disease (AD).

Description of the Invention

The present invention makes use of antigen presentations that result in enhanced exposure and stabilization of a preferred antigen conformation, which ultimately results in antibodies with unique properties.

In one embodiment of the invention, an antibody is provided including any functionally equivalent antibody or functional parts thereof, or, more particularly, a monoclonal antibody including any functionally equivalent antibody or functional parts thereof, which has been raised against a supramolecular antigenic construct comprising an antigenic peptide corresponding to the amino acid sequence of the .beta.-amyloid peptide, particularly of .beta.-amyloid peptide A.beta..sub.1-15, A.beta..sub.1-16 and A.beta..sub.1-16(.DELTA.14), modified with a hydrophobic moiety such as, for example, palmitic acid or a hydrophilic moiety such as, for example, polyethylene glycol (PEG) or a combination of both, wherein said hydrophobic and hydrophilic moiety, respectively, is covalently bound to each of the termini of the antigenic peptide through at least one, particularly one or two amino acids such as, for example, lysine, glutamic acid and cystein or any other suitable amino acid or amino acid analogue capable of serving as a connecting device for coupling the hydrophobic and hydrophilic moiety to the peptide fragment. When a PEG is used as the hydrophilic moiety, the free PEG termini are covalently bound to phosphatidylethanolamine or any other compound suitable to function as the anchoring element, for example, to embed the antigenic construct in the bilayer of a liposome.

In another embodiment of the invention, an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof is provided, which recognized the native conformation of amyloid in that it specifically binds to amyloid oligomers and fibers, but not to not linearized amyloid species.

In a further embodiment of the invention, an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof according to the present invention and as described herein before, is provided which antibody or fragment binds to an A.beta. monomer with a binding affinity of at least about 1.times.10.sup.-6 to at least about 1.times.10.sup.-8, particularly of at least about 1.times.10.sup.-6 to at least about 1.times.10.sup.-7, more particularly of at least about 1.times.10.sup.-7 to at least about 1.times.10.sup.-8, even more particularly of at least about 1.times.10.sup.-7 to at least about 4.times.10.sup.-7, but, preferably, does not show any significant cross-reactivity with amyloid precursor protein (APP)

In another embodiment of the invention, an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof according to the present invention and as described herein before, is provided which antibody or fragment binds to an A.beta. fiber, fibril or filament with a binding affinity of at least about 1.times.10.sup.-7 to at least about 1.times.10.sup.-9, particularly of at least about 1.times.10.sup.-7 to at least about 1.times.10.sup.-8, more particularly of at least about 1.times.10.sup.-8 to at least about 1.times.10.sup.-9, even more particularly of at least about 1.times.10.sup.-8 to at least about 5.times.10.sup.-8, but, preferably, does not show any significant cross-reactivity with amyloid precursor protein (APP).

In another embodiment, the antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof according to the present invention and as described herein before exhibits an binding affinity to an A.beta. fiber, fibril or filament which is at least 5 times, particularly at least 10 times, more particularly at least 15 times, higher than the binding affinity to an A.beta. monomer.

The antibodies according to the invention are capable of inhibiting, in vitro and in vivo, the aggregation of amyloidogenic monomeric peptides, specifically .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, into high molecular polymeric amyloid fibrils or filaments.

In a specific embodiment of the invention an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody, upon co-incubation with amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, inhibits the aggregation of the A.beta. monomers into high molecular polymeric fibrils.

In a further embodiment of the invention an antibody is provided, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody, upon co-incubation, particularly upon co-incubation at a molar concentration ratio of up to 1:100, more particularly at a molar concentration ratio of between 1:30 and 1:100, but especially at a molar concentration ratio of 1:100, with amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, inhibits the aggregation of the A.beta. monomers into high molecular polymeric fibrils. In particular, said inhibition amounts to at least 50%, particularly to at least 65%, more particularly to at least 75%, even more particularly to at least 80%, but especially to at least 85%-90%, or more as compared to the respective amyloid peptide monomers incubated in buffer (control).

In particular, the co-incubation of the antibody according to the invention with amyloid monomeric peptides is carried out for 24 hours to 60 hours, particularly for 30 hours to 50 hours, more particularly for 48 hours at a temperature of between 28.degree. C. and 40.degree. C., particularly of between 32.degree. C. and 38.degree. C., more particularly at 37.degree. C.

In another embodiment the present invention provides an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof which antibody, upon co-incubation for 48 hours at 37.degree. C. at a molar concentration ratio of 1:100 with an amyloid monomeric peptide, specifically a .beta.-amyloid monomeric peptide such as, for example, A.beta. monomeric peptide 1-39; 1-40, 1-41, 1-42, or 1-43, but especially a A.beta..sub.1-42 monomeric peptide, is capable of inhibiting the aggregation of the amyloid monomers, particularly the aggregation of .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially of the A.beta..sub.1-42 monomeric peptide into high molecular polymeric fibrils or filaments by at least 85%, particularly by at least 89% and more particularly by at least 95% as compared to the respective amyloid peptide monomers incubated in buffer (control).

In a specific embodiment, the invention provides an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which exhibits high specificity to A.beta..sub.1-42 monomeric peptides but shows essentially no or only minor cross-reactivity to A.beta..sub.1-38, A.beta..sub.1-39, A.beta..sub.1-40, and/or A.beta..sub.1-41 monomeric peptides, particularly an antibody, but especially a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody is up to 100 fold, particularly 50 to 100 fold, more particularly 80 to 100 fold, but especially 100 fold more sensitive to amyloid peptide A.beta..sub.1-42 as compared to A.beta..sub.1-38, A.beta..sub.1-39, A.beta..sub.1-40, A.beta..sub.1-41 and up to 1000 fold, particularly 500 to 1000 fold, more particularly 800 to 1000 fold, but especially 1000 fold more sensitive to amyloid peptide A.beta..sub.1-42 as compared to A.beta..sub.1-38, and thus capable of inhibiting, in vitro and in vivo, the aggregation of amyloidogenic monomeric peptides, but especially of amyloid peptide A.beta..sub.1-42

In another specific embodiment of the invention an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which has a high binding sensitivity to amyloid peptide A.beta..sub.1-42 and is capable of detecting A.beta..sub.1-42 fibers in a concentration of down to at least 0.001 .mu.g, but particularly in a concentration range of between 0.5 .mu.g and 0.001 .mu.g, more particularly between 0.1 .mu.g and 0.001 .mu.g, but especially in a concentration of 0.001 .mu.g.

In a very specific embodiment of the invention an antibody is provided, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody is capable of detecting A.beta..sub.1-42 fibers down to a minimal concentration of 0.001 .mu.g and A.beta..sub.1-40 fibers down to a minimal concentration of 0.1 .mu.g and A.beta..sub.1-38 fibers down to a minimal concentration of 1 .mu.g amount of fibers.

Binding of the antibodies according to the invention and as described herein before to amyloidogenic monomeric peptides but, particularly, to the amyloid form (1-42) leads to inhibition of the aggregation of monomeric amyloidogenic peptides to high molecular fibrils or filaments. Through the inhibition of the aggregation of amyloidogenic monomeric peptides the antibodies according to the present invention are capable of preventing or slowing down the formation of amyloid plaques, particularly the amyloid form (1-42), which is know to become insoluble by change of secondary conformation and to be the major part of amyloid plaques in brains of diseased animals or humans.

The aggregation inhibition potential of the antibody according to the invention may be determined by any suitable method known in the art, particularly by density-gradient ultracentrifugation followed by a SDS-PAGE sedimentation analysis on a preformed gradient and/or by a thioflavin T (Th-T) fluorescent assay.

The present invention further provides antibodies which, upon co-incubation with preformed high molecular polymeric amyloid fibrils or filaments formed by the aggregation of amyloid monomeric peptides, specifically .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, are capable of disaggregating said high molecular polymeric amyloid fibrils or filaments.

In another embodiment of the invention an antibody is provided, but especially a monoclonal antibody including any functionally equivalent antibody or functional parts thereof, which antibody, upon co-incubation at a molar concentration ratio of up to 1:100, more particularly at a molar concentration ratio of between 1:30 and 1:100, but especially at a molar concentration ratio of 1:100, with preformed high molecular polymeric amyloid fibrils or filaments formed by the aggregation of amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, is capable of disaggregating the preformed polymeric fibrils or filaments by at least 35%, particularly by at least 40%, more particularly by at least 50%, even more particularly by at least 60%, but especially by at least 70% or more.

In particular, the antibody according to the invention is co-incubated with amyloid preformed high molecular polymeric amyloid fibrils or filaments for 12 hours to 36 hours, particularly for 18 hours to 30 hours, more particularly for 24 hours at a temperature of between 28.degree. C. and 40.degree. C., particularly of between 32.degree. C. and 38.degree. C., more particularly at 37.degree. C.

In a specific embodiment the present invention provides an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody, upon co-incubation for 24 hours at 37.degree. C. at a molar concentration ratio of 1:100 with preformed high molecular polymeric amyloid fibrils or filaments formed by the aggregation of amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, is capable of disaggregating said preformed high molecular polymeric amyloid fibrils or filaments by at least 35%, particularly by at least 40%, more particularly by at least 50%, even more particularly by at least 60%, but especially by at least 70% or more as compared to the respective preformed amyloid polymeric fibrils or filaments incubated with a control vehicle (amyloid alone) (control).

The disaggregation potential of the antibody according to the invention may be determined by any suitable method known in the art, particularly by density-gradient ultracentrifugation followed by a SDS-PAGE sedimentation analysis on a preformed gradient and/or by a thioflavin T (Th-T) fluorescent assay.

The present invention further provides antibodies or functional parts thereof which are conformationally sensitive.

In a further embodiment of the invention an antibody is provided, but especially a monoclonal antibody including any functionally equivalent antibody or functional parts thereof, which antibody, upon co-incubation with preformed high molecular polymeric amyloid fibrils or filaments formed by the aggregation of amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, is capable of inducing a transition of the .beta.-sheet conformation towards an .alpha.-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 Val12 of the A.beta. protein, which leads to an increase of the random coil conformation at the expense of the .beta.-sheet conformation and an improved solubilization of the preformed high molecular polymeric amyloid fibrils or filaments. In particular the decrease of the .beta.-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).

In particular, the antibody according to the invention is co-incubated with amyloid preformed high molecular polymeric amyloid fibrils or filaments for 12 hours to 36 hours, particularly for 18 hours to 30 hours, more particularly for 24 hours at a temperature of between 28.degree. C. and 40.degree. C., particularly of between 32.degree. C. and 38.degree. C., more particularly at 37.degree. C.

In particular, the present invention provides an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody, upon co-incubation for 24 hours at 37.degree. C. at a molar concentration ratio of 1:100 with preformed high molecular polymeric amyloid fibrils or filaments formed by the aggregation of amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, is capable of inducing a transition of the .beta.-sheet conformation towards an .alpha.-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 Val12 of the A.beta. protein, which leads to an increase of the random coil conformation at the expense of the .beta.-sheet conformation, with the latter being reduced by at least 30%, particularly by at least 35%, and more particularly by at least 40% and more as compared to the respective preformed amyloid polymeric fibrils or filaments incubated in buffer (control).

The antibody's potential in inducing a conformational transition may be determined by any suitable method known in the art, particularly by solid state 13C NMR spectroscopy but, in particular, by measuring the integral intensities of the conformations of Val 12 C.beta. in the A.beta. peptide, particularly the A.beta. peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially in the A.beta..sub.1-42 monomeric peptide.

Through the disaggregation of amyloidogenic polymeric fibrils or filaments the antibodies according to the present invention are capable of preventing or slowing down the formation of amyloid plaques, which leads to an alleviation of the symptoms associated with the disease and a delay or reversal of its progression.

Accordingly, it is a further embodiment of the invention to provide an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof as described herein before, which antibody is capable of decreasing the total amount of A.beta. in the brain of an animal, particularly a mammal, but especially a human suffering from a disease or condition leading to increased concentration of A.beta. in the brain.

In another embodiment of the invention an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof as described herein before is provided, which antibody is capable of disrupting plaques thus decreasing the plaque load in the brain of an animal, particularly a mammal, but especially a human suffering from a disease or condition leading to an increased plaque load in the brain. The antibody according to the invention including any functionally equivalent antibody or functional parts thereof decreases the plaque load in the brain by at least 20%, particularly by at least 25%, more particularly by at least 30%, even more particularly more than 30%.

In still another embodiment of the invention an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof as described herein before is provided, which antibody is capable of solubilizing plaques leading to a reduction of the amount of plaques in the brain of an animal, particularly a mammal, but especially a human suffering from a disease or condition leading to an increased plaque load in the brain. The antibody according to the invention including any functionally equivalent antibody or functional parts thereof reduces the amount of plaques in the brain by at least 10%, particularly by at least 15%, more particularly by at least 20%.

It is to be understood that the antibody according to the invention can exhibit one, two or more of the specific properties described herein before in various combinations.

For example, in one embodiment, the present invention provides antibodies, but especially monoclonal antibodies including any functionally equivalent antibody or functional parts thereof, which antibodies are bifunctional in that they exhibit both an aggregation inhibition property as well as a disaggregation property as defined herein before, particularly paired with a high degree of conformational sensitivity.

In still another embodiment of the invention a bifunctional antibody is provided including any functionally equivalent antibody or functional parts thereof, but especially a bifunctional monoclonal antibody including any functionally equivalent antibody or functional parts thereof, which antibody, upon co-incubation with amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, inhibits the aggregation of the A.beta. 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 .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, is capable of disaggregating the preformed polymeric fibrils or filaments.

In a specific embodiment of the invention the co-incubation of the bifunctional antibody according to the invention, but especially of the bifunctional monoclonal antibody according to the invention with amyloid monomeric peptides and preformed high molecular polymeric amyloid fibrils or filaments, respectively, takes place at a molar concentration ratio of up to 1:100, particularly at a ratio of between 1:30 and 1:100, and more particularly at a ration of 1:100.

In particular, the co-incubation of the antibody according to the invention with amyloid monomeric peptides is carried out for 24 hours to 60 hours, particularly for 30 hours to 50 hours, more particularly for 48 hours at a temperature of between 28.degree. C. and 40.degree. C., particularly of between 32.degree. C. and 38.degree. C., more particularly at 37.degree. C., whereas the co-incubation with amyloid preformed high molecular polymeric amyloid fibrils or filaments is carried out for 12 hours to 36 hours, particularly for 18 hours to 30 hours, more particularly for 24 hours at a temperature of between 28.degree. C. and 40.degree. C., particularly of between 32.degree. C. and 38.degree. C., more particularly at 37.degree. C.

In still another specific embodiment of the invention the bifunctional antibody according to the invention, particularly the bifunctional monoclonal antibody according to the invention, including any functionally equivalent antibody or functional parts thereof, is capable of disaggregating the preformed polymeric fibrils or filaments by at least 10%, particularly by at least 25%, more particularly by at least 35%, even more particularly by at least 50%, but especially by at least 60-70% or more.

In still another specific embodiment of the invention the bifunctional antibody according to the invention, particularly the bifunctional monoclonal antibody according to the invention, including any functionally equivalent antibody or functional parts thereof, inhibits the aggregation of amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides by at least 50%, particularly by at least 65%, more particularly by at least 75%, even more particularly by at least 80%, but especially by at least 85-90%, or more as compared to the respective amyloid peptide monomers incubated in buffer (control).

In particular, the present invention provides an antibody, particularly a bifunctional antibody, but especially a monoclonal antibody, particularly a bifunctional monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody mediates inhibition of polymerization of amyloid monomeric peptides, specifically .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides and/or induces solubilization of preformed high molecular polymeric amyloid fibrils or filaments formed by the aggregation of amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, through specific and direct binding of the antibody to the A.beta. fibers, which leads to a transition of secondary conformation.

The present invention further provides an antibody, particularly a bifunctional antibody, but especially a monoclonal antibody, particularly a bifunctional monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody directly and specifically binds to .beta.-amyloid fibers such as, for example, fibers comprising A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially to fibers comprising A.beta..sub.1-42 monomeric peptides and/or induces solubilization of preformed high molecular polymeric amyloid fibrils or filaments formed by the aggregation of amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, by targeting and specifically binding to an epitope within an epitopic region of the .beta.-amyloid protein, particularly an epitopic region of the A.beta. polypeptide confined by amino acid residues aa.sub.n-aa.sub.m with n being an integer between 2 and 16, particularly between 5 and 16, more particularly between 8 and 16, even more particularly between 10 and 16 and m being an integer between 3 and 25, particularly between 3 and 23, particularly between 3 and 20, particularly between 3 and 17, particularly between 6 and 17, more particularly between 9 and 17, even more particularly between 11 and 17, wherein n and m cannot be identical numbers and n must always be a smaller number than m, with the difference between n and m.gtoreq.2.

In a specific embodiment of the invention, n is an integer between 13 and 15, but especially 14 and m is an integer between 22 and 24, but especially 23.

The binding of the antibody according to the invention may induce a conformational transition in said protein, particularly a transition of the .alpha.-sheet conformation towards an .alpha.-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, particularly in the environment of Val12 of the A.beta. protein.

In a further embodiment, the invention provides an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody incorporates at least one of the properties mentioned herein before and selected from the group consisting of aggregation inhibition, disaggregation, induction of conformational transition, recognition of and direct binding to an epitope, particularly a conformational discontinuous epitope in the 14-23, particularly in the 14-20 region, preventing or slowing down the formation of amyloid plaques, decreasing the total amount of soluble A.beta. in the brain, decreasing the plaque load in the brain, reducing the amount of plaques in the brain, retaining or increasing cognitive memory capacity, but especially a combination of two or more of said properties.

In specific embodiment, the invention relates to an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody incorporates at least 2, particularly at least 3, more particularly at least 4, even more particularly at least 5, 6, 7 or 8, but especially all of the above mentioned properties.

In a specific embodiment, the invention provides an antibody, particularly a bifunctional antibody, but especially a monoclonal antibody, particularly a bifunctional monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which exhibits high specificity to A.beta..sub.1-42 monomeric peptides but shows essentially no or only minor cross-reactivity to A.beta..sub.1-38, A.beta..sub.1-39, A.beta..sub.1-40, and/or A.beta..sub.1-41 monomeric peptides, particularly an antibody, but especially a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody is up to 100 fold, particularly 50 to 100 fold, more particularly 80 to 100 fold, but especially 100 fold more sensitive to amyloid peptide A.beta..sub.1-42 as compared to A.beta..sub.1-38, A.beta..sub.1-39, A.beta..sub.1-40, A.beta..sub.1-41 and up to 1000 fold, particularly 500 to 1000 fold, more particularly 800 to 1000 fold, but especially 1000 fold more sensitive to amyloid peptide A.beta..sub.1-42 as compared to A.beta..sub.1-38, and thus capable of inhibiting, in vitro and in vivo, the aggregation of amyloidogenic monomeric peptides, but especially of amyloid peptide A.beta..sub.1-42

In another specific embodiment of the invention an antibody, particularly a bifunctional antibody, but especially a monoclonal antibody, particularly a bifunctional monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which has a high binding affinity to amyloid peptide A.beta..sub.1-42 and is capable of detecting A.beta..sub.1-42 fibers in a concentration of down to at least 0.001 .mu.g, but particularly in a concentration range of between 0.5 .mu.g and 0.001 .mu.g, more particularly between 0.1 .mu.g and 0.001 .mu.g, but especially in a concentration of 0.001 .mu.g.

In a very specific embodiment of the invention an antibody is provided, particularly a bifunctional antibody, but especially a monoclonal antibody, particularly a bifunctional monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody is capable of detecting A.beta..sub.1-42 fibers down to a minimal concentration of 0.001 .mu.g and A.beta..sub.1-40 fibers down to a minimal concentration of 0.1 .mu.g and A.beta..sub.1-38 fibers down to a minimal concentration of 1 .mu.g amount of fibers.

In one specific aspect, the invention relates to an antibody or a fragment thereof, which recognizes and binds to at least one distinct binding site, particularly to a least two distinct binding sites on the .beta.-amyloid protein.

In a specific embodiment, the invention relates to an antibody including any functionally equivalent antibody or functional parts thereof which antibody recognizes and binds to at least one distinct binding site, particularly to at least two distinct binding sites on the .beta.-amyloid protein wherein the said at least one or said at least two distinct binding sites comprise at least one amino acid residue and at least two consecutive amino acid residues, respectively, predominantly involved in the binding of the antibody, wherein, in a specific embodiment of the invention, the at least one residue comprising the first distinct binding site is Leu and the at least two consecutive amino acid residues, comprising the second distinct binding site, are -Phe-Phe- embedded within the following core sequence:

-Xaa1-Xaa2-Xaa3-Leu-Xaa4-Phe-Phe-Xaa5-Xaa6-Xaa7- (SEQ ID NO: 19) wherein Xaa.sub.1 is an amino acid residue selected from the group comprising His, Asn, Gln Lys, and Arg; Xaa.sub.2 is an amino acid residue selected from the group comprising Asn and Gln; Xaa.sub.3 is an amino acid residue selected from the group comprising Lys, His, Asn, Gln and Arg Xaa.sub.4 is an amino acid residue selected from the group comprising Ala, Val, Leu, norleucine, Met, Phe, and Ile; Xaa.sub.5 is an amino acid residue selected from the group comprising Ala, Val, Leu, Ser and Ile; Xaa.sub.6 is an amino acid residue selected from the group comprising Glu and Asp, Xaa.sub.7 is an amino acid residue selected from the group comprising Glu and Asp.

In particular, an antibody or a fragment thereof is provided, which recognizes and binds to at least one distinct binding site, particularly to a least two distinct binding sites on the .beta.-amyloid protein wherein the said at least one or said at least two distinct binding sites comprise at least one amino acid residue and at least two consecutive amino acid residues, respectively, predominantly involved in the binding of the antibody, wherein, in a specific embodiment of the invention, the at least one residue constituting the first distinct binding site is Leu and the at least two consecutive amino acid residues, constituting the second distinct binding site, are -Phe-Phe- embedded within the following core sequence:

-Xaa1-Xaa2-Xaa3-Leu-Xaa4-Phe-Phe-Xaa5-Xaa6-Xaa7- (SEQ ID NO: 19) wherein Xaa.sub.1 is an amino acid residue selected from the group comprising His, Asn, Gln Lys, and Arg; Xaa.sub.2 is an amino acid residue selected from the group comprising Asn and Gln; Xaa.sub.3 is an amino acid residue selected from the group comprising Lys, His, Asn, Gln and Arg Xaa.sub.4 is an amino acid residue selected from the group comprising Ala, Val, Leu, norleucine, Met, Phe, and Ile; Xaa.sub.5 is an amino acid residue selected from the group comprising Ala, Val, Leu, Ser and Ile; Xaa.sub.6 is an amino acid residue selected from the group comprising Glu and Asp, Xaa.sub.7 is an amino acid residue selected from the group comprising Glu and Asp.

In another embodiment of the invention, an antibody or a fragment thereof is provided, wherein

Xaa.sub.1 is His or Arg, but particularly His;

Xaa.sub.2 is Gln or Asn, but particularly Gln;

Xaa.sub.3 is Lys or Arg, but particularly Lys

Xaa.sub.4 is Val or Leu, but particularly Val;

Xaa.sub.5 is Ala or Val, but particularly Ala;

Xaa.sub.6 is Glu or Asp, but particularly Glu; and

Xaa.sub.7 is Asp or Glu, but particularly Asp.

In another aspect, the invention relates to an antibody or a fragment thereof, which recognizes and binds to at least one distinct binding site, particularly to a least two distinct binding sites, more particularly to at least three distinct binding sites on the .beta.-amyloid protein, wherein said one or the at least two or the at least three distinct binding sites each comprise at least one, particularly at least two consecutive amino acid residues predominantly involved in the binding of the antibody.

In particular, the antibody or a fragment thereof according to the invention binds to at least two distinct binding sites on the .beta.-amyloid protein, wherein said at least two distinct binding sites each comprise at least two consecutive amino acid residues predominantly involved in the binding of the antibody, wherein said at least two distinct binding sites are located in close proximity to each other on the antigen, separated by at least one amino acid residue not involved in antibody binding or to a significantly smaller extent as compared to said at least two consecutive amino acid residues, thus forming a conformational discontinuous epitope.

In another embodiment of the invention, an antibody or a fragment thereof according to the invention is provided, which recognizes and binds to at least one distinct binding site, particularly to at least two distinct binding sites, more particularly to at least three distinct binding sites on the .beta.-amyloid protein wherein said distinct binding sites comprise at least one and at least two consecutive amino acid residues, respectively, predominantly involved in the binding of the antibody, wherein the at least one and the at least two consecutive amino acids, which are separated by at least one amino acid residue not involved in antibody binding or to a significantly smaller extent as compared to the amino acid residues predominantly involved in the binding of the antibody, are -His- and -Lys-Leu-, respectively, embedded within the following core sequence:

-His-Xaa2-Lys-Leu-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8- (SEQ ID NO: 13) wherein

Xaa2 is an amino acid residue selected from the group comprising Asn and Gln;

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

Xaa4 is an amino acid residue selected from the group comprising Ala, Val, Leu, norleucine, Met, Phe, and Ile

Xaa5 is an amino acid residue selected from the group comprising Ala, Val, Leu, norleucine, Met, Phe, and Ile

Xaa6 is an amino acid residue selected from the group comprising Ala, Val, Leu, Ser and Ile;

Xaa7 is an amino acid residue selected from the group comprising Glu and Asp,

Xaa8 is an amino acid residue selected from the group comprising Glu and Asp

and wherein said amino acid residues Xaa2, Xaa3, Xaa6, Xaa7, Xaa8, are not involved in antibody binding or to a significantly smaller extent as compared to the -His- and the -Lys-Leu- binding site.

In another embodiment, an antibody or a fragment thereof is provided, which recognizes and binds to at least one distinct binding site, particularly to a least two distinct binding sites, more particularly to at least three distinct binding sites on the .beta.-amyloid protein wherein said distinct binding sites comprise at least one and at least two consecutive amino acid residues, respectively, predominantly involved in the binding of the antibody, wherein the at least two consecutive amino acid residues representing a first binding site are -Phe-Phe- and the at least one amino acid residue is -His- embedded within the following core sequence:

-Xaa1-His-Xaa3-Xaa4-Xaa5-Xaa6-Phe-Phe-Xaa7-Xaa8-Xaa9- (SEQ ID NO: 14), wherein Xaa.sub.1 is an amino acid residue selected from the group comprising His, Asn, Gln, Lys and Arg Xaa.sub.3 is an amino acid residue selected from the group comprising Asn and Gln Xaa.sub.4 is an amino acid residue selected from the group comprising His, Asn, Gln, Lys and Arg Xaa.sub.5 is an amino acid residue selected from the group comprising Ala, Val, Leu, Ser and Ile; Xaa.sub.6 is an amino acid residue selected from the group comprising Ala, Val, Leu and Ile Xaa.sub.7 is an amino acid residue selected from the group comprising Ala, Val, Leu and Ile Xaa.sub.8 is an amino acid residue selected from the group comprising Glu and Asp, Xaa.sub.9 is an amino acid residue selected from the group comprising Glu and Asp, and wherein said amino acid residues Xaa.sub.1, Xaa.sub.3, Xaa.sub.6, Xaa.sub.7, Xaa.sub.8 and Xaa.sub.9, are not involved in antibody binding or to a significantly smaller extent as compared to the His and the -Phe-Phe- binding site.

In a specific embodiment of the invention, the first of at least two consecutive amino acid residues predominantly involved in the binding of the antibody involve -Lys- and -Leu-, and the second of the at least two consecutive amino acid residues involve -Phe-Phe- embedded within the following core sequence:

-Xaa1-Xaa2-Lys-Leu-Xaa4-Phe-Phe-Xaa5-Xaa6-Xaa7- (SEQ ID NO: 15), wherein Xaa.sub.1 is an amino acid residue selected from the group comprising His, Asn, Gln Lys, and Arg; Xaa.sub.2 is an amino acid residue selected from the group comprising Asn and Gln; Xaa.sub.4 is an amino acid residue selected from the group comprising Ala, Val, Leu, norleucine, Met, Phe, and Ile; Xaa.sub.5 is an amino acid residue selected from the group comprising Ala, Val, Leu, Ser and Ile; Xaa.sub.6 is an amino acid residue selected from the group comprising Glu and Asp, Xaa.sub.7 is an amino acid residue selected from the group comprising Glu and Asp, and wherein said amino acid residues Xaa.sub.2, Xaa.sub.3, Xaa.sub.4, Xaa.sub.5, Xaa.sub.6, Xaa.sub.7 are not involved in antibody binding or to a significantly smaller extent as compared to the -Lys-Leu and the -Phe-Phe- binding site.

In another embodiment of the invention, an antibody or a fragment thereof is provided, wherein

Xaa.sub.1 is His or Arg, but particularly His;

Xaa.sub.2 is Gln or Asn, but particularly Gln;

Xaa.sub.4 is Val or Leu, but particularly Val;

Xaa.sub.5 is Ala or Val, but particularly Ala;

Xaa.sub.6 is Glu or Asp, but particularly Glu; and

Xaa.sub.7 is Asp or Glu, but particularly Asp.

In a further embodiment of the invention, the antibody or a fragment thereof according to the invention binds to at least three distinct binding sites on the .beta.-amyloid protein wherein said at least three distinct binding sites comprise at least one amino acid residue and at least two consecutive amino acid residues, respectively, which residues are predominantly involved in the binding of the antibody, wherein said at least three distinct binding sites are located in close proximity to each other on the antigen, separated by at least one amino acid residue not involved in antibody binding or to a significantly smaller extent as compared to said at least one amino acid residue and said at least two consecutive amino acid residues, respectively, thus forming a conformational discontinuous epitope.

In a specific embodiment of the invention, the first of the at least two consecutive amino acid residues predominantly involved in the binding of the antibody involve -Lys-Leu-, and the second of the at least two consecutive amino acid residues involve -Phe-Phe-, and the third at least one amino residue involves -His- embedded within the following core sequence:

-His-Xaa2-Lys-Leu-Xaa4-Phe-Phe-Xaa5-Xaa6-Xaa7- (SEQ ID NO: 16) wherein Xaa.sub.2 is an amino acid residue selected from the group comprising Asn and Gln; Xaa.sub.4 is an amino acid residue selected from the group comprising Ala, Val, Leu, norleucine, Met, Phe, and Ile; Xaa.sub.5 is an amino acid residue selected from the group comprising Ala, Val, Leu, Ser and Ile; Xaa.sub.6 is an amino acid residue selected from the group comprising Glu and Asp, Xaa.sub.7 is an amino acid residue selected from the group comprising Glu and Asp, and wherein said amino acid residues Xaa.sub.2, Xaa.sub.3, Xaa.sub.4, Xaa.sub.5, Xaa.sub.6, Xaa.sub.7 are not involved in antibody binding or to a significantly smaller extent as compared to the -His-, the -Lys-Leu, and the -Phe-Phe- binding site.

In another embodiment of the invention, an antibody or a fragment thereof is provided, wherein

Xaa.sub.2 is Gln or Asn, but particularly Gln;

Xaa.sub.4 is Val or Leu, but particularly Val;

Xaa.sub.5 is Ala or Val, but particularly Ala;

Xaa.sub.6 is Glu or Asp, but particularly Glu; and

Xaa.sub.7 is Glu or Asp, but particularly Asp.

In a specific embodiment of the invention, the first of the at least two consecutive amino acid residues predominantly involved in the binding of the antibody involve -Lys-Leu-, and the second of the at least two consecutive amino acid residues involve -Phe-Phe-, and the third at least one amino residue involves -Asp- embedded within the following core sequence:

-Xaa1-Xaa2-Lys-Leu-Xaa4-Phe-Phe-Xaa5-Xaa6-Asp- (SEQ ID NO: 17) wherein Xaa.sub.1 is an amino acid residue selected from the group comprising His, Asn, Gln Lys, and Arg; Xaa.sub.2 is an amino acid residue selected from the group comprising Asn and Gln; Xaa.sub.4 is an amino acid residue selected from the group comprising Ala, Val, Leu, norleucine, Met, Phe, and Ile; Xaa.sub.5 is an amino acid residue selected from the group comprising Ala, Val, Leu, Ser and Ile; Xaa.sub.6 is an amino acid residue selected from the group comprising Glu and Asp, and wherein said amino acid residues Xaa.sub.2, Xaa.sub.3, Xaa.sub.4, Xaa.sub.5, Xaa.sub.6, Xaa.sub.7 are not involved in antibody binding or to a significantly smaller extent as compared to the -Asp-, the -Lys-Leu, and the -Phe-Phe- binding site.

In another embodiment of the invention, an antibody or a fragment thereof is provided, wherein

Xaa.sub.1 is His or Arg, but particularly His;

Xaa.sub.2 is Gln or Asn, but particularly Gln;

Xaa.sub.4 is Val or Leu, but particularly Val;

Xaa.sub.5 is Ala or Val, but particularly Ala; and

Xaa.sub.6 is Glu or Asp, but particularly Glu

In a further specific embodiment of the invention, an antibody or a fragment thereof according to the invention is provided, which binds to 4 distinct binding sites on the .beta.-amyloid protein wherein said 4 distinct binding sites comprise one amino acid residue and two consecutive amino acid residues, respectively, which residues are predominantly involved in the binding of the antibody, wherein said 4 distinct binding sites are located in close proximity to each other on the antigen, separated by at least one amino acid residue not involved in antibody binding or to a significantly smaller extent as compared to said one amino acid residue and said two consecutive amino acid residues of the 4 distinct binding sites thus forming a conformational discontinuous epitope.

In particular, the first of the two consecutive amino acid residues predominantly involved in the binding of the antibody are -Lys-Leu-, and the second of the at least two consecutive amino acid residues are -Phe-Phe-, the first of the single amino residues is -His- and the second of the single amino residues is -Asp- embedded within the following core sequence:

-His-Xaa2-Lys-Leu-Xaa4-Phe-Phe-Xaa5-Xaa6-Asp- (SEQ ID NO: 18) wherein Xaa.sub.2 is an amino acid residue selected from the group comprising Asn and Gln; Xaa.sub.4 is an amino acid residue selected from the group comprising Ala, Val, Leu, norleucine, Met, Phe, and Ile; Xaa.sub.5 is an amino acid residue selected from the group comprising Ala, Val, Leu, Ser and Ile; Xaa.sub.6 is an amino acid residue selected from the group comprising Glu and Asp, and wherein said amino acid residues Xaa.sub.2, Xaa.sub.3, Xaa.sub.4, Xaa.sub.5, Xaa.sub.6, Xaa.sub.7 are not involved in antibody binding or to a significantly smaller extent as compared to the -His-, -Asp-, the -Lys-Leu, and the -Phe-Phe- binding site.

In a specific embodiment of the invention, the recognition and binding sites as defined herein before are forming a conformational discontinuous epitope localized in a region of the .beta.-amyloid protein between amino acid residue 12 to 24, particularly between residues 14 to 23, more particularly between amino acid residues 14 and 20, wherein the three distinct recognition and binding sites comprising 1 and 2 amino acid residues, respectively, are located at position 16, 17, and at position 19 and 20, and at position 14, respectively, which residues are predominantly involved in the binding of the .beta.-amyloid protein and wherein said three distinct recognition and binding sites are separated by one amino acid residue located at position 15 and 18, respectively, which amino acids are not involved in the binding of the antigen or, at least, to a substantially smaller extent.

In a specific embodiment, said consecutive amino acid residues, particularly -Lys-Leu- at position 16 and 17 and -Phe-Phe- at position 19 and 20, which are predominantly involved in the binding of the .beta.-amyloid protein, are embedded into the following core region -- see Original Patent.

In a further specific embodiment, said consecutive amino acid residues, particularly -Lys- at position 16, -Leu- at position 17 and -Phe-Phe- at position 19 and 20, and -His- at position 14, which are predominantly involved in the binding of the .beta.-amyloid protein are embedded into the following core region -- see Original Patent.

In a specific embodiment of the invention, the antibody according to the invention is raised against an antigen fragment which does not contain said distinct binding site.

This shift in the epitopic region may have at least partially been caused by the use of a supramolecular antigenic construct comprising an antigenic peptide corresponding to the amino acid sequence of the .beta.-amyloid peptide, particularly of .beta.-amyloid peptide A.beta..sub.1-16, modified with a hydrophilic moiety such as, for example, polyethylene glycol (PEG), wherein said hydrophilic moiety is covalently bound to each of the termini of the antigenic peptide through at least one, particularly one or two amino acids such as, for example, lysine, glutamic acid and cystein or any other suitable amino acid or amino acid analogue capable of serving as a connecting device for coupling the hydrophilic moiety to the peptide fragment, as described herein below in the immunization process. When a PEG is used as the hydrophilic moiety, the free PEG termini are covalently bound to phosphatidylethanolamine or any other compound suitable to function as the anchoring element, for example, to embed the antigenic construct in the bilayer of a liposome as described herein.

Also the use of lipid A as part of the immunization protocol may have contributed to a shift in the epitopic region.

In a specific embodiment of the invention, an antibody is provided, which comprises the Light Chain Variable Region (LCVR) of SEQ ID NO: 7.

In another specific embodiment, the invention relates to the Light Chain Variable Region (LCVR) of SEQ ID NO: 7.

In still another specific embodiment of the invention, an antibody is provided, which comprises the Heavy Chain Variable Region (HCVR) of SEQ ID NO: 8.

In a further specific embodiment, the invention relates to the Heavy Chain Variable Region (HCVR) of SEQ ID NO: 8.

In one embodiment, the invention relates to an antibody, which comprises both the heavy chain variable region of SEQ ID NO: 8 and the light chain variable region of SEQ ID NO: 7.

Also part of the invention is an antibody comprising a Light Chain Variable Region (LCVR) or Heavy Chain Variable Region (HCVR) or both, a Light Chain Variable Region (LCVR) and a Heavy Chain Variable Region (HCVR) that is homolgous to any of the peptides provided in SEQ ID NO: 7 and 8, respectively.

In particular, the invention relates to an antibody or a fragment thereof, according to the present invention and as described herein before wherein the Light Chain Variable Region (LCVR) has an amino acid sequence that is 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the sequence given in SEQ ID NO: 7.

Further, the invention relates to an antibody or a fragment thereof, according to the present invention and as described herein before wherein the Heavy Chain Variable Region (HCVR) has an amino acid sequence that is 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the sequence given in SEQ ID NO: 8.

Further, the invention relates to an antibody or a fragment thereof, according to the present invention and as described herein before wherein the Light Chain Variable Region (LCVR) and the Heavy Chain Variable Region (HCVR) together have an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the sequence given in SEQ ID NOS: 7 and 8.

In another specific embodiment, the invention relates to the light chain variable region which has an amino acid sequence that is 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the sequence given in SEQ ID NO: 7.

In a further specific embodiment, the invention relates to the heavy chain variable region which has an amino acid sequence that is 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the sequence given in SEQ ID NO: 8.

In another embodiment of the invention, a polynucleotide is provided comprising a nucleotide sequence encoding the antibody according to the invention as described herein before.

In particular, the invention relates to a polynucleotide comprising a nucleotide sequence encoding an antibody according to the invention comprising a) at least the nucleotide sequence of the light chain variable region of SEQ ID NO: 9 b) a nucleotide sequence that differ from the nucleotide sequence of (a) in codon sequence due to the degeneracy of the genetic code c) the complementary sequence to (a) and (b) or d) a fragment of a nucleotide sequence of (a), (b) or (c) comprising a contiguous stretch of nucleotides selected from the group consisting of at least 20 contiguous nucleotides, at least 25 contiguous nucleotides, at least 30 contiguous nucleotides, at least 35 contiguous nucleotides, at least 40 contiguous nucleotides, at least 45 contiguous nucleotides, and at least 50 contiguous nucleotides.

In another embodiment, the invention relates to a polynucleotide comprising a nucleotide sequence encoding an antibody according to the invention comprising a) at least the nucleotide sequence of the light chain of SEQ ID NO: 10 b) a nucleotide sequence that differ from the nucleotide sequence of (a) in codon sequence due to the degeneracy of the genetic code c) the complementary sequence to (a) and (b) or d) a fragment of a nucleotide sequence of (a), (b) or (c) comprising a contiguous stretch of nucleotides selected from the group consisting of at least 20 contiguous nucleotides, at least 25 contiguous nucleotides, at least 30 contiguous nucleotides, at least 35 contiguous nucleotides, at least 40 contiguous nucleotides, at least 45 contiguous nucleotides, and at least 50 contiguous nucleotides.

In still another embodiment, the invention relates to a polynucleotide comprising a nucleotide sequence encoding an antibody according to the invention comprising a) at least the nucleotide sequence of the heavy chain variable region of SEQ ID NO: 11. b) a nucleotide sequence that differ from the nucleotide sequence of (a) in codon sequence due to the degeneracy of the genetic code c) the complementary sequence to (a) and (b) or d) a fragment of a nucleotide sequence of (a), (b) or (c) comprising a contiguous stretch of nucleotides selected from the group consisting of at least 20 contiguous nucleotides, at least 25 contiguous nucleotides, at least 30 contiguous nucleotides, at least 35 contiguous nucleotides, at least 40 contiguous nucleotides, at least 45 contiguous nucleotides, and at least 50 contiguous nucleotides.

In still another embodiment, the invention relates to a polynucleotide comprising a nucleotide sequence encoding an antibody according to the invention comprising a) at least the nucleotide sequence of the heavy chain of SEQ ID NO: 12 or to the complementary sequence b) a nucleotide sequence that differ from the nucleotide sequence of (a) in codon sequence due to the degeneracy of the genetic code c) the complementary sequence to (a) and (b) or d) a fragment of a nucleotide sequence of (a), (b) or (c) comprising a contiguous stretch of nucleotides selected from the group consisting of at least 20 contiguous nucleotides, at least 25 contiguous nucleotides, at least 30 contiguous nucleotides, at least 35 contiguous nucleotides, at least 40 contiguous nucleotides, at least 45 contiguous nucleotides, and at least 50 contiguous nucleotides.

Also comprised by the invention is a polynucleotide comprising a) the nucleotide sequence of SEQ ID NO: 9 encoding the light chain variable region b) a nucleotide sequence that differ from the nucleotide sequence of (a) in codon sequence due to the degeneracy of the genetic code c) the complementary sequence to (a) and (b) or d) a fragment of a nucleotide sequence of (a), (b) or (c) comprising a contiguous stretch of nucleotides selected from the group consisting of at least 20 contiguous nucleotides, at least 25 contiguous nucleotides, at least 30 contiguous nucleotides, at least 35 contiguous nucleotides, at least 40 contiguous nucleotides, at least 45 contiguous nucleotides, and at least 50 contiguous nucleotides.

Also comprised by the invention is a polynucleotide comprising a) the nucleotide sequence of SEQ ID NO: 10 encoding the light chain. b) a nucleotide sequence that differ from the nucleotide sequence of (a) in codon sequence due to the degeneracy of the genetic code c) the complementary sequence to (a) and (b) or d) a fragment of a nucleotide sequence of (a), (b) or (c) comprising a contiguous stretch of nucleotides selected from the group consisting of at least 20 contiguous nucleotides, at least 25 contiguous nucleotides, at least 30 contiguous nucleotides, at least 35 contiguous nucleotides, at least 40 contiguous nucleotides, at least 45 contiguous nucleotides, and at least 50 contiguous nucleotides.

Also comprised by the invention is a polynucleotide comprising a) the nucleotide sequence of SEQ ID NO: 11 encoding the heavy chain variable region b) a nucleotide sequence that differ from the nucleotide sequence of (a) in codon sequence due to the degeneracy of the genetic code c) the complementary sequence to (a) and (b) or d) a fragment of a nucleotide sequence of (a), (b) or (c) comprising a contiguous stretch of nucleotides selected from the group consisting of at least 20 contiguous nucleotides, at least 25 contiguous nucleotides, at least 30 contiguous nucleotides, at least 35 contiguous nucleotides, at least 40 contiguous nucleotides, at least 45 contiguous nucleotides, and at least 50 contiguous nucleotides.

Also comprised by the invention is a polynucleotide comprising a) the nucleotide sequence of SEQ ID NO: 12 encoding the heavy chain b) a nucleotide sequence that differ from the nucleotide sequence of (a) in codon sequence due to the degeneracy of the genetic code c) the complementary sequence to (a) and (b) or d) a fragment of a nucleotide sequence of (a), (b) or (c) comprising a contiguous stretch of nucleotides selected from the group consisting of at least 20 contiguous nucleotides, at least 25 contiguous nucleotides, at least 30 contiguous nucleotides, at least 35 contiguous nucleotides, at least 40 contiguous nucleotides, at least 45 contiguous nucleotides, and at least 50 contiguous nucleotides.

In a further embodiment, the invention relates to any nucleotide sequence that hybridizes to a) a nucleotide sequence according to the invention and as given in SEQ ID NOS: 9, 10, 11 and 12, respectively, b) a nucleotide sequence that differ from the nucleotide sequence of (a) in codon sequence due to the degeneracy of the genetic code c) the complementary sequence to (a) and (b) or d) a fragment of a nucleotide sequence of (a), (b) or (c) comprising a contiguous stretch of nucleotides selected from the group consisting of at least 20 contiguous nucleotides, at least 25 contiguous nucleotides, at least 30 contiguous nucleotides, at least 35 contiguous nucleotides, at least 40 contiguous nucleotides, at least 45 contiguous nucleotides, and at least 50 contiguous nucleotides.

In particular, the invention relates to any nucleotide sequence that hybridizes under conventional hybridization conditions, particularly under stringent hybridization conditions, to a nucleotide sequence according to the invention and as given in SEQ ID NOS: 9, 10, 11 and 12, respectively, particularly to the complementary strand thereof.

In a further embodiment, the invention relates to any nucleotide sequence that hybridizes to a nucleotide sequence according to the present invention and as given in SEQ ID NOS: 9, 10, 11 and 12, respectively, particularly to the complementary strand thereof, under conventional hybridization conditions at which 5.times.SSPE, 1% SDS, 1.times.Denhardts solution is used as a solution and/or hybridization temperatures are between 35.degree. C. and 70.degree. C., preferably 65.degree. C. After hybridization, washing is preferably carried out first with 2.times.SSC, 1% SDS and subsequently with 0.2.times.SSC at temperatures between 35.degree. C. and 70.degree. C., preferably at 65.degree. C. (regarding the definition of SSPE, SSC and Denhardts solution (see Sambrook et al. loc. cit.).

In particular, the invention relates to any nucleotide sequence that hybridizes to a nucleotide sequence according to the present invention and as given in SEQ ID NOS: 9, 10, 11 and 12, respectively, particularly to the complementary strand thereof, under stringent hybridization conditions as for instance described in Sambrook et al, supra, more particularly under stringent hybridization conditions where hybridization and washing occurs at 65.degree. C. as indicated above.

In a specific embodiment the present invention provides an antibody, particularly a bifunctional antibody, but especially a monoclonal antibody, particularly a bifunctional monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody has the characteristic properties of an antibody produced by a hybridoma cell line selected from the group consisting of FP 12H3, FP 12H3-C2, and FP 12H3-G2 deposited on Dec. 1, 2005 and Dec. 9, 2005, respectively, as DSM ACC2752, DSM ACC 2750 and DSM ACC2751, respectively.

More particularly, the invention relates to an antibody including any functionally equivalent antibody or functional parts thereof produced by hybridoma cell line FP 12H3, deposited on Dec. 1, 2005 and Dec. 9, 2005, respectively as DSM ACC2752.

More particularly, the invention relates to a monoclonal antibody including any functionally equivalent antibody or functional parts thereof produced by hybridoma cell line FP 12H3-C2, deposited on Dec. 1, 2005 and Dec. 9, 2005, respectively as DSM ACC2750.

More particularly, the invention relates to a monoclonal antibody including any functionally equivalent antibody or functional parts thereof produced by hybridoma cell line FP 12H3-C2, deposited on Dec. 1, 2005 and Dec. 9, 2005, respectively as DSM ACC2751.

In another specific embodiment the present invention provides an antibody, particularly a bifunctional antibody, but especially a monoclonal antibody, particularly a bifunctional monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody has the characteristic properties of an antibody produced by hybridoma cell line ET 7E3 deposited on Dec. 8, 2005 as DSM ACC2755.

More particularly, the invention relates to a monoclonal antibody including any functionally equivalent antibody or functional parts thereof produced by hybridoma cell line ET 7E3, deposited on Dec. 8, 2005 as DSM ACC2755.

In another specific embodiment the present invention provides an antibody, particularly a bifunctional antibody, but especially a monoclonal antibody, particularly a bifunctional monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody has the characteristic properties of an antibody produced by hybridoma cell line EJ 7H3 deposited on Dec. 8, 2005 as DSM ACC2756.

More particularly, the invention relates to a monoclonal antibody including any functionally equivalent antibody or functional parts thereof produced by hybridoma cell line EJ 7H3, deposited on Dec. 8, 2005 as DSM ACC2756.

It is another object of the present invention to provide methods and compositions comprising an antibody according to the invention and as described herein before for the prevention and/or therapeutic treatment and/or alleviation of the effects of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including 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), including diseases or conditions characterized by a loss of cognitive memory capacity such as, for example, mild cognitive impairment (MCI), including diseases or conditions characterized by a loss of cognitive memory capacity such as, for example, mild cognitive impairment (MCI), Lewy body dementia, Down's syndrome, hereditary cerebral hemorrhage with amyloidosis (Dutch type); the Guam Parkinson-Dementia complex; as well as other diseases and conditions 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), inclusion-body myositis (IBM), Adult Onset Diabetis; senile cardiac amyloidosis; endocrine tumors, and others, including macular degeneration, for example, by passively immunizing a human or animal with an antibody according to the invention and as described herein before.

Another object of the present invention is to provide a method of using a monoclonal antibody and/or a functional part thereof according to the invention and compositions comprising an antibody according to the invention and as described herein before for diagnosing and therapeutic intervention of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including 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), including diseases or conditions characterized by a loss of cognitive memory capacity such as, for example, mild cognitive impairment (MCI), including diseases or conditions characterized by a loss of cognitive memory capacity such as, for example, mild cognitive impairment (MCI), 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), inclusion-body myositis (IBM), Adult Onset Diabetis; senile cardiac amyloidosis; endocrine tumors, and others, including macular degeneration.

In particular, it is an object of the present invention to provide a method of using a monoclonal antibody and/or a functional part thereof according to the invention and compositions comprising an antibody, particularly a bispecific or bi-effective antibody but especially a bispecific or bi-effective monoclonal antibody according to the invention and as described herein before for reducing and preventing the occurrence of neurological disorders, including but not limited to Alzheimer's Disease.

The compositions according to the present invention comprise an antibody, particularly a bispecific or bi-effective antibody, according to the invention and as described herein before including any functionally equivalent antibody or functional parts thereof, particularly in a therapeutically effective amount or, more particularly, a monoclonal antibody, especially a bispecific or bi-effective monoclonal antibody, according to the invention and as described herein before including any functionally equivalent antibody or functional parts thereof, particularly in a therapeutically effective amount and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

Particularly, the composition according to the present invention comprises an antibody, particularly a monoclonal antibody including any functionally equivalent antibody or functional parts thereof which antibody is capable of inhibiting the aggregation of amyloid monomeric peptides, specifically .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, into high molecular polymeric amyloid fibrils or filaments and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

In a further embodiment of the invention a composition is provided comprising an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody, upon co-incubation, particularly upon co-incubation at a molar concentration ratio of up to 1:100, more particularly at a molar concentration ratio of between 1:30 and 1:100, but especially at a molar concentration ratio of 1:100, with amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, inhibits the aggregation of the A.beta. monomers into high molecular polymeric fibrils or filaments. In particular, said inhibition amounts to at least 50%, particularly to at least 65%, more particularly to at least 75%, even more particularly to at least 80%, but especially to at least 85%-90%, or more as compared to the respective amyloid peptide monomers incubated in buffer (control) and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

In particular, the co-incubation of the antibody according to the invention with amyloid monomeric peptides is carried out for 48 hours at a temperature of 37.degree. C.

The aggregation inhibition potential of the antibody according to the invention may be determined by density-gradient ultracentrifugation followed by a SDS-PAGE sedimentation analysis on a preformed gradient and/or by a thioflavin T (Th-T) fluorescent assay.

The present invention further provides a composition comprising an antibody which is capable of disaggregating high molecular polymeric amyloid fibrils or filaments formed by the aggregation of amyloid monomeric peptides, specifically .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

Through the disaggregation of amyloidogenic polymeric fibrils or filaments the antibodies according to the present invention are capable of preventing or slowing down the formation of amyloid plaques which leads to an alleviation of the symptoms associated with the disease and a delay or reversal of its progression.

In another embodiment of the invention a composition is provided comprising an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody, upon co-incubation at a molar concentration ratio of between 1:30 and 1:100, particularly at a ratio of 1:100, with preformed high molecular polymeric amyloid fibrils or filaments formed by the aggregation of amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, particularly upon co-incubation for 24 hours at a temperature of 37.degree. C., is capable of disaggregating the preformed polymeric fibrils or filaments by at least 35%, particularly by at least 40%, more particularly by at least 50%, even more particularly by at least 60%, but especially by at least 70% or more and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

The disaggregation potential of the antibody according to the invention may be determined by density-gradient ultracentrifugation followed by a SDS-PAGE sedimentation analysis on a preformed gradient and/or by a thioflavin T (Th-T) fluorescent assay.

The present invention further provides a composition comprising an antibody or functional parts thereof which antibody is conformationally sensitive, particularly in an effective amount, more particularly in a therapeutically effective amount.

In a further embodiment of the invention composition is provided comprising an antibody is provided, but especially a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody, upon co-incubation with preformed high molecular polymeric amyloid fibrils or filaments formed by the aggregation of amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, particularly upon co-incubation for 24 hours at a temperature of 37.degree. C., is capable of inducing a transition of the .beta.-sheet conformation towards an .alpha.-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 Val12 of the A.beta. protein, which leads to an increase of the random coil conformation at the expense of the .beta.-sheet conformation and an improved solubilization of the preformed high molecular polymeric amyloid fibrils or filaments and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient. In particular the decrease of the .beta.-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).

The antibody's potential in inducing a transition in the secondary structure is determined by solid state 13C NMR spectroscopy but, in particular, by measuring the integral intensities of the conformations of Val 12 C.beta. in the A.beta..sub.1-42 peptide.

The present invention further provides a composition comprising an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, particularly in a therapeutically effective amount, which antibody is bifunctional in that it exhibits both an aggregation inhibition property as well as a disaggregation property as defined herein before, preferably paired with a high degree of conformational sensitivity and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

In still another embodiment of the invention a composition is provided comprising a bifunctional antibody, but especially a bifunctional monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody, upon co-incubation with amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, inhibits the aggregation of the A.beta. monomers into high molecular polymeric fibrils and, in addition, upon co-incubation with preformed high molecular polymeric amyloid fibrils or filaments formed by the aggregation of amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, is capable of disaggregating the preformed polymeric fibrils or filaments and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

In a specific embodiment of the invention the co-incubation of the bifunctional antibody according to the invention, but especially of the bifunctional monoclonal antibody according to the invention with amyloid monomeric peptides and preformed high molecular polymeric amyloid fibrils or filaments, respectively, takes place at a molar concentration ratio of up to 1:100, particularly at a ratio of between 1:30 and 1:100, and more particularly at a ration of 1:100.

In a further specific embodiment of the invention co-incubation with amyloid monomeric peptides and preformed high molecular polymeric amyloid fibrils or filaments is done for 48 hours and 24 hours, respectively, at a temperature of 37.degree. C.

In still another specific embodiment of the invention a composition is provided comprising a bifunctional antibody according to the invention, but especially a bifunctional monoclonal antibody according to the invention which is capable of disaggregating the preformed polymeric fibrils or filaments by at least 10%, particularly by at least 25%, more particularly by at least 35%, even more particularly by at least 50%, but especially by at least 60-70% or more and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

In still another specific embodiment of the invention a composition is provided comprising a bifunctional antibody according to the invention, but especially a bifunctional monoclonal antibody according to the invention inhibits the aggregation of amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides by at least 50%, particularly by at least 65%, more particularly by at least 75%, even more particularly by at least 80%, but especially by at least 85-90%, or more as compared to the respective amyloid peptide monomers incubated in buffer (control) and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

In particular, the present invention provides a composition comprising an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody mediates the inhibition of polymerization of amyloid monomeric peptides, specifically .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially a A.beta..sub.1-42 monomeric peptides and/or induces solubilization of preformed high molecular polymeric amyloid fibrils or filaments formed by the aggregation of amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, through specific and direct binding of the antibody to the A.beta. fibers, which leads to a transition of secondary conformation and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

The present invention further provides a composition comprising an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody directly and specifically binds to .beta.-amyloid fibers such as, for example, fibers comprising A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially to fibers comprising A.beta..sub.1-42 monomeric peptides and/or induces solubilization of preformed high molecular polymeric amyloid fibrils or filaments formed by the aggregation of amyloid monomeric peptides, particularly .beta.-amyloid monomeric peptides such as, for example, A.beta. monomeric peptides 1-39; 1-40, 1-41, 1-42, or 1-43, but especially A.beta..sub.1-42 monomeric peptides, by targeting and specifically binding to an epitopic region of the .beta.-amyloid protein, particularly an epitopic region of the A.beta. polypeptide confined by amino acid residues aa.sub.n-aa.sub.m with n being an integer between 2 and 15, particularly between 5 and 15, more particularly between 8 and 15, even more particularly between 10 and 15 and m being an integer between 3 and 17, particularly between 6 and 17, more particularly between 9 and 17, even more particularly between 11 and 17, wherein n and m cannot be identical numbers and n must always be a smaller number than m, with the difference between n and m.gtoreq.2 and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

The binding of the antibody according to the invention may induce a conformational transition in said protein, particularly a transition of the .beta.-sheet conformation towards an .alpha.-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 Val12 of the A.beta. protein.

In a further embodiment, the invention provides a composition comprising an antibody, particularly a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody incorporates at least one of the properties mentioned herein before, that is aggregation inhibition, disaggregation, induction of conformational transition, recognition of and direct binding to the 4-16 and/or the 14 to 23, but particularly the 14 to 20 epitopic region, but especially a combination of two or more of said properties and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

In a specific embodiment, the invention provides a composition comprising an antibody, particularly a bifunctional antibody, but especially a monoclonal antibody, particularly a bifunctional monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which exhibits high specificity to A.beta..sub.1-42 monomeric peptides but shows essentially no or only minor cross-reactivity to A.beta..sub.1-38, A.beta..sub.1-39, A.beta..sub.1-40, and/or A.beta..sub.1-41 monomeric peptides, particularly an antibody, but especially a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody is up to 100 fold, particularly 50 to 100 fold, more particularly 80 to 100 fold, but especially 100 fold more sensitive to amyloid peptide A.beta..sub.1-42 as compared to A.beta..sub.1-38, A.beta..sub.1-39, A.beta..sub.1-40, A.beta..sub.1-41 and up to 1000 fold, particularly 500 to 1000 fold, more particularly 800 to 1000 fold, but especially 1000 fold more sensitive to amyloid peptide A.beta..sub.1-42 as compared to A.beta..sub.1-38, and thus capable of inhibiting, in vitro and in vivo, the aggregation of amyloidogenic monomeric peptides, but especially of amyloid peptide A.beta..sub.1-42 and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

In another specific embodiment of the invention a composition is provided comprising an antibody, particularly a bifunctional antibody, but especially a monoclonal antibody, particularly a bifunctional monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which has a high binding sensitivity to amyloid peptide A.beta..sub.1-42 and is capable of detecting A.beta..sub.1-42 fibers in a concentration of down to at least 0.001 .mu.g, but particularly in a concentration range of between 0.5 .mu.g and 0.001 .mu.g, more particularly between 0.1 .mu.g and 0.001 .mu.g, but especially in a concentration of 0.001 .mu.g and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

In a very specific embodiment of the invention a composition comprising an antibody is provided, particularly a bifunctional antibody, but especially a monoclonal antibody, particularly a bifunctional monoclonal antibody, including any functionally equivalent antibody or functional parts thereof, which antibody is capable of detecting A.beta..sub.1-42 fibers down to a minimal concentration of 0.001 .mu.g and A.beta..sub.1-40 fibers down to a minimal concentration of 0.1 .mu.g and A.beta..sub.1-38 fibers down to a minimal concentration of 1 .mu.g amount of fibers and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

In a specific embodiment the present invention relates to a composition comprising a monoclonal antibody including any functionally equivalent antibody or functional parts thereof which antibody has the characteristic properties of an antibody produced by a hybridoma cell line selected from the group consisting of FP 12H3, FP 12H3-C2, and FP 12H3-G2 deposited on Dec. 1, 2005 and Dec. 9, 2005, respectively, as DSM ACC2752, DSM ACC2750 and DSM ACC2751, and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

More particularly, the invention relates to a composition comprising a monoclonal antibody including any functionally equivalent antibody or functional parts thereof produced by hybridoma cell line FP 12H3, deposited on Dec. 1, 2005 and Dec. 9, 2005, respectively as DSM ACC2752 and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

More particularly, the invention relates to a composition comprising a monoclonal antibody including any functionally equivalent antibody or functional parts thereof produced by hybridoma cell line FP 12H3-C2, deposited on Dec. 1, 2005 and Dec. 9, 2005, respectively as DSM ACC2750 and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient The invention further relates to a composition comprising a monoclonal antibody including any functionally equivalent antibody or functional parts thereof produced by hybridoma cell line FP 12H3-G2, deposited on Dec. 1, 2005 and Dec. 9, 2005, respectively as DSM ACC2751 and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

In another specific embodiment the present invention relates to a composition comprising a monoclonal antibody including any functionally equivalent antibody or functional parts thereof which antibody has the characteristic properties of an antibody produced by hybridoma cell line ET 7E3 deposited on Dec. 8, 2005 as DSM ACC2755 and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

The invention further relates to a composition comprising a monoclonal antibody including any functionally equivalent antibody or functional parts thereof produced by hybridoma cell line ET 7E3, deposited on Dec. 8, 2005 as DSM ACC2755 and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient

In another specific embodiment the present invention relates to a composition comprising a monoclonal antibody including any functionally equivalent antibody or functional parts thereof which antibody has the characteristic properties of an antibody produced by hybridoma cell line EJ 7H3 deposited on Dec. 8, 2005 as DSM ACC2756 and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

The invention further relates to a composition comprising a monoclonal antibody including any functionally equivalent antibody or functional parts thereof produced by hybridoma cell line EJ 7H3, deposited on Dec. 8, 2005 as DSM ACC2756 and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

The antibody, particularly the monoclonal antibody according to the invention including any functionally equivalent antibody or functional parts thereof may be administered in combination with other biologically active substances and procedures for the treatment of diseases. The other biologically active substances may be part of the same composition already comprising the antibody according to the invention, in form of a mixture, wherein the antibody and the other biologically active substance are intermixed in or with the same pharmaceutically acceptable solvent and/or carrier or may be provided separately as part of a separate composition, which may be offered separately or together in form a kit of parts.

The antibody, particularly the monoclonal antibody according to the invention including any functionally equivalent antibody or functional parts thereof may be administered at the same time with the other biologically active substance or substances, intermittently or sequentially. For example, the monoclonal antibody according to the invention including any functionally equivalent antibody or functional parts thereof may be administered simultaneously with a first additional biologically active substance or sequentially after or before administration of the antibody. If an application scheme is chosen where more than one additional biologically active substance are administered together with the at least one antibody according to the invention, the compounds or substances may partially be administered simultaneously, partially sequentially in various combinations.

It is another object of the present invention to provide for mixtures of antibodies comprising at least one antibody according to the present invention and, optionally, one or more further biologically active substances, as well as to methods of using individual antibodies, or mixtures thereof including compositions comprising said antibodies or mixtures of antibodies for the prevention and/or therapeutic treatment and/or alleviation of the effects of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including 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), including diseases or conditions characterized by a loss of cognitive memory capacity such as, for example, mild cognitive impairment (MCI), 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), inclusion-body myositis (IBM), Adult Onset Diabetis; senile cardiac amyloidosis; endocrine tumors, and others, including macular degeneration.

The mixtures according to the invention may comprise, in addition to an antibody according to the invention, a biologically active substance such as, for example, known compounds used in the medication of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including amyloidosis, a group of diseases and disorders associated with amyloid or amyloid-like protein such as the A.beta. protein involved in Alzheimer's disease.

In another embodiment of the invention, the other biologically active substance or compound may also be a therapeutic agent that may be used in the treatment of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including amyloidosis caused by amyloid .beta. or may be used in the medication of other neurological disorders.

The other biologically active substance or compound may exert its biological effect by the same or a similar mechanism as the antibody according to the invention or by an unrelated mechanism of action or by a multiplicity of related and/or unrelated mechanisms of action.

Generally, the other biologically active compound may include neutron-transmission enhancers, psychotherapeutic drugs, acterylcholine esterase inhibitors, calcium-channel blockers, biogenic amines, benzodiazepine tranquilizers, acetylcholine synthesis, storage or release enhancers, acetylcholine postsynaptic receptor agonists, monoamine oxidase-A or -B inhibitors, N-methyl-D-aspartate glutamate receptor antagonists, non-steroidal anti-inflammatory drugs, antioxidants, and serotonergic receptor antagonists.

In particular, the mixture according to the invention may comprise at least one other biologically active compound selected from the group consisting of compounds against oxidative stress, anti-apoptotic compounds, metal chelators, inhibitors of DNA repair such as pirenzepine and metabolites, 3-amino-1-propanesulfonic acid (3APS), 1,3-propanedisulfonate (1,3PDS), secretase activators, .beta.- and .gamma.-secretase inhibitors, tau proteins, neurotransmitter, .beta.-sheet breakers, anti-inflammatory molecules, or cholinesterase inhibitors (ChEIs) such as tacrine, rivastigmine, donepezil, and/or galantamine and other drugs 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.

In a further embodiment, the mixtures according to the invention may comprise niacin or memantine together with an antibody according to the present invention and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

In still another embodiment of the invention mixtures are provided that comprise "atypical antipsychotics" such as, for example clozapine, ziprasidone, risperidone, aripiprazole or olanzapine for the treatment of positive and negative psychotic symptoms including hallucinations, delusions, thought disorders (manifested by marked incoherence, derailment, tangentiality), and bizarre or disorganized behavior, as well as anhedonia, flattened affect, apathy, and social withdrawal, together with an antibody according to the invention and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.

In a specific embodiment of the invention, the compositions and mixtures according to the invention and as described herein before comprise the antibody and the biologically active substance, respectively, in a therapeutically effective amount.

Other compounds that can be suitably used in mixtures in combination with the antibody according to the present invention are, for example, described in WO 2004/058258 (see especially pages 16 and 17) including therapeutic drug targets (page 36-39), alkanesulfonic acids and alkanolsulfuric acids (pages 39-51), cholinesterase inhibitors (pages 51-56), NMDA receptor antagonists (pages 56-58), estrogens (pages 58-59), non-steroidal anti-inflammatory drugs (pages 60-61), antioxidants (pages 61-62), peroxisome proliferators-activated receptor (PPAR) agonists (pages 63-67), cholesterol-lowering agents (pages 68-75); amyloid inhibitors (pages 75-77), amyloid formation inhibitors (pages 77-78), metal chelators (pages 78-79), anti-psychotics and anti-depressants (pages 80-82), nutritional supplements (pages 83-89) and compounds increasing the availability of biologically active substances in the brain (see pages 89-93) and prodrugs (pages 93 and 94), which document is incorporated herein by reference.

Further provided is a method for producing an antibody, particularly a method for producing a monoclonal antibody, including any functionally equivalent antibody or functional parts thereof according to the invention, which method comprises raising antibodies but particularly monoclonal antibodies against a supramolecular antigenic construct comprising an antigenic peptide corresponding to the amino acid sequence of the .beta.-amyloid peptide, particularly of .beta.-amyloid peptide A.beta..sub.1-15, A.beta..sub.1-16 and A.beta..sub.1-16(.DELTA.14), modified with hydrophobic moieties such as, for example, palmitic acid or a hydrophilic moiety such as, for example, polyethylene glycol (PEG) or a combination of both, wherein said hydrophobic and hydrophilic moiety, respectively, is covalently bound to each terminus through at least one, particularly one or two, amino acids such as, for example, lysine or any other suitable amino acid or amino acid analogue capable of serving as a coupling device or linker molecule for the coupling of said hydrophobic and hydrophilic moiety such as, for example, glutamic acid or cystein.

Also part of the invention is the use of a monoclonal antibody and/or a functional part thereof according to the invention and as described herein before and/or a pharmaceutical composition, or a mixture comprising said antibody, for the preparation of a medicament for treating or alleviating the effects of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including 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 and diseases or conditions characterized by a loss of cognitive memory capacity such as, for example, mild cognitive impairment (MCI), 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), inclusion-body myositis (IBM), Adult Onset Diabetis; senile cardiac amyloidosis; endocrine tumors, and others, including macular degeneration.

In another embodiment of the present invention a method is provided for the preparation of a pharmaceutical composition using an antibody according to the invention and/or a functional part thereof but especially a monoclonal antibody and/or a functional part thereof or a functionally equivalent antibody, for use in treating or alleviating the effects of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including 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) and diseases or conditions characterized by a loss of cognitive memory capacity such as, for example, mild cognitive impairment (MCI), 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 Diabetis; senile cardiac amyloidosis; endocrine tumors, and others, including macular degeneration comprising formulating an antibody according to the invention in a pharmaceutically acceptable form.

The antibodies and/or functional parts thereof but especially the monoclonal antibodies and/or functional parts thereof or a functionally equivalent antibody and the compositions and mixtures comprising said antibody according to the present invention may be used for the preparation of a medicament for preventing, treating or alleviating the effects of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including amyloidosis, a group of diseases and disorders associated with amyloid plaque formation including secondary amyloidosis and age-related amyloidosis including, but not limited to, neurological disorders such as Alzheimer's Disease (AD), including diseases or conditions characterized by a loss of cognitive memory capacity such as, for example, mild cognitive impairment (MCI), 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), inclusion-body myositis (IBM), Adult Onset Diabetis; senile cardiac amyloidosis; endocrine tumors, and others, including macular degeneration.

In a further embodiment of the invention a method is provided for reducing the plaque load in the brain of an animal, particularly a mammal, but especially a human suffering from a disease or condition leading to an increased plaque load in the brain comprising administering to an animal, particularly a mammal, more particularly a human in need of such a treatment, a therapeutically effective amount of an antibody and/or a functional part thereof but especially of the monoclonal antibody and/or a functional part thereof or of a functionally equivalent antibody according to the invention and as described herein before, or a composition or a mixture comprising said antibody.

In particular, the plaque load is reduced by at least 20%, particularly by at least 25%, more particularly by at least 30%, even more particularly by more than 30%.

In a further embodiment of the invention a method for reducing the amount of plaques in the brain of an animal, particularly a mammal, but especially a human suffering from a disease or condition leading to an increased plaque load in the brain comprising administering to an animal, particularly a mammal, more particularly a human in need of such a treatment, a therapeutically effective amount of an antibody and/or a functional part thereof but especially of the monoclonal antibody and/or a functional part thereof or of a functionally equivalent antibody according to the invention and as described herein before, or a composition or a mixture comprising said antibody.

In particular, the amount of plaques in the brain is reduced by at least 10%, particularly by at least 15%, more particularly by more than 15%.

In still another embodiment of the invention a method for decreasing the total amount of soluble A.beta. in the brain of an animal, particularly a mammal, but especially a human suffering from a disease or condition leading to increased concentrations of soluble A.beta. in the brain comprising administering to an animal, particularly a mammal, more particularly a human in need of such a treatment, a therapeutically effective amount of an antibody and/or a functional part thereof but especially of the monoclonal antibody and/or a functional part thereof or of a functionally equivalent antibody according to the invention and as described herein before, or a composition or a mixture comprising said antibody.

It is an objective of the present invention to provide a method for preventing, treating or alleviating the effects of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including amyloidosis, a group of diseases and disorders associated with amyloid plaque formation including secondary amyloidosis and age-related amyloidosis including, but not limited to, neurological disorders such as Alzheimer's Disease (AD), including diseases or conditions characterized by a loss of cognitive memory capacity such as, for example, mild cognitive impairment (MCI), 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), inclusion-body myositis (IBM), Adult Onset Diabetis; senile cardiac amyloidosis; endocrine tumors, and others, including macular degeneration, by administering an antibody, but particularly a monoclonal antibody or a composition or mixture comprising such an antibody according to the invention to an animal or a human affected by such a disorder comprising administering to an animal, particularly a mammal, more particularly a human in need of such a treatment, a therapeutically effective amount of an antibody and/or a functional part thereof but especially of the monoclonal antibody and/or a functional part thereof or of a functionally equivalent antibody according to the invention and as described herein before, or a composition or a mixture comprising said antibody.

In a specific embodiment the invention provides a method for retaining or increasing cognitive memory capacity of an animal, particularly a mammal or a human suffering from memory impairment by administering to an animal, particularly a mammal or a human in need of such a treatment, an antibody, but particularly a monoclonal antibody according to the invention or a composition or mixture comprising such an antibody according to the invention and as described herein before.

In another embodiment of the present invention a method is provided for the preparation of a pharmaceutical composition using an antibody according to the invention and/or a functional part thereof but especially a monoclonal antibody and/or a functional part thereof or a functionally equivalent antibody for preventing, treating or alleviating the effects of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including 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), including diseases or conditions characterized by a loss of cognitive memory capacity such as, for example, mild cognitive impairment (MCI), 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), inclusion-body myositis (IBM), Adult Onset Diabetis; senile cardiac amyloidosis; endocrine tumors, and others, including macular degeneration.

In a specific embodiment the invention provides a method for the preparation of a pharmaceutical composition using an antibody according to the invention and/or a functional part thereof but especially a monoclonal antibody and/or a functional part thereof or a functionally equivalent antibody for retaining or increasing cognitive memory capacity of an animal, particularly a mammal or a human, suffering from memory impairment by administering to an animal, particularly a mammal or a human, an antibody, but particularly a monoclonal antibody or a composition or mixture comprising such an antibody according to the invention and as described herein before.

Claim 1 of 15 Claims

1. A monoclonal antibody which specifically binds to monomeric, oligomeric or polymeric forms of amyloid-.beta., wherein: (i) the CDR 1 of the light chain variable region of the monoclonal antibody has the amino acid sequence of SEQ ID NO: 23; (ii) the CDR2 of the light chain variable region of the monoclonal antibody has the amino acid sequence of SEQ ID NO: 24: (iii) the CDR3 of the light chain variable region of the monoclonal antibody has the amino acid sequence of SEQ ID NO: 25: (iv) the CDR1 of the heavy chain variable region of the monoclonal antibody has the amino acid sequence of SEQ ID NO: 26: (v) the CDR2 of the heavy chain variable region of the monoclonal antibody has the amino acid sequence of SEQ ID NO: 27; and (vi) the CDR3 of the heavy chain variable region of the monoclonal antibody has the amino acid sequence of SEQ ID NO: 28.

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