Treatment of neurological disorders using anti-Abeta antibodies

Abstract

Provided are antibodies that bind to human beta amyloid peptide, methods for detecting, measuring, and treating amyloidogenic disorders with the antibodies, pharmaceutical compositions comprising the antibodies, and methods of manufacture. The present disclosure relates to antibodies (and antibody fragments) that specifically bind to Aβ, methods for making such antibodies and antibody fragments and related nucleic acids, methods for treating patients with Aβ-related neurological disorders, pharmaceutical formulations and compositions of antibodies that exhibit high affinity binding to Aβ for prophylactic and / or therapeutic use, e.g., treating, reducing the risk of, or delaying the onset of amyloidogenic disease, preventing, reducing, or inhibiting markers of amyloidogenic disease, e.g., amyloid plaques, and improving cognition.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 481,631, filed January 26, 2023, U.S. Provisional Application No. 63 / 597,868, filed November 10, 2023, U.S. Provisional Application No. 63 / 597,861, filed November 10, 2023, and U.S. Provisional Application No. 63 / 618,045, filed January 5, 2024, each of which is incorporated by reference in its entirety.

[0002] Sequence Listing A computer-readable form of the Sequence Listing is being submitted with this application via electronic submission and is incorporated herein by reference in its entirety. This Sequence Listing is contained in file name "20-1030-WO2_SeqList", created on January 4, 2024, and is 124,150 bytes in size.

[0003] The present disclosure relates to anti-amyloid beta (Aβ) antibodies, as well as compositions and methods of use thereof. [Background technology]

[0004] Alzheimer's disease (AD) is a progressive disease that leads to senile dementia. The disease is generally classified as late-onset, which occurs at an advanced age (over 65 years of age), or early-onset, which occurs long before old age, i.e., between 35 and 60 years of age. While the pathology of the disease appears to be the same in both types of disease, abnormalities tend to be more severe and widespread in cases beginning earlier. The disease is characterized by at least two types of lesions in the brain: neurofibrillary tangles and senile plaques. Neurofibrillary tangles are intracellular deposits of microtubule-associated tau protein, consisting of two filaments twisted around each other in pairs. Senile plaques (i.e., amyloid plaques) are areas of disorganized neuropil spanning up to 150 μm with an extracellular amyloid deposit in the center, visible by microscopic analysis of brain tissue sections. The accumulation of amyloid plaques in the brain has also been associated with Down's syndrome and other cognitive disorders.

[0005] The primary component of plaques is a peptide called amyloid beta (Aβ or Abeta), or β-amyloid peptide. The Aβ peptide is a 4-kDa internal fragment of a larger transmembrane glycoprotein called amyloid precursor protein (APP), consisting of 39–43 amino acids. As a result of proteolytic processing of APP by different secretases, Aβ is found in both a short form, primarily 40 amino acids long, and a long form ranging from 42–43 amino acids long. Part of the hydrophobic transmembrane domain of APP is found in the carboxy terminus of Aβ, which may explain the ability of Aβ to aggregate into plaques, especially in the long form. The accumulation of amyloid plaques in the brain ultimately leads to neuronal death. This type of physical symptoms associated with the deterioration of neuronal function characterizes Alzheimer's disease.

[0006] Monoclonal antibodies (mAbs) targeting amyloid beta have been clinically demonstrated to reduce amyloid plaque burden in patients. The FDA granted accelerated approval to the anti-amyloid beta antibodies aducanumab and lecanemab. The FDA approval of these antibodies was based on the antibodies demonstrating amyloid beta reduction on PET imaging, a surrogate endpoint deemed relatively likely to predict clinical benefit. Indeed, clinical trials of anti-amyloid beta antibodies, including aducanumab and lecanemab, have shown that plaque reduction is associated with a slowdown in cognitive decline in Alzheimer's disease. However, these treatments have limited efficacy and / or treatment-related side effects. Furthermore, these antibodies require intravenous administration and / or frequent, high-dose subcutaneous administration, placing a burden on patients and caregivers. Therefore, there remains a need for effective and safe anti-amyloid beta antibodies for the treatment of Alzheimer's disease, particularly those that are effective when administered subcutaneously as part of a relatively infrequent dosing regimen. Summary of the Invention [Means for solving the problem]

[0007] The present disclosure relates to antibodies (and antibody fragments) that specifically bind to Aβ, methods for making such antibodies and antibody fragments and related nucleic acids, methods for treating patients with Aβ-related neurological disorders, pharmaceutical formulations and compositions of antibodies that exhibit high-affinity binding to Aβ for prophylactic and / or therapeutic use, e.g., to treat, reduce the risk of, or delay the onset of amyloidogenic disease, prevent, reduce, or inhibit markers of amyloidogenic disease, e.g., amyloid plaques, and improve cognition. The present disclosure further relates to methods for detecting amyloid plaques in patients undergoing treatment for amyloidogenic disease and methods for measuring the effectiveness of treatment. The present disclosure is based, at least in part, on the identification and characterization of monoclonal antibodies that specifically bind to Aβ peptide and are effective in reducing plaque burden and neutralizing soluble Aβ species associated with amyloidogenic disorders.

[0008] In a first aspect, the present disclosure provides a method of treating Alzheimer's disease in a subject in need thereof. In one aspect, the present disclosure provides a method of treating Alzheimer's disease in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid β antibody or antigen-binding fragment thereof about once every 3 to 5 weeks. In another aspect, the present disclosure provides a method of reducing amyloid plaques in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid β antibody or antigen-binding fragment thereof about once every 3 to 5 weeks. In another aspect, the present disclosure provides a method of converting a subject from amyloid-positive to amyloid-negative, the method comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid β antibody or antigen-binding fragment thereof about once every 3 to 5 weeks. The method comprises administering to the subject about 65 mg to about 200 mg of the antibody or antigen-binding fragment thereof about once every 3 to 5 weeks.

[0009] In one embodiment, the present disclosure provides a method for treating Alzheimer's disease in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid β antibody or antigen-binding fragment thereof about once every four weeks. In another embodiment, the present disclosure provides a method for reducing amyloid plaques in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid β antibody or antigen-binding fragment thereof about once every four weeks. In another embodiment, the present disclosure provides a method for converting a subject from amyloid-positive to amyloid-negative, the method comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid β antibody or antigen-binding fragment thereof about once every four weeks.

[0010] In some embodiments, the anti-amyloid β antibody or antigen-binding fragment thereof binds to an epitope located within the N-terminus of the Aβ peptide, wherein the epitope comprises at least one amino acid selected from amino acids 1 to 10 of the Aβ peptide. In some embodiments, the anti-amyloid β antibody or antigen-binding fragment thereof binds to an epitope comprising at least one amino acid selected from amino acids 1 to 7 of the Aβ peptide.

[0011] In some embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof binds to amyloid beta with an apparent KD of about 5 nM or less. 1-42 In some embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof binds to amyloid beta protofibrils with an apparent KD of about 1 nM or less. 1-42 In some embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof binds to amyloid beta protofibrils with an apparent KD of about 10 nM or less. 1-28 Binds to monomers.

[0012] In some embodiments, the method comprises administering about 20 mg to about 100 mg of an anti-amyloid β antibody or antigen-binding fragment thereof. In some embodiments, the method comprises administering about 100 mg to about 200 mg of an anti-amyloid β antibody or antigen-binding fragment thereof. In some embodiments, the method comprises administering about 45 mg of an anti-amyloid β antibody or antigen-binding fragment thereof. In some embodiments, the method comprises administering about 70 mg of an anti-amyloid β antibody or antigen-binding fragment thereof. In some embodiments, the method comprises administering about 200 mg of an anti-amyloid β antibody or antigen-binding fragment thereof. In some embodiments, the anti-amyloid β antibody is administered about once every four weeks.

[0013] In some embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof is administered as a pharmaceutical composition comprising the anti-amyloid beta antibody or antigen-binding fragment thereof and a pharmaceutically acceptable diluent. In some embodiments, the pharmaceutically effective amount of the anti-amyloid beta antibody or antigen-binding fragment thereof comprises about 45 mg. In some embodiments, the pharmaceutically effective amount of the anti-amyloid beta antibody or antigen-binding fragment thereof comprises about 70 mg. In some embodiments, the pharmaceutically effective amount of the anti-amyloid beta antibody or antigen-binding fragment thereof comprises about 200 mg. In some embodiments, administration is about once every four weeks.

[0014] In some embodiments, administration is intravenous or subcutaneous. In some embodiments, administration is subcutaneous.

[0015] In some embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3, and a light chain variable region comprising light chain CDR1, CDR2, and CDR3; heavy chain CDR1 comprises the amino acid sequence of one of SEQ ID NOs: 16, 19, or 20; the heavy chain CDR2 comprises the amino acid sequence of one of SEQ ID NOs: 20, 21, 22, or 23; the heavy chain CDR3 comprises the amino acid sequence of one of SEQ ID NOs: 18, 24, or 25; light chain CDR1 comprises the amino acid sequence of one of SEQ ID NOs: 26, 29, 31, or 32; the light chain CDR2 comprises the amino acid sequence of one of SEQ ID NOs: 33, 34, 35, or 36; the light chain CDR3 comprises the amino acid sequence of one of SEQ ID NOs: 28, 38, or 39.

[0016] In some embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises: heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 16; a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 20; a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 18; a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 29; a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 34, and A light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 38.

[0017] In some embodiments, the heavy chain variable region, excluding the CDRs, is at least 98% identical to the amino acid sequence of SEQ ID NO: 3, and the light chain variable region, excluding the CDRs, is at least 98% identical to the amino acid sequence of SEQ ID NO: 9. In one embodiment of the first aspect, the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 3, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 9.

[0018] In some embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises: heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 16; a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 20; a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 18; a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 29; a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 33, and A light chain CDR3 comprising the amino acid sequence of SEQ ID NO:28.

[0019] In some embodiments, the heavy chain variable region, excluding the CDRs, is at least 98% identical to the amino acid sequence of SEQ ID NO: 3, and the light chain variable region, excluding the CDRs, is at least 98% identical to the amino acid sequence of SEQ ID NO: 8. In some embodiments, the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 3, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 8.

[0020] In some embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises: heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 19; a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 21; a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 24; a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 29; a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 34, and A light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 38.

[0021] In some embodiments, the heavy chain variable region, excluding the CDRs, is at least 98% identical to the amino acid sequence of SEQ ID NO: 4, and the light chain variable region, excluding the CDRs, is at least 98% identical to the amino acid sequence of SEQ ID NO: 9. In some embodiments, the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 4, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 9.

[0022] In some embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises: heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 19; a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 21; a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 25; a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 29; a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 38;

[0023] In some embodiments, the heavy chain variable region, excluding the CDRs, is at least 98% identical to the amino acid sequence of SEQ ID NO: 5, and the light chain variable region, excluding the CDRs, is at least 98% identical to the amino acid sequence of SEQ ID NO: 9. In some embodiments, the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 5, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 9.

[0024] In some embodiments, the anti-amyloid beta antibody is a humanized IgG 1. In one embodiment of the first aspect, the anti-amyloid beta antibody is a whole antibody, a chimeric antibody, a CDR-grafted antibody, or a recombinant antibody.

[0025] In some embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof further comprises a heavy chain constant region comprising an amino acid sequence at least 95% identical to SEQ ID NO: 40 and / or a light chain constant region comprising an amino acid sequence at least 95% identical to SEQ ID NO: 41.

[0026] In some embodiments, the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without the C-terminal lysine, and a light chain of SEQ ID NO: 102. In some embodiments, the anti-amyloid beta antibody is h2731.

[0027] Thus, in various aspects, the present disclosure relates to methods utilizing antibodies or fragments thereof that specifically bind to Aβ peptides. In some embodiments, the antibodies and fragments comprise a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3, and a light chain variable region comprising light chain CDR1, CDR2, and CDR3, where the heavy chain CDR1, CDR2, and CDR3 and the light chain CDR1, CDR2, and CDR3 are set forth for one of the antibodies in Table 1. Furthermore, the antibodies or fragments of the present disclosure can have a heavy chain variable region set forth for one of the antibodies in Table 1 and a light chain variable region set forth for one of the antibodies in Table 1.

[0028] In another aspect, the present disclosure provides a method of treating Alzheimer's disease in a subject, the method comprising subcutaneously administering to the subject about 20 mg to about 200 mg of an anti-amyloid β antibody about once every four weeks, the anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102. In another aspect, the present disclosure provides a method of treating Alzheimer's disease in a subject, the method comprising subcutaneously administering to the subject about 45 mg of an anti-amyloid β antibody about once every four weeks, the anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102. In another aspect, the disclosure provides a method of treating Alzheimer's disease in a subject, the method comprising subcutaneously administering to the subject about 70 mg of an anti-amyloid beta antibody about once every four weeks, the anti-amyloid beta antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102. In another aspect, the disclosure provides a method of treating Alzheimer's disease in a subject, the method comprising subcutaneously administering to the subject about 200 mg of an anti-amyloid beta antibody about once every four weeks, the anti-amyloid beta antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102.

[0029] In another aspect, the present disclosure provides a method for reducing amyloid plaques in a subject, the method comprising subcutaneously administering to the subject about 20 mg to about 200 mg of an anti-amyloid β antibody about once every four weeks, the anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102. In another aspect, the present disclosure provides a method for reducing amyloid plaques in a subject, the method comprising subcutaneously administering to the subject about 45 mg of an anti-amyloid β antibody about once every four weeks, the anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102. In another aspect, the disclosure provides a method of reducing amyloid plaques in a subject, the method comprising subcutaneously administering to the subject about 70 mg of an anti-amyloid beta antibody about once every four weeks, the anti-amyloid beta antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102. In another aspect, the disclosure provides a method of reducing amyloid plaques in a subject, the method comprising subcutaneously administering to the subject about 200 mg of an anti-amyloid beta antibody about once every four weeks, the anti-amyloid beta antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102.

[0030] In another aspect, the disclosure provides a method of treating Alzheimer's disease in a subject, the method comprising administering to the subject a C ave The method comprises subcutaneously administering a dose of an anti-Aβ antibody sufficient to obtain a value, wherein the anti-amyloid β antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102.

[0031] In another aspect, the disclosure provides a method of treating Alzheimer's disease in a subject, the method comprising administering to the subject an AUC 0-tauThe method comprises subcutaneously administering a dose of an anti-Aβ antibody sufficient to obtain a value, wherein the anti-amyloid β antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102.

[0032] In some embodiments, the maximum concentration (C) of an anti-amyloid beta antibody or antigen-binding fragment thereof in a subject over an administration interval is max ) is about 30 μg / mL to about 60 μg / mL. In some embodiments, the C of the anti-amyloid β antibody or antigen-binding fragment thereof in the subject max In some embodiments, the C value of the anti-amyloid β antibody or antigen-binding fragment thereof in the subject is about 50 μg / mL to about 60 μg / mL. max In some embodiments, the C value does not exceed about 60 μg / mL. max The value is serum C max In some embodiments, C max The value is plasma C max value.

[0033] In some embodiments, the average concentration (C) of an anti-amyloid beta antibody or antigen-binding fragment thereof in a subject over an administration interval is ave In some embodiments, the C value of the anti-amyloid β antibody or antigen-binding fragment thereof in the subject is about 20 μg / mL to about 40 μg / mL. ave In some embodiments, the C value of the anti-amyloid β antibody or antigen-binding fragment thereof in the subject is about 30 μg / mL to about 40 μg / mL. ave In some embodiments, the C value does not exceed about 40 μg / mL. ave The value is serum C max In some embodiments, C ave The value is plasma C max value.

[0034] In some embodiments, the area under the concentration-time curve (AUC) is measured relative to the administration interval of an anti-amyloid beta antibody or antigen-binding fragment thereof in a subject. 0-tauIn some embodiments, the AUC of the anti-amyloid β antibody or antigen-binding fragment thereof in the subject is about 15,000 hr*ug / mL to about 30,000 hr*ug / mL. 0-tau In some embodiments, the AUC of the anti-amyloid β antibody or antigen-binding fragment thereof in the subject is about 20,000 hr*ug / mL to about 30,000 hr*ug / mL. 0-tau In some embodiments, the AUC 0-tau Values ​​are serum AUC 0-tau In some embodiments, the AUC 0-tau Values ​​are plasma AUC 0-tau value.

[0035] In some embodiments, the subject's amyloid plaques (i.e., cerebral amyloid beta plaques) are reduced. In some embodiments, the method further comprises reducing the subject's amyloid plaques.

[0036] In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of at least about 30 to about 70 centiloids. In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of about 45 to about 80 centiloids. In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of about 50 to about 85 centiloids.

[0037] In some embodiments, the reduction in cerebral amyloid beta plaques comprises at least about a 40% to about a 90% reduction. In some embodiments, the reduction in cerebral amyloid beta plaques comprises about a 60% to about a 100% reduction. In some embodiments, the reduction in cerebral amyloid beta plaques comprises about a 65% to about a 100% reduction.

[0038] In some embodiments, the reduction in cerebral amyloid plaques is a reduction compared to a baseline (e.g., a pre-treatment value). In some embodiments, the reduction in cerebral amyloid beta plaques is a reduction compared to the subject before administration of the anti-amyloid beta antibody.

[0039] In some embodiments, the reduction of cerebral amyloid beta plaques is achieved after 6 months of treatment. In some embodiments, the reduction of cerebral amyloid beta plaques is achieved after about 12 months of treatment. In some embodiments, the reduction of cerebral amyloid beta plaques is achieved after about 18 months of treatment.

[0040] In some embodiments, reduction of cerebral amyloid-beta plaques is assessed by positron emission tomography (PET).

[0041] In some embodiments, the subject is converted from amyloid positive to amyloid negative. In some embodiments, treatment comprises increasing the likelihood of converting a subject from amyloid positive to amyloid negative.

[0042] In some embodiments, the treatment comprises about a 10% to about 40% chance of converting the control from amyloid positive to amyloid negative. In some embodiments, the treatment comprises about a 30% to about 60% chance of converting the control from amyloid positive to amyloid negative. In some embodiments, the treatment comprises about a 40% to about 80% chance of converting the control from amyloid positive to amyloid negative.

[0043] In some embodiments, the likelihood of converting the control from amyloid positive to amyloid negative is about 6 months after treatment. In some embodiments, the likelihood of converting the control from amyloid positive to amyloid negative is about 12 months after treatment. In some embodiments, the likelihood of converting the control from amyloid positive to amyloid negative is about 18 months after treatment.

[0044] In some embodiments, treating comprises slowing, stopping, or reversing the decline in cognitive function. In some embodiments, treating comprises slowing the decline in cognitive function. In some embodiments, cognitive function is measured by at least one of the following: CRD-SB, ADAS-Cog14, ADCOMS, and ADCS MCI-ADL. In some embodiments, cognitive function is measured by ADCOMS.

[0045] In another aspect, the present disclosure provides a method for modulating a biomarker in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid β antibody about once every 3 to 5 weeks, the anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102. In another aspect, the present disclosure provides a method for increasing the Aβ42 / 40 ratio in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid β antibody about once every 3 to 5 weeks, the anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102. In another aspect, the present disclosure provides a method for reducing phospho-tau levels in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of the anti-amyloid beta antibody about once every 3 to 5 weeks, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102.

[0046] In some embodiments, the administration is intravenous or subcutaneous. In some embodiments, the administration is a subcutaneous injection. In some embodiments, the administration is a single subcutaneous injection.

[0047] In some embodiments, the biomarker in the subject is modulated. In some embodiments, the biomarker in the subject is modulated compared to baseline. In some embodiments, the method further comprises detecting the biomarker in a sample taken from the subject. In some embodiments, the method further comprises quantifying the biomarker in a sample taken from the subject.

[0048] In some embodiments, the biomarker comprises the ratio of Aβ42 / 40 in the subject. In some embodiments, the ratio of Aβ42 / 40 in the subject. In some embodiments, the ratio of Aβ42 / 40 in the subject is increased by at least 25%. In some embodiments, the ratio of Aβ42 / 40 in the subject is increased by at least 50%. In some embodiments, the ratio of Aβ42 / 40 in the subject is increased by about 25% to about 100%. In some embodiments, the ratio of Aβ42 / 40 in the subject is increased by about 50% to about 100%.

[0049] In some embodiments, the biomarker comprises a phospho-tau level. In some embodiments, the phospho-tau level comprises at least one of the following: a p181-tau level, a p212-tau level, a p217-tau level, a p231-tau level, and a p235-tau level. In some embodiments, the phospho-tau level comprises a p181-tau level. In some embodiments, the phospho-tau level comprises a p212-tau level. In some embodiments, the phospho-tau level comprises a p217-tau level. In some embodiments, the phospho-tau level comprises a p231-tau level. In some embodiments, the phospho-tau level comprises a p235-tau level. In some embodiments, the phospho-tau level is decreased. In some embodiments, the phospho-tau level is decreased by at least about 10%. In some embodiments, the phospho-tau level is decreased by about 10% to about 30%. In some embodiments, the phospho-tau level is decreased by at least about 20% to about 30%.

[0050] In some embodiments, the sample comprises blood or a portion thereof taken from the subject. In some embodiments, the sample comprises plasma taken from the subject. In some embodiments, the sample comprises serum taken from the subject. In some embodiments, the sample comprises cerebrospinal fluid ("CSF") taken from the subject.

[0051] In some embodiments, the method comprises a risk of ARIA-E of less than about 45%. In some embodiments, the method comprises a risk of ARIA-E of between about 25% and about 45%. In some embodiments, the method comprises a risk of ARIA-E of less than about 75%. In some embodiments, the method comprises a risk of ARIA-E of between about 50% and about 75%. In some embodiments, the method comprises a risk of symptomatic ARIA-E of less than about 15%. In some embodiments, the method comprises a risk of symptomatic ARIA-E of less than about 30%. In some embodiments, the risk of ARIA-E is a risk of severe ARIA-E. In some embodiments, the risk of ARIA-E is a risk after about 6 months of treatment. In some embodiments, the risk of ARIA-E is a risk after about 12 months of treatment. In some embodiments, the risk of ARIA-E is a risk after about 18 months of treatment. In some embodiments, the subject does not experience symptomatic ARIA-E during treatment.

[0052] In some embodiments, the method includes a risk of ARIA-H of less than about 35%. In some embodiments, the method includes a risk of ARIA-H of about 10% to about 35%. In some embodiments, the risk of ARIA-H is a risk of severe ARIA-H. In some embodiments, the risk of ARIA-H is a risk after about 6 months of treatment. In some embodiments, the subject does not experience symptomatic ARIA-H during treatment.

[0053] In some embodiments, ARIA is assessed by magnetic resonance imaging (“MRI”).

[0054] In some embodiments, the subject is an APOE4 homozygous subject. In some embodiments, the subject is an APOE4 heterozygous subject or an APOE4 non-carrier. In some embodiments, the method further comprises determining the subject's APOE4 status before administering.

[0055] In some embodiments, the duration of treatment is at least 6 months, in some embodiments, the duration of treatment is at least 12 months, in some embodiments, the duration of treatment is at least 18 months.

[0056] In some embodiments, administration is performed using a syringe, hi some embodiments, administration is performed using an autoinjector.

[0057] In some embodiments, the subject is a mammal, hi some embodiments, the subject is a human.

[0058] In various aspects, the present disclosure relates to pharmaceutical compositions comprising an anti-amyloid beta antibody or antigen-binding fragment described herein for treating Alzheimer's disease in a subject. In various aspects, the treatment comprises administering to the subject about 20 mg to about 200 mg of the antibody or antigen-binding fragment thereof approximately once every 3 to 5 weeks. In several aspects, the intermediate doses described herein can be used for subcutaneous administration. In several embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable excipient for administration, including, for example, subcutaneous administration.

[0059] In various aspects, the present disclosure relates to pharmaceutical compositions comprising an anti-amyloid beta antibody or antigen-binding fragment for reducing amyloid plaques in a subject. In various aspects, treating a subject involves administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof approximately once every 3 to 5 weeks. In several aspects, the intermediate doses described herein can be used for subcutaneous administration. In several embodiments, the pharmaceutical composition includes a pharmaceutically acceptable excipient for administration, including, for example, subcutaneous administration.

[0060] In various aspects, the present disclosure relates to pharmaceutical compositions comprising the anti-amyloid beta antibodies or antigen-binding fragments described herein for converting a subject from amyloid-positive to amyloid-negative. In various aspects, treating a subject involves administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof approximately once every 3 to 5 weeks. In several aspects, the intermediate doses described herein can be used for subcutaneous administration. In several embodiments, the pharmaceutical composition includes a pharmaceutically acceptable excipient for administration, including, for example, subcutaneous administration.

[0061] In various pharmaceutical composition embodiments, administration includes, for example, subcutaneously administering to a subject about 45 mg of an anti-amyloid beta antibody about once every four weeks, subcutaneously administering to a subject about 70 mg of an anti-amyloid beta antibody about once every four weeks, or subcutaneously administering to a subject about 200 mg of an anti-amyloid beta antibody about once every four weeks.

[0062] In various embodiments, the present disclosure relates to the use of an anti-amyloid beta antibody or antigen-binding fragment described herein for the manufacture of a medicament for treating Alzheimer's disease in a subject. In various embodiments, the medicament comprises administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks. In several embodiments, the intermediate doses described herein can be used for subcutaneous administration.

[0063] In various embodiments, the present disclosure relates to the use of an anti-amyloid beta antibody or antigen-binding fragment described herein for the manufacture of a medicament for reducing amyloid plaques in a subject. In various embodiments, the medicament is for administering to a subject about 20 mg to about 200 mg of the anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks. In several embodiments, the intermediate doses described herein can be used for subcutaneous administration.

[0064] In various embodiments, the present disclosure relates to the use of an anti-amyloid beta antibody or antigen-binding fragment described herein for the manufacture of a medicament for converting a subject from amyloid-positive to amyloid-negative. In various embodiments, the medicament is for administering to the subject about 20 mg to about 200 mg of the anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks. In several embodiments, the intermediate doses described herein can be used for subcutaneous administration.

[0065] In several embodiments of the use of an anti-amyloid beta antibody or antigen-binding fragment thereof, administration includes, for example, subcutaneously administering to a subject about 45 mg of an anti-amyloid beta antibody or binding fragment about once every four weeks, subcutaneously administering to a subject about 70 mg of an anti-amyloid beta antibody or binding fragment about once every four weeks, or subcutaneously administering to a subject about 200 mg of an anti-amyloid beta antibody or binding fragment about once every four weeks. [Brief explanation of the drawings]

[0066] [Figure 1]

[0023] Figure 1 shows an alignment of three different versions of VL designed by incorporating human germline framework residues into the bapineuzumab (hBP) VL sequence. No canonical or interface residues were changed.

[0067] [Figure 2] 1 shows a graph of a competitive ELISA assay for 4918, 4917, 4921, 3818, 49 human 3, 2931 and a bapineuzumab control to determine the IC50 ratio relative to bapineuzumab (hBP).

[0068] [Figure 3] 1 shows a graph of a competitive ELISA assay for 2926, 2831, 2927, 2726, 2731, 2826, and a bapineuzumab control to determine the IC50 ratio relative to bapineuzumab (hBP).

[0069] [Figure 4] 1 shows a graph of a competitive ELISA assay for 2727, 2931 and a bapineuzumab control to determine the IC50 ratio relative to bapineuzumab (hBP).

[0070] [Figure 5] [Figure 5A] Graphs of competitive ELISA assays for 2931, 2731, and bapineuzumab. [Figure 5B] Graphs of competitive ELISA assays for 2726, 2831, and bapineuzumab (B).

[0071] [Figure 6A] BIAcore sensorgrams of binding of h2726 (A) to Aβ1-28 at analyte concentrations from 100 nM to 0.39 nM (2-fold serial dilutions) are shown. [Figure 6B] BIAcore sensorgrams of binding of h2731 (B) to Aβ1-28 at analyte concentrations from 100 nM to 0.39 nM (2-fold serial dilutions) are shown. [Figure 6C] BIAcore sensorgrams of binding of h2831(C) to Aβ1-28 at analyte concentrations from 100 nM to 0.39 nM (2-fold serial dilutions) are shown. [Figure 6D] BIAcore sensorgrams of the binding of 2931(D) to Aβ1-28 at analyte concentrations from 100 nM to 0.39 nM (2-fold serial dilutions) are shown.

[0072] [Figure 7]Figure 1 shows BIAcore sensorgrams comparing the binding properties of humanized antibodies (PB-0569 (aducanumab), PB-0573 (h2726), PB-0574 (h2731), PB-0575 (h2831), PB-0576 (h2931)) to recombinant Abeta 1-42 (Aβ1-42) fibrils.

[0073] [Figure 8] 1 shows that h2931 binds to soluble Aβ oligomers with high relative affinity.

[0074] [Figure 9] Graphs evaluating the Aβ fibril binding activity of 2726, 2731, 2831, and 2931 relative to the aducanumab control are shown. The antibodies were titrated in a fixed concentration of Aβ fibrils (left panel), or Aβ fibrils were titrated in a fixed concentration of antibody (right panel). Both showed substantially better binding to 2726, 2731, 2831, and 2931 than aducanumab.

[0075] [Figure 10] Figure 1 shows Aβ binding in AD brains. Binding to tissue Aβ pathology appears to be similar among the h2726, h2731, h2831, and h2931 antibodies. Example images stained with 0.3 μg / ml of the four antibodies h2726, h2731, h2831, and h2931 demonstrate the pattern of staining in two AD brains with different amounts of Aβ pathology (AD11-97 and AD13-75). For each brain, images are from the same region of the section, showing relatively similar intensity and distribution of lesions for all four antibodies. Staining with aducanumab was consistently the weakest (scale bar: 500 μm).

[0076] [Figure 11] 1 shows Aβ binding in control AD ​​brains. A human IgG isotype control antibody did not produce staining in AD brains. As shown in these examples, AD sections incubated with 1 μg / ml of human IgG isotype did not have any staining (scale bar: 500 μm).

[0077] [Figure 12] Quantification of Aβ binding in AD brains is shown. Quantification of Aβ pathology staining in AD tissues revealed similar binding among the h2726, h2731, h2831, and h2931 antibodies. Sections from four AD brains were incubated with antibodies h2726, h2731, h2831, and h2931, as well as aducanumab at the following concentrations: 0.03, 0.1, 0.3, 1, 3, and 9 μg / ml. After imaging the sections, the percentage of stained tissue area was determined morphometrically using Halo® imaging analysis software. Each graph compares measurements in AD brains obtained with five antibodies. The four graphs consistently demonstrate similar binding profiles for the h2726, h2731, h2831, and h2931 antibodies. Measurements obtained with aducanumab were significantly lower.

[0078] [Figure 13] Figure 1 shows Aβ binding in AD brain. hBP binds strongly and dose-dependently to tissue Aβ lesions. Images are from the same region of a section (brain AD13-75) with similar lesion distribution. hBP showed increasing staining with concentration, and its binding to Aβ lesions was higher than that of BAN2401 or aducanumab at each concentration (scale bar: 500 μm).

[0079] [Figure 14] [Figure 14A] Individual results from ex vivo phagocytosis studies of h2931 and aducanumab in APP.PS1 Tg mouse tissue with primary mouse microglia are shown. Both h2931 and aducanumab demonstrate highly significant reductions in Aβ1-42 relative to the isotype control. [Figure 14B] Pooled results from ex vivo phagocytosis studies of h2931 and aducanumab in APP.PS1 Tg mouse tissue with primary mouse microglia are shown. Both h2931 and aducanumab demonstrate highly significant reductions in Aβ1-42 relative to the isotype control.

[0080] [Figure 15] [Figure 15A] Graph showing reduced soluble oligomer binding to neurites of rat hippocampal neurons treated with increasing concentrations of h2726, h2731, h2831, and h2931 compared to isotype control, normalized by + / -Aβ addition. Spots per neuron are shown (40 fields per well). [Figure 15B] Graph showing reduced soluble oligomer binding to neurites of rat hippocampal neurons treated with increasing concentrations of h2726, h2731, h2831, and h2931 compared to isotype control, normalized by + / -Aβ addition. Total spot counts are shown (40 fields per well).

[0081] [Figure 16] Graphs depicting the percentage of Aβ spots per neuron with increasing concentrations of 2726, 2731, 2831 and 2931, normalized to the addition of + / - Aβ, are shown.

[0082] [Figure 17] 1 shows an alignment of the bapineuzumab variable heavy chain sequence with four sequences of the disclosure: 2726, 2731, 2831, and 2931. The CDRs are shown in bold.

[0083] [Figure 18] 1 shows the alignment of the bapineuzumab light chain sequence with four (variable light chain) sequences of the disclosure: 2726, 2731, 2831, and 2931. The CDRs are shown in bold.

[0084] [Figure 19A] Figure 1 shows a CDR table listing the variable heavy and light chain CDR sequences for the antibodies of this disclosure. Heavy chain CDRs are indicated. [Figure 19B] Figure 1 shows a CDR table listing the variable heavy and light chain CDR sequences for the antibodies of this disclosure. Light chain CDRs are indicated.

[0085] [Figure 20][Figure 20A] Graph showing the potency of antibodies for binding to heterogeneously aggregated Aβ42 species by competitive ELISA. Shown are h2931, h2731, and a bapineuzumab control. [Figure 20B] Graph showing the potency of antibodies for binding to heterogeneously aggregated Aβ42 species by competitive ELISA. Shown are h2831, h2726, and a bapineuzumab control.

[0086] [Figure 21] 1 shows a graph measuring the direct binding and relative affinity of antibodies to fibrillar Aβ42 by ELISA.

[0087] [Figure 22] 1 shows a graph measuring antibody dose response of Aβ plaque area binding measured as percent positive tissue by immunohistochemical staining in AD brains.

[0088] [Figure 23] Quantification of soluble Aβ binding to rat hippocampal neurons in the presence of antibodies.

[0089] [Figure 24] Figure 1 shows the results of an ex vivo phagocytosis study of h2731 in AD tissue using primary mouse microglia. h2731 showed highly significant reduction of Aβ1-42, indicating that the antibody robustly promoted phagocytosis and clearance of these species.

[0090] [Figure 25] [Figure 25A] Confirms the presence of pyroglutamic acid 3Aβ (AβpE3-42) in AD tissue used in ex vivo phagocytosis assays (A) and shows similar binding patterns for pyroglutamic acid 3Aβ and h2931 (A and B). [Figure 25B] Confirms the presence of pyroglutamic acid 3Aβ (AβpE3-42) in AD tissue used in ex vivo phagocytosis assays (A) and shows similar binding patterns for pyroglutamic acid 3Aβ and h2931 (A and B).

[0091] [Figure 26] [Figure 26A] Ex vivo phagocytosis studies of h2931 and h2731 in AD tissue using primary mouse microglia are shown. Both h2931 and h2731 showed highly significant reductions in pyroglutamate 3Aβ (AβpE3-42), indicating that both antibodies robustly promoted phagocytosis and clearance of these species. [Figure 26B] Ex vivo phagocytosis studies of h2931 and h2731 in AD tissue using primary mouse microglia are shown. Both h2931 and h2731 showed highly significant reductions in pyroglutamate 3Aβ (AβpE3-42), indicating that both antibodies robustly promoted phagocytosis and clearance of these species.

[0092] [Figure 27] This shows that h2731 binds to the N-terminus of Aβ1-42 but not to AβpE3-42.

[0093] [Figure 28] [Figure 28A] Antibodies of the present invention induce phagocytosis of Aβ1-42 protofibrils in THP-1 human monocytes in vitro. [Figure 28B] Antibodies of the present invention induce phagocytosis of Aβ1-42 protofibrils in THP-1 human monocytes in vitro.

[0094] [Figure 29A] 1 shows the distribution pattern of Aβ1-XX measured by N-terminal anti-Ab antibody compared with AβpE3-42 in human AD brain tissue. [Figure 29B] 1 shows the distribution pattern of Aβ1-XX measured by N-terminal anti-Ab antibody compared with AβpE3-42 in human AD brain tissue. [Figure 29C] Quantification of the percent area covered by Aβ1-XX compared to AβpE3-42 in human AD brain tissue is shown.

[0095] [Figure 30]Localization of h2731 to Aβ plaques, localization of anti-AβpE3-42 antibody signals to Aβ plaques, and co-localization of h2731 and anti-AβpE3-42 antibody signals to Aβ plaques are shown.

[0096] [Figure 31] [Figure 31A] Anti-Aβ antibody h2731 promotes AβpE3-42 clearance from AD brain tissue ex vivo in a dose-dependent manner with greater potency than aducanumab. [Figure 31B] Anti-Aβ antibody h2731 promotes AβpE3-42 clearance from AD brain tissue ex vivo in a dose-dependent manner with greater potency than aducanumab.

[0097] [Figure 32] [Figure 32A] Concentration dependence of h2731 and aducanumab clearance of AβpE3-42 from AD brain tissue. [Figure 32B] The effect of h2731 is microglia-dependent.

[0098] [Figure 33] The predicted CNS exposure of h2731 and aducanumab will be compared with repeated doses.

[0099] [Figure 34] 1 shows that anti-Aβ antibody h2731 promotes clearance of Aβp E3-42-containing plaques in ex vivo AD brain tissue.

[0100] [Figure 35] 1 is a schematic diagram of the clinical trial design for a single ascending dose study of h2731 in healthy volunteers and subjects with Alzheimer's disease, showing certain inclusion criteria, dosages, and certain evaluation schedules.

[0101] [Figure 36] 1 is a schematic diagram of the clinical trial design for a multiple ascending dose study of h2731 in subjects with Alzheimer's disease, showing certain inclusion criteria, dosages, and certain evaluation schedules.

[0102] [Figure 37]

[0023] Figure 1 shows details of an open-label extension study of h2731 in some Alzheimer's disease subjects enrolled in either the single ascending dose study of Example 19 or the multiple ascending dose study of Example 20. Certain dosages and certain evaluation schedules are shown. DETAILED DESCRIPTION OF THE INVENTION

[0103] Monoclonal antibodies (mAbs) targeting the N-terminus of amyloid beta (Aβ) have been clinically demonstrated to reduce amyloid plaque burden, and one such antibody, aducanumab, has shown that significant reductions in plaque burden are associated with slowed cognitive decline in Alzheimer's disease (AD). Preclinical studies have also shown that monoclonal antibodies (mAbs) targeting N-terminal epitopes of Aβ induce antibody-dependent microglia-mediated Aβ plaque clearance and neutralization of soluble toxic Aβ oligomers both in vitro and in vivo. It has been hypothesized that administration of mAbs targeting N-terminal Aβ may slow disease progression in AD patients through Aβ plaque clearance and neutralization of soluble Aβ aggregation.

[0104] The Aβ antibody bapineuzumab (hBP) is a humanized antibody developed from the parent murine antibody 3D6. According to various aspects of the present disclosure, a multifaceted approach was applied to construct a superior antibody against hBP. The human nature of hBP was analyzed and it was determined that humanization of the light chain could be optimized.

[0105] Protein sequences in the PDB database [Deshpande et al., 2005] were searched to find structures that could provide a rough structural model for hBP. The crystal structure of hBP fab PDB code 4HIX [Miles, et al., 2013] was utilized for both the Vh and Vk structures because it had acceptable resolution and an exact sequence match with hBP Vh and Vk while retaining the same canonical structure of the loops.

[0106] IMGT / DomainGapAlignment was performed on the hBP VL as the input sequence, and the human germline VK gene sequence IGHV2-30*02 was identified as the closest to the hBP VL. The framework of hBP VL shares a high degree of sequence similarity with the corresponding framework region of IGHV2-30*02. Therefore, the framework region of IGHV2-30*02 VL was selected as a guidance sequence for further optimization of the hBP framework region. Additional residues in CDR-L2 that do not directly contact the antigen according to the hBP 3D structure were also changed to the germline sequence, resulting in the following changes:

[0107] Three different versions of the VL were designed by incorporating human germline framework residues into the hBP VL sequence. Canonical or interface residues were not changed. P15L was also tested in one version of the variable light chain, based on the structural observation that P15 is located at a turn and the germline gene has a Leu at this position.

[0108] Based on observations of the 3D structure, substitutions were designed at several residues in the CDRs and frameworks of the light and heavy chains. Mutant VL and VH versions were generated and tested for binding in the first round of rational design. Mutations that showed improved binding were combined in a second round of rational design. Additionally, new mutations guided by further analysis of the structure were also incorporated into the design.

[0109] Accordingly, the present disclosure provides antibodies (and antibody fragments), nucleic acids encoding such antibodies and antibody fragments, and methods of making such antibodies and antibody fragments, pharmaceutical compositions, and methods of preventing or treating amyloidogenic diseases, reducing the risk of or delaying the onset of amyloidogenic diseases, improving cognition in subjects with conditions associated with amyloidogenic diseases, inhibiting the formation of Aβ plaques in a subject, reducing Aβ plaques in the brain of a subject, inhibiting or reducing amyloid plaques in subjects at risk of developing amyloidogenic diseases, detecting amyloid plaques, and measuring the effectiveness of treatment in subjects being treated for amyloidogenic diseases, including Alzheimer's disease, etc., as described herein. The present disclosure is based, at least in part, on the characterization of a genus of monoclonal antibodies that are effective in binding to beta amyloid protein (Aβ) (e.g., binding soluble and / or aggregated Aβ), mediating phagocytosis (e.g., of aggregated Aβ), reducing plaque burden and / or reducing neurofibrotic dystrophy (e.g., in a patient), and neutralizing soluble toxic Aβ species. The antibodies and fragments of the present disclosure exhibit higher binding strength (affinity and / or avidity) for pathological fibrillar Aβ than current reported experimental therapies, as well as higher affinity for soluble toxic Aβ forms. These antibodies may enable more convenient administration strategies and improve patient access.

[0110] Before describing certain aspects of the present disclosure in more detail, some terms will be defined.

[0111] definition The term "antibody" includes intact antibodies and binding fragments thereof. Typically, fragments compete with the resulting intact antibody for specific binding to a target. Fragments include separate heavy and light chains, Fab, Fab', F(ab')2, F(ab)c, Fv, and single-domain antibodies. Single (variable) domain antibodies include VH regions separated from their VL partners (or vice versa) in conventional antibodies (Ward et al., 1989, Nature 341:544-546), as well as VH regions (sometimes known as VHHs) from species, such as camelids or cartilaginous fish (e.g., nurse sharks), in which the VH region is not associated with the VL region (see, e.g., WO9404678). Single-domain antibodies in which one chain is separated from its natural partner are sometimes known as Dabs, and single-domain antibodies from camelids or cartilaginous fish are sometimes known as nanobodies. Constant regions or portions of constant regions may or may not be present in single domain antibodies. For example, natural single variable region antibodies from camelids contain VHH variable regions and CH2 and CH3 constant regions. Single domain antibodies can be subject to humanization using approaches similar to conventional antibodies. Dab-type antibodies are typically derived from antibodies of human origin. Nanobody-type antibodies are of camelid or shark origin and can be subject to humanization. Fragments can be generated by recombinant DNA techniques or by enzymatic or chemical separation of intact immunoglobulins. The term "antibody" also includes bispecific antibodies. Bispecific or bifunctional antibodies are artificial hybrid antibodies having two different heavy / light chain pairs and two different binding sites (see, e.g., Songsivilai and Lachmann, Clin. Exp. Immunol., 79:315-321 (1990); Kostelny et al., J. Immunol., 148:1547-53 (1992)).

[0112] The variable region of an immunoglobulin light or heavy chain (sometimes referred to herein as a "light chain variable domain" ("VL domain") or "heavy chain variable domain" ("VH domain"), respectively) consists of three "complementarity-determining regions" or "CDRs" interrupted by three. The framework regions function to align the CDRs for specific binding to an epitope of an antigen. The CDRs contain the amino acid residues of an antibody primarily responsible for antigen binding. From the amino to carboxyl terminus, both VL and VH domains comprise the following framework (FR) and CDR regions: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. CDR1, 2, and 3 of the VL domain are sometimes referred to herein as CDR-L1, CDR-L2, and CDR-L3, respectively. CDR1, 2, and 3 of the VH domain are sometimes referred to herein as CDR-H1, CDR-H2, and CDR-H3, respectively. When the present application discloses a VL sequence having R as the C-terminal residue, R can alternatively be considered to be the N-terminal residue of the light chain constant region, and therefore, the present application should also be understood to disclose a VL sequence that does not have a C-terminal R.

[0113] The assignment of amino acids to each VL and VH domain follows conventional definitions of CDRs, including those of Kabat (Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, MD, 1987 and 1991)), Chothia (Chothia & Lesk, J. Mol. Biol. 196:901-917, 1987; Chothia et al., Nature 342:878-883, 1989), the Chothia-Kabat CDR complex in which CDR-H1 is a complex of the Chothia and Kabat CDRs, the AbM definition used by Oxford Molecular's antibody modeling software, and the contact definition of Martin et al. (bioinfo.org.uk / abs) (see Table A). Kabat provides a widely used numbering system (Kabat numbering) that assigns the same number to corresponding residues between different heavy chains or different light chains. When an antibody is said to contain a CDR according to a particular definition of CDR (e.g., Kabat), that definition specifies the minimum number of CDR residues present in the antibody (i.e., Kabat CDRs). This does not exclude the presence of other residues that fall within other conventional CDR definitions but are outside the specified definition. For example, antibodies containing CDRs defined by Kabat include, among other possibilities, antibodies in which the CDRs contain Kabat CDR residues but no other CDR residues, and antibodies in which the CDR H1 is a composite Chothia-Kabat CDR H1 and the other CDRs contain Kabat CDR residues but no additional CDR residues according to other definitions. [Table A]

[0114] In some embodiments, the CDRs of the humanized antibodies of the invention are of a definition selected from the group consisting of Kabat, Chothia, Kabat / Chothia complex, AbM, and Contact.

[0115] One or several amino acids at the amino or carboxy termini of the light and / or heavy chains, such as the C-terminal lysine of the heavy chain, can be deleted or derivatized in some or all of the molecules. Substitutions in the constant region can be made to reduce or increase effector functions such as complement-mediated cytotoxicity or ADCC (e.g., Winter et al., U.S. Pat. No. 5,624,821; Tso et al., U.S. Pat. No. 5,834,597; and Lazar et al., Proc. Natl. Acad. Sci. USA 103:4005, 2006) or to extend half-life in humans (see, e.g., Hinton et al., J. Biol. Chem. 279:6213, 2004). Exemplary substitutions include Gln at position 250 and / or Leu at position 428 (EU numbering is used for the constant region in this paragraph) to extend antibody half-life. Substitutions at any or all of positions 234, 235, 236, and / or 237 reduce affinity for Fcγ receptors, particularly FcγRI receptors (see, e.g., US Pat. No. 6,624,821 ). Alanine substitutions at positions 234, 235, and 237 of human IgG1 can be used to reduce effector function. Some antibodies have alanine substitutions at positions 234, 235, and 237 of human IgG1 to reduce effector function. Optionally, positions 234, 236, and / or 237 of human IgG2 are substituted with alanine, and position 235 is substituted with glutamine (see, e.g., US Pat. No. 5,624,821 ). Some antibodies utilize mutations at one or more of positions 241, 264, 265, 270, 296, 297, 322, 329, and 331 of human IgG1 according to EU numbering. Some antibodies utilize mutations at one or more of positions 318, 320, and 322 according to EU numbering of human IgG1. Some antibodies utilize substitutions at positions 234 and / or 235 with alanine and / or 329 with glycine. Some antibodies utilize substitutions at positions 234 and 235 with alanine. Some antibodies are of the isotype human IgG2 or IgG4.As an example, the C-terminal lysine of the antibody heavy chain constant regions described herein is optional, such that the sequence can be considered with or without the C-terminal lysine.

[0116] The term "humanized immunoglobulin" or "humanized antibody" refers to an immunoglobulin or antibody comprising at least one humanized immunoglobulin or antibody chain (i.e., at least one humanized light or heavy chain). The term "humanized immunoglobulin chain" or "humanized antibody chain" (i.e., "humanized immunoglobulin light chain" or "humanized immunoglobulin heavy chain") refers to an immunoglobulin or antibody chain (i.e., light chain or heavy chain, respectively) having a variable region comprising variable framework regions substantially derived from a human immunoglobulin or antibody and complementarity-determining regions (CDRs) substantially derived from a non-human immunoglobulin or antibody (e.g., at least one CDR, preferably two CDRs, more preferably three CDRs), and further comprising a constant region (e.g., at least one constant region or portion thereof in the case of a light chain, and preferably three constant regions in the case of a heavy chain). The term "humanized variable region" (e.g., "humanized light chain variable region" or "humanized heavy chain variable region") refers to a variable region comprising a variable framework region substantially derived from a human immunoglobulin or antibody and a complementarity determining region (CDR) substantially derived from a non-human immunoglobulin or antibody. As used herein, "excluding the CDRs" refers to a portion of an antibody that does not include the amino acids of the CDRs, e.g., framework regions and antibody constant regions.

[0117] Thus, regions or residues of a humanized immunoglobulin or antibody, or regions or residues of a humanized immunoglobulin or antibody chain, are substantially identical to corresponding regions or residues of one or more naturally occurring human immunoglobulin sequences, possibly excluding the CDRs. The term "corresponding region" or "corresponding residue" refers to a region or residue on a second amino acid or nucleotide sequence that occupies the same (i.e., equivalent) position as a region or residue on a first amino acid or nucleotide sequence, when the first and second sequences are optimally aligned for comparison purposes.

[0118] The term "epitope" or "antigenic determinant" refers to a site on an antigen to which an antibody binds. Epitopes can be formed from adjacent amino acids or nonadjacent amino acids juxtaposed by one or more tertiary folding of a protein. Epitopes formed from adjacent amino acids are typically retained upon exposure to denaturing solvents, whereas epitopes formed by tertiary folding are typically lost upon treatment with denaturing solvents. An epitope typically contains at least three, more usually at least five, or 8-10 amino acids in a unique spatial conformation. When an epitope is said to be within a range of amino acid residues of a protein (e.g., within residues 1-6 of Aβ), the range includes the residues that define its boundaries. Certain residues within this range may contribute to the epitope, while others may not. The residues that form an epitope may or may not be adjacent to each other. Similarly, when an antibody binds to an epitope found within a particular range of amino acids, the antibody need not contact every amino acid residue within that range, and the residues of the epitope contacted by the antibody may or may not be adjacent to each other. Methods for determining the spatial conformation of epitopes include, for example, X-ray crystallography and two-dimensional nuclear magnetic resonance. See, e.g., Epitope Mapping Protocols, in Methods in Molecular Biology, Vol. 66, Glenn E. Morris, Ed. (1996).

[0119] Antibodies that recognize the same epitope can be identified in a simple immunoassay, i.e., a competitive binding assay, which shows the ability of one antibody to block or compete with the binding of another antibody to a target antigen. Competitive binding is determined by an assay in which the immunoglobulin under test inhibits the specific binding of a reference antibody to a common antigen, such as Aβ. There are many types of competitive binding assays, such as solid-phase direct or indirect radioimmunoassays (RIA), solid-phase direct or indirect enzyme immunoassays (EIA), sandwich competition assays (Stahli et al., Methods in Enzymology 9:242 (1983)); solid-phase direct biotin-avidin EIA (see Kirkland et al., J. Immunol. 137:3614 (1986)), solid-phase direct label assays, solid-phase direct label sandwich assays (see Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Press (1988)), solid-phase direct label RIA using I-125 labels (see Morel et al., Mol. Immunol. 25(1):7 (1988)), solid-phase direct biotin-avidin EIA (Cheung et al., Virology 176:546 (1990)), and direct label RIA. (Moldenhauer et al., Scand. J. Immunol. 32:77 (1990)) is known. Generally, such assays involve the use of purified antigen bound to a solid surface or cells bearing either an unlabeled test immunoglobulin or a labeled reference immunoglobulin. Competitive inhibition is measured by determining the amount of label bound to the solid surface or cells in the presence of the test immunoglobulin. Typically, the test immunoglobulin is present in excess. Typically, the presence of an excess of competing antibody inhibits specific binding of the reference antibody to the common antigen by at least 50-55%, 55-60%, 60-65%, 65-70%, 70-75%, or more.

[0120] Competition between antibodies is determined by an assay in which the antibody being tested inhibits the specific binding of a reference antibody (e.g., 3D6, aducanumab, bapineuzumab) to a common antigen (see, e.g., Junghans et al., Cancer Res. 50:1495, 1990). A test antibody competes with a reference antibody when an excess of the test antibody (e.g., at least 2-fold, 5-fold, 10-fold, 20-fold, or 100-fold) inhibits binding of the reference antibody by at least 50%, preferably 75%, 90%, or 99%, as measured in a competitive binding assay. Antibodies identified by competitive assays (competing antibodies) include antibodies that bind to the same epitope as the reference antibody and antibodies that bind to an adjacent epitope sufficiently close to the epitope bound by the reference antibody to create steric hindrance.

[0121] An antibody epitope can also be defined by X-ray crystallography of the antibody bound to its antigen to identify contact residues. Alternatively, two antibodies have the same epitope if all amino acid mutations in the antigen that reduce or eliminate binding of one antibody also reduce or eliminate binding of the other. Two antibodies have overlapping epitopes if some amino acid mutations that reduce or eliminate binding of one antibody also reduce or eliminate binding of the other.

[0122] Epitopes are also recognized by immune cells, such as B cells and / or T cells. Cellular recognition of epitopes is 3 It can be determined by in vitro assays that measure antigen-dependent proliferation as judged by H-thymidine incorporation, by cytokine secretion, by antibody secretion, or by antigen-dependent killing (cytotoxic T lymphocyte assay).

[0123] Exemplary epitopes or antigenic determinants may be found within the human amyloid precursor protein (APP), preferably within the Aβ peptide of APP. APP has multiple isoforms, e.g., APP 695 ,APP 751 , and APP 770 The amino acids in APP are770 Numbers are assigned according to the sequence of the isoform (see, eg, GenBank Accession No. P05067, also set forth as SEQ ID NO: 85).

[0124] Aβ (also referred to herein as beta amyloid peptide and A-beta) peptides are approximately 4 kDa internal fragments of APP consisting of 39-43 amino acids (Aβ39, Aβ40, Aβ41, Aβ42, and Aβ43). For example, Aβ40 consists of residues 672-711 of APP, and Aβ42 consists of residues 673-713 of APP. As a result of proteolytic processing of APP by different secretases in vivo or in situ, Aβ is found in both a 40 amino acid long "short form" and a "long form" ranging from 42-43 amino acids in length. Preferred epitopes or antigenic determinants described herein are located within the N-terminus of the Aβ peptide and include residues within amino acids 1-10 of Aβ, preferably residues 1-3, 1-4, 1-5, 1-6, 1-7, or 3-7 of Aβ42. Additional mentioned epitopes or antigenic determinants include residues 2-4, 5, 6, 7, or 8 of Aβ, residues 3-5, 6, 7, 8, or 9 of Aβ, or residues 4-7, 8, 9, or 10 of Aβ42.

[0125] "Soluble" or "dissociated" Aβ refers to Aβ species that are monomeric, aggregated, or oligomeric, with or without association with other proteins and lipids, and remain in solution (supernatant) after centrifugation at 100,000 × g. "Insoluble" Aβ refers to aggregated Aβ species, with or without amyloid (beta sheets), that do not remain in solution after centrifugation at 100,000 × g, e.g., Aβ held together by non-covalent bonds. Aβ (e.g., Aβ42) is believed to aggregate, at least in part, due to the presence of hydrophobic residues at the C-terminus of the peptide (part of the transmembrane domain of APP). One method for preparing soluble Aβ is to dissolve lyophilized peptide in solvent-free DMSO with sonication. The resulting solution is centrifuged to remove insoluble particulates.

[0126] "Specific binding" of an antibody means that the antibody exhibits appreciable affinity for the antigen or preferred epitope and preferably does not exhibit significant cross-reactivity. "Appreciable" or preferred binding requires at least 10 6 , 10 7 , 10 8 , 10 9 M -1 , or 10 10 M -1 This includes binding with an affinity of 10 7 M -1 Greater affinity, preferably 10 8 M -1 Greater affinities are more preferred. Values ​​intermediate to the values ​​recited herein are also intended to be within the scope of the present disclosure, and preferred binding affinities are within a range of affinities, e.g., 10 6 ~10 10 M -1 , preferably 10 7 ~10 10 M -1 , more preferably 10 8 ~10 10 M -1 An antibody that "does not exhibit significant cross-reactivity" is one that does not appreciably bind to undesired entities (e.g., undesired proteinaceous entities). For example, an antibody that specifically binds to Aβ appreciably binds to Aβ but does not significantly react with non-Aβ proteins or peptides (e.g., non-Aβ proteins or peptides contained in plaques). An antibody specific for a preferred epitope does not significantly cross-react with, for example, distant epitopes on the same protein or peptide. Specific binding can be determined according to any art-recognized means for determining such binding. Preferably, specific binding is determined according to Scatchard analysis and / or competitive binding assays.

[0127] Binding fragments can be produced by recombinant DNA techniques, or by enzymatic or chemical separation of intact immunoglobulins. Binding fragments include Fab, Fab', F(ab')2, Fabc, Fv, single chain, and single chain antibodies.

[0128] The term "patient" includes human and other mammalian subjects receiving either prophylactic or therapeutic treatment. In some embodiments, the term "individual" is used interchangeably with "patient."

[0129] The term "effective dose" or "effective administration amount" is defined as an amount sufficient to achieve, or at least partially achieve, the desired effect. The term "therapeutically effective dose" is defined as an amount sufficient to cure or at least partially arrest the disease and its complications in patients already suffering from the disease. Amounts effective for this use will depend on the severity of the infection and the general state of the patient's own immune system.

[0130] The term "treatment" as used herein is defined as the application or administration of a therapeutic agent to a patient having a disease, a symptom of a disease, or a predisposition to a disease, or to an isolated tissue or cell line from said patient, with the intent to cure, heal, alleviate, mitigate, alter, relieve, ameliorate, improve, or affect the disease, symptom of a disease, or predisposition to a disease.

[0131] The term "amyloidogenic disease" includes any disease associated with (or caused by) the formation or deposition of insoluble amyloid fibrils or amyloid plaques. Exemplary amyloidogenic diseases include, but are not limited to, systemic amyloidosis, Alzheimer's disease, maturity-onset diabetes of the young, Parkinson's disease, Huntington's disease, frontotemporal dementia, Down's syndrome, mild cognitive impairment, prion-associated transmissible spongiform encephalopathies (kuru and Creutzfeldt-Jakob disease in humans, and scrapie and BSE in sheep and cattle, respectively), and the like. Different amyloidogenic diseases are defined or characterized by the nature of the polypeptide component of the deposited fibrils. For example, in subjects or patients with Alzheimer's disease, beta-amyloid protein (e.g., wild-type, variant, or truncated beta-amyloid protein) is the characteristic polypeptide component of amyloid deposits. Thus, Alzheimer's disease, for example, is an example of a "disease characterized by or associated with the deposition of Aβ" in the brain of a subject or patient. The terms "β-amyloid protein," "β-amyloid peptide," "β-amyloid," "Aβ," and "Aβ peptide" are used interchangeably herein.

[0132] A subject is at increased risk of a disease if they have at least one known risk factor (e.g., genetic, biochemical, familial, situational exposure) that places the individual with the risk factor at a statistically significantly higher risk of developing the disease than an individual without the risk factor.

[0133] The term "symptom" refers to subjective evidence of disease, such as a change in gait, as perceived by the patient. "Sign" refers to objective evidence of disease as observed by a physician.

[0134] Statistical significance means p<0.05.

[0135] "Half-life (t1 / 2)" refers to the time required for the concentration of an antigen-binding polypeptide to reach half of its original value. The serum half-life of a protein can be measured by pharmacokinetic studies as described by Kim et al. (Eur. J. of Immuno. 24:542, 1994). According to this method, a radiolabeled protein is intravenously injected into mice, and its plasma concentration is measured periodically as a function of time, for example, from about 3 minutes to about 72 hours after injection. Other methods for pharmacokinetic analysis and determination of the half-life of a molecule are well known to those skilled in the art. Details can be found in Kenneth, A. et al.: Chemical Stability of Pharmaceuticals: A Handbook for Pharmacists and Peters et al., Pharmacokinetic analyses: A Practical Approach (1996).

[0136] "Clearance (CL)" refers to the volume of plasma that irreversibly removes a protein per unit time. Clearance is calculated as dose / AUC (AUC: area under the curve or area under the plasma drug concentration-time curve). Clearance can also be calculated by dividing the drug excretion rate by the drug's plasma concentration (excretion rate = CL * concentration).

[0137] "Mean residence time (MRT)" is the average time an antigen-binding polypeptide remains in the body before being irreversibly eliminated. It was calculated as MRT = AUMC / AUC.

[0138] The "steady-state concentration" (Css) is the concentration reached as a result of continued drug administration when the drug elimination rate equals the drug administration rate. Css fluctuates between peak and trough levels and is measured in micrograms / ml.

[0139] As used herein, "baseline" refers to the value of a parameter before or at the time of administration of a pharmaceutical composition of the invention, including, for example, the value of a given biomarker or the status of a subject before the first administration of an antibody of the present disclosure.

[0140] "Amyloid negative" means that the subject does not have observable cerebral amyloid beta plaques using positron emission tomography ("PET"), including, but not limited to, subjects with a centiloid value of zero.

[0141] "Amyloid positive" means that the subject has cerebral amyloid beta plaques observable using PET.

[0142] As used herein, "ARIA risk" or "risk of ARIA" refers to the probability that a subject will develop an amyloid-related imaging abnormality observable by MRI. Total ARIA risk includes the risk of developing observable ARIA-E and / or observable ARIA-H. In contrast, "risk of ARIA-E" refers only to the probability that a subject will develop MRI-observable ARIA-E, regardless of whether the subject will also develop observable ARIA-H, and "risk of ARIA-H" refers only to the probability that a subject will develop MRI-observable ARIA-E, regardless of whether the subject will also develop observable ARIA-E. ARIA risk can be assessed at baseline (before administration of an anti-amyloid beta antibody of the present disclosure) or, alternatively, during or after treatment.

[0143] Treatment regimen Prophylactic use: A pharmaceutical composition or medicament is administered to a patient susceptible to or otherwise at risk of Alzheimer's disease or an amyloidogenic disease in an amount sufficient to eliminate or reduce the risk, lessen the severity, or delay the onset of the disease, including the biochemical, histological, and / or behavioral symptoms of the disease, its complications, and intermediate pathological phenotypes present during the development of the disease. A patient's susceptibility or risk of developing an amyloidogenic disease can be determined, for example, from genetic markers, biochemical markers, unspecified genetic risk, or other means. In therapeutic use, a composition or medicament is administered to a patient suspected of or already suffering from such a disease in an amount sufficient to cure or at least partially halt the symptoms of the disease (biochemical, histological, and / or behavioral), including its complications and intermediate pathological phenotypes in the development of the disease.

[0144] In some embodiments, administration of an agent reduces or eliminates cognitive impairment in patients who have not yet developed the cognitive pathology characteristic of Alzheimer's disease or other amyloidogenic diseases. An amount adequate to achieve therapeutic or prophylactic treatment is defined as a therapeutically or prophylactically effective dose. In both prophylactic and therapeutic regimens, agents are typically administered in several doses until a sufficient immune response is achieved, where an "immune response" or "immunological response" includes the development of a humoral (antibody-mediated) and / or cellular (mediated by antigen-specific T cells or their secretory products) response to an antigen in the recipient subject. Such a response can be an active response, i.e., induced by administration of an immunogen, or a passive response, i.e., induced by administration of immunoglobulins or antibodies or primed T cells.

[0145] In some embodiments, the antibody is administered multiple times. The interval between single administrations can be weekly, monthly, or yearly. In some embodiments, the single dose can be administered once or twice a week, about once or twice every two weeks, about once or twice every three weeks, about once or twice every four weeks, about once or twice every five weeks, or about once or twice every six weeks. In one specific embodiment, the antibody is administered subcutaneously about once every four weeks.

[0146] The intervals may also be irregular, as determined by measuring the patient's blood levels of antibodies against Aβ. In some methods, dosage is adjusted to achieve a plasma concentration of the antibody of 1-1000 μg / ml, and in some methods, 25-300 μg / ml. Alternatively, the antibody can be administered as a sustained-release formulation, in which case less frequent administration is required. Dosage and frequency vary depending on the half-life of the antibody in the patient. Generally, human antibodies exhibit the longest half-life, followed by humanized antibodies, chimeric antibodies, and non-human antibodies.

[0147] In some embodiments, patients with Alzheimer's disease who are administered a pharmaceutically effective amount of an anti-Aβ antibody (or antigen-binding fragment thereof) described herein are treated with a reduction in amyloid plaque burden as measured by PET imaging.

[0148] In another embodiment, the antibody can be administered in a fixed amount at each administration.For example, a fixed amount of about 65 mg, or about 70 mg, or about 75 mg, or about 195 mg, or about 200 mg can be administered to an individual at one time.In one specific embodiment, about 70 mg of the antibody is subcutaneously administered about once every four weeks.In one specific embodiment, about 200 mg of the antibody is subcutaneously administered about once every four weeks.

[0149] In another specific embodiment, about 45 mg of h2731 is subcutaneously administered to an individual about once every four weeks. In another specific embodiment, about 70 mg of h2731 is subcutaneously administered to an individual about once every four weeks. In another specific embodiment, about 200 mg of h2731 is subcutaneously administered to an individual about once every four weeks.

[0150] In another specific embodiment, about 45 mg of h2726 is subcutaneously administered to an individual about once every four weeks. In another specific embodiment, about 70 mg of h2726 is subcutaneously administered to an individual about once every four weeks. In another specific embodiment, about 200 mg of h2726 is subcutaneously administered to an individual about once every four weeks.

[0151] In another specific embodiment, about 45 mg of h2726 is subcutaneously administered to an individual about once every four weeks. In another specific embodiment, about 70 mg of h2831 is subcutaneously administered to an individual about once every four weeks. In another specific embodiment, about 200 mg of h2831 is subcutaneously administered to an individual about once every four weeks.

[0152] In another specific embodiment, about 45 mg of h2726 is subcutaneously administered to an individual about once every four weeks. In another specific embodiment, about 70 mg of h2931 is subcutaneously administered to an individual about once every four weeks. In another specific embodiment, about 200 mg of h2931 is subcutaneously administered to an individual about once every four weeks.

[0153] The dosage and frequency of administration can vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic applications, compositions containing the antibodies or cocktails of the present invention are administered to patients not yet in a disease state to enhance the patient's resistance. Such an amount is defined as a "prophylactically effective dose." In this use, the exact amount also depends on the patient's health status and general immune status. Relatively low dosages are administered at relatively infrequent intervals over a long period of time. Some patients continue to receive treatment for the rest of their lives.

[0154] In therapeutic applications, relatively high dosages at relatively short intervals may be required until the progression of the disease is reduced or halted, and preferably until the patient shows partial or complete improvement in disease symptoms, after which the patient may be placed on a prophylactic regimen.

[0155] Administration: Therapeutic agents can be administered by parenteral, topical, intravenous, oral, subcutaneous, intraarterial, intracranial, intraperitoneal, intranasal, intraocular, or intramuscular means for prophylactic and / or therapeutic treatment. Intramuscular injections are most typically performed in the arm or leg muscles. In some methods, agents are injected directly into specific tissues where deposits have accumulated, e.g., intracranially. Intramuscular injection or intravenous infusion is preferred for administration of antibodies. In some methods, certain therapeutic antibodies are injected directly intracranially. In some methods, antibodies are administered as sustained-release compositions or devices. In some embodiments, antibodies are administered subcutaneously using an autoinjector device.

[0156] Dosing regimen The present disclosure provides methods for treating neurological disorders (e.g., Alzheimer's disease), comprising administering a composition comprising an anti-amyloid beta antibody. For example, in some embodiments, the present disclosure provides methods for treating Alzheimer's disease in a subject, comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks. In some embodiments, the present disclosure provides methods for treating Alzheimer's disease in a subject, comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 4 weeks. In exemplary embodiments, administration is subcutaneous.

[0157] The present disclosure also provides a method for reducing amyloid plaques in a subject. For example, in some embodiments, the present disclosure provides a method for reducing amyloid plaques in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid β antibody or antigen-binding fragment thereof about once every 3 to 5 weeks. In some embodiments, the present disclosure provides a method for reducing amyloid plaques in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid β antibody or antigen-binding fragment thereof about once every 4 weeks. In some embodiments, the administration is subcutaneous.

[0158] The present disclosure further provides a method for converting a subject from amyloid-positive to amyloid-negative. For example, in some embodiments, the present disclosure provides a method for converting a subject from amyloid-positive to amyloid-negative, the method comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks. In some embodiments, the present disclosure provides a method for converting a subject from amyloid-positive to amyloid-negative, the method comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 4 weeks. In an exemplary embodiment, the administration is subcutaneous.

[0159] In an exemplary embodiment, the method comprises administering to the subject about 45 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof about once every four weeks. In an exemplary embodiment, the method comprises administering to the subject about 70 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof about once every four weeks. In an exemplary embodiment, the method comprises administering to the subject about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof about once every four weeks. In an exemplary embodiment, the administration is subcutaneous.

[0160] In some embodiments, the method comprises administering to the subject a pharmaceutically effective amount of an anti-amyloid beta antibody or antigen-binding fragment thereof. For example, in some embodiments, the method comprises administering to the subject up to about 200 mg (e.g., up to about 180 mg, up to about 160 mg, up to about 140 mg, up to about 120 mg, up to about 100 mg, up to about 70 mg, or up to about 50 mg). In some embodiments, the method comprises administering to the subject about 20 mg to about 200 mg (e.g., about 30 mg to about 200 mg, about 40 mg to about 200 mg, about 50 mg to about 200 mg, about 60 mg to about 200 mg, about 70 mg to about 200 mg, about 80 mg to about 200 mg, about 90 mg to about 200 mg, about 100 mg to about 200 mg, about 120 mg to about 200 mg, about 140 mg to about 200 mg, about 160 mg to about 200 mg, about 180 mg to about 200 mg, or about 190 mg to about 200 mg) of an anti-amyloid beta antibody or antigen-binding fragment thereof. For example, in some embodiments, the method comprises administering to the subject about 160 mg to about 200 mg (e.g., about 170 mg to about 200 mg, about 180 mg to about 200 mg, or about 190 mg to about 200 mg) of an anti-amyloid beta antibody or antigen-binding fragment thereof.

[0161] In some embodiments, the method comprises administering to the subject about 20 mg to about 140 mg (e.g., about 30 mg to about 140 mg, about 40 mg to about 140 mg, about 50 mg to about 140 mg, about 60 mg to about 140 mg, about 70 mg to about 140 mg, about 80 mg to about 140 mg, about 90 mg to about 140 mg, about 100 mg to about 140 mg, about 110 mg to about 140 mg, about 120 mg to about 140 mg, or about 130 mg to about 140 mg) of an anti-amyloid beta antibody or antigen-binding fragment thereof. In some embodiments, the method comprises administering to the subject about 20 mg to about 120 mg (e.g., about 30 mg to about 120 mg, about 40 mg to about 120 mg, about 50 mg to about 120 mg, about 60 mg to about 120 mg, about 70 mg to about 120 mg, about 80 mg to about 120 mg, about 90 mg to about 120 mg, about 100 mg to about 120 mg, or about 110 mg to about 120 mg) of an anti-amyloid beta antibody or antigen-binding fragment thereof. In some embodiments, the method comprises administering to the subject about 20 mg to about 100 mg (e.g., about 30 mg to about 100 mg, about 40 mg to about 100 mg, about 50 mg to about 100 mg, about 60 mg to about 100 mg, about 70 mg to about 100 mg, about 80 mg to about 100 mg, or about 90 mg to about 100 mg) of an anti-amyloid β antibody or antigen-binding fragment thereof. In some embodiments, the method comprises administering to the subject about 20 mg to about 80 mg (e.g., about 30 mg to about 80 mg, about 40 mg to about 80 mg, about 50 mg to about 80 mg, about 60 mg to about 80 mg, or about 70 mg to about 80 mg) of an anti-amyloid β antibody or antigen-binding fragment thereof. In some embodiments, the method comprises administering to the subject about 20 mg to about 60 mg (e.g., about 30 mg to about 60 mg, about 40 mg to about 60 mg, or about 50 mg to about 60 mg) of an anti-amyloid beta antibody or antigen-binding fragment thereof.

[0162] In an exemplary embodiment, the method comprises administering to the subject about 40 mg to about 50 mg of an anti-amyloid β antibody or antigen-binding fragment thereof. In an exemplary embodiment, the method comprises administering to the subject about 65 mg to about 75 mg of an anti-amyloid β antibody or antigen-binding fragment thereof. In an exemplary embodiment, the method comprises administering to the subject about 195 mg to about 205 mg of an anti-amyloid β antibody or antigen-binding fragment thereof.

[0163] In some embodiments, the method comprises administering to the subject about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120 mg, about 125 mg, about 130 mg, about 135 mg, about 140 mg, about 145 mg, about 150 mg, about 155 mg, about 160 mg, about 165 mg, about 170 mg, about 175 mg, about 180 mg, about 185 mg, about 190 mg, about 195 mg, or about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof. In some embodiments, the method comprises administering to the subject about 45 mg, about 70 mg, or about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof.

[0164] In an exemplary embodiment, the method comprises administering to the subject about 45 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof. In an exemplary embodiment, the method comprises administering to the subject about 70 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof. In an exemplary embodiment, the method comprises administering to the subject about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof.

[0165] In some embodiments, the method comprises administering an anti-amyloid beta antibody or antigen-binding fragment thereof about once every four weeks. In some embodiments, the method comprises administering an anti-amyloid beta antibody or antigen-binding fragment thereof about once every month. In some embodiments, the method comprises administering an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 28 days.

[0166] In some embodiments, the method comprises administering an anti-amyloid beta antibody or antigen-binding fragment thereof as a single dose about once every 3 to 5 weeks. In some embodiments, the method comprises administering an anti-amyloid beta antibody or antigen-binding fragment thereof as a single dose about once every 4 weeks. In some embodiments, the method comprises administering an anti-amyloid beta antibody or antigen-binding fragment thereof as a single dose about once every 4 weeks. In some embodiments, the method comprises administering an anti-amyloid beta antibody or antigen-binding fragment thereof as a single dose about once per month. In some embodiments, the method comprises administering an anti-amyloid beta antibody or antigen-binding fragment thereof as a single dose about once every 28 days.

[0167] In some embodiments, the method comprises administering the anti-amyloid beta antibody or antigen-binding fragment thereof as an injection. In some embodiments, the method comprises administering the anti-amyloid beta antibody or antigen-binding fragment thereof as a parenteral injection. In some embodiments, the method comprises administering the anti-amyloid beta antibody or antigen-binding fragment thereof via intravenous or subcutaneous injection. In exemplary embodiments, the method comprises administering the anti-amyloid beta antibody or antigen-binding fragment thereof as a subcutaneous injection. In some embodiments, the method comprises administering the anti-amyloid beta antibody or antigen-binding fragment thereof via a syringe. In some embodiments, the method comprises administering the anti-amyloid beta antibody or antigen-binding fragment thereof via an autoinjector.

[0168] Anti-Aβ antibody The anti-amyloid beta antibody or fragment comprises heavy chain and light chain CDRs from one of the constructs identified herein as h2726, h2731, h2831, h2931, h2926, h4921, h2828, h2929, h3818G, h2927, h49k3G, h4917G h2727, and h4918G. Certain monoclonal antibodies of the present disclosure may bind to an epitope within residues 1-6 of Aβ (the first N-terminal residue of native Aβ is designated 1). Some monoclonal antibodies bind to an epitope within amino acids 1-6, some bind to an epitope within 1-5, and some bind to an epitope within 1-4. Some antibodies bind to an epitope within amino acids 1-3, 2-5, 3-5, 2-4, 2-5, 2-6, 3-5, or 3-6. When an antibody is said to bind to an epitope within specific residues, e.g., Aβ1-6, it means that the antibody specifically binds to a polypeptide containing at least one of the specified residues (i.e., in this example, at least one amino acid selected from Aβ amino acids 1-6). Such an antibody does not necessarily contact every residue within Aβ1-6. In some embodiments, the antibody binds to an epitope comprising at least one amino acid from amino acids selected from amino acids 1-10 of the Aβ peptide. In another aspect, the antibody binds to an epitope comprising at least one amino acid from amino acids 1-7 of the Aβ peptide. Additional amino acids of the epitope can be outside the region of amino acids 1-10 or amino acids 1-7.

[0169] In another embodiment, the anti-amyloid beta antibody or fragment thereof comprises a heavy chain variable region having heavy chain CDR1, CDR2, and CDR3, and a light chain variable region comprising light chain CDR1, CDR2, and CDR3, from the construct shown in Table 1A. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5]

[0170] In another embodiment, an anti-amyloid beta antibody or fragment of the present disclosure comprises a heavy chain variable region (VH) as shown for one of the constructs in Table 1. An anti-amyloid beta antibody or fragment may also comprise a light chain variable region (VL) as shown for one of the constructs in Table 1A.

[0171] An alignment of the CDRs for each of the heavy and light chain sequences identified in Table 1A and the CDRs from bapineuzumab ("Bapi," "hBP") is shown in Figures 19A and 19B. In one embodiment, the present disclosure relates to an antibody or fragment thereof comprising heavy chain CDR1, CDR2, and CDR3, wherein CDR1 may be selected from any one of SEQ ID NOs: 16, 19, and 20; CDR2 may be selected from any one of SEQ ID NOs: 17, 20, 21, 22, and 23; and CDR3 may be selected from any one of SEQ ID NOs: 18, 24, and 25. Furthermore, the anti-amyloid beta antibody or fragment thereof comprises light chain CDR1, CDR2, and CDR3, wherein CDR1 may be selected from any one of SEQ ID NOs: 26, 29, 31, and 32, CDR2 may be selected from any one of SEQ ID NOs: 27, 33, 34, and 35, and CDR3 may be selected from any one of SEQ ID NOs: 28, 38, and 39. In each of these embodiments, the heavy chain CDRs and light chain CDRs, in combination, are not simultaneously SEQ ID NOs: 16, 17, 18, 26, 27, and 28.

[0172] Analysis of the protein modeling information of the antibodies described above identified two changes in the CDRs that, among others, contributed to the increased avidity / affinity properties of the antibodies of the present disclosure: CDR-L1: S32Y (Ser to Tyr at position 32), and CDR-H2: G55S (Gly to Ser at position 55)

[0173] Anti-Aβ antibodies with Tyr at position 32 of CDR-L1 and Ser at position 55 of CDR-H2 that bind to the same epitope as the antibodies listed herein are expected to have the same properties as the listed identified antibodies (see Table 1A and Figures 19A and 19B). Disclosed antibodies that do not have Tyr at position 32 of CDR-L1 and Ser at position 55 of CDR-H2 can be modified to have Tyr at position 32 of CDR-L1 and Ser at position 55 of CDR-H2 and would be expected to confer similar binding properties to such antibodies identified herein.

[0174] Examples of CDR-L1 with Tyr at position 32 include SEQ ID NOs: 29 and 31. Examples of CDR-H2 with Ser at position 55 include SEQ ID NOs: 20 and 21.

[0175] By way of example, antibodies comprising a CDR-L1 with a Tyr at position 32 and a CDR-H2 with a Ser at position 55 include antibodies having the CDRs h2726, h2731, h2727, h2826, h2831, h2926, h2927, h2931, and h2929 (see Table 1A). Further such antibodies include those comprising LC CDRs 1, 2, and 3 and HC CDRs 1, 2, and 3 as shown in Table 1B below. [Table 1-6]

[0176] Given the binding characteristics identified for the antibodies identified herein, consensus sequences that are predicted to provide similar binding characteristics can be identified. For example, in embodiments of the present disclosure, an antibody or binding fragment thereof that specifically binds to Aβ peptide can include a heavy chain variable region having heavy chain CDR1, CDR2, and CDR3, and a light chain variable region having light chain CDR1, CDR2, and CDR3, as follows: heavy chain CDR1 comprises the amino acid sequence GFTFSNX1GMS, where X1 is Y or F (SEQ ID NO: 88); the heavy chain CDR2 comprises the amino acid sequence SX1RSGSGRTYYSDNVKG, where X1 is I or V (SEQ ID NO: 89); the heavy chain CDR3 comprises the amino acid sequence YDHYX1GX2SDY, where X1 is S or T and X2 is S or T (SEQ ID NO: 90); light chain CDR1 comprises the amino acid sequence KSSQSLLDYDGKTYLN (SEQ ID NO: 91); The light chain CDR2 comprises the amino acid sequence X1VX2NRDX3, where X1 is K or R, X2 is S or T, and X3 is S or T (SEQ ID NO: 92). The light chain CDR3 comprises the amino acid sequence WQGTHFPRX1, where X1 is S or T (SEQ ID NO: 93).

[0177] In some embodiments, the light chain CDR3 comprises WQGTHFPRX1FX2, wherein X1 is S or T and X2 is F or Y (SEQ ID NO: 94).

[0178] Similar consensus sequences that may be expected to provide similar binding characteristics to the antibodies described herein include a heavy chain variable region having a heavy chain CDR1, CDR2, and CDR3, and a light chain variable region having a light chain CDR1, CDR2, and CDR3 as follows: heavy chain CDR1 comprises the amino acid sequence GFTFX1NX2GMS, where X1 is S or A and X2 is Y or F (SEQ ID NO: 95); heavy chain CDR2 comprises the amino acid sequence SX1RSGX2X3RTYYSDNVKG, where X1 is I or V, X2 is S or G, and X3 is S or G (SEQ ID NO: 96); the heavy chain CDR3 comprises the amino acid sequence YDHYX1GX2SDY, where X1 is S or T and X2 is S or T (SEQ ID NO: 90); The light chain CDR1 comprises the amino acid sequence X1SSQSLX2DX3DGKTYLN, where X1 is K or R, X2 is V, M, or L, and X3 is Y, T, or S (SEQ ID NO: 97). The light chain CDR2 comprises the amino acid sequence X1VX2NRX3X4, where X1 is K or R, X2 is S or T, X3 is E or D, and X4 is iS or T (SEQ ID NO: 98). The light chain CDR3 comprises the amino acid sequence WQGX1HFPRX2, where X1 is S or T and X2 is S or T (SEQ ID NO: 99).

[0179] In some embodiments, the light chain CDR3 comprises WQGTHFPRX1FX2X3, where X1 is S or T, X2 is S or T, and X3 is F or Y (SEQ ID NO: 100).

[0180] Furthermore, the light and heavy chain variable regions can be at least 75% identical to the light and heavy chain variable regions shown in Table 1A. For example, the light and heavy chain variable regions can be 75% identical, 80% identical, 85% identical, 90% identical, 95% identical, 96% identical, 97% identical, 98% identical, 99% identical, or 100% identical to the VH and / or VL sequences identified in Table 1A. In various embodiments, any sequence variation in the VH and VL can occur outside of the CDRs, such that the VH and VL sequences of the present disclosure include the CDRs identified in Table 1A, but regions of the VH and VL sequences outside the CDRs (e.g., regions excluding the CDRs) can be at least 75% identical to regions outside the CDRs of the VH and VL sequences of Table 1A.

[0181] For example, an anti-amyloid beta antibody or fragment of the present disclosure may comprise a heavy chain variable region, excluding the CDRs, that is at least 95% identical to one of SEQ ID NOs: 3, 4, 5, 6, and 7, and a light chain variable region, excluding the CDRs, that is at least 95% identical to one of SEQ ID NOs: 8, 9, 10, 11, 12, 13, 14, and 15.

[0182] The antibodies and fragments of the present disclosure may also comprise a heavy chain constant region that is at least 75% identical to SEQ ID NO: 40. For example, the heavy chain constant region can be 75% identical, 80% identical, 85% identical, 90% identical, 95% identical, 96% identical, 97% identical, 98% identical, 99% identical, or 100% identical to SEQ ID NO: 40.

[0183] The antibodies and fragments of the present disclosure may also comprise a light chain constant region that is at least 75% identical to SEQ ID NO: 41. For example, the light chain constant region can be 75% identical, 80% identical, 85% identical, 90% identical, 95% identical, 96% identical, 97% identical, 98% identical, 99% identical, or 100% identical to SEQ ID NO: 41.

[0184] Variant antibodies or fragments that are less than 100% identical to the sequences set forth in Table 1A (plus any constant region) may differ from the anti-Aβ antibodies of Table 1A by as few as 1-15 amino acid residues, as few as 1-10 amino acid residues, e.g., 6-10, as few as 5, as few as 4, 3, 2, or 1 amino acid residue. A "conservative amino acid substitution" is one in which an amino acid residue is replaced with an amino acid residue having a side chain with a similar charge. Families of amino acid residues with side chains with similar charges are defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Alternatively, mutations can be introduced randomly along all or part of a coding sequence, such as by saturation mutagenesis, and the resulting mutants can be screened for biological activity to identify variants that retain activity (e.g., the ability to bind to an Aβ polypeptide).

[0185] For example, it is possible to introduce mutations only within the framework region(s) of an antibody molecule (i.e., within the region(s) excluding the CDRs). The introduced mutations may be silent or neutral missense mutations, i.e., have no or little effect on the antibody's ability to bind to an antigen. These types of mutations may be useful for optimizing codon usage or improving hybridoma antibody production. Alternatively, non-neutral missense mutations may alter the antibody's ability to bind to an antigen. One skilled in the art can design and test mutant molecules with desired properties, such as no change in antigen binding activity or altered binding activity (e.g., improved antigen binding activity or altered antibody specificity). Following mutagenesis, the encoded protein can be routinely expressed, and the functional and / or biological activity of the encoded protein (e.g., the ability to immunospecifically bind to at least one epitope of an Aβ polypeptide) can be determined using the techniques described herein or by routine modification techniques well known in the art. The anti-amyloid beta antibody h2731 has several physicochemical properties that make it suitable for use in the methods of the present disclosure, including, for example, the properties set forth in Tables 2 and 3. Additional properties, including pharmacokinetic parameters, are discussed herein. [Table 2-1]

[0186] Table 2A shows the Aβ 1-42 fibrils, Aβ 1-42 Aggregates and Aβ pE3-42 IC of h2731 binding to various amyloid β species, including fibrils 50 and / or EC 50 For example, see U.S. Patent No. 11,440,953.

[0187] IC for h2731 using an assay based on competition (inhibition) of labeled antibody binding to antigen-coated plates 50 Then, IC 50IC for bapineuzumab (hBP) 50 Divide by the half-maximal inhibitory concentration (IC) shown in column 1 of Table 2A. 50 ) ratio was obtained. The ratio of 0.61 indicates that h2731 is the primary Aβ 1-42 Using a similar competition assay, h2731 demonstrated superior performance compared to hBP in binding to fibrils, with an IC of 5.024 μg / mL. 50 values ​​(column 2 of Table 2A), which is comparable to the IC observed with hBP. 50 several times lower than the value.

[0188] Aβ 1-42 and Aβ pE3-42 Direct binding of h2731 to fibrils was also assessed by ELISA (Table 2A, columns 3 and 4), demonstrating Aβ 1-42 EC of h2731 on fibrils 50 The value was 36.71 ng / mL. h2731 demonstrated a strong affinity for fibrils and a significantly higher avidity than aducanumab. Furthermore, it showed a 3-fold increase in assay signal (OD490) and a 15- to 20-fold lower estimated EC 50 demonstrated increased overall binding and relative avidity of h2731 for fibrillar Aβ compared to aducanumab.

[0189] However, h2731 binds with high apparent affinity to the N-terminus of full-length Aβ but not to pyroglutamic acid-modified Aβ (Aβ pE3-42 h2731 does not specifically bind to erythrocytes. It has a median effective concentration (EC ) of 8.1 ng / mL (54 pM). 50 ) and unmodified N-terminus (Aβ 1-42 h2731 bound to fibrillar Aβ species with up to 100 ng / ml of Aβ pE3-42 showed no detectable binding to [Table 2-2]

[0190] Table 2B shows recombinant Aβ 1-42 Fibrils and Aβ 1-28We provide additional amyloid-beta binding data for h2731, including data on the binding kinetics of h2731 to monomers. As shown in column 2 of Table 3A, h2731 binds Aβ 1-42 Fibrils and Aβ 1-28 The association constant for each monomer is 3.72 × 10 5 M -1 s -1 and 1.19 × 10 5 M -1 s -1 While aducanumab binds to Aβ fibrils with a faster association rate (ka), the much slower dissociation rate (kd) of h2731 of the present disclosure resulted in a higher measured avidity (i.e., lower KD*) than aducanumab.

[0191] Off-rate data (kd) are for h2731 shown in column 3 of Table 3A and for Aβ 1-42 Fibrils and Aβ 1-28 The kd value of h2731 relative to the monomer is 2.62 × 10 -5 s -1 and 5.95 x 10 -4 s -1 The enhanced relative avidity of h2731 of the present disclosure for fibrillar Aβ observed by ELISA is confirmed by the SPR equilibrium binding kinetics (column 4 of Table 3A), which indicates that Aβ 1-28 It showed avidity (apparent KD) 5-11 times higher than aducanumab, including a binding affinity of 4-7 nM to the monomer.

[0192] In additional embodiments, the methods of the present disclosure may utilize one or more of several different anti-amyloid beta antibodies or fragments thereof. In particular, antibodies suitable for use in the methods of the present disclosure have the physiochemical and pharmacological properties discussed herein that allow for therapeutically effective administration using monthly subcutaneous administration. Further exemplary anti-amyloid beta antibodies suitable for use in the methods of the present disclosure include those described in U.S. Patent No. 11,440,953, which is incorporated herein by reference in its entirety.

[0193] In each of the aforementioned embodiments, the anti-amyloid beta antibody or fragment of the present disclosure may be a humanized antibody as described herein. For example, the antibody may be a human IgG1 antibody. Furthermore, the antibody may be a complete antibody, a chimeric antibody, a CDR-grafted antibody, or a recombinant antibody. The antibody fragment may be Fab, Fab', F(ab')2, Fabc, or Fv. Fragments may be produced by recombinant DNA techniques or by enzymatic or chemical separation of intact immunoglobulins.

[0194] The anti-amyloid beta antibodies or binding fragments, variants, or derivatives disclosed herein bind to Aβ or a fragment or variant thereof in an amount of 5×10 -2 seconds -1 , 10 -2 seconds -1 , 5×10 -3 seconds -1 , or 10 -3 seconds -1 In certain embodiments, the antibodies of the present disclosure can be said to bind with an off rate (k(off)) of 5×10 4 seconds -1 , 10 -4 seconds -1 , 5×10 -5 seconds -1 , or 10 -5 seconds -1 , 5×10 -6 seconds -1 , 10 -6 seconds -1 , 5×10 -7 seconds -1 , or 10 -7 seconds -1 It can be said to bind to Aβ or a fragment or variant thereof with an off rate (k(off)) that is:

[0195] The antibodies or antigen-binding fragments, variants, or derivatives disclosed herein bind to a target polypeptide (e.g., Aβ) disclosed herein, or a fragment or variant thereof, in an amount of 10 3 M-1 sec-1, 5×10 3 M-1 sec-1, 10 4 M-1 sec-1, or 5 x 104 In certain embodiments, the antibodies of the present disclosure bind to a target polypeptide (e.g., Aβ), or a fragment or variant thereof, disclosed herein with an on rate (k(on)) of 10 or more. 5 M-1 sec-1, 5×10 5 M-1 sec-1, 10 6 M-1 sec-1, or 5 x 10 6 M-1 seconds - 1 or 10 7 It can be said to bind with an on-rate (k(on)) of M-1 sec-1 or greater.

[0196] The anti-Aβ antibodies, or antigen-binding fragments, variants, or derivatives thereof described herein may also be described or specified in terms of their binding affinity to Aβ. Binding affinities include those with a binding affinity of 5×10 -2 M, 10 -2 M, 5 x 10 -3 M, 10 -3 M, 5 x 10 -4 M, 10 -4 M, 5 x 10 -5 M, 10 -5 M, 5 x 10 -6 M, 10 -6 M, 5 x 10 -7 M, 10 -7 M, 5 x 10 -8 M, 10 -8 M, 5 x 10 -9 M, 10 -9 M, 5 x 10 -10 M, 10 -10 M, 5 x 10 -11 M, 10 -11 M, 5 x 10 -12 M, 10 -12 M, 5 x 10 -13 M, 10 -13 M, 5 x 10 -14 M, 10 -14 M, 5 x 10 -15 M, or 10 -15 These may include those with a dissociation constant or Kd less than M.

[0197] The agents of the present disclosure can optionally be administered in combination with other agents that are at least partially effective in treating amyloidogenic diseases. In the case of Alzheimer's disease and Down's syndrome, where amyloid deposits occur in the brain, the agents of the present disclosure can also be administered in combination with other agents that increase passage of the agents of the present disclosure across the blood-brain barrier.

[0198] Expression of recombinant antibodies The present disclosure also relates to recombinant polynucleotides encoding antibodies that, when expressed, contain the heavy and light chain CDRs of the antibodies of the present disclosure. Exemplary polynucleotides (e.g., SEQ ID NOs: 42-69) for expressing polypeptide chains containing the heavy and light chain CDRs of monoclonal antibodies are provided herein, which encode variable light and heavy chain polypeptides and their CDRs according to SEQ ID NOs: 3-39. Due to codon degeneracy, other polynucleotide sequences can readily be substituted for these sequences.

[0199] Humanized and human antibodies are typically produced by recombinant expression. Nucleic acids encoding humanized light and heavy chain variable regions may be linked to constant regions and inserted into an expression vector. The light and heavy chains may be cloned into the same or different expression vectors. The DNA segments encoding immunoglobulin chains are operably linked to control sequences within the expression vector(s) that ensure the expression of immunoglobulin polypeptides. Expression control sequences include, but are not limited to, promoters (e.g., naturally associated or heterologous promoters), signal sequences, enhancer elements, and transcription termination sequences. Preferably, the expression control sequences are eukaryotic promoter systems in vectors capable of transforming or transfecting eukaryotic host cells. Once the vector is incorporated into an appropriate host, the host is maintained under conditions suitable for high-level expression of the nucleotide sequences and the collection and purification of cross-reactive antibodies.

[0200] These expression vectors are typically replicable in the host organisms either as episomes or as an integral part of the host chromosomal DNA. Commonly, expression vectors contain selection markers (e.g., ampicillin-resistance, hygromycin-resistance, tetracycline-resistance, or neomycin-resistance) to permit detection of cells transformed with the desired DNA sequences.

[0201] One prokaryotic host useful for cloning the polynucleotides of the present disclosure is E. coli. Other microbial hosts suitable for use include bacilli, such as Bacillus subtilis, and other enterobacteriaceae, such as Salmonella, Serratia, and various Pseudomonas species. Expression vectors can also be made in these prokaryotic hosts, which typically contain expression control sequences compatible with the host cell (e.g., an origin of replication). Additionally, there are numerous well-known promoters, such as the lactose promoter system, the tryptophan (trp) promoter system, the beta-lactamase promoter system, or promoter systems derived from phage lambda. The promoters typically control expression, optionally via an operator sequence, and contain ribosome binding site sequences for initiating and completing transcription and translation.

[0202] Other microorganisms, such as yeast, are also useful for expression. Saccharomyces is a preferred yeast host, and appropriate vectors having expression control sequences (e.g., promoters), origins of replication, termination sequences, etc., are used as desired. Typical promoters include 3-phosphoglycerate kinase and other glycolytic enzymes. Inducible yeast promoters include promoters from alcohol dehydrogenase, isocytochrome C, and enzymes responsible for maltose and galactose utilization, among others. Additionally, plants (e.g., rice, tobacco) are useful for expression.

[0203] Mammalian tissue cell cultures can also be used to express and produce the polypeptides of the present disclosure (e.g., polynucleotides encoding immunoglobulins or fragments thereof). Eukaryotic cells can be particularly useful, as numerous suitable host cell lines capable of secreting heterologous proteins (e.g., intact immunoglobulins) have been developed in the art, including CHO cell lines, various Cos cell lines, HeLa cells, and preferably myeloma cell lines, or transformed B cells or hybridomas. Preferably, the cells are non-human. Expression vectors for these cells can include expression control sequences such as an origin of replication, a promoter, and an enhancer, as well as necessary processing information sites such as ribosome binding sites, RNA splice sites, polyadenylation sites, and transcription terminator sequences. Preferred expression control sequences are promoters derived from immunoglobulin genes, SV40, adenovirus, bovine papilloma virus, cytomegalovirus, etc.

[0204] The antibody coding sequence can be incorporated into a transgene for introduction into the genome of a transgenic animal and subsequent expression in the milk of the transgenic animal. Suitable transgenes include light and / or heavy chain coding sequences in operably linked to a promoter and enhancer derived from a mammary gland-specific gene such as casein or beta-lactoglobulin.

[0205] Vectors containing polynucleotide sequences of interest (e.g., heavy and light chain coding sequences and expression control sequences) can be transferred into host cells by well-known methods, which vary depending on the type of cellular host. For example, calcium chloride transfection is commonly utilized for prokaryotic cells, while calcium phosphate treatment, electroporation, lipofection, biolistics, or viral-based transfection can be used for other cellular hosts. Other methods used to transform mammalian cells include the use of polybrene, protoplast fusion, liposomes, electroporation, and microinjection (see generally, Sambrook et al., supra). When producing transgenic animals, transgenes can be microinjected into fermented egg cells or integrated into the genome of embryonic stem cells, the nuclei of which can be transferred into enucleated egg cells.

[0206] When the heavy and light chains are cloned into separate expression vectors, the vectors are co-transfected to obtain expression and assembly of intact immunoglobulins. After expression, whole antibodies of the present disclosure, their dimers, individual light and heavy chains, or other immunoglobulin forms can be purified according to standard techniques in the art, including ammonium sulfate precipitation, affinity columns (e.g., Protein A), column chromatography, HPLC purification, gel electrophoresis, and the like. For pharmaceutical uses, substantially pure immunoglobulins having at least about 90-95% homogeneity are preferred, with 98-99% or more homogeneity being most preferred.

[0207] Increasing the copy number of an expression vector containing a polynucleotide sequence of interest is desirable as a way to increase antibody or antibody fragment production. Several methods for genetically engineering cells for this purpose, followed by selection of the best cells, are known in the art. These methods often include an "amplification" step to increase the copy number of the incorporated expression vector and thereby improve the yield obtained for the desired protein. Amplification methods have been previously reported, for example, by Bebbington and Hentschel (DNA Cloning Volume III (IRL Press, 1987)). Any of several selectable markers, often in the form of nucleic acid sequences encoding enzymes involved in the metabolism of host cells and essential for their survival under certain culture conditions, can be operably linked to the expression vector, thereby allowing the desired protein to be promoted upon selection for the selectable marker. Cells selected for high copy number may be subjected to further amplification methods if the protein titer is not increased to an acceptable level. Such methods may involve subjecting cells to certain toxic drugs that inhibit the selectable marker (e.g., methotrexate and dihydrofolate reductase, methionine sulfoximine and glutamine synthase, multidrug resistance / adriamycin). Such inhibition allows selection of cell populations with increased expression levels of this marker, which often also leads to increased expression levels of operably linked expression cassettes. The vector copy number in individual cells subjected to the amplification method is increased until a plateau of protein production is reached, preferably at least about 100 mg / ml / 10 6 The clones grown through such selection and amplification are then screened for titer / yield to select the best clones, which are then further evaluated. From such titration and screening, it is common to identify one or a few clones, which are then used alone for subsequent production of one or more desired proteins.

[0208] Pharmaceutical Composition In some embodiments, the anti-Aβ antibody or fragment thereof is administered as part of a pharmaceutical composition. Several methods are known for preparing pharmaceutical compositions containing an anti-Aβ antibody, or an antigen-binding fragment, variant, or derivative thereof, for a subject in need thereof. In some embodiments, the anti-Aβ antibody or antigen-binding fragment thereof is formulated for parenteral administration. In an exemplary embodiment, the anti-Aβ antibody or antigen-binding fragment thereof is formulated for subcutaneous injection.

[0209] For purposes of this disclosure, a pharmaceutically effective amount of an anti-Aβ antibody, or antigen-binding fragment, variant, or derivative thereof, means an amount sufficient to achieve effective binding to the target and provide a benefit, e.g., reducing cerebral amyloid plaques without affecting vascular amyloid, or minimizing the occurrence of microhemorrhages during chronic administration of the anti-Aβ antibody or antigen-binding fragment thereof. In some embodiments, the anti-Aβ antibody, or antigen-binding fragment, variant, or derivative thereof, in an effective amount, crosses the blood-brain barrier and reduces cerebral amyloid plaques.

[0210] The amount of anti-Aβ antibody, or fragment, variant, or derivative thereof, that is combined with the carrier materials to produce a single dosage form may vary depending on the subject being treated and the particular method of administration. Dosage regimens may also be adjusted to provide the optimum desired response (e.g., a therapeutic or prophylactic response).

[0211] The present disclosure provides several pharmaceutically effective amounts of anti-Aβ antibodies or antigen-binding fragments thereof (e.g., about 20 mg to about 200 mg, as well as additional amounts and / or ranges disclosed herein). Accordingly, the present disclosure provides for the use of pharmaceutical compositions comprising these amounts in the methods disclosed herein. Such pharmaceutically effective amounts can be administered as a single dose, multiple doses, or by infusion over an established period of time. In exemplary embodiments, these pharmaceutical compositions are administered as a single dose. In exemplary embodiments, these pharmaceutical compositions are administered as a single subcutaneous injection.

[0212] For example, in some embodiments, the present disclosure provides a method of treating Alzheimer's disease in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of a pharmaceutical composition comprising an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks.

[0213] In some embodiments, the present disclosure provides a method for reducing amyloid plaques in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of a pharmaceutical composition comprising an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks.

[0214] In some embodiments, the present disclosure provides a method for converting a subject from amyloid-positive to amyloid-negative, the method comprising administering to the subject about 20 mg to about 200 mg of a pharmaceutical composition comprising an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks.

[0215] For example, in some embodiments, the present disclosure provides a method of treating Alzheimer's disease in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of a pharmaceutical composition comprising an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks.

[0216] In some embodiments, the present disclosure provides a method for reducing amyloid plaques in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of a pharmaceutical composition comprising an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks.

[0217] In some embodiments, the present disclosure provides a method for converting a subject from amyloid-positive to amyloid-negative, the method comprising administering to the subject about 20 mg to about 200 mg of a pharmaceutical composition comprising an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks.

[0218] In some embodiments, the method comprises administering to the subject about 40 mg to about 50 mg of a pharmaceutical composition comprising an anti-amyloid β antibody or antigen-binding fragment thereof. In some embodiments, the method comprises administering to the subject about 65 mg to about 75 mg of a pharmaceutical composition comprising an anti-amyloid β antibody or antigen-binding fragment thereof. In an exemplary embodiment, the method comprises administering to the subject about 195 mg to about 205 mg of a pharmaceutical composition comprising an anti-amyloid β antibody or antigen-binding fragment thereof.

[0219] In an exemplary embodiment, the method comprises administering to the subject about 45 mg of a pharmaceutical composition comprising an anti-amyloid beta antibody or antigen-binding fragment thereof. In an exemplary embodiment, the method comprises administering to the subject about 70 mg of a pharmaceutical composition comprising an anti-amyloid beta antibody or antigen-binding fragment thereof. In an exemplary embodiment, the method comprises administering to the subject about 200 mg of a pharmaceutical composition comprising an anti-amyloid beta antibody or antigen-binding fragment thereof. In an exemplary embodiment, these pharmaceutical compositions are administered as a single subcutaneous injection approximately once every 3 to 5 weeks (once every 4 weeks).

[0220] Several methods are known for preparing and administering anti-Aβ antibodies, or antigen-binding fragments, variants, or derivatives thereof, to subjects in need thereof. The route of administration of anti-Aβ antibodies, or antigen-binding fragments, variants, or derivatives thereof, can be, for example, peripheral, oral, central (e.g., intrathecal, intracranial), parenteral, by inhalation, or topical.

[0221] As discussed herein, anti-Aβ antibodies, or antigen-binding fragments, variants, or derivatives thereof, can be formulated to facilitate administration and promote stability of the active agent. In certain embodiments, pharmaceutical compositions according to the present disclosure comprise a pharmaceutically acceptable, non-toxic, sterile carrier, such as saline, non-toxic buffers, or preservatives. For purposes of this application, a pharmaceutically effective amount of an anti-Aβ antibody, or antigen-binding fragment, variant, or derivative thereof, shall be taken to mean an amount sufficient to achieve effective target binding and benefit, e.g., reduce cerebral amyloid plaques without affecting vascular amyloid, or minimize the occurrence of microhemorrhages during long-term administration of the anti-Aβ antibody or antigen-binding fragment thereof. In some embodiments, an effective amount of an anti-Aβ antibody, or antigen-binding fragment, variant, or derivative thereof, crosses the blood-brain barrier and reduces cerebral amyloid plaques.

[0222] The pharmaceutical compositions used in this disclosure comprise pharmaceutically acceptable carriers including ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, saturated vegetable fatty acids such as glycine, sorbic acid, potassium sorbate, protamine sulfate, partial glyceride mixtures such as water, salts, or electrolytes, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and wool fat.

[0223] Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, etc. In many cases, isotonic agents, for example, sugars, polyalcohols, or salts, can be included in the composition. Prolonged absorption of injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, aluminum monostearate and gelatin.

[0224] Parenteral formulations can be a single bolus dose, an infusion, or a loading bolus dose followed by a maintenance dose. These compositions can be administered at specific fixed or variable intervals, for example, once daily, or essentially "as needed."

[0225] Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions, or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's solution, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (e.g., those based on Ringer's dextrose), and the like. Preservatives and other additives, such as antibacterial agents, antioxidants, chelating agents, and inert gases, may also be present. Additionally, the pharmaceutical compositions of the present disclosure may contain additional agents, such as dopamine or psychotropic drugs, depending on the intended use of the pharmaceutical composition.

[0226] The amount of anti-Aβ antibody, or its fragment, variant, or derivative, combined with the carrier material to produce a single dosage form can vary depending on the host being treated and the specific method of administration. The composition can be administered as a single dose, multiple doses, or by infusion over an established period of time. The dosage regimen can be adjusted to provide the optimal desired response (e.g., therapeutic or prophylactic response).

[0227] As used herein, the term "peripheral administration" includes intravenous, intraarterial, intraperitoneal, intramuscular, subcutaneous, intranasal, intraocular / intravitreal, rectal, or intravaginal administration. While all of these administration forms are expressly contemplated as being within the scope of this disclosure, an example of a form for administration would be a solution for injection, particularly for subcutaneous, intravenous, or intraarterial injection or infusion. Pharmaceutical compositions suitable for injection may include buffers, surfactants, optionally stabilizers, and the like. Preparations for peripheral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Preservatives and other additives may also be present, such as antibacterial agents, antioxidants, chelating agents, and inert gases.

[0228] Therapeutic compositions of the present disclosure are typically substantially pure from undesired contaminants. This means that the agent is typically at least 50% w / w pure of interfering proteins and other contaminants resulting from its production or purification, but does not exclude the possibility that the agent may be combined with an excess of pharmaceutically acceptable carrier(s) or other carriers intended to facilitate its use. In some cases, the monoclonal antibody (or other therapeutic agent) is at least 60%, 70%, 80%, 90%, 95%, or 99% w / w pure of interfering proteins and contaminants resulting from its production or purification.

[0229] Pharmacokinetic endpoints The present disclosure provides dosing regimens for anti-amyloid beta antibodies designed to achieve a drug exposure profile in a subject that is suitable for amyloid plaque clearance and / or treatment of a neurodegenerative disease (e.g., Alzheimer's disease). To this end, the present disclosure provides methods for administering anti-amyloid beta antibodies to achieve specific pharmacokinetic endpoints in a subject, including, for example, values ​​of the following parameters that are suitable for plaque clearance and / or treatment of the disease: mean concentration over the dosing interval (C ave ), steady-state concentration over the dosing interval (C SS ), maximum concentration over the dosing interval (C max ), time zero to infinity AUC 0-∞The area under the concentration-time curve for the first 24 h and the area under the concentration-time curve for the dosing interval (AUC 0-tau In various embodiments, these pharmacokinetic endpoints may be assessed in several bodily fluids collected from the subject, including, for example, whole blood, serum, plasma, and / or CSF.

[0230] Maximum drug concentration (C max ) Accordingly, in another aspect, the present disclosure provides a method of treating Alzheimer's disease in a subject, the method comprising administering to the subject a C max value (steady state C max In another aspect, the present disclosure provides a method of reducing amyloid plaques in a subject, the method comprising subcutaneously administering to the subject a dose of an anti-Aβ antibody sufficient to achieve a C value of about 30 μg / mL to about 60 μg / mL. max In another aspect, the present disclosure provides a method for converting a subject from amyloid-positive to amyloid-negative, the method comprising administering to the subject a dose of an anti-Aβ antibody sufficient to obtain a C value of about 30 μg / mL to about 60 μg / mL. max The method comprises subcutaneously administering a dose of an anti-Aβ antibody sufficient to achieve a value of 100%. In an exemplary embodiment, the anti-Aβ antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102.

[0231] In some embodiments, treating includes increasing the C of an anti-amyloid beta, anti-amyloid beta antibody, or antigen-binding fragment thereof, in a subject. max to about 30 μg / mL to about 60 μg / mL (e.g., about 35 μg / mL to about 60 μg / mL, about 40 μg / mL to about 60 μg / mL, about 45 μg / mL to about 60 μg / mL, about 30 μg / mL to about 55 μg / mL, about 35 μg / mL to about 55 μg / mL, about 30 μg / mL to about 50 μg / mL, or about 35 μg / mL to about 50 μg / mL). In some embodiments, max Values ​​are based on steady-state serum C max In some embodiments, C maxValues ​​are measured using steady-state plasma C max value.

[0232] Mean drug concentration (C ave ) In another aspect, the present disclosure provides a method of treating Alzheimer's disease in a subject, the method comprising administering to the subject a serum C of about 20 μg / mL to about 40 μg / mL. ave In another aspect, the present disclosure provides a method for reducing amyloid plaques in a subject, the method comprising subcutaneously administering to the subject a dose of an anti-Aβ antibody sufficient to achieve a serum C value of about 20 μg / mL to about 40 μg / mL. ave In another aspect, the present disclosure provides a method for converting a subject from amyloid-positive to amyloid-negative, the method comprising administering to the subject a dose of an anti-Aβ antibody sufficient to achieve a serum C value of about 20 μg / mL to about 40 μg / mL. ave In an exemplary embodiment, the anti-Aβ antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102. In some embodiments, treating comprises increasing the C of an anti-amyloid beta antibody or antigen-binding fragment thereof in a subject. max to about 20 μg / mL to about 40 μg / mL (e.g., about 23 μg / mL to about 40 μg / mL, about 25 μg / mL to about 40 μg / mL, about 28 μg / mL to about 40 μg / mL, about 30 μg / mL to about 40 μg / mL, about 35 μg / mL to about 40 μg / mL, about 20 μg / mL to about 38 μg / mL, about 23 μg / mL to about 38 μg / mL, about 25 μg / mL to about 38 μg / mL, about 28 μg / mL to about 38 μg / mL, about 20 μg / mL to about 35 μg / mL, about 20 μg / mL to about 30 μg / mL, or about 25 μg / mL to about 30 μg / mL). In some embodiments, ave Values ​​are based on steady-state serum C max In some embodiments, C ave Values ​​are measured using steady-state plasma C max value.

[0233] Area under the concentration-time curve (AUC 0-tau ) In another aspect, the disclosure provides a method of treating Alzheimer's disease in a subject, the method comprising administering to the subject an area under the concentration-time curve (AUC) over a dosing interval of about 15,000 hr*ug / mL to about 30,000 hr*ug / mL. 0-tau In another aspect, the present disclosure provides a method of reducing amyloid plaques in a subject, the method comprising subcutaneously administering to the subject a dose of an anti-Aβ antibody sufficient to achieve an AUC of about 15,000 hr*ug / mL to about 30,000 hr*ug / mL. 0-tau In another aspect, the present disclosure provides a method for converting a subject from amyloid-positive to amyloid-negative, the method comprising subcutaneously administering to the subject a dose of an anti-Aβ antibody sufficient to achieve an AUC value of about 15,000 hr*ug / mL to about 30,000 hr*ug / mL. 0-tau The method comprises subcutaneously administering a dose of an anti-Aβ antibody sufficient to achieve a value of 100%. In an exemplary embodiment, the anti-Aβ antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102. In some embodiments, treating comprises increasing the AUC of an anti-amyloid beta antibody or antigen-binding fragment thereof in a subject. 0-tau value (steady-state AUC 0-tau value) of about 15,000 hr*ug / mL to about 30,000 hr*ug / mL (e.g., 16,000 hr*ug / mL to about 30,000 hr*ug / mL, 18,000 hr*ug / mL to about 30,000 hr*ug / mL, 20,000 hr*ug / mL to about 30,000 hr*ug / mL, 22,000 hr*ug / mL to about 30,000 hr*ug / mL, or 25,000 hr*ug / mL to about 30,000 hr*ug / mL). In some embodiments, treating includes achieving an AUC of an anti-amyloid beta antibody or antigen-binding fragment thereof in the subject. 0-tau value (steady-state AUC 0-tauvalue) of about 15,000 hr*ug / mL to about 25,000 hr*ug / mL (e.g., 16,000 hr*ug / mL to about 25,000 hr*ug / mL, 18,000 hr*ug / mL to about 25,000 hr*ug / mL, 20,000 hr*ug / mL to about 25,000 hr*ug / mL, or 22,000 hr*ug / mL to about 25,000 hr*ug / mL). 0-tau Values ​​are steady-state serum AUC 0-tau In some embodiments, the AUC 0-tau Values ​​are steady-state plasma AUC 0-tau value.

[0234] Amyloid plaque clearance The present disclosure further provides the use of anti-amyloid-beta antibodies for reducing amyloid plaques in a subject. Reduction of amyloid plaques has been shown to correlate with slowing of cognitive decline during treatment with anti-amyloid-beta antibodies. See, for example, M. Shi, et al., Impact of Anti-amyloid-beta Monoclonal Antibodies on the Pathology and Clinical Profile of Alzheimer's Disease: A Focus on Aducanumab and Lecanemab. 14 FRONT.AGING NEUROSCI.1 (2022); C.H. van Dyck, et al., Lecanemab in Early Alzheimer's Disease 388 N.ENGL.J.MED.9 (2023). Indeed, the FDA granted accelerated approval to both aducanumab and lecanemab based on plaque reduction data from clinical trials.

[0235] Thus, in another aspect, the present disclosure provides a method of treating Alzheimer's disease in a subject having amyloid plaques, the method comprising: (a) administering to the subject approximately once every four weeks a composition comprising approximately 20 mg to approximately 200 mg of an anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine and a light chain of SEQ ID NO: 102; (b) reducing amyloid plaques in said subject.

[0236] The detection of cerebral amyloid plaques is carried out by methods known to those skilled in the art. In some embodiments, the method further comprises assessing amyloid by positron emission tomography (PET) imaging. In some embodiments, amyloid reduction is determined by PET. PET imaging agents are known to those skilled in the art and include 18F-florbetapir, florbetaben F18, and flutemetamol F18. In some embodiments, amyloid plaques measured by PET are quantified by the composite standardized uptake value ratio (SUVR). In some embodiments, amyloid plaques measured by PET are calculated using the centiloid scale. In some embodiments, changes in amyloid plaque burden are measured by the change in SUVR over time. In some embodiments, changes in amyloid plaque burden are measured by the change in centiloid over time. Navitsky M, Joshi AD, Kennedy I, et al., Standardization of amyloid quantitation with florbetapir standardized uptake value ratios to the Centiloid scale, Alzheimers Dement 2018 14:1565-71 and Oshi AD, Pontecorvo MJ, Lu M, et al., A Semiautomated Method for Quantification of F 18 Florbetapir PET Images, J Nucl See Med.2015;56(11):1736-41.

[0237] Thus, in another aspect, the present disclosure provides a method of reducing amyloid plaques in a subject, the method comprising: (a) administering to the subject about 20 mg to about 200 mg (e.g., about 45 mg, about 70 mg, about 150 mg, or about 200 mg) of an anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine and a light chain of SEQ ID NO: 102, once every four weeks; (b) reducing amyloid plaques in the subject as measured by PET.

[0238] In another aspect, the disclosure provides a method of treating Alzheimer's disease in a subject, the method comprising: (a) observing a first amyloid plaque value obtained from a first PET scan of the subject; (b) subcutaneously administering to the subject about 20 mg to about 200 mg (e.g., about 45 mg, about 70 mg, about 150 mg, or about 200 mg) of an anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine and a light chain of SEQ ID NO: 102, once every 3 to 5 weeks; (c) observing a second amyloid plaque value obtained from a second PET scan of the subject; and (d) comparing the first amyloid plaque value with the second amyloid plaque value, thereby observing a reduction in amyloid plaques in the subject.

[0239] In another aspect, the present disclosure provides a method of treating Alzheimer's disease in a subject having amyloid plaques, the method comprising: (a) performing a first PET scan of the subject, thereby observing a first amyloid plaque level; (b) administering to the subject about 20 mg to about 200 mg (e.g., about 45 mg, about 70 mg, about 150 mg, or about 200 mg) of an anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine and a light chain of SEQ ID NO: 102, once every 3 to 5 weeks; (c) performing a second PET scan of the subject, thereby observing a second amyloid plaque level; (d) comparing the first amyloid plaque value with the second amyloid plaque value, thereby observing a reduction in amyloid plaques in the subject.

[0240] In another aspect, the present disclosure provides a method of reducing amyloid plaques in a subject, the method comprising: (a) observing a first amyloid plaque value obtained from a first PET scan of the subject; (b) subcutaneously administering to the subject about 20 mg to about 200 mg (e.g., about 45 mg, about 70 mg, about 150 mg, or about 200 mg) of an anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine and a light chain of SEQ ID NO: 102, once every 3 to 5 weeks; (c) observing a second amyloid plaque value obtained from a second PET scan of the subject; and (d) comparing the first amyloid plaque value with the second amyloid plaque value, thereby observing a reduction in amyloid plaques in the subject.

[0241] In another aspect, the present disclosure provides a method of reducing amyloid plaques in a subject having amyloid plaques, the method comprising: (a) performing a first PET scan of the subject, thereby observing a first amyloid plaque level; (b) administering to the subject about 20 mg to about 200 mg (e.g., about 45 mg, about 70 mg, about 150 mg, or about 200 mg) of an anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine and a light chain of SEQ ID NO: 102, once every 3 to 5 weeks; (c) performing a second PET scan of the subject, thereby observing a second amyloid plaque level; (d) comparing the first amyloid plaque value with the second amyloid plaque value, thereby observing a reduction in amyloid plaques in the subject.

[0242] In some embodiments, treating comprises reducing the subject's amyloid beta plaques (i.e., cerebral amyloid beta plaques). In some embodiments, the subject's treatment results in a reduction of cerebral amyloid beta plaques. In some embodiments, the subject achieves a reduction in cerebral amyloid beta plaques as assessed by PET.

[0243] In some embodiments, the treatment results in a patient achieving a reduction in cerebral amyloid beta plaques. In some embodiments, the patient achieves a reduction in cerebral amyloid beta plaques as assessed by positron emission tomography (PET). In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of at least about 10 centiloids (e.g., at least about 15 centiloids, at least about 20 centiloids, at least about 25 centiloids, or at least about 30 centiloids) compared to baseline. In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of at least about 30 centiloids (e.g., at least about 35 centiloids, at least about 40 centiloids, at least about 45 centiloids, at least about 50 centiloids) compared to baseline. In some embodiments, the reduction in cerebral amyloid beta plaques is achieved after 6 months (e.g., after about 24 weeks) of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques is achieved after about 12 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques is achieved after about 18 months of treatment.

[0244] In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of at least about 10 centiloids (e.g., at least about 15 centiloids, at least about 20 centiloids, at least about 25 centiloids, or at least about 30 centiloids). In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of at least about 30 centiloids (e.g., at least about 35 centiloids, at least about 40 centiloids, at least about 45 centiloids, at least about 50 centiloids, at least about 55 centiloids, or at least about 60 centiloids) after 6 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques is achieved after about 6 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques is achieved after about 12 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques is achieved after 18 months of treatment.

[0245] In some embodiments, the reduction of cerebral amyloid beta plaques comprises a reduction of about 10 to about 90 centiloids (e.g., about 20 to about 90 centiloids, about 30 to about 90 centiloids, about 40 to about 90 centiloids, or about 50 to about 90 centiloids). In some embodiments, the reduction of cerebral amyloid beta plaques comprises a reduction of about 10 to about 80 centiloids (e.g., about 20 to about 80 centiloids, about 30 to about 80 centiloids, about 40 to about 80 centiloids, or about 50 to about 80 centiloids). In some embodiments, the reduction of cerebral amyloid beta plaques comprises a reduction of about 10 to about 70 centiloids (e.g., about 20 to about 70 centiloids, about 30 to about 70 centiloids, about 40 to about 70 centiloids, or about 50 to about 70 centiloids). In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of about 10 to about 60 centiloids (e.g., about 20 to about 60 centiloids, about 30 to about 60 centiloids, about 40 to about 60 centiloids, or about 50 to about 60 centiloids). In some embodiments, the reduction in cerebral amyloid beta plaques is achieved after about 6 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques is achieved after about 12 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques is achieved after 18 months of treatment.

[0246] In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of at least about 10 centiloids (e.g., at least about 15 centiloids, at least about 20 centiloids, at least about 25 centiloids, or at least about 30 centiloids) after about 6 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of at least about 30 centiloids (e.g., at least about 35 centiloids, at least about 40 centiloids, at least about 45 centiloids, or at least about 50 centiloids) after about 6 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, or about 95 centiloids. In some embodiments, the reduction in cerebral amyloid beta plaques is achieved after about 6 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques is achieved after about 12 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques is achieved after 18 months of treatment.

[0247] In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of about 10 to about 80 centiloids (e.g., about 20 to about 80 centiloids, about 30 to about 80 centiloids, about 10 to about 60 centiloids, or about 10 to about 50 centiloids) after about 6 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of about 30 to about 70 centiloids (e.g., about 40 to about 70 centiloids, about 50 to about 70 centiloids, about 30 to about 60 centiloids, or about 30 to about 50 centiloids) after about 6 months of treatment.

[0248] In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of at least about 25 centiloids (e.g., at least about 35 centiloids, at least about 40 centiloids, at least about 45 centiloids, or at least about 50 centiloids) after about 12 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of at least about 40 centiloids (e.g., at least about 45 centiloids, at least about 50 centiloids, at least about 55 centiloids, or at least about 60 centiloids) after about 12 months of treatment.

[0249] In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of about 20 to about 90 centiloids (e.g., about 30 to about 90 centiloids, about 40 to about 90 centiloids, about 20 to about 80 centiloids, or about 20 to about 70 centiloids) after about 12 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of about 45 to about 80 centiloids (e.g., about 50 to about 80 centiloids, about 55 to about 80 centiloids, about 45 to about 75 centiloids, or about 45 to about 70 centiloids) after about 12 months of treatment.

[0250] In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of at least about 35 centiloids (e.g., at least about 40 centiloids, at least about 45 centiloids, at least about 50 centiloids, or at least about 55 centiloids) after about 18 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of at least about 50 centiloids (e.g., at least about 55 centiloids, at least about 60 centiloids, at least about 65 centiloids, or at least about 70 centiloids) after about 18 months of treatment.

[0251] In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of about 30 to about 100 centiloids (e.g., about 40 to about 100 centiloids, about 50 to about 100 centiloids, about 30 to about 95 centiloids, or about 30 to about 90 centiloids) after about 18 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques comprises a reduction of about 50 to about 85 centiloids (e.g., about 65 to about 85 centiloids, about 70 to about 85 centiloids, about 50 to about 80 centiloids, or about 50 to about 75 centiloids) after about 18 months of treatment.

[0252] In some embodiments, treatment results in reducing a patient's cerebral amyloid beta plaques include a reduction of at least 20% (e.g., at least 22%, at least 25%, at least 27%, at least 30%, at least 32%, at least 35%, or at least 37%) compared to baseline, or at least 40% (e.g., at least 41%, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, or at least 50%) compared to baseline.

[0253] In some embodiments, treating comprises reducing cerebral amyloid beta plaques. In some embodiments, reducing cerebral amyloid beta plaques comprises a reduction of at least about 30% (e.g., at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, or at least about 70%). In some embodiments, reducing cerebral amyloid beta plaques comprises a reduction of about 30% to about 100% (e.g., about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, or about 70% to about 100%). In some embodiments, reducing cerebral amyloid beta plaques in a subject comprises a reduction of about 30% to about 90% (e.g., about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, or about 70% to about 90%). In some embodiments, the reduction in cerebral amyloid beta plaques comprises about a 30% to about 80% reduction (e.g., about a 40% to about 80%, about a 50% to about 80%, about a 50% to about 80%, or about a 60% to about 80%). In some embodiments, the reduction in cerebral amyloid beta plaques comprises about a 30% to about 70% reduction (e.g., about a 40% to about 70% reduction or about a 50% to about 70%). In some embodiments, the reduction in cerebral amyloid beta plaques comprises about a 25%, about a 30%, about a 35%, about a 40%, about a 45%, about a 50%, about a 55%, about a 60%, about a 65%, about a 70%, about a 75%, about a 80%, about a 85%, about a 90%, about a 95%, or about a 100% reduction. In some embodiments, the reduction in cerebral amyloid beta plaques is achieved after about 6 months of treatment. In some embodiments, reduction in cerebral amyloid beta plaques is achieved after about 12 months of treatment, hi some embodiments, reduction in cerebral amyloid beta plaques is achieved after 18 months of treatment.

[0254] In some embodiments, the reduction in cerebral amyloid beta plaques comprises at least about a 20% (e.g., at least about a 25%, at least about a 30%, at least about a 35%, at least about a 40%, at least about a 45%, or at least about a 50%) reduction after about 6 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques comprises about a 20% to about a 90% (e.g., about a 30% to about a 90%, about a 40% to about a 90%, or about a 50% to about a 90%) reduction after about 6 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques comprises about a 30% to about a 70% (e.g., about a 35% to about a 70%, about a 40% to about a 70%, about a 30% to about a 65%, or about a 30% to about a 60%) reduction after about 6 months of treatment.

[0255] In some embodiments, the reduction in cerebral amyloid beta plaques comprises at least about 30% (e.g., at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, or at least about 60%) reduction after about 12 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques comprises about 30% to about 100% (e.g., about 40% to about 100%, about 50% to about 100%, or about 50% to about 90%) reduction after about 12 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques comprises about 60% to about 100% (e.g., about 65% to about 100%, about 70% to about 100%, about 65% to about 95%, or about 65% to about 90%) reduction after about 12 months of treatment.

[0256] In some embodiments, the reduction in cerebral amyloid beta plaques comprises at least about 40% (e.g., at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, or at least about 70%) reduction after about 18 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques comprises about 40% to about 100% (e.g., about 50% to about 100%, about 60% to about 100%, or about 50% to about 90%) reduction after about 18 months of treatment. In some embodiments, the reduction in cerebral amyloid beta plaques in a subject comprises about 65% to about 100% (e.g., about 70% to about 100%, about 75% to about 100%, about 65% to about 95%, or about 65% to about 90%) reduction after about 18 months of treatment.

[0257] In some embodiments, a treatment outcome in reducing a patient's cerebral amyloid beta plaques comprises a reduction of at least 0.05 PET standardized updated value ratio ("SUVr") units (e.g., at least 0.10 PET SUVr units, at least 0.15 PET SUVr units, at least 0.20 PET SUVr units, or at least 0.25 PET SUVr units) compared to baseline. In some embodiments, a reduction in cerebral amyloid beta plaques comprises a reduction of at least 0.25 PET SUVr units (e.g., at least 0.30 PET SUVr units, at least 0.35 PET SUVr units, or at least 0.40 PET SUVr units, or at least 0.45 PET SUVr units) compared to baseline. In some embodiments, the reduction in brain amyloid beta plaques comprises a reduction of at least 0.50 PET SUVr units (e.g., at least 0.55 PET SUVr units, at least 0.60 PET SUVr units, or at least 0.65 PET SUVr units, or at least 0.70 PET SUVr units) compared to baseline.

[0258] In some embodiments, reduction of cerebral amyloid beta plaque is achieved after about 6 months (e.g., after about 24 weeks) of treatment. In some embodiments, reduction of cerebral amyloid beta plaque occurs after about 12 months (e.g., after about 48 weeks) of treatment. In some embodiments, reduction of cerebral amyloid beta plaque occurs after 18 months (e.g., after about 72 weeks) of treatment.

[0259] In some embodiments, the treatment outcome in the patient is achieving amyloid-negative status, hi some embodiments, the patient is converted from amyloid-positive status to amyloid-negative status.

[0260] In another aspect, the present disclosure provides a method for converting a subject from amyloid positive to amyloid negative, the method comprising: (a) administering to the subject approximately once every four weeks a composition comprising approximately 20 mg to approximately 200 mg of an anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine and a light chain of SEQ ID NO: 102; (b) reducing amyloid plaques in the subject such that the subject is amyloid-negative as determined by PET.

[0261] In some embodiments, treating comprises converting at least about 10% (e.g., at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 45%, or at least about 50%) of subjects from amyloid-positive status to amyloid-negative status. In some embodiments, treating comprises converting about 10% to about 90% (e.g., about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 10% to about 80%, about 10% to about 70%, or about 10% to about 60%) of subjects from amyloid-positive status to amyloid-negative status. In some embodiments, treating comprises converting about 30% to about 80% (e.g., about 40% to about 80%, about 50% to about 80%, about 30% to about 70%, or about 30% to about 60%) of subjects from amyloid-positive status to amyloid-negative status. In some embodiments, amyloid-positive and / or amyloid-negative status is assessed by PET. In some embodiments, achievement of amyloid-negative status occurs after about 6 months (e.g., about 24 weeks) of treatment. In some embodiments, achievement of amyloid-negative status occurs after about 12 months (e.g., about 48 weeks) of treatment. In some embodiments, achievement of amyloid-negative status occurs after about 18 months (e.g., about 72 weeks) of treatment.

[0262] In some embodiments, treating comprises converting at least about 5% (e.g., at least about 10%, at least about 15%, at least about 20%, or at least about 25%) of subjects from amyloid-positive status to amyloid-negative status after about 6 months of treatment. In some embodiments, treating comprises converting about 5% to about 50% (e.g., about 10% to about 50%, about 20% to about 50%, about 10% to about 45%, or about 10% to about 40%) of subjects from amyloid-positive status to amyloid-negative status after about 6 months of treatment. In some embodiments, treating comprises converting about 10% to about 40% (e.g., about 15% to about 40%, about 20% to about 40%, about 10% to about 35%, or about 10% to about 30%) of subjects from amyloid-positive status to amyloid-negative status after about 6 months of treatment.

[0263] In some embodiments, treating comprises converting at least about 15% (e.g., at least about 20%, at least about 25%, at least about 30%, or at least about 35%) of subjects from amyloid-positive status to amyloid-negative status after about 12 months. In some embodiments, treating comprises converting about 15% to about 80% (e.g., about 20% to about 80%, about 30% to about 80%, about 15% to about 75%, or about 15% to about 70%) of subjects from amyloid-positive status to amyloid-negative status after about 12 months. In some embodiments, treating comprises converting about 30% to about 60% (e.g., about 35% to about 60%, about 40% to about 60%, about 30% to about 55%, or about 30% to about 50%) of subjects from amyloid-positive status to amyloid-negative status after about 12 months.

[0264] In some embodiments, treating comprises converting at least about 25% (e.g., at least about 30%, at least about 35%, at least about 40%, or at least about 45%) of subjects from amyloid-positive status to amyloid-negative status after about 18 months. In some embodiments, treating comprises converting about 25% to about 100% (e.g., about 30% to about 100%, about 40% to about 100%, about 25% to about 95%, or about 25% to about 90%) of subjects from amyloid-positive status to amyloid-negative status after about 18 months. In some embodiments, treating comprises converting about 40% to about 80% (e.g., about 45% to about 80%, about 50% to about 80%, about 40% to about 75%, or about 40% to about 70%) of subjects from amyloid-positive status to amyloid-negative status after about 18 months.

[0265] In some embodiments, the subject's brain amyloid beta plaques are measured about 3 months, 6 months, 12 months, and / or 18 months after the start of treatment. In some embodiments, the subject's brain amyloid beta plaques are measured about once a month, about once every 3 months, about once every 6 months, or about once a year after the start of treatment.

[0266] cognitive decline Cognitive decline is a hallmark of neurodegenerative diseases, including Alzheimer's disease. The early stages of cognitive decline in Alzheimer's disease may manifest as forgetfulness. However, as the disease progresses, the effects of cognitive decline more broadly impact a patient's life and behavior, executive function, language ability, and visuospatial processing. This, in turn, can lead to problems with decision-making, problem-solving, and independent living. Finally, Alzheimer's disease can lead to significant cognitive decline and dementia, leading to impairments in basic memory retention and simple daily activities.

[0267] Recent evidence has demonstrated an association between reduction in brain amyloid burden and slowing of cognitive decline in patients with Alzheimer's disease. See, e.g., Y. Zhang, et al., Amyloid β-based therapy for Alzheimer's disease: challenges, successes, and future, 8 SIGNAL TRANSDUCTION AND TARGETED THERAPY 248 (2023). For example, a phase 2 clinical trial of donanemab demonstrated both a reduction in brain amyloid burden and a slowing of cognitive decline. More recently, a phase 3 clinical trial of lecanemab reported both a reduction in brain amyloid plaques and a slowing of cognitive decline. In light of this recent clinical evidence, a causal relationship between amyloid plaque removal and a slowing of cognitive decline in patients is becoming clear.

[0268] In some embodiments, treating Alzheimer's disease can include slowing, halting, and / or reversing cognitive decline in a subject. In some embodiments, treating includes slowing, halting, and / or reversing cognitive decline. In another embodiment, treating includes a slowing (e.g., slowing or halting) in cognitive decline. In another embodiment, treating includes slowing cognitive decline. In another embodiment, treating includes halting cognitive decline. In another embodiment, treating includes reversing cognitive decline.

[0269] Various cognitive assessment tools are available and can be used in conjunction with the methods of the present disclosure, including, for example, the Mini-Mental State Examination (MMSE), Alzheimer's Disease Composite Score (ADCOMS), Alzheimer's Disease Assessment Scale-Cognitive (ADAS-COG) (including, for example, the 14-item Alzheimer's Disease Assessment Scale-Cognitive (ADAS-Cog14)), Activities of Daily Living for Mild Cognitive Impairment (ADCS-ADL-MCI), Clinician Interview Impression (CIBI), Neurological Examination Battery (NTB), Disability Assessment of Dementia (DAD), Clinical Dementia Rating Box (CDR-SB), and Neuropsychiatric Impression (NPI). In some embodiments, treating comprises slowing, halting, and / or reversing cognitive decline as assessed using one of these cognitive assessment tools. In another embodiment, the methods of the present disclosure further comprise monitoring the subject with at least one of these cognitive assessment tools.

[0270] In some embodiments, cognitive function is measured by at least one of the following: CRD-SB, ADAS-Cog14, ADCOMS, and ADCS MCI-ADL. In some embodiments, cognitive function is measured using the CRD-SB. In some embodiments, cognitive function is measured using the ADAS-Cog14. In some embodiments, cognitive function is measured using the ADCOMS. In some embodiments, cognitive function is measured using the ADCS MCI-ADL.

[0271] In some embodiments, cognitive function is measured on multiple occasions, such as before dosing and at 4 weeks, 16 weeks, 6 months, and / or 1 year after dosing. In some embodiments, cognitive function is measured about 3 months, 6 months, 12 months, and / or 18 months after treatment begins. In some embodiments, cognitive function is measured about monthly, about every 3 months, about every 6 months, or yearly after treatment begins.

[0272] Biomarker Regulation In another aspect, the present disclosure provides a method for modulating a biomarker in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of a composition comprising an anti-amyloid β antibody, approximately once every 3 to 5 weeks, wherein the anti-amyloid β antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102. In another embodiment, the present disclosure provides a method for modulating a biomarker in a subject, the method comprising administering to the subject about 100 mg to about 200 mg of a composition comprising an anti-amyloid β antibody, approximately twice every 3 to 5 weeks (approximately once every 2 weeks), wherein the anti-amyloid β antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102. In another embodiment, the present disclosure provides a method for modulating a biomarker in a subject, the method comprising administering to the subject about 100 mg to about 200 mg of a composition comprising an anti-amyloid beta antibody about twice every four weeks (about once every two weeks), wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102.

[0273] Amyloid beta ratio In some embodiments, the biomarker comprises the ratio of Aβ42 / 40 in the subject. 42 / Aβ 40 Ratio (e.g., amyloid beta in CSF and / or blood) 40 against amyloid beta 42 The ratio of Aβ to β has been demonstrated to be associated with well-established indicators of AD, including amyloid PET and CSF biomarkers. 42 / Aβ 40A higher Aβ42 / Aβ1-40 ratio (e.g., Aβ42 / Aβ1-40 ratio <0.150) corresponds to a higher amyloid plaque burden. See, for example, C. Delaby, et al., "The Aβ1-42 / Aβ1-40 ratio in CSF is more strongly associated with tau markers and clinical progression than Aβ1-42 alone," 14 ALZHEIMER'S RESEARCH & THERAPY 20 (2022); X. Chang, et al., "A Review of Application of Aβ42 / Aβ40 Ratio in Diagnosis and Prognosis of Alzheimer's Disease," 90 J. ALZHEIMER'S DISEASE 495 (2002). For example, total plasma Aβ 42 / Aβ 40 The ratio has shown value in identifying individuals with mild cognitive impairment (MCI), predicting progression to dementia, and detecting underlying AD pathology as revealed by FDG-PET, amyloid-PET, and CSF biomarkers. See, e.g., V. Perez-Grijalba, et al., "Plasma Aβ42 / 40 Ratio Detects Early Stages of Alzheimer's Disease and Correlates with CSF and Neuroimaging Biomarkers in the AB255 Study," 6 J. PREVENTION OF ALZHEIMER'S DISEASE 34, (2019). Therefore, Aβ 42 / Aβ 40 Changes in the ratio can be useful in following an individual throughout the course of treatment, e.g., Aβ 42 / Aβ 40 An increase in the ratio may indicate a reduction in amyloid plaque burden during treatment.

[0274] Furthermore, an emerging trend in the treatment of Alzheimer's disease (AD) is a shift in the treatment target population from those with dementia or MCI to cognitively healthy individuals at risk for AD. Identifying this population of individuals at risk is challenging when beta-amyloid (Aβ) positivity is the primary criterion for eligibility in clinical trials. Using this metric, the screening failure rate (SRF) rises to over 70% in this population. For example, JD Doecke, et al., Total Aβ 42 / Aβ 40 See ratio in plasma predicts amyloid-PET status, independent of clinical AD diagnosis.94 NEROLOGY 1580 (2020). Aβ 42 / Aβ 40 The ratio can be useful in identifying this population and tracking them throughout treatment (eg, prophylactic treatment).

[0275] Accordingly, the present disclosure provides a method for modulating the Aβ42 / 40 ratio in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of a composition comprising an anti-amyloid β antibody about once every 3 to 5 weeks, wherein the anti-amyloid β antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102. In some embodiments, the modulation comprises an increase in the Aβ42 / 40 ratio in the subject.

[0276] In another aspect, the present disclosure provides a method for increasing the Aβ42 / 40 ratio in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of a composition comprising an anti-amyloid beta antibody about once every 3 to 5 weeks, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102.

[0277] In another aspect, the present disclosure provides a method for increasing the Aβ42 / 40 ratio in a subject, the method comprising administering to the subject about 100 mg to about 200 mg of a composition comprising an anti-amyloid beta antibody twice every about 3 to 5 weeks (e.g., about once every 2 weeks), wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102.

[0278] 1. A method for increasing the ratio of Aβ42 / 40 in a subject, the method comprising: (a) administering to the subject about 20 mg to about 200 mg of a composition comprising an anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine and a light chain of SEQ ID NO: 102, approximately once every 3 to 5 weeks; (b) determining an Aβ42 / 40 ratio value from a sample collected from the subject, wherein the Aβ42 / 40 ratio value indicates an increased Aβ42 / 40 ratio in the subject.

[0279] 1. A method for increasing the ratio of Aβ42 / 40 in a subject, the method comprising: (a) administering to the subject about 100 mg to about 200 mg of a composition comprising an anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine and a light chain of SEQ ID NO: 102, twice every about 3 to 5 weeks; (b) determining an Aβ42 / 40 ratio value from a sample collected from the subject, wherein the Aβ42 / 40 ratio value indicates an increased Aβ42 / 40 ratio in the subject.

[0280] In some embodiments, the Aβ42 / 40 ratio value increases by at least about 10% (e.g., about 15%, about 20%, about 25%, about 30%, about 35%, or about 40%). In some embodiments, the Aβ42 / 40 ratio value increases by about 10% to about 150% (e.g., about 20% to about 150%, about 30% to about 150%, about 10% to about 120%, about 20% to about 120%, about 30% to about 120%, about 10% to about 100%, or about 20% to about 100%). In some embodiments, the Aβ42 / 40 ratio value is increased by about 25% to about 100% (e.g., about 30% to about 100%, about 35% to about 100%, about 40% to about 100%, about 25% to about 90%, about 25% to about 80%, about 35% to about 90%, or about 35% to about 80%). In some embodiments, the Aβ42 / 40 ratio value is increased compared to baseline.

[0281] Phospho-tau Phospho-tau (p-tau) species have emerged as the most promising biomarker for Alzheimer's disease. See, e.g., S. Janelidze, et al., Head-to-head comparison of 10 plasma phospho-tau assays in prodromal Alzheimer's disease. 19BRAIN 1591-1601 (2023). Hyperphosphorylation of tau is a hallmark of Alzheimer's disease pathology, leading to the self-aggregation of p-tau bundles in affected subjects. See, e.g., C.-X. Gong, K. Iqbal, Hyperphosphorylation of Microtubule-Associated Protein Tau: A Promising Therapeutic Target for Alzheimer's Disease, 15CURRENT MED.CHEM.2331 (2009). Phospho-tau levels (e.g., in blood) correlate with amyloid beta (Aβ) pathology and disease severity, as well as established cerebrospinal fluid (CSF) and neuroimaging biomarkers. See, e.g., Kac, PR, et al. Diagnostic value of serum versus plasma phospho-tau for Alzheimer's disease. 14 ALZ RES THERAPY 65 (2022). Phospho-tau further distinguishes biomarker-positive AD dementia from other dementias and Aβ-negative controls, thereby demonstrating the specificity of AD versus non-AD neurodegenerative disease. Thus, p-tau levels can inform clinical diagnosis and eligibility for treatment, including treatment with anti-amyloid beta antibodies. Furthermore, because p-tau species can be measured in blood samples and, in contrast to cerebrospinal fluid, do not require lumbar puncture for acquisition, p-tau species have the potential to help expand access to AD diagnosis worldwide.See, for example, F. Gonzalez-Ortiz, et al., Plasma phospho-tau in Alzheimer's disease: towards diagnostic and therapeutic trial applications. 18 MOL. NEURODEGENERATION 18 (2023).

[0282] Several phospho-tau species, including p181-tau, p212-tau, p217-tau, p231-tau, and p235-tau, have been identified as biomarkers for Alzheimer's disease. For example, plasma levels of p-tau181, p-tau217, and p-tau231 have been shown to correlate with in vivo pathological features and autopsy diagnosis. Some of these p-tau species are highly accurate in detecting cerebral amyloidosis and predicting whether patients will progress to cognitive impairment and neurodegeneration. Furthermore, p-tau levels have been shown to change in subjects receiving anti-amyloid-β therapy and correlate with amyloid clearance. See, e.g., F. Gonzalez-Ortiz (2023).

[0283] In another aspect, the present disclosure provides a method for modulating the amount of phospho-tau in a subject, the method comprising administering to the subject about 20 mg to about 200 mg of a composition comprising an anti-amyloid β antibody, the anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102, about once every 3 to 5 weeks. In some aspects, the present disclosure provides a method for modulating the amount of phospho-tau in a subject, the method comprising administering to the subject about 100 mg to about 200 mg of a composition comprising an anti-amyloid β antibody, the anti-amyloid β antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102, about twice every 3 to 5 weeks (about once every 2 weeks). In some embodiments, the modulation comprises an increase in the amount of phospho-tau in the subject.

[0284] Thus, in some embodiments, the biomarker comprises a phospho-tau value. In some embodiments, the phospho-tau value comprises at least one of the following: a p181-tau value, a p212-tau value, a p217-tau value, a p231-tau value, and a p235-tau value. In some embodiments, the phospho-tau value comprises a p181-tau value. In some embodiments, the phospho-tau value comprises a p212-tau value. In some embodiments, the phospho-tau value comprises a p217-tau value. In some embodiments, the phospho-tau value comprises a p231-tau value. In some embodiments, the phospho-tau value comprises a p235-tau value.

[0285] In some embodiments, phospho-tau levels are decreased by about 5% to about 50% (e.g., about 10% to about 50%, about 15% to about 50%, about 20% to about 50%, about 10% to about 45%, about 10% to about 40%, or about 10% to about 35%). In some embodiments, phospho-tau levels are decreased by about 10% to about 30% (e.g., about 15% to about 30%, about 20% to about 30%, about 10% to about 25%, about 15% to about 25%, or about 20% to about 30%). In some embodiments, phospho-tau levels are decreased compared to baseline.

[0286] In some embodiments, p181-tau levels are decreased by about 5% to about 50% (e.g., about 10% to about 50%, about 15% to about 50%, about 20% to about 50%, about 10% to about 45%, about 10% to about 40%, or about 10% to about 35%). In some embodiments, p181-tau levels are decreased by about 10% to about 30% (e.g., about 15% to about 30%, about 20% to about 30%, about 10% to about 25%, about 15% to about 25%, or about 20% to about 30%). In some embodiments, p181-tau levels are decreased compared to baseline. In some embodiments, p217-tau levels are decreased by about 5% to about 50% (e.g., about 10% to about 50%, about 15% to about 50%, about 20% to about 50%, about 10% to about 45%, about 10% to about 40%, or about 10% to about 35%). In some embodiments, p217-tau levels are decreased by about 10% to about 30% (e.g., about 15% to about 30%, about 20% to about 30%, about 10% to about 25%, about 15% to about 25%, or about 20% to about 30%). In some embodiments, p217-tau levels are decreased compared to baseline.

[0287] Amyloid-related imaging abnormalities (ARIA) Treatment with amyloid-beta-targeting passive immunotherapies (e.g., aducanumab (AduHelm), lecanemab, donanemab, and gantenerumab) carries a significant risk of amyloid-related imaging abnormalities (ARIA). See, for example, M. Filippi, et al., Amyloid-Related Imaging Abnormalities and β-Amyloid-Targeting Antibodies: A Systematic Review. 79 JAMA NEUROL. 291 (2022). ARIA is the most common side effect of anti-amyloid-beta antibodies and can be classified as ARIA-E (cerebral edema with disruption of the tight endothelial junctions of the blood-brain barrier and subsequent fluid accumulation) and ARIA-H (cerebral microhemorrhages (mH), small bleeding on the brain often accompanied by hemosideria). ARIA-E may be associated with acute neuroinflammation and a preponderance of perivascular clearance systems, while ARIA-H may be associated with weakening of vascular amyloid clearance and subsequence and / or small blood vessel rupture. See, e.g., H. Hampel, et al., Amyloid-related imaging abnormalities (ARIA): radiological, biological and clinical characteristics, 146BRAIN4414 (2023).

[0288] Although often asymptomatic and detected only via MRI, in some cases, ARIA can be symptomatic. For example, ARIA has been shown to include numerous side effects, such as headache, worsening confusion, dizziness, visual disturbances, nausea, and seizures. Furthermore, at least one death associated with ARIA-E during aducanumab treatment and at least one death due to ARIA-H during donanemab treatment have been reported to date. See, for example, C.G. Withington & R.S. Turner, Amyloid-Related Imaging Abnormalities With Anti-amyloid Antibodies for the Treatment of Dementia Due to Alzheimer's Disease, 13 Frontiers in Neurology 1 (2022). The risk of ARIA-H increases with age and cerebrovascular disease, and ARIA rates are generally higher in ApoE4 homozygous patients (ApoE4 carriers) compared with either ApoE4 non-carriers or ApoE4 heterozygous patients. Furthermore, an increased risk of ARIA-E was observed at the start of treatment, corresponding to higher doses and >4 microbleeds on baseline MRI.

[0289] In some embodiments, the treatment involves a risk of ARIA-E that is less than about 70% (e.g., less than about 65%, less than about 60%, less than about 55%, or less than about 50%). In some embodiments, the treatment involves a risk of ARIA-E that is less than about 45% (e.g., less than about 40%, less than about 35%, less than about 30%, less than about 25%, or less than about 20%). In some embodiments, the treatment involves a risk of ARIA-E that is less than about 15% (e.g., less than about 14%, less than about 13%, less than about 12%, less than about 11%, or less than about 10%).

[0290] In some embodiments, treatment results in fewer than about 45% of subjects experiencing symptomatic ARIA-E (e.g., fewer than about 40%, less than about 35%, less than about 30%, less than about 25%, or less than about 20%). In some embodiments, treatment results in fewer than about 15% of subjects experiencing symptomatic ARIA-E (e.g., fewer than about 14%, less than about 13%, less than about 12%, less than about 11%, or less than about 10%). In some embodiments, treatment results in fewer than about 10% of subjects experiencing symptomatic ARIA-E (e.g., fewer than about 9%, less than about 8%, less than about 7%, less than about 6%, or less than about 5%).

[0291] In some embodiments, the risk of ARIA-E is a risk of severe ARIA-E. In some embodiments, the risk of ARIA-E is a risk of moderate+ ARIA-E. In some embodiments, the risk of ARIA-E is a risk of moderate ARIA-E. In some embodiments, the risk of ARIA-E is a risk of mild+ ARIA-E. In some embodiments, the risk of ARIA-E is a risk of mild ARIA-E. In some embodiments, the risk of ARIA-E includes a risk of FLAIR hyperintensity at multiple locations, each FLAIR location having a size of 5-10 cm. In some embodiments, the risk of ARIA-E includes a risk of FLAIR hyperintensity at one location, the FLAIR location having a size of 5-10 cm. In some embodiments, the risk of ARIA-E includes a risk of FLAIR hyperintensity at multiple locations, each FLAIR location having a size of less than 5 cm, and each FLAIR location being confined to the sulci, cortex, and / or subcortical white matter. In some embodiments, the risk of ARIA-E includes the risk of FLAIR hyperintensity at one location, the FLAIR location having a size of less than 5 cm, and each FLAIR location being restricted to the sulci, cortex, and / or subcortical white matter.

[0292] In some embodiments, the patient is an APOE4 homozygous patient. In some embodiments, the treatment involves reducing the risk of ARIA-E in APOE4 homozygous patients to less than about 70% (e.g., less than about 65%, less than about 60%, less than about 55%, or less than about 50%). In some embodiments, the treatment involves reducing the risk of ARIA-E in APOE4 homozygous patients to less than about 40% (e.g., less than about 35%, less than about 30%, or less than about 25%).

[0293] In some embodiments, the subject is an APOE4 homozygous subject, and the treatment involves reducing the risk of ARIA-E in the APOE4 homozygous subject to less than about 75% (e.g., less than about 70%, less than about 65%, less than about 60%, less than about 55%, or less than about 50%). In some embodiments, the subject is an APOE4 homozygous subject, and the treatment involves reducing the risk of symptomatic ARIA-E in the APOE4 homozygous subject to less than about 30% (e.g., less than about 25%, less than about 20%, or less than about 15%).

[0294] In some embodiments, the patient is an APOE4 heterozygous patient or an APOE4 negative patient. In some embodiments, the treatment involves reducing the risk of ARIA-E in APOE4 heterozygous patients or APOE4 negative patients to less than about 40% (e.g., less than about 35%, less than about 30%, less than about 25%, or less than about 20%). In some embodiments, the treatment involves reducing the risk of ARIA-E in APOE4 heterozygous patients or APOE4 negative patients to less than about 15% (e.g., less than about 14%, less than about 13%, less than about 12%, less than about 11%, or less than about 10%).

[0295] In some embodiments, the subject is an APOE4 heterozygous subject or an APOE4 negative subject, and the treatment involves reducing the risk of ARIA-E in an APOE4 homozygous subject or an APOE4 negative subject to less than about 45% (e.g., less than about 40%, less than about 35%, less than about 30%, less than about 25%, or less than about 20%). In some embodiments, the subject is an APOE4 heterozygous subject or an APOE4 negative subject, and the treatment involves reducing the risk of symptomatic ARIA-E in an APOE4 heterozygous subject or an APOE4 negative subject to less than about 15% (e.g., less than about 14%, less than about 13%, less than about 12%, less than about 11%, or less than about 10%). In some embodiments, the risk of ARIA-E is after about 6 months (e.g., about 24 weeks) of treatment. In some embodiments, the risk of ARIA-E is after about 12 months (e.g., about 48 weeks) of treatment. In some embodiments, the risk of ARIA-E is the risk after about 18 months (eg, about 72 weeks) of treatment.

[0296] In some embodiments, the treatment involves a risk of ARIA-H of less than about 25% (e.g., less than about 22%, less than about 20%, less than about 18%, or less than about 16%). In some embodiments, the treatment involves a risk of ARIA-H of less than about 15% (e.g., less than about 14%, less than about 13%, less than about 12%, less than about 11%, or less than about 10%).

[0297] In some embodiments, the risk of ARIA-H is a risk of severe ARIA-H. In some embodiments, the risk of ARIA-H is a risk of moderate ARIA-H. In some embodiments, the risk of ARIA-H is a risk of mild ARIA-H. In some embodiments, the risk of ARIA-H comprises a risk of no more than four new microhemorrhages and / or a risk of no more than one superficial hemosiderosis lesion area. In some embodiments, the risk of ARIA-H comprises a risk of no more than nine new microhemorrhages and / or no more than two superficial hemosiderosis lesion area.

[0298] In some embodiments, the risk of ARIA-H is the risk after about 6 months (e.g., about 24 weeks) of treatment. In some embodiments, the risk of ARIA-H is the risk after about 12 months (e.g., about 48 weeks) of treatment. In some embodiments, the risk of ARIA-H is the risk after about 18 months (e.g., about 72 weeks) of treatment.

[0299] In some embodiments, treatment results in fewer than about 45% of subjects experiencing ARIA-E (e.g., fewer than about 40%, fewer than about 35%, fewer than about 30%, fewer than about 25%, or fewer than about 20%). In some embodiments, treatment results in fewer than about 15% of subjects experiencing ARIA-E (e.g., fewer than about 14%, fewer than about 13%, fewer than about 12%, fewer than about 11%, or fewer than about 10%). In some embodiments, treatment results in fewer than about 10% of subjects experiencing ARIA-E (e.g., fewer than about 9%, fewer than about 8%, fewer than about 7%, fewer than about 6%, or fewer than about 5%).

[0300] In some embodiments, treatment results in less than about 15% of subjects (e.g., less than about 14%, less than about 13%, less than about 12%, less than about 11%, less than about 10%, less than about 9%, less than about 8%, less than about 7%, less than about 6%, or less than about 5%) experiencing symptomatic ARIA-E.

[0301] In some embodiments, treatment results in less than about 25% of subjects (e.g., less than about 22%, less than about 20%, less than about 18%, or less than about 16%) experiencing ARIA-H. In some embodiments, treatment results in less than about 15% of subjects (e.g., less than about 14%, less than about 13%, less than about 12%, less than about 11%, or less than about 10%, less than about 9%, less than about 8%, less than about 7%, less than about 6%, or less than about 5%) experiencing ARIA-H.

[0302] In some embodiments, the ARIA-E and / or ARIA-H levels (e.g., % of subjects experiencing ARIA-E and / or ARIA E) are the levels of ARIA-E and / or ARIA H after about 6 months (e.g., about 24 weeks) of treatment. In some embodiments, the ARIA-E and / or ARIA-H levels are the levels of ARIA-E and / or ARIA H after about 12 months (e.g., about 48 weeks) of treatment. In some embodiments, the ARIA-E and / or ARIA-H are the levels of ARIA-E and / or ARIA H after about 18 months (e.g., 72 weeks) of treatment.

[0303] In some embodiments, the patient does not experience symptomatic ARIA as assessed by magnetic resonance imaging (MRI). In some embodiments, the patient does not experience symptomatic ARIA-E as assessed by MRI. In some embodiments, the patient does not experience symptomatic ARIA-H as assessed by MRI. In some embodiments, the patient is an APOE4 heterozygous patient or an APOE4 negative patient. In some embodiments, the patient is an APOE4 homozygous patient.

[0304] Treatable patients The present disclosure is also directed to the treatment of Alzheimer's disease and other amyloidogenic diseases by administration of the disclosed antibodies, fragments, and pharmaceutical compositions that produce a beneficial therapeutic response in a patient (e.g., inducing phagocytosis of Aβ, reducing plaque burden, inhibiting plaque formation, reducing neuritic dystrophy, neutralizing soluble and toxic Aβ species, improving cognitive function, and / or reversing, treating, or preventing cognitive decline), e.g., for the prevention or treatment of amyloidogenic diseases. The present disclosure also relates to the use of the disclosed antibodies and fragments in the manufacture of a medicament for the treatment or prevention of amyloidogenic diseases.

[0305] In one embodiment, the present disclosure provides a method for preventing or treating a disease associated with amyloid deposition of Aβ in a patient. In one embodiment, the amyloid deposits are in the brain or other CNS regions. Such diseases include Alzheimer's disease, Down's syndrome, age-related macular degeneration (AMD), and cognitive impairment. The latter may occur with or without other characteristics of amyloidogenic disease. Some methods of the present disclosure involve administering to a patient an effective dosage of an antibody that specifically binds to a component of amyloid deposits. Such methods are useful for preventing or treating Alzheimer's disease in human patients.

[0306] The methods can be used in both asymptomatic patients and patients currently exhibiting symptoms of the disease. The antibodies used in such methods can be humanized, human, or fragments thereof (e.g., antigen-binding fragments), and can be monoclonal or polyclonal, as described herein. In yet another aspect, the disclosure features administering an antibody prepared from a human immunized with Aβ peptide, where the human can be the patient to be treated with the antibody.

[0307] In another aspect, the present disclosure features administering an antibody with a pharmaceutical carrier as a pharmaceutical composition. Alternatively, the antibody can be administered to a patient by administering a polynucleotide encoding at least one antibody chain. The polynucleotide is expressed in the patient to produce the antibody chain. Optionally, the polynucleotide encodes heavy and light chains of the antibody. The polynucleotide is expressed in the patient to produce the heavy and light chains. In an exemplary embodiment, the patient is monitored for levels of the administered antibody in the patient's blood.

[0308] Patients suitable for treatment include individuals at risk for the disease but who do not show symptoms, as well as those currently showing symptoms. In the case of Alzheimer's disease, anyone who lives long enough is potentially at risk for Alzheimer's disease. Therefore, the present method involves prophylactic administration to the general population without the need for any assessment of the subject's risk. The present method is particularly useful for individuals with a known genetic risk for Alzheimer's disease. Such individuals include those with relatives who have experienced the disease and those whose risk is determined by analysis of genetic or biochemical markers. Genetic markers of risk for Alzheimer's disease include mutations in the APP gene, particularly mutations at positions 717, 670, and 671, known as the Hardy and Swedish mutations, respectively. Other risk markers include mutations in the presenilin genes, PS1 and PS2, and ApoE4, a family history of AD, hypercholesterolemia, or atherosclerosis. Individuals currently suffering from Alzheimer's disease can be recognized by the presence of characteristic dementia and the risk factors listed above. Furthermore, several diagnostic tests are available to identify individuals with AD. These include measuring CSF tau and Aβ42 levels. Elevated tau and decreased Aβ42 levels indicate the presence of AD. Individuals suffering from Alzheimer's disease can also be diagnosed by the ADRDA criteria, as discussed in the Examples section.

[0309] Treatment of asymptomatic patients can begin at any age (e.g., 10, 20, 30). However, it is not necessary to begin treatment until the patient reaches 40, 50, 60, or 70 years of age. Treatment usually involves multiple doses over a period of time. Treatment can be monitored by assaying antibody levels over time. If the response decreases, booster doses are indicated. For potential Down syndrome patients, treatment can begin by administering a therapeutic agent to the mother before birth or shortly after birth.

[0310] APOE4 status Subjects carrying the ε4 allele of apolipoprotein E (APOE4) are at increased risk of ARIA during treatment with anti-amyloid therapy. See, for example, C. Dagostin, et al., Efficacy of anti-amyloid-β monoclonal antibody therapy in early Alzheimer's disease: a systematic review and meta-analysis. NEOLOGICAL SCIENCES (2023). Subjects homozygous for APOE4 are at highest risk, due in part to increased amounts of aggregated β-amyloid in cerebral microvasculature. Furthermore, ongoing research suggests that APOE4 carriers experience greater perivascular Aβ clearance and increased vascular permeability, resulting in extravascular fluid and red blood cell expulsion, resulting in higher rates of ARIA-E and ARIA-H during treatment. See, for example, M. Roytman, et al., Amyloid-Related Imaging Abnomalities: An Update. 220 Am. J. ROENTGENOLOGY 562 (2023).

[0311] In some embodiments, the subject is an APOE4 heterozygous subject or an APOE4 negative subject. In some embodiments, the subject is an APOE4 homozygous subject. In some embodiments, the subject is an APOE4 heterozygous subject. In some embodiments, the subject is an APOE4 negative subject (non-carrier).

[0312] In vivo detection In another aspect, the present disclosure provides a method for detecting amyloid plaques and deposits in patients who have or are at risk of developing an amyloidogenic disease. Such a method is useful for diagnosing or confirming amyloidogenic disease or susceptibility to it. For example, the method can be used in patients with dementia symptoms, and the observation of abnormal amyloid deposits is likely to indicate Alzheimer's disease. The method can also be used in asymptomatic patients. The presence of abnormal amyloid deposits indicates susceptibility to future symptomatic disease.

[0313] In some embodiments, the method comprises administering to a subject / patient an antibody or fragment thereof of the present disclosure and detecting the antibody or fragment thereof bound to Aβ.

[0314] The antibody and / or antibody fragment thereof can be administered by any suitable means that results in delivery to the tissue to be visualized, for example, by intravenous injection into the patient's body or directly into the brain by intracranial injection. The administered dose of the antibody and / or fragment thereof can include a therapeutic dose, a subtherapeutic dose, or a supratherapeutic dose. In some embodiments, the antibody or fragment thereof is labeled, including a fluorescent label, a paramagnetic label, or a radioactive label. The choice of label depends on the detection means. For example, a fluorescent label is suitable for visual detection. The use of a paramagnetic label is suitable for tomographic detection without surgical intervention. In some embodiments, the radioactive label is detected using positron emission tomography (PET) or single photon emission computed tomography (SPECT).

[0315] In another aspect, the present disclosure provides a method for measuring the effectiveness of treatment in a subject receiving treatment for an amyloidogenic disease. In some embodiments, a first level of amyloid plaques in a subject is measured before treatment by administering an antibody or fragment thereof of the present disclosure, and a first amount of the antibody or fragment thereof bound to Aβ in the subject is detected. The subject can then be treated, followed by measuring a second level of amyloid plaques in the subject and detecting the antibody or fragment thereof bound to Aβ in the subject. In some embodiments, a decrease in the level of amyloid plaques indicates a positive response to treatment, and in some embodiments, no change in the level of amyloid plaques or a slight increase in amyloid plaques indicates a positive response to treatment. In some embodiments, the level of amyloid plaques can be measured using the methods for detecting amyloid plaques described herein.

[0316] In some embodiments, a diagnosis of an amyloidogenic disease can be made, for example, by comparing the number, size, and / or intensity of labeled loci from a first level (i.e., baseline) to a second subsequent level of amyloid plaques measured in a subject, where an increase over time indicates disease progression, no change, and a decrease in amyloid plaques or intensity over time indicates remission.

[0317] Detection of cerebral amyloid plaques is performed by methods known to those skilled in the art. In some embodiments, amyloid plaque burden is measured in patients by positron emission tomography (PET) imaging. PET imaging agents are known to those skilled in the art and include 18F-florbetapir, florbetaben F18, and flutemetamol F18. In some embodiments, amyloid plaques measured by PET are quantified by the composite standardized uptake value ratio (SUVR). In some embodiments, amyloid plaques measured by PET are calculated using the centiloid scale. In some embodiments, changes in amyloid plaque burden are measured by the change in SUVR over time. In some embodiments, changes in amyloid plaque burden are measured by the change in centiloid over time. Navitsky M, Joshi AD, Kennedy I, et al., Standardization of amyloid quantitation with florbetapir standardized uptake value ratios to the Centiloid scale, Alzheimers Dement 2018 14:1565-71 and Oshi AD, Pontecorvo MJ, Lu M, et al., A Semiautomated Method for Quantification of F 18 Florbetapir PET Images, J Nucl See Med.2015;56(11):1736-41.

[0318] use In various embodiments, the present disclosure relates to pharmaceutical compositions comprising an anti-amyloid beta antibody or antigen-binding fragment described herein for treating Alzheimer's disease in a subject. In various embodiments, the treatment comprises administering to the subject about 20 mg to about 200 mg of the antibody or antigen-binding fragment thereof approximately once every 3 to 5 weeks. In several embodiments, the intermediate doses described herein can be used for subcutaneous administration.

[0319] In various embodiments, the present disclosure relates to pharmaceutical compositions comprising an anti-amyloid beta antibody or antigen-binding fragment for reducing amyloid plaques in a subject. In various embodiments, treating the subject involves administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof approximately once every 3 to 5 weeks. In several embodiments, the intermediate doses described herein can be used for subcutaneous administration.

[0320] In various embodiments, the present disclosure relates to pharmaceutical compositions comprising an anti-amyloid beta antibody or antigen-binding fragment described herein for converting a subject from amyloid-positive to amyloid-negative. In various embodiments, treating a subject comprises administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks, or about once every 4 weeks. In several embodiments, the intermediate doses described herein can be used for subcutaneous administration.

[0321] In various pharmaceutical composition embodiments, administering comprises, for example, subcutaneously administering to a subject about 45 mg of an anti-amyloid beta antibody about once every four weeks, subcutaneously administering to a subject about 70 mg of an anti-amyloid beta antibody about once every four weeks, or subcutaneously administering to a subject about 200 mg of an anti-amyloid beta antibody about once every four weeks. In several embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable excipient for administration, including, for example, subcutaneous administration.

[0322] In various embodiments, the present disclosure relates to the use of an anti-amyloid beta antibody or antigen-binding fragment described herein for the manufacture of a medicament for treating Alzheimer's disease in a subject. In various embodiments, the medicament is for administering to the subject about 20 mg to about 200 mg of the anti-amyloid beta antibody or antigen-binding fragment approximately once every 3 to 5 weeks. In several embodiments, the intermediate doses described herein can be used for subcutaneous administration.

[0323] In various embodiments, the present disclosure relates to the use of an anti-amyloid beta antibody or antigen-binding fragment described herein for the manufacture of a medicament for reducing amyloid plaques in a subject. In various embodiments, the medicament is for administering to a subject about 20 mg to about 200 mg of the anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks. In several embodiments, the intermediate doses described herein can be used for subcutaneous administration.

[0324] In various embodiments, the present disclosure relates to the use of an anti-amyloid beta antibody or antigen-binding fragment described herein for the manufacture of a medicament for converting a subject from amyloid-positive to amyloid-negative. In various embodiments, the medicament is for administering to the subject about 20 mg to about 200 mg of the anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks. In several embodiments, the intermediate doses described herein can be used for subcutaneous administration.

[0325] In several embodiments of the use of an anti-amyloid beta antibody or antigen-binding fragment thereof, administration includes, for example, subcutaneously administering to a subject about 45 mg of an anti-amyloid beta antibody or binding fragment about once every four weeks, subcutaneously administering to a subject about 70 mg of an anti-amyloid beta antibody or binding fragment about once every four weeks, or subcutaneously administering to a subject about 200 mg of an anti-amyloid beta antibody or binding fragment about once every four weeks.

[0326] The present disclosure is further illustrated by the following non-limiting examples.

[0327] SEQ ID NO: 40: huIgG1 constant ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0328] SEQ ID NO: 41: hu kappa stationary RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0329] SEQ ID NO: 42: h2726_VH (variable heavy chain) nucleotide sequence GAAGTGCAGCTTCTGGAGAGCGGGGGCGGCCTGGTGCAGCCGGGCGGATCCCTGAGACTGTCCTGTGCCGCGTCCGGTTTTACCTTCTCCAACTACGGAATGTCATGGGTCCGCCAAGCACCCGGAAAGGGATTGGAATGGGTGGCTTCGATCCGGTCCGGCTCGGGACGGACCTACT ACTCCGATAACGTCAAGGGCAGATTCACTATTAGCCGGGACAACAGCAAGAATACCCTGTACCTCCAAATGAACTCCCTGAGGGCCGAGGACACCGCCGTGTATTACTGCGTGCGCTACGACCACTACTCGGGTTCCTCTGATTACTGGGGACAGGGGACCCTCGTGACTGTGTCAAGC

[0330] SEQ ID NO: 43: h2726_VL (variable light chain) nucleotide sequence GATGTCGTGATGACCCAGTCACCACTGTCCCTTCCTGTGACTCCCGGAGAACCGGCGTCCATTTCGTGCAAGAGCAGCCAGTCCCTGCTCGATTATGACGGAAAGACCTACCTGAACTGGTTGCTCCAAAAGCCTGGCAGAGCCCCCAGAGACTGATCTACAAAGTG TCCAACAGGGACTCGGGCGTGCCGGACCGCTTCGGGGTCCGGTTCCGGTACCGACTTTACGCTGAAGATCTCACGGGTGGAAGCCGAGGACGTGGGAGTGTACTACTGTTGGCAGGGCACTCACTTCCCGCGGACCTTCGGACAAGGCACCAAGGTCGAGATCAAG

[0331] SEQ ID NO: 44: h2931_VH (variable heavy chain) nucleotide sequence GAAGTGCAGCTCCTGGAGTCCGGGGGTGGACTGGTGCAGCCCGGGGGCAGCCTGAGGCTGAGCTGCGCCGCGTCAGGATTCACCTTCTCCAACTTCGGAATGTCCTGGGTCAGACAGGCCCGGGAAAGGGCCTTGAATGGGTGGCTAGCGTGCGCTCCGGTTCCGGACGGACCTACT ACTCGGACAACGTGAAGGGCCGGTTTACTATCTCCCGGGACAATTCGAAGAACACCCTGTACCTCCAAATGAACTCCTTGCGCGCCGAGGATACCGCAGTGTATTACTGCGTGCGCTACGACCACTACTCTGGCACTAGCGATTACTGGGCCAGGGAACTCTGGTCACCGTGTCGTCA

[0332] SEQ ID NO: 45: h2931_VL (variable light chain) nucleotide sequence GATGTCGTGATGACTCAGTCACCTCTGTCCCTGCCTGTGACCCTTGGGGAACCCGCCTCGATCTCGTGCAAGAGCTCCCAGAGCCTGCTCGACTATGATGGAAAGACCTACCTGAACTGGTTGCTCCAAAGCCGGCCAGAGCCCCCAGAGGCTGATCTACCGCGTG ACCAACCGCGACACCGGGGTGCCGGACCGGTTCTCCGGATCCGGCAGCGGCACTGACTTCACCCTGAAAATTTCCAGAGTGGAAGCCGAGGACGTGGGAGTGTACTACTGTTGGCAGGGTACTCACTTTCCACGGTCCTTCGGTCAAGGAACCAAGGTCGAGATCAAG

[0333] SEQ ID NO: 46: h2731_VH (variable heavy chain) nucleotide sequence GAAGTGCAGCTTCTGGAGAGCGGGGGCGGCCTGGTGCAGCCGGGCGGATCCCTGAGACTGTCCTGTGCCGCGTCCGGTTTTACCTTCTCCAACTACGGAATGTCATGGGTCCGCCAAGCACCCGGAAAGGGATTGGAATGGGTGGCTTCGATCCGGTCCGGCTCGGGACGGACCTACT ACTCCGATAACGTCAAGGGCAGATTCACTATTAGCCGGGACAACAGCAAGAATACCCTGTACCTCCAAATGAACTCCCTGAGGGCCGAGGACACCGCCGTGTATTACTGCGTGCGCTACGACCACTACTCGGGTTCCTCTGATTACTGGGGACAGGGGACCCTCGTGACTGTGTCAAGC

[0334] SEQ ID NO: 47: h2731_VL (variable light chain) nucleotide sequence GATGTCGTGATGACTCAGTCACCTCTGTCCCTGCCTGTGACCCTTGGGGAACCCGCCTCGATCTCGTGCAAGAGCTCCCAGAGCCTGCTCGACTATGATGGAAAGACCTACCTGAACTGGTTGCTCCAAAGCCGGCCAGAGCCCCCAGAGGCTGATCTACCGCGTG ACCAACCGCGACACCGGGGTGCCGGACCGGTTCTCCGGATCCGGCAGCGGCACTGACTTCACCCTGAAAATTTCCAGAGTGGAAGCCGAGGACGTGGGAGTGTACTACTGTTGGCAGGGTACTCACTTTCCACGGTCCTTCGGTCAAGGAACCAAGGTCGAGATCAAG

[0335] SEQ ID NO: 48: h2831_VH (variable heavy chain) nucleotide sequence GAAGTGCAGCTGCTGGAGTCTGGCGGCGGACTGGTGCAGCCCGGGGGATCCCTGCGGCTTTCCTGCGCCGCATCCGGCTTCACCTTTTCAAACTTCGGAATGTCGTGGGTCAGACAGGCCCCGGGAAAGGGTCTGGAATGGGTGGCCTCAGTGCGGTCCGGATCGGGTAGAACCTACT ACAGCGATAACGTGAAGGGCCGGTTCACGATCTCCCGCGACAACTCCAAGAACACCCTGTACTTGCAAATGAATAGCCTCAGGGCTGAGGATACCGCGGTCTACTACTGTGTGCGCTATGACCACTACACTGGAACTAGCGACTACTGGGCCAGGGGGACCCTCGTGACTGTGTCGTCC

[0336] SEQ ID NO: 49: h2831_VL (variable light chain) nucleotide sequence GATGTCGTGATGACTCAGTCACCTCTGTCCCTGCCTGTGACCCTTGGGGAACCCGCCTCGATCTCGTGCAAGAGCTCCCAGAGCCTGCTCGACTATGATGGAAAGACCTACCTGAACTGGTTGCTCCAAAGCCGGCCAGAGCCCCCAGAGGCTGATCTACCGCGTG ACCAACCGCGACACCGGGGTGCCGGACCGGTTCTCCGGATCCGGCAGCGGCACTGACTTCACCCTGAAAATTTCCAGAGTGGAAGCCGAGGACGTGGGAGTGTACTACTGTTGGCAGGGTACTCACTTTCCACGGTCCTTCGGTCAAGGAACCAAGGTCGAGATCAAG

[0337] SEQ ID NO: 50: h2926_VH (variable heavy chain) nucleotide sequence GAAGTGCAGCTCCTGGAGTCCGGGGGTGGACTGGTGCAGCCCGGGGGCAGCCTGAGGCTGAGCTGCGCCGCGTCAGGATTCACCTTCTCCAACTTCGGAATGTCCTGGGTCAGACAGGCCCGGGAAAGGGCCTTGAATGGGTGGCTAGCGTGCGCTCCGGTTCCGGACGGACCTACT ACTCGGACAACGTGAAGGGCCGGTTTACTATCTCCCGGGACAATTCGAAGAACACCCTGTACCTCCAAATGAACTCCTTGCGCGCCGAGGATACCGCAGTGTATTACTGCGTGCGCTACGACCACTACTCTGGCACTAGCGATTACTGGGCCAGGGAACTCTGGTCACCGTGTCGTCA

[0338] SEQ ID NO: 51: h2926_VL (variable light chain) nucleotide sequence GATGTCGTGATGACCCAGTCACCACTGTCCCTTCCTGTGACTCCCGGAGAACCGGCGTCCATTTCGTGCAAGAGCAGCCAGTCCCTGCTCGATTATGACGGAAAGACCTACCTGAACTGGTTGCTCCAAAAGCCTGGGCAGAGCCCCCAGAGACTGATCTACAAAGTGTCCAACAGGGACTCGGGCGTGCCGGACCGCTTCTCGGGGTCCGGTTCCGGTACCGACTTTACGCTGAAGATCTCACGGGTGGAAGCCGAGGACGTGGGAGTGTACTACTGTTGGCAGGGCACTCACTTCCCGCGGACCTTCGGACAAGGCACCAAGGTCGAGATCAAG

[0339] Sequence number 52: h4921G VH (Variable heavy chain) nucleotide sequence GAGGTGCAGCTGCTGGAGTCGGGGGGGGGACTCGTGCAGCCCGGGGGCTCCCTGAGACTCTCTTGTGCCGCCTCCGGCTTCACTTTTTCAAACTTCGGAATGTCCTGGGTCCGCCAAGCACCGGGAAAGGGTCTGGAATGGGTCGCCAGCGTGCGGTCCGGCGGCGGACGGACTTACTACTCCGACAACGTGAAGGGCCGGTTCACCATCTCAAGGGATAACTCCAAGAATACTCTGTACTTGCAAATGAACTCGCTGCGCGCTGAAGATACCGCGGTGTACTATTGCGTGCGCTACGACCACTACTCCGGTACCAGCGACTACTGGGGACAGGGAACCCTTGTGACCGTGTCGAGC

[0340] Sequence number 53: h4921G VL (Variable light chain) nucleotide sequence GATGTCGTGATGACTCAGTCGCCCCTCTCCCTGCCTGTGACTCTGGGGGAACCCGCGTCCATTTCGTGCAAGAGCAGCCAGTCCCTGTTGGACTCAGACGGAAAGACCTACCTTAACTGGCTGCTGCAAAAGCCAGGACAGAGCCCGCAGAGGCTGATCTACCGCGTG ACCAACCGGGATACGGGAGTGCCGGACAGATTCAGCGGCTCGGGTTCCGGCACCGACTTCACCCTCAAAATCTCCCGCGTCGAGGCCGAGGACGTGGGCGTGTATTACTGTTGGCAGGGAACCCACTTTCCTCGGACCTTCGGTCAAGGGACTAAGGTCGAAATCAAG

[0341] SEQ ID NO: 54: h2826 VH (variable heavy chain) nucleotide sequence GAAGTGCAGCTGCTGGAGTCTGGCGGCGGACTGGTGCAGCCCGGGGGATCCCTGCGGCTTTCCTGCGCCGCATCCGGCTTCACCTTTTCAAACTTCGGAATGTCGTGGGTCAGACAGGCCCCGGGAAAGGGTCTGGAATGGGTGGCCTCAGTGCGGTCCGGATCGGGTAGAACCTACT ACAGCGATAACGTGAAGGGCCGGTTCACGATCTCCCGCGACAACTCCAAGAACACCCTGTACTTGCAAATGAATAGCCTCAGGGCTGAGGATACCGCGGTCTACTACTGTGTGCGCTATGACCACTACACTGGAACTAGCGACTACTGGGCCAGGGGGACCCTCGTGACTGTGTCGTCC

[0342] SEQ ID NO: 55: h2826 VL (variable light chain) nucleotide sequence GATGTCGTGATGACCCAGTCACCACTGTCCCTTCCTGTGACTCCCGGAGAACCGGCGTCCATTTCGTGCAAGAGCAGCCAGTCCCTGCTCGATTATGACGGAAAGACCTACCTGAACTGGTTGCTCCAAAAGCCTGGCAGAGCCCCCAGAGACTGATCTACAAAGTG TCCAACAGGGACTCGGGCGTGCCGGACCGCTTCGGGGTCCGGTTCCGGTACCGACTTTACGCTGAAGATCTCACGGGTGGAAGCCGAGGACGTGGGAGTGTACTACTGTTGGCAGGGCACTCACTTCCCGCGGACCTTCGGACAAGGCACCAAGGTCGAGATCAAG

[0343] SEQ ID NO: 56: h2929 VH (variable heavy chain) nucleotide sequence GAAGTGCAGCTCCTGGAGTCCGGGGGTGGACTGGTGCAGCCCGGGGGCAGCCTGAGGCTGAGCTGCGCCGCGTCAGGATTCACCTTCTCCAACTTCGGAATGTCCTGGGTCAGACAGGCCCGGGAAAGGGCCTTGAATGGGTGGCTAGCGTGCGCTCCGGTTCCGGACGGACCTACT ACTCGGACAACGTGAAGGGCCGGTTTACTATCTCCCGGGACAATTCGAAGAACACCCTGTACCTCCAAATGAACTCCTTGCGCGCCGAGGATACCGCAGTGTATTACTGCGTGCGCTACGACCACTACTCTGGCACTAGCGATTACTGGGCCAGGGAACTCTGGTCACCGTGTCGTCA

[0344] SEQ ID NO: 57: h2929 VL (variable light chain) nucleotide sequence GATGTCGTGATGACCCAAAGCCCCCTGTCCCTCCCTGTGACTCCTGGAGAGCCGGCGTCCATTTCCTGCCGGTCAAGCCAGTCCTTGGTGGACTACGACGGAAAGACCTACCTCAACTGGCTGCTGCAGCGCCCCGGGCAGTCGCCGCAGCGGCTTATCTACAAAGTGTCCAACCGCGACTCGGGCGTGCCGGATAGGTTTTCGGGTTCCGGAAGCGGCACCGACTTCACCCTGAAAATCTCCAGAGTGGAAGCCGAGGACGTGGGAGTGTACTACTGTTGGCAGGGTTCTCACTTCCCACGGTCATATGGCCAAGGGACTAAGGTCGAAATCAAG

[0345] SEQ ID NO: 58: h3818G VH (Variable Heavy Chain) Nucleotide Sequence GAAGTGCAGCTCCTGGAGTCCGGCGGTGGACTGGTGCAGCCGGGCGGATCCCTGAGACTGTCCTGCGCCGCGTCGGGCTTTACTTTCGCAAATTACGGCATGAGCTGGGTCAGACAGGCCCCCGGGAAGGGTCTGGAATGGGTGGCCAGCGTCCGGAGCGGGGGATCCCGGACCTATTACTCCGACAACGTGAAGGGCCGCTTCACCATCTCAAGGGACAACTCCAAGAACACCCTGTACTTGCAAATGAACAGCCTTCGGGCTGAGGATACTGCCGTGTACTACTGCGTGCGCTACGACCACTACTCCGGATCCTCGGATTACTGGGGACAGGGAACCCTCGTGACCGTGTCATCG

[0346] SEQ ID NO: 59: h3818G VL (Variable Light Chain) Nucleotide Sequence GATGTCGTGATGACTCAGTCGCCCCTCTCCCTGCCTGTGACTCTGGGGGAACCCGCGTCCATTTCGTGCAAGAGCAGCCAGTCCCTGATGGACACCGACGAAAGACCTACCTTAACTGGCTGCTGCAAAAGCCAGGACAGAGCCCGCAGAGGCTGATCTACAAAGTG TCAAACCGGGAGTCCGGAGTGCCGGACAGATTCAGCGGCTCGGGTTCCGGCACCGACTTCACCCTCAAAATCTCCCGCGTCGAGGCCGAGGACGTGGGCGTGTATTACTGTTGGCAGGGAACCCACTTTCCTCGGACCTTCGGTCAAGGGACTAAGGTCGAAATCAAG

[0347] SEQ ID NO: 60: h2927 VH (variable heavy chain) nucleotide sequence GAAGTGCAGCTCCTGGAGTCCGGGGGTGGACTGGTGCAGCCCGGGGGCAGCCTGAGGCTGAGCTGCGCCGCGTCAGGATTCACCTTCTCCAACTTCGGAATGTCCTGGGTCAGACAGGCCCGGGAAAGGGCCTTGAATGGGTGGCTAGCGTGCGCTCCGGTTCCGGACGGACCTACT ACTCGGACAACGTGAAGGGCCGGTTTACTATCTCCCGGGACAATTCGAAGAACACCCTGTACCTCCAAATGAACTCCTTGCGCGCCGAGGATACCGCAGTGTATTACTGCGTGCGCTACGACCACTACTCTGGCACTAGCGATTACTGGGCCAGGGAACTCTGGTCACCGTGTCGTCA

[0348] SEQ ID NO: 61: h2927 VL (variable light chain) nucleotide sequence GATGTCGTGATGACTCAGTCACCGCTCTCCCTCCCTGTGACCCCGGGCGAACCAGCGTCGATCTCCTGCAAGAGCAGCCAATCATTGCTGGACTACGACGGAAAGACCTATCTTAACTGGCTGCTGCAGAAGCCCGGGCAGAGCCCGCAGGCCTGATCTACAAAGTG TCCAACAGAGACTCCGGAGTGCCTGATAGGTTCTCGGGTTCCGGCTCCGGTACCGACTTCACTCTGAAAATTTCCCGGGTGGAAGCCGAGGACGTGGGAGTGTACTACTGTTGGCAGGGCACCCACTTCCCCCGGTCGTTTGGACAAGGGACCAAGGTCGAGATCAAG

[0349] SEQ ID NO: 62: h49K3G VH (variable heavy chain) nucleotide sequence GAGGTGCAGCTGCTGGAGTCGGGGGGGGGACTCGTGCAGCCCGGGGGCTCCCTGAGACTCTCTTGTGCCGCCTCCGGCTTCACTTTTTCAAACTTCGGAATGTCCTGGGTCCGCCAAGCACCGGGAAAGGGTCTGGAATGGGTCGCCAGCGTGCGGTCCGGCGGCGGACGGACTTACT ACTCCGACAACGTGAAGGGCCGGTTCACCATCTCAAGGGATAACTCCAAGAATACTCTGTACTTGCAAATGAACTCGCTGCGCGCTGAAGATACCGCGGTGTACTATTGCGTGCGCTACGACCACTACTCCGGTACCAGCGACTACTGGGGACAGGGAACCCTTGTGACCGTGTCGAGC

[0350] SEQ ID NO: 63: h49K3G VL (variable light chain) nucleotide sequence GATGTCGTGATGACTCAGTCGCCCCTCTCCCTGCCTGTGACTCTGGGGGAACCCGCGTCCATTTCGTGCAAGAGCAGCCAGTCCCTGTTGGACTCAGACGGAAAGACCTACCTTAACTGGCTGCTGCAAAAGCCAGGACAGAGCCCGCAGAGGCTGATCTACAAAGTGTCAAACCGGGATTCCGGAGTGCCGGACAGATTCAGCGGCTCGGGTTCCGGCACCGACTTCACCCTCAAAATCTCCCGCGTCGAGGCCGAGGACGTGGGCGTGTATTACTGTTGGCAGGGAACCCACTTTCCTCGGACCTTCGGTCAAGGGACTAAGGTCGAAATCAAG

[0351] Accession number 64: h4917G VH (Variable heavy chain) nucleotide sequence GAGGTGCAGCTGCTGGAGTCGGGGGGGGGACTCGTGCAGCCCGGGGGCTCCCTGAGACTCTCTTGTGCCGCCTCCGGCTTCACTTTTTCAAACTTCGGAATGTCCTGGGTCCGCCAAGCACCGGGAAAGGGTCTGGAATGGGTCGCCAGCGTGCGGTCCGGCGGCGGACGGACTTACTACTCCGACAACGTGAAGGGCCGGTTCACCATCTCAAGGGATAACTCCAAGAATACTCTGTACTTGCAAATGAACTCGCTGCGCGCTGAAGATACCGCGGTGTACTATTGCGTGCGCTACGACCACTACTCCGGTACCAGCGACTACTGGGGACAGGGAACCCTTGTGACCGTGTCGAGC

[0352] Accession number 65: h4917G VL (Variable light chain) nucleotide sequence GATGTCGTGATGACTCAGTCGCCCCTCTCCCTGCCTGTGACTCTGGGGGAACCCGCGTCCATTTCGTGCAAGAGCAGCCAGTCCCTGTTGGACTCAGACGGAAAGACCTACCTTAACTGGCTGCTGCAAAAGCCAGGACAGAGCCCGCAGAGGCTGATCTACAAAGTG ACCAACCGGGAGTCCGGAGTGCCGGACAGATTCAGCGGCTCGGGTTCCGGCACCGACTTCACCCTCAAAATCTCCCGCGTCGAGGCCGAGGACGTGGGCGTGTATTACTGTTGGCAGGGAACCCACTTTCCTCGGTCATTCGGTCAAGGGACTAAGGTCGAAATCAAG

[0353] SEQ ID NO: 66: h2727 VH (variable heavy chain) nucleotide sequence GAAGTGCAGCTTCTGGAGAGCGGGGGCGGCCTGGTGCAGCCGGGCGGATCCCTGAGACTGTCCTGTGCCGCGTCCGGTTTTACCTTCTCCAACTACGGAATGTCATGGGTCCGCCAAGCACCCGGAAAGGGATTGGAATGGGTGGCTTCGATCCGGTCCGGCTCGGGACGGACCTACT ACTCCGATAACGTCAAGGGCAGATTCACTATTAGCCGGGACAACAGCAAGAATACCCTGTACCTCCAAATGAACTCCCTGAGGGCCGAGGACACCGCCGTGTATTACTGCGTGCGCTACGACCACTACTCGGGTTCCTCTGATTACTGGGGACAGGGGACCCTCGTGACTGTGTCAAGC

[0354] SEQ ID NO: 67: h2727 VL (variable light chain) nucleotide sequence GATGTCGTGATGACTCAGTCACCGCTCTCCCTCCCTGTGACCCCGGGCGAACCAGCGTCGATCTCCTGCAAGAGCAGCCAATCATTGCTGGACTACGACGGAAAGACCTATCTTAACTGGCTGCTGCAGAAGCCCGGGCAGAGCCCGCAGGCCTGATCTACAAAGTG TCCAACAGAGACTCCGGAGTGCCTGATAGGTTCTCGGGTTCCGGCTCCGGTACCGACTTCACTCTGAAAATTTCCCGGGTGGAAGCCGAGGACGTGGGAGTGTACTACTGTTGGCAGGGCACCCACTTCCCCCGGTCGTTTGGACAAGGGACCAAGGTCGAGATCAAG

[0355] SEQ ID NO: 68: h4918G VH (variable heavy chain) nucleotide sequence GAGGTGCAGCTGCTGGAGTCGGGGGGGGGACTCGTGCAGCCCGGGGGCTCCCTGAGACTCTCTTGTGCCGCCTCCGGCTTCACTTTTTCAAACTTCGGAATGTCCTGGGTCCGCCAAGCACCGGGAAAGGGTCTGGAATGGGTCGCCAGCGTGCGGTCCGGCGGCGGACGGACTTACT ACTCCGACAACGTGAAGGGCCGGTTCACCATCTCAAGGGATAACTCCAAGAATACTCTGTACTTGCAAATGAACTCGCTGCGCGCTGAAGATACCGCGGTGTACTATTGCGTGCGCTACGACCACTACTCCGGTACCAGCGACTACTGGGGACAGGGAACCCTTGTGACCGTGTCGAGC

[0356] SEQ ID NO: 69: h4918G VL (variable light chain) nucleotide sequence GATGTCGTGATGACTCAGTCGCCCCTCTCCCTGCCTGTGACTCTGGGGGAACCCGCGTCCATTTCGTGCAAGAGCAGCCAGTCCCTGATGGACACCGACGAAAGACCTACCTTAACTGGCTGCTGCAAAAGCCAGGACAGAGCCCGCAGAGGCTGATCTACAAAGTG TCAAACCGGGAGTCCGGAGTGCCGGACAGATTCAGCGGCTCGGGTTCCGGCACCGACTTCACCCTCAAAATCTCCCGCGTCGAGGCCGAGGACGTGGGCGTGTATTACTGTTGGCAGGGAACCCACTTTCCTCGGACCTTCGGTCAAGGGACTAAGGTCGAAATCAAG

[0357] SEQ ID NO: 70: Aducanumab heavy chain: QVQLVESGGGVVQPGRSLRLSCAASGFAFSSYGMHWVRQAPGKGLEWVAVIWFDGTKKYYTDSVKGRFTISRDNSKNTLYLQMNTLRAEDTAVYYCARDRGIGARRGPYYMDV WGKGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSC DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0358] SEQ ID NO: 71: Aducanumab light chain: DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0359] SEQ ID NO: 72: Bapineuzumab HC (heavy chain) EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYGMSWVRQAPGKGLEWVASIRSGGGRTYYSDNVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVRYDHYSGSSDYWGQG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0360] SEQ ID NO: 73: Bapineuzumab VH (variable heavy chain) EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYGMSWVRQAPGKGLEWVASIRSGGGRTYYSDNVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVRYDHYSGSSDYWGQGTLVTVSS

[0361] SEQ ID NO: 16: VH CDR1 GFTFSNYGMS

[0362] SEQ ID NO: 17: VH CDR2 SIRSGGGRTYYSNDYNVKG

[0363] SEQ ID NO: 18: VH CDR3 YDHYSGSSDY

[0364] SEQ ID NO: 77: Bapineuzumab LC (light chain) DVVMTQSPLSLPVTPGEPASISCKSSQSLLDSDGKTYLNWLLQKPGQSPQRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPRTFGQGTKV EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0365] SEQ ID NO: 78: Bapineuzumab VL (variable light chain) DVVMTQSPLSLPVTPGEPASISCKSSQSLLDSDGKTYLNWLLQKPGQSPQRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPRTFGQGTKVEIK

[0366] SEQ ID NO: 26: VL CDR1 KSSQSLLDSDGKTYLN

[0367] SEQ ID NO: 27: VL CDR2 LVSKLDS

[0368] SEQ ID NO: 28: VL CDR3 WQGTHFPRT

[0369] SEQ ID NO: 82: Gantenerumab HC amino acid sequence: QVELVESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAINASGTRTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGKGNTHKPYGYVRYFD VWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0370] SEQ ID NO: 83: Gantenerumab LC amino acid sequence: DIVLTQSPATLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGVPARFSGSGSGTDFTLTISSLEPEDFATYYCLQIYNMPITFGQGTKVEI KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0371] SEQ ID NO: 84: Amyloid beta (Aβ) 1-42: DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA

[0372] SEQ ID NO: 85: Amyloid beta (Aβ) precursor protein: MLPGLALLLLAAWTARALEVPTDGNAGLLAEPQIAMFCGRLNMHMNVQNGKWDSDPSGTKTCIDTKEGILQYCQEVYPELQITNVVEANQPVTIQNWCKRGRKQCKTHPHFVIPYRCLVGEFVSDALLVPDKCKFLHQERMDVCETHLHWHTVAKETCSEKSTNLHDYGMLLPCGIDKFRGVEFVCCPLAEE SDNVDSADAEEDDSDVWWGGADTDYADGSEDKVVEVAEEEEVAEVEEEEADDDEDDEDGDEVEEEAEEPYEEATERTTSIATTTTTTTESVEEVVREVCSEQAETGPCRAMISRWYFDVTEGKCAPFFYGGCGGNRNNFDTEEYCMAVCGSAMSQSLLKTTQEPLARDPVKLPTTAASTPDAVDKYLETPGDE NEHAHFQKAKERLEAKHRERMSQVMREWEEAERQAKNLPKADKKAVIQHFQEKVESLEQEAANERQQLVETHMARVEAMLNDRRRLALENYITALQAVPPRPRHVFNMLKKYVRAEQKDRQHTLKHFEHVRMVDPKKAAQIRSQVMTHLRVIYERMNQSLSLLYNVPAVAEEIQDEVDELLQKEQNYSDDVL ANMISEPRISYGNDALMPSLTETKTTVELLPVNGEFSLDDLQPWHSFGADSVPANTENENEVEPVDARPAADRGLTTRPGSGLTNIKTEEISEVKMDAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIATVIVITLVMLKKKQYTSIHHGVVEVDAAVTPEERHLSKMQQNGYENPTYKFFEQMQN

[0373] SEQ ID NO: 86: huIgG1 constant nucleotide sequence GCCAGCACTAAGGGGCCTAGCGTCTTTCCGCTGGCCCCGTCCTCCAAGTCCACTTCGGGTGGAACCGCGGCACTGGGGTGCCTCGTGAAGGACTACTTCCCCGAGCCGGTCACCGTGTCCTGGAACTCGGGAGCCCTGACCTCCGGAGTGCATACTTTCCCTGCGGTGCTGCAGTCCTCCGGGCTCTACTCGCTGTCAAGCGTGGTCACCGTCCCGAGCTCATCCCTGGGTACTCAGACCTACATTTGCAACGTGAACCACAAACCTTCCAACACCAAGGTCGACAAGAAAGTGGAGCCTAAGAGCTGCGACAAGACCCACACCTGTCCCCCGTGTCCCGCCCCTGAGCTGCTGGGCGGCCCCAGCGTGTTCCTCTTCCCGCCTAAGCCGAAGGACACTCTGATGATCTCGAGAACCCCTGAAGTGACCTGTGTGGTGGTGGATGTGTCCCACGAGGATCCGGAAGTGAAGTTCAATTGGTACGTGGACGGAGTGGAAGTCCATAACGCCAAGACCAAGCCCCGCGAGGAACAGTACAACTCAACTTACCGGGTGGTGTCAGTGCTGACCGTGCTGCACCAAGATTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTCTCCAACAAGGCGCTGCCGGCCCCCATTGAAAAGACCATCAGCAAGGCTAAGGGCCAGCCCCGGGAACCACAGGTCTACACCTTGCCCCCTTCCCGGGAGGAAATGACCAAGAACCAAGTGTCGCTGACGTGCCTGGTCAAGGGCTTTTATCCATCTGACATCGCCGTGGAGTGGGAAAGCAACGGCCAGCCGGAAAACAACTACAAGACTACCCCGCCTGTGCTGGACTCCGACGGCTCGTTCTTCCTGTATTCCAAGCTCACCGTGGATAAGTCCAGATGGCAGCAGGGCAATGTGTTCAGCTGCAGCGTGATGCATGAGGCCCTGCACAACCACTACACTCAGAAATCACTGTCCCTTTCCCCCGGAAAGTAA

[0374] SEQ ID NO: 87: hu kappa constant nucleotide sequence CGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAG AGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA

[0375] SEQ ID NO: 101: h2731 complete heavy chain amino acid sequence EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYGMSWVRQAPGKGLEWVASIRSGSGRTYYSDNVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVRYDHYSGSSDYWGQG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0376] SEQ ID NO: 102: h2731 complete light chain amino acid sequence DVVMTQSPLSLPVTLGEPASISCKSSQSLLDYDGKTYLNWLLQKPGQSPQRLIYRVTNRDTGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPRSFGQGTKV EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0377] SEQ ID NO: 103: h2731 complete heavy chain nucleotide sequence

[0378] Accession number 104: Complete light chain nucleotide acid sequence GATGTTGTGATGACCCAGTCCCCACTCTCTTTGCCCGTTACCCTTGGAGAACCTGCCTCCATCTCTTGCAAGTCAAGTCAGAGCCTCTTAGATTACGATGGAAAGACATATTTGAATTGGTTGCTGCAGAAGCCAGGCCAGTCTCCACAGCGCCTAATCTATCGGGTGACCAACCGGGACACTGGAGTCCCTGACAGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATGTGGGAGTTTATTATTGCTGGCAAGGCACACATTTTCCGCGCTCTTTCGGACAGGGGACCAAGGTGGAAATAAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGCTTAAGTCCGGAACTGCTAGCGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGAAACTCCCAGGAGAGCGTCACAGAGCAGGACAGCAAAGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT

Example

[0379] The following examples are included to illustrate the modes disclosed herein. Certain aspects of the following examples are described in terms of techniques and procedures discovered or contemplated by the inventors to work well in the embodiments disclosed herein. In light of this disclosure and the general level of skill in the art, those skilled in the art will recognize that the following examples are illustrative only, and that numerous changes, modifications, and variations can be employed without departing from the scope of the present disclosure.

[0380] As used in these experiments, "aducanumab" or "Adu" refers to the antibody having a heavy chain of SEQ ID NO: 70 and a light chain of SEQ ID NO: 71, and described in U.S. Patent Publication No. US2015 / 0315267 and PCT Publication No. WO2014 / 089500.

[0381] "BAN-2401" and "gantenerumab" as used in these experiments refer to the antibody having a heavy chain of SEQ ID NO: 82 and a light chain of SEQ ID NO: 83, as described, for example, in European Patent No. EP1960428B1.

[0382] In the following methods, antibody binding profiles against aggregated or fibrillar Aβ are characterized by ELISA, surface plasmon resonance (SPR), and immunohistochemistry (IHC). The ability to mediate phagocytic plaque clearance is assessed ex vivo using primary mouse microglia in APP / PS1 transgenic mouse brains and AD brains by immunofluorescence, ELISA, and MSD quantification, and neutralization of Aβ oligomer neuronal binding is assessed in rat primary hippocampal cultures.

[0383] Results presented herein: Compared to other N-terminal Aβ antibody therapies (bapineuzumab, aducanumab), the disclosed mAb demonstrated higher apparent affinity for aggregated and fibrillar Aβ in competitive or standard binding ELISAs. The improved avidity of the disclosed mAb for fibrillar Aβ was confirmed by SPR equilibrium binding kinetics, which demonstrated 5-11-fold higher avidity than aducanumab due to slower off-rate kinetics. IHC dose-response assessment in frozen human AD brain sections demonstrated higher apparent affinity and plaque area binding than aducanumab, regardless of the individual AD donor tissue tested. In ex vivo activity assays, the disclosed mAb significantly promoted microglial phagocytosis-mediated Aβ plaque reduction in APP / PS1 mouse tissue and concentration-dependently blocked the binding of soluble Aβ oligomers to rat primary neurons. In an ex vivo functional assay using human AD brains, the present mAb was shown to significantly promote the clearance of pyroglutamylated Aβ, a post-translationally modified component of senile plaques.

[0384] Example 1. Design of Aβ antibodies The Aβ antibody bapineuzumab (hBP) is a humanized antibody developed from the parent murine antibody 3D6. Herein, a multifaceted approach was applied to construct a superior antibody against hBP. The humanity of hBP was analyzed, and it was determined that humanization of the light chain could be optimized.

[0385] Protein sequences in the PDB database [Deshpande et al., 2005] were searched to find structures that could provide a rough structural model for hBP. The crystal structure of hBP fab PDB code 4HIX [Miles, et al., 2013] was utilized for both the Vh and Vk structures because it had acceptable resolution (2.2 Å) and an exact sequence match with hBP Vh and Vk, while retaining the same canonical structure for the loops.

[0386] IMGT / DomainGapAlignment was performed on the hBP VL as the input sequence. The human germline VK gene sequence IGHV2-30*02 best matches the hBP VL. The framework of hBP VL shares a high degree of sequence similarity with the corresponding framework region of IGHV2-30*02. Therefore, the framework region of IGHV2-30*02 VL was selected as a guidance sequence for further optimization of the hBP framework region. In addition, three residues in CDR-L2 that do not directly contact the antigen according to the hBP 3D structure were also changed to the germline sequence, resulting in the following changes: L50K, K53N, and L54R (Kabat).

[0387] Three different versions of VL were designed by incorporating human germline framework residues into the hBP VL sequence. No canonical or interface residues were changed. An alignment of the designed VK versions is shown in Figure 1.

[0388] Based on the structural observation that P15 is located at a turn and the germline gene has a Leu at this position, P15L was tested in one version of the variable light chain.

[0389] Based on the 3D structure observations, substitutions were designed at several residues in the CDRs and frameworks of the light and heavy chains. A total of 31 variant VL and VH versions of the light chain and 32 heavy chains were generated and tested for binding in the first round of rational design. Mutations that showed improved binding were combined in the second round of rational design. Additionally, new mutations guided by further structural analysis were also incorporated into the design.

[0390] Rational design-based mutagenesis was performed on the following positions within CDR-H1: T28, S30, N31, Y32, and G33 (Kabat). For CDR-H2 positions, I51, G53, G54, T57, S60, D61, and N62 (Kabat) were also mutated. CDR-H3 positions D96, H97, S99, S100a, and Y102 (Kabat) were subjected to rational mutagenesis.

[0391] For the variable light chain, multiple substitutions were attempted at CDR-L1 positions K24, L27c, D27d, and S27e (Kabat). Light chain CDR-L2 positions K53 and L54 (Kabat) were subjected to directed and limited mutagenesis. The CDR-L3 position was not subjected to substitutions.

[0392] Selected positions within the framework regions were also subjected to rational mutagenesis of the heavy and light chains.

[0393] Fifty-seven additional heavy chain and 33 light chain variants were designed and analyzed with the aid of Atum GPSpro software, which analyzes a database of human variable heavy and light chains and suggests query sequence-specific changes based on computer learning.

[0394] For the variable heavy domain, several substitutions were designed and analyzed at positions A24, S25, G26, F27, T28, F29, S30, N31, Y32, G33, and M34 (Kabat). Most of these positions were within CDR-H1. Similarly, many of the CDR-H2 residues were subjected to mutagenesis, including positions A49, S50, I51, R52, S52a, G53, G54, G55, R56, T57, Y58, Y59, S60, D61, N62, V63, and K64 (Kabat). Additionally, multiple substitutions of amino acids within CDR-H3 were made, for example, at positions V93, R94, Y95, D96, H97, Y98, S99, G100, S100a, S100b, D101, and Y102 (Kabat).

[0395] Multiple substitutions were also designed at variable light chain CDR-L1 positions K24, S25, S26, Q27, S27a, L27b, L27c, D27d, S27e, D28, G29, K30, T31, Y32, L33, and N34 (Kabat). For CDR-L2, mutagenesis was performed at positions L50, V51, S52, K53, L54, D55, and S56 (Kabat). Most of the CDR-L3 positions, e.g., Q90, G91, T92, H93, F94, P95, R96, and T97 (Kabat), were also rationally substituted with multiple amino acids.

[0396] All variant antibodies obtained from rational and GPSpro design were analyzed for expression, melting temperature (Tm), affinity, and avidity. Eight antibodies from the rational design and six from the computational training campaign were selected for further analysis based on the assays described above.

[0397] Example 2. IC by competitive ELISA assay 50 Determination of ratio. Using an assay based on the competition (inhibition) of binding of labeled antibodies to antigen-coated plates, the IC of antibodies of the disclosure 50 It was decided that:

[0398] To generate fibrils, Aβ1-42 polypeptide, previously treated with HFIP (hexafluoroisopropanol) and dried, was resuspended in DMSO to 5 mM and then further diluted to 100 μM with 10 mM HCl. The sample was incubated at 37°C for 24 hours and then centrifuged to separate soluble and fibrillar species. The pellet was resuspended to the original volume in 1x D-PBS and sonicated before use.

[0399] Plates were coated with 0.5 mg / ml fibrillar Aβ42 and blocked, e.g., with 1% BSA / PBS. Seven 3-fold dilutions of hBP starting at 150 μg / ml (final concentration 75 μg / ml) and four 3-fold dilutions of test antibodies starting at 20 μg / ml (final concentration 10 μg / ml) prepared in 0.1% BSA / PBS were added to wells in triplicate at 50 μl per well. 50 μl of hBP-biotin prepared in 0.1% BSA / PBS at 0.75 μg / ml (final concentration 0.35 μg / ml) was added to all wells, and the plates were incubated for 2 hours at room temperature and then washed three times with TTBS. Next, 100 μl of GE Streptavidin-HRP diluted 1 / 10,000 was added and incubated for 30 minutes. The plates were then washed six times with TTBS. Thermo Fisher o-phenylenediamine dihydrochloride (OPD) substrate was freshly prepared according to the manufacturer's instructions, and 100 μl was added per well. Reactions were incubated for 15 minutes, and the reaction was stopped with 50 μl of 2NH2SO4. Samples were read at 490 nM on a Spectromax. Figures 2, 3, and 4 show graphs of the competitive ELISA assays for 4918, 4917, 4921, 3818, 493, 2931, and bapineuzumab control (Figure 2), 2926, 2831, 2927, 2726, 2731, 2826, and bapineuzumab control (Figure 3), and 2727, 2929, and bapineuzumab control (Figure 4). The IC for each test antibody was 0.01. 50 IC for hBP 50 Divide by the half-maximal inhibitory concentration (IC 50 ) ratio is obtained. A ratio less than 1 indicates better performance than hBP. See Table 3A. [Table 3-1]

[0400] Example 3. Determining potency of monoclonal antibodies by competitive ELISA The binding potency of certain disclosed monoclonal antibodies and hBP was measured by their ability to compete with biotinylated bapineuzumab for binding to aggregated Aβ42 and assessed by competitive ELISA. 1 mg of Aβ42 was added to 1 ml of diH2O, vortexed vigorously, and placed on a nutator at room temperature for 48 hours. Plates were coated with 0.5 mg / ml of heterozygous Aβ42 aggregate mixture and blocked, e.g., with 1% BSA / PBS. Seven 3-fold dilutions of hBP starting at 150 μg / ml (75 μg / ml after dilution with hBP-biotin) and four 3-fold dilutions of test antibodies starting at 20 μg / ml (10 μg / ml after dilution with hBP-biotin) were added to triplicate wells at 50 μl per well. 50 μl of hBP-biotin at 0.75 μg / ml (0.35 μg / ml after dilution) was added to all wells, and the plate was incubated for 2 hours at room temperature and then washed three times with TTBS. Next, 100 μl of GE Streptavidin HRP diluted 1 / 10,000 was added and incubated for 30 minutes. The plate was washed six times with TTBS. Thermo Fisher o-phenylenediamine dihydrochloride (OPD) substrate was freshly prepared according to the manufacturer's instructions and 100 μl was added per well. The reaction was incubated for 15 minutes, and the reaction was stopped with 50 μl of 2NH2SO4. Samples were read at 490 nM on a Spectromax. Figure 5A shows the graph of the competitive ELISA assay for 2931, 2731, and the bapineuzumab control. Figure 5B shows the graph of the competitive ELISA assay for 2726, 2831, and the bapineuzumab control. Figure 20A shows graphs of the competitive ELISA assay for 2931, 2731, and the bapineuzumab control (data shown in Table 3B, columns 1-2). Figure 20B shows graphs of the competitive ELISA assay for 2831, 2726, and the bapineuzumab control (data shown in Table 3B, columns 4-5). For Figures 20A and 20B, the curves and resulting IC50 estimates represent nonlinear three-parameter least-squares fits of the data. Individual points are the means of triplicate samples (coefficient of variation <20%). [Table 3-2]

[0401] The results showed that antibodies 2931, 2731, 2726, and 2831 had higher potency than hBP, with IC values ​​approximately 2-4 times lower than hBP. 50 Indicates that the value is shown.

[0402] Example 4. Characterization of humanized mAb or Fab by BIAcore The binding properties of humanized antibodies or humanized antigen-binding fragments (Fab) were compared with those of recombinant Aβ. 1-42 For comparison with fibrils, analysis was performed using a BIAcore T200 (GE Life Sciences).

[0403] To generate fibrils, Aβ was pretreated with HFIP (hexafluoroisopropanol) and dried. 1-42 The polypeptides were resuspended in DMSO to 5 mM and then further diluted to 100 μM with 10 mM HCl. Samples were incubated at 37° C. for 24 hours and then centrifuged to separate soluble and fibrillar species. The pellets were resuspended to the original volume in D-PBS and sonicated before use.

[0404] Fibrils were immobilized on a sensor chip CM5 (GE Healthcare Life Sciences) via amine coupling to a level ensuring maximum binding of approximately 100 RU of analyte. Various concentrations of antibody or Fab (ranging from 1 nM to 100 nM) were passed over the coupled ligand at 30 μL / min in running buffer (HBS + 0.05% P-20, 1 mg / mL BSA) with an association time of 300 s and a dissociation time of 1200 s. Regeneration of the chip surface was achieved with two short injections of 10 mM glycine-HCl at pH 1.7. Data were blank-subtracted for both the sensor containing no ligand and a 0 nM analyte concentration. Analysis was performed using a global 1:1 fit with the bulk refractive index set to zero RU using BIAcore Insight Evaluation software (v2.0). Off-rate data (k diss, kd) are shown in Table 4 (Fab) and Table 6 (antibody).

[0405] Similarly, smaller dissociation constants can be seen for the Fabs and antibodies h2726, h2731, h2831, and h2931 compared to aducanumab, which showed a significantly larger dissociation constant. [Table 4]

[0406] Example 5. Characterization of apparent humanized mAb affinity by BIAcore Aβ 1-28 Determination of the binding affinity of anti-Aβ candidates to β-glucan (Bachem, Torrance, CA) was performed using a Biacore T200. Anti-human Fc antibodies were immobilized on a CM3 sensor chip (GE Healthcare Life Sciences) via amine coupling and used to capture Aβ antibodies.

[0407] Various concentrations of Aβ 1-28 Analyte (concentrations ranging from 100 nM to 0.39 nM, serially diluted 2-fold at each dilution step) was passed over the captured ligand in running buffer (HBS + 0.05% P-20, 1 mg / mL BSA) at 50 μl / min with an association time of 240 s and a dissociation time of 900 s. Data were blank-subtracted for both runs of an unrelated sensor containing no ligand and buffer containing 0 nM analyte. Analysis was performed using a global 1:1 fit with Biacore Evaluation software (v3.0).

[0408] The apparent dissociation constants (KD) are shown in Table 5, and the mAbs of the present disclosure bind to Aβ 1-28 The binding affinity to the monomer was 4 to 7 nM. The sensorgrams of binding at concentrations of 0.39 nM to 100 nM are shown in Figure 6A (h2726), Figure 6B (h2731), Figure 6C (h2831), and Figure 6D (h2931). [Table 5]

[0409] Example 6. Characterization of apparent humanized mAb affinity by BIAcore The binding properties of humanized antibodies were compared with those of recombinant Aβ 1-42 For comparison with fibrils, analysis was performed using a BIAcore T200.

[0410] To generate fibrils, Aβ was pretreated with HFIP (hexafluoroisopropanol) and dried. 1-42 The polypeptides were resuspended in DMSO to 5 mM and then further diluted to 100 μM with 10 mM HCl. Samples were incubated at 37° C. for 24 hours and then centrifuged to separate soluble and fibrillar species. The pellets were resuspended to the original volume in 1×D-PBS and sonicated before use.

[0411] Fibrils were immobilized on a Sensor Chip CM5 (GE Healthcare Life Sciences) via amine coupling to a level ensuring maximum binding of approximately 50 RU of analyte. Various concentrations of antibody (ranging from 0.411 nM to 100 nM) were passed over the coupled ligand at 30 μL / min in running buffer (HBS + 0.05% P-20, 1 mg / mL BSA) with a 300 s association time and a 1200 s dissociation time. Regeneration of the chip surface was achieved with two short injections of 10 mM glycine-HCl at pH 1.7. Data were blank-subtracted for both sensors containing no ligand and 0 nM analyte concentration. Analysis was performed using a global 1:1 fit with the bulk refractive index set to zero RU using BIAcore Insight Evaluation software (v2.0). Apparent dissociation constants (KD) are shown in Table 6, and a comparative sensorgram of binding at 100 nM is shown in Figure 7. [Table 6]

[0412] The enhanced relative avidity of the monoclonal antibodies of the present disclosure for fibrillar Aβ observed by ELISA was confirmed by SPR equilibrium binding kinetics (Table 6), which showed a 5-11-fold higher avidity (apparent KD) than aducanumab.

[0413] This is explained by the different kinetic binding profiles observed in the SPR sensorgrams (Figure 7): aducanumab binds to Aβ fibrils with a faster association rate (ka), while the much slower dissociation rate (kd) of the monoclonal antibodies of the present disclosure resulted in a higher measured avidity (i.e., a lower KD*) than aducanumab.

[0414] Example 7. Aβ fibril binding by ELISA Aβ of certain monoclonal antibodies of the present disclosure and aducanumab 1-42 and Aβ pE3-42 Direct binding to fibrils was assessed by ELISA. Aβ was pretreated with HFIP (hexafluoroisopropanol) and dried to generate fibrils. 1-42 or Aβ pE3-42 The polypeptides were resuspended in DMSO to 5 mM and then further diluted to 100 μM with 10 mM HCl. Samples were incubated at 37° C. for 24 hours and then centrifuged to separate soluble and fibrillar species. The pellets were resuspended to the original volume in 1×D-PBS and sonicated before use.

[0415] Aβ fibrils were coated overnight at room temperature at 1.0 μg / ml or 2.5 μg / ml in PBS. Plates were blocked with 1% BSA / PBS for 1 hour. Antibodies were serially diluted from 10 μg / ml to 4.8 ng / ml in 0.1% BSA-PBS and 0.1% Tween® 20, and 100 μl of each dilution was added in duplicate to each antibody and incubated for 2 hours at room temperature. Plates were washed four times with TBS / Tween® 20, and 100 μl of goat anti-human IgG HRP (Jackson ImmunoResearch Laboratories, Inc., West Grove, PA or Invitrogen, Carlsbad, CA) was added at a 1 / 5000 dilution to each well and incubated for 1 hour at room temperature. Plates were washed six times in TBS / Tween® 20, and Thermo Fisher o-phenylenediamine dihydrochloride (OPD) tablets and ThermoFisher substrate buffer were prepared according to the manufacturer's instructions. 100 μl of substrate was added and incubated for 15 minutes. The reaction was stopped with 50 μl of H2SO4. Plates were read at 490 nm on a Molecular Devices Spectromax. Figures 9A and 21. For Figure 21, the curves and resulting EC 50 The estimates represent a nonlinear three-parameter least-squares fit of the data (data shown in Table 7). [Table 7]

[0416] Plates were coated overnight at room temperature with dilutions of Aβ fibrils ranging from 10 μg / ml to 4.8 ng / ml in PBS. Plates were blocked with 1% BSA / PBS for 1 hour. Antibodies at 2 μg / ml in 0.1% BSA / PBS 0.1% Tween® 20 were added in duplicate to appropriate wells and incubated for 2 hours at room temperature. Plates were washed four times with TBS / Tween® 20, and then 100 μl of Jackson goat anti-human IgG HRP 1 / 5000 dilution was added to each well and incubated for 1 hour at room temperature. Plates were washed six times with TBS / Tween® 20, and Thermo Fisher o-phenylenediamine dihydrochloride (OPD) tablets and ThermoFisher substrate buffer were prepared according to the manufacturer's instructions. 100 μl of substrate was added and incubated for 15 minutes. The reaction was stopped with 50 μl of H2SO4. The plate was read at 490 nm on a Molecular Devices Spectromax. Figure 9B, right panel.

[0417] Antibodies h2726, h2731, h2831, and h2931 all showed strong affinity for fibrils, with the best and worst performing antibodies within 25% of each other. Furthermore, all four of these antibodies showed significantly higher avidity than aducanumab. Figure 21 shows a 3-fold increase in assay signal (OD490) and a 15- to 20-fold lower estimated EC 50 demonstrated increased overall binding and relative avidity of h2726, h2731, h2831, and h2931 mAbs against fibrillar Aβ compared to aducanumab.

[0418] Example 8. h2931 binding of Aβ oligomers by ELISA Direct binding of h2931 to Aβ oligomers was assessed by ELISA. To generate oligomers, lyophilized biotinylated and unlabeled Aβ (Bachem) were first solubilized at 1 mg / mL in 1,1,1,3,3,3-hexafluoroisopropanol (HFIP, Sigma), respectively. HFIP was allowed to evaporate from the samples overnight at room temperature in a fume hood. Aliquots were then centrifuged in a SpeedVac at room temperature, and all liquid was removed to generate 250 μg aliquots of HFIP films, which were stored at -80 °C until further use.

[0419] Oligomers were prepared by solubilizing 250 μg of biotinylated and unlabeled Aβ HFIP pellets in dry DMSO (Sigma) to a final concentration of 5 mM. For the unlabeled:biotinylated mixture, samples were combined in a 9:1 ratio (unlabeled:biotinylated) in a sterile 1.5 mL low-binding microcentrifuge tube (Axygen). The DMSO-solubilized sample was then diluted to 100 μM in cold phenol-free neurobasal medium (Invitrogen) and incubated at 4°C for 24 hours. After incubation, oligomers were separated from larger insoluble material via centrifugation at 14,000 g for 15 minutes. The top 90% of the supernatant was carefully removed, placed in a new sterile low-binding microcentrifuge tube, and stored on ice until use.

[0420] Each preparation, 2.5 μg / mL in PBS, was coated onto a Costar ELISA high-binding plate at 100 μl per well overnight at room temperature. The plate was aspirated, and then 200 μl of 1% BSA in PBS was added to each well and incubated for 1 hour at room temperature. h2931 mAb was prepared at a starting concentration of 10 μg / ml in 0.1% BSA / PBS 0.1% Tween® 20 buffer and serially diluted 7-fold (1:2 each time). Samples were incubated for 2 hours at room temperature. The plate was washed four times with TBS 0.1% Tween® 20. Goat anti-human (H+L) HRP (Jackson Immunoresearch, PA) was diluted 1 / 5000 in 0.1% BSA / PBS 0.1% Tween® 20, added at 100 μl / well, and incubated for 1 hour at room temperature. Plates were washed four times and o-phenylenediamine dihydrochloride tablets (ThermoFisher) were prepared according to the manufacturer's instructions. 100 μl was added per well and incubated at room temperature for 15 minutes. The reaction was stopped by adding 50 μl of H2SO4 and samples were read at 490 nM on a Molecular Devices SpectroMax. Curves and resulting EC 50 Estimates represent a nonlinear three-parameter least-squares fit of the data using GraphPad Prism software.

[0421] mAb h2931 was shown to bind soluble oligomers with high relative affinity and had an estimated EC 50 The concentration was 23 ng / mL, or 0.15 nM. Figure 8.

[0422] Example 9. Binding of anti-Aβ antibodies in AD brains Tissue Samples. Frozen human AD brain samples were obtained from Banner Sun Health Research Institute, Sun City, AZ. These tissues were derived from donors identified as having abundant Aβ pathology and classified according to the institution's Braak system (Table 8). Additionally, quality control was performed internally on all tissue blocks to confirm their pathology levels and distribution. [Table 8]

[0423] Tissue sectioning and fixation. Unfixed frozen brain tissue samples were embedded in Tissue-Tek OCT (Sakura Finetek) in cryomolds immersed in a mixture of 2-methylbutane and dry ice slurry (-60°C) and then stored at -80°C until sectioning. Serial 10-μm-thick frozen sections were prepared using a Leica 3050S cryostat. Sections were thaw-mounted directly onto positively charged glass slides and stored at -20°C until use. Prior to the immunohistochemistry (IHC) procedure, slides were immersed in 10% neutral buffered formalin solution for 10 minutes at 4°C, rinsed with PBS, and then incubated in glucose oxidase solution (20 mM beta-D(+) glucose, 2 mM sodium azide, and 2 units / mL glucose oxidase in 1x PBS) at 37°C for 1 hour. Slides were rinsed three times for 5 minutes in PBS and then transferred to a staining rack for processing in an automated stainer.

[0424] Antibody biotinylation. Humanized IgG antibodies were biotinylated using a non-covalent method by incubating with biotin-conjugated goat anti-human monovalent Fab fragments (Jackson ImmunoResearch) at a 1:4 ratio for 1 hour at room temperature. Excess unbound Fab was absorbed by preincubating with human serum for an additional hour before use. The freshly prepared antibodies were then loaded into the stainer and immediately applied to the tissue sections.

[0425] Immunostaining. Staining was performed in an automated Leica Bond Rx stainer (Leica Biosystems) using a Bond Research kit (DS980, Leica Biosystems) and an avidin-biotin-amplified immunoperoxidase detection system. Each biotinylated anti-Aβ antibody, or a human IgG control, was added to the sections at the specified concentrations for 1 hour, and staining was visualized using an avidin-biotin amplification system (ABC Elite Standard, PK-6100, Vector Laboratories). Nuclear hematoxylin counterstain was then applied to the sections, which were then dehydrated in an ascending alcohol series, removed in xylene, coverslipped, and air-dried.

[0426] Histology Imaging. Stained slides were digitally imaged using a Hamamatsu NanoZoomer 2.0HT slide scanner (Hamamatsu Corporation), and images were captured in .ndpi file format using NanoZoomer Digital Pathology software (NDP.scan, version 2.7.25). Images included in this report were captured directly from NDP.view and transferred without highlighting. For morphometry, digitized slides were analyzed using Halo software (V2.1.1537) to measure the percentage of stained tissue, and results were plotted using GraphPad Prism 8.

[0427] Results for h2726, h2731, h2831, h2931, and aducanumab. Four humanized anti-Aβ antibodies of the present disclosure, h2726, h2731, h2831, and h2931, as well as aducanumab, were added to all four AD brains at increasing concentrations: 0.03, 0.1, 0.3, 1, 3, and 9 μg / ml. As shown in Figure 10 (0.3 μg / ml), AD brain sections incubated with these antibodies displayed immunoreactive structures typical of Aβ pathology in AD. While AD13-75 and AD14-11 brains had a high density of Aβ plaques, lesions in AD11-97 and AD15-19 brains were relatively sparse. In each brain, the staining produced by the four antibodies h2726, h2731, h2831, and h2931 at a particular concentration was comparable in intensity and distribution. Aducanumab staining was the weakest across samples and concentrations. As illustrated in Figure 11, all four brain sections incubated with 1 μg / ml or 9 μg / ml of a control human IgG isotype were free of pathological staining.

[0428] The graphs in Figures 12 and 22 plot quantification of staining by the five antibodies in all four AD brains. Measurement of the percentage of tissue surface area occupied by stained pathology confirms that the four antibodies, h2726, h2731, h2831, and h2931, have similar binding levels in each AD brain at all concentrations tested. Correspondingly, the data in Table 9 show that the area under the curve and EC50 values ​​for the four antibodies remain comparable in each brain. Values ​​obtained with aducanumab were consistently low among AD brains throughout the concentration range tested.

[0429] Figure 22, notably, demonstrates that 10 mg / kg aducanumab resulted in greater plaque area binding (as a percentage of positively stained tissue) than aducanumab, at antibody concentrations estimated to be clinically relevant exposures in cerebrospinal fluid. Similar plaque area staining was observed at the highest concentration tested, suggesting saturation of binding at this level. [Table 9]

[0430] Results with bapineuzumab (hBP). Sections from brain AD13-75 were incubated with the humanized antibody hBP, as well as aducanumab and BAN2401 at increasing concentrations: 0.03, 0.1, 0.3, 1, 3, and 9 μg / ml. The level of staining with hBP increased in a dose-dependent manner, as seen with antibodies h2726, h2731, h2831, and h2931. Furthermore, as shown in Figure 13, hBP staining was more intense than aducanumab and BAN2401 at all concentrations tested.

[0431] Example 10. (Aβ 1-42 and Aβ pE3-42 ) Ex vivo phagocytosis assay to determine plaque clearance In the early stages of AD, microglial function is neuroprotective, acting not only to remove apoptotic cells and pathological protein aggregates but also to form a barrier around plaques, limiting their proliferation and the spread of synaptotoxic Aβ oligomers. Ex vivo phagocytosis assays quantify antibody-mediated microglial clearance responses.

[0432] Generation of primary microglial cultures: To dissect brain tissue from newborn mice, P1 pups are quickly decapitated with sterile scissors. Meninges are removed, and the forebrains are immediately immersed in 1–5 ml of dissection medium (e.g., high-glucose DMEM containing 20% ​​FBS, P / S) on ice until the desired number of pup brains have been dissected. To minimize cell damage, it is preferable to limit the total procedure time to less than 10 min.

[0433] Using a 22G needle followed by a 25G needle, the tissue was carefully aspirated twice consecutively using a new sterile pipette. The sample was centrifuged at 2,500 x g for 5 minutes at 4°C. The supernatant was carefully aspirated, and 5 ml of fresh growth medium (high-glucose DMEM, 10% FBS, P / S, and 25 ng / ml recombinant mouse GM-CSF) was added to the cell pellet. The cell pellet was pipetted up and down approximately 10 times using a 10 ml sterile pipette to separate the pellet.

[0434] A cell strainer (100 μm pores) was placed over a new 50 ml conical tube, and the material was dispensed through the cell strainer into the conical tube. The cell strainer was rinsed with 4–5 ml of fresh medium, followed by centrifugation at 200 × g for 5 minutes at 4°C.

[0435] Cells were plated at a density of two mouse brains per T-75 plastic culture flask. The supernatant was carefully aspirated, and 3 ml of fresh growth medium (high-glucose DMEM, 10% FBS, P / S, and 25 ng / ml recombinant mouse GM-CSF) was added to each cell pellet in a 10 ml sterile pellet. Resuspend by pipetting up and down 10 times using a 10 ml pipette. One sterile T-75 flask was prepared by adding 6 ml of growth medium (high-glucose DMEM, 10% FBS, P / S, and 25 ng / ml recombinant mouse granulocyte-monocyte colony-stimulating factor) to each flask, followed by the addition of 6 ml of resuspended cell pellet in a 37°C, 5% CO2 incubator to obtain a final volume of 12 ml.

[0436] The flasks were incubated for 5 days without disturbance to allow the cells to adhere. On day 5, the culture medium was replaced with 12 ml of fresh growth medium (high-glucose DMEM, 10% FBS, P / S, and 25 ng / ml recombinant mouse GM-CSF) in each flask. Approximately 10% of the plated mixed cells adhered and proliferated on the plastic surface. The medium was replaced twice weekly (every 3-4 days) to achieve confluence. These changes were made very carefully, without touching the bottom of the flask where the cells were attached.

[0437] After 7–11 days, flasks were rotated at 200 rpm for 2 h at 37°C using a Lab-line orbital shaker with a 19 mm orbit. The cell suspension was centrifuged at 200 × g and resuspended in assay medium (hybridoma-serum-free medium H-SFM [Life Technologies] + 1% FBS, glutamine, P / S, and 5 ng / ml recombinant mouse GM-CSF).

[0438] Ex vivo assay. Cryostat sections (10 μm thick, using a wide blade) of APP / PS1 mouse or human AD brain (postmortem interval, less than 3 h) were "thaw-mounted" onto polylysine-coated circular glass coverslips and placed in wells of a 24-well tissue culture plate (CT-30C OT-20C). Tissue samples could be warmed between sections using a thumb or by lowering the OT to -12°C. Coverslips were washed twice with assay medium. Antibodies (control or against Aβ) were added at 2x concentrations in assay medium (final 20 μg / ml) in a volume of 250 μl for 1 h in a tissue culture incubator.

[0439] Microglial cells were then seeded in 250 μl of assay medium at a final density of 800,000 cells / ml (1,600,000 cells / ml stock). Cultures were maintained in a humidified incubator at 37°C in an atmosphere of 5% CO for 72 hours.

[0440] Total Aβ (Aβ 1-42) Quantification of Aβ levels. The medium was carefully aspirated and subsequently washed with ice-cold PBS. 100 μl of 8 M urea was added, and the tissue was resuspended by pipetting and scraped down with the pipette tip. The suspension was then frozen at -20°C until ready for analysis. The suspension was thawed on ice and centrifuged at 16,000 × g for 20 minutes at 4°C before being diluted and analyzed using the V-PLEX Total Aβ42 Peptide (4G8) Kit (Meso Scale Discovery). The results are shown in Figures 14A, 14B, and 24. Figures 14A and 24 show Aβ levels per brain section, and Figure 14B shows the same data as a scatter plot by treatment (data from Figure 14B are shown in Table 10, and data from Figure 24 are shown in Table 11). h2731, h2931, and aducanumab demonstrated highly significant reductions in Aβ plaque species relative to the isotype control. [Table 10] [Table 11]

[0441] Pyroglutamic acid-3Aβ (Aβ pE3-42 ) Quantification of N-terminally truncated pyroglutamic acid-modified Aβ (e.g., Aβ pE3-42 ) has been described as a component of mature senile plaques in AD brains (Saido et al., Neuron 14, 1995). It was unclear whether pyroglutamic acid modification of N-terminal Aβ affects the binding of N-terminal antibodies such as h2731 and the other antibodies described herein. Similarly, it was unclear whether these antibodies inhibit Aβ. pE3-42 It was unclear whether IL-1 has the ability to promote phagocytosis-mediated clearance of IL-1.

[0442] The presence of pyroglutamic acid-3Aβ in the AD brains used in the ex vivo experiments and its similar staining pattern compared to h2931 were confirmed by immunohistochemistry (Figures 25A and 25B). To demonstrate the clearance of pyroglutamic acid-3Aβ, a commercially available ELISA method was used to measure its clearance during ex vivo phagocytosis. The suspension collected according to the above method was thawed on ice and centrifuged at 16,000 × g for 20 minutes at 4 °C, then diluted and analyzed using a commercially available ELISA kit (Amyloid Beta N3pE Aβ, IBL America). Aβ pE3-42 The ELISA assay detects unmodified Aβ 1-42 Compared to Aβ pE3-42 It is highly specific for (data not shown).

[0443] The results are shown in Figures 26A and 26B (data are shown in Tables 12 and 13, respectively). These results show the levels of pyroglutamate-3Aβ in brain sections after treatment with the indicated antibody, h2931 in Figure 26A and h2731 in Figure 26B. Each was compared to a healthy control and to an AD brain treated with an IgG1 isotype control. Sections from different AD brains were used for each treatment. Both h2731 and h2931 show highly significant reductions in pyroglutamate-3Aβ relative to the isotype control.

[0444] Figures 24 and 26B taken together show that when the anti-Aβ antibody of the present invention (e.g., h2731) was incubated with primary mouse microglia on brain tissue sections from AD patients, it inhibited Aβ 1-42 and Aβ pE3-42 These results demonstrate the efficacy of these antibodies in the treatment of Aβ in human pathological settings. 1-42 and Aβ pE3-42 It has been shown to remove both

[0445] N-terminally targeted anti-Aβ antibodies promoted significant microglia-mediated clearance of Aβ plaque species, including pyroglutamate-modified Aβ, in brain tissue from AD patients. These data support further development of the antibodies of this invention as subcutaneous antibody immunotherapy for Alzheimer's disease. [Table 12] [Table 13]

[0446] Example 11. Blocking oligomers in hippocampal binding assays Aβ binding assay in rat hippocampal neurons

[0447] E18 primary rat hippocampal neurons were cultured as described by Zago et al. (J. Neurosci. 22 February 2012, 32(8)2696-2702). Soluble Aβ was preincubated with or without antibodies on days 14-21 of culture to block neurite binding to the primary neurons.

[0448] Fresh unlabeled biotinylated or (9:1) unlabeled:biotinylated soluble Aβ was prepared one day in advance and incubated overnight at 4° C. Before use, Aβ was spun down at 14,000 RPM for 15 minutes.

[0449] Dilutions of Aβ and antibodies were prepared in NeuroBasal-phenol red-free (NB-NPR) or NbActiv4-NPR medium at half the final volume (2x the final treatment concentration). After combining, the mixture was mixed 3-4 times and then pre-incubated at 37°C for 30 minutes.

[0450] Immediately before the binding assay, neurons were rinsed with pre-warmed NB-NPR (150 µL / well). The buffer was aspirated, and then 60 µL / well of antibody / Aβ treatment was added to the cells. The cells were then incubated at 37 °C under normal incubator conditions (5% CO2, 9% O2) for 30–40 min.

[0451] Neurons were washed twice with 150 μL / well of NB-NPR and then fixed with 4% paraformaldehyde in 1× DPBS for 20 min at room temperature.

[0452] Cells were permeabilized with 0.1% Triton® X-100 in 1×DPBS for 5 min and then blocked with 10% normal goat serum (NGS) for 1 h at room temperature (RT).

[0453] Samples were incubated with microtubule-associated protein 2 (MAP2) and neuronal nucleus protein (NeuN) primary antibodies in 100 μL / well of 1x DPBS containing 1% BSA and 1% NGS overnight at 4°C. The next day, samples were rinsed twice with 150 μL / well of 1x DPBS for 5 minutes each. Secondary antibodies were added in 100 μL / well of 1x DPBS, 1% BSA, and 1% NGS for 1 hour at room temperature.

[0454] High-content imaging (HCI) analysis was performed to quantify soluble Aβ neurite-associated spots using an Operetta HCI CLS instrument (Perkin Elmer; modified Neurote Outgrowth algorithm; 40x HO objective; 25–40 fields per well in a microplate format, n = 3 per condition). MAP2 and NeuN neuronal markers were used to trace each neurite tree and count the number of cell bodies per optical field (e.g., microtubule-associated protein 2 (Abcam; Cambridge, UK) and NeuN (EMD Millipore) primary antibodies, followed by AlexaFluor (Thermo Fisher Scientific) secondary detection antibodies). Neuritic Aβ spots were visualized using various monoclonal and polyclonal Aβ antibodies (e.g., mouse monoclonal anti-Aβ antibody MabN254 (EMD Millipore)), followed by AlexaFluor (Thermo Fisher Scientific) secondary detection antibodies). Detection was performed using a secondary detection antibody from Biosciences Scientific or streptavidin-AF488 for biotinylated Aβ material. Figures 15A and 15B show that increasing concentrations of anti-Aβ antibody reduced the number of spots per neuron, indicating activity against Aβ. Figure 23 shows that h2731 effectively blocked the binding of soluble Aβ aggregates to rat hippocampal synapses (Aβ42 spots per neuron) in a concentration-dependent manner. The effect of h2731 was detected at a mAb:Aβ42 molar ratio as low as 1:500 (p<0.05), reaching >90% block of binding at a 1:50 molar ratio (p<0.001) compared to Aβ42 alone (no mAb preincubation). Data are shown in Table 14. [Table 14]

[0455] Example 12. Anti-Aβ antibodies that bind to native and modified Aβ species Cryostat sections of human AD brains were thaw-mounted onto poly-D-lysine-coated coverslips, placed in 24-well tissue culture plates, and incubated with the test antibodies for 1 hour at 37°C and 5% CO2. Primary mouse microglial cells were then seeded at 800,000 cells / ml, and the cultures were maintained at 37°C and 5% CO2 for 72 hours. The medium was carefully aspirated, and the sections were washed with PBS. The sections were resuspended in 8 M urea and analyzed for Aβ. pE3-42 (Immuno-Biological Laboratories, Minneapolis, MN) by ELISA or for Aβ 1-42 Aβ was quantified by MSD (Meso Scale Diagnostics, Rockland, MD). pE3-42 The ELISA kit specifically detects the pE3-42 species, which has no detectable signal with full-length Aβ.

[0456] Figure 27 shows that h2731 binds with high apparent affinity to the N-terminus of full-length Aβ, but not to pyroglutamic acid-modified Aβ (Aβ pE3-42 h2731 was shown to not directly bind to α-glucan. 50 ) and unmodified N-terminus (Aβ 1-42 h2731 bound to fibrillar Aβ species with up to 100 ng / ml of Aβ pE3-42 showed no detectable binding to

[0457] Example 13. In vitro phagocytosis-mediated clearance - Aβ 1-42 THP-1 human monocyte-mediated uptake of protofibrils Aβ containing the S26C mutation 1-42Synthetic protofibrils were produced as described by Paranjape et al., ACS Chem. Neurosci. 2012, 3, 302-311. Briefly, Aβ peptides were dissolved at 1 mM in 100% hexafluoroisopropanol (HFIP) (Sigma-Aldrich, St. Louis, MO), aliquoted into sterile microcentrifuge tubes, and allowed to evaporate uncapped overnight at room temperature in a fume hood. The next day, the aliquots were vacuum centrifuged to remove residual HFIP and stored in a desiccant at -20 °C. Some Aβ peptides were treated with 100% trifluoroacetic acid and vacuum centrifuged prior to HFIP treatment. Aβ oligomers and fibrils obtained directly from the lyophilized aliquots were prepared by resuspending the lyophilized Aβ peptide aliquots in sterile anhydrous dimethyl sulfoxide (DMSO) (Sigma-Aldrich, St. Louis, MO) at 5 mM. For oligomer preparations, samples were diluted to 100 μM in sterile, ice-cold, phenol red-free Ham's F-12 cell culture medium with L-glutamine (F-12, Bioworld, Dublin, OH) and incubated for 24 h at 4 °C. For fibril preparations, samples were diluted to 100 μM with 10 mM HCl and incubated for 24 h at 37 °C. The Aβ concentration in these preparations was based on dry peptide weight.

[0458] Mature protofibrils were conjugated to pHrodo Red maleimide (Thermo Fisher) and then used in an in vitro phagocytosis-mediated clearance assay.

[0459] Antibodies at concentrations of 6.25, 3.13, 1.56, 0.78, 0.39, 0.20, 0.098, and 0.049 μg / ml were added to pHrodo-Aβ 1-42 After preincubation with protofibrils for 30 min at room temperature, THP-1 phagocytic cells were added. After 3 h of incubation at 37°C and 5% CO, antibody-mediated phagocytosis-mediated clearance was assessed by measuring the cellular pHrodo signal via flow cytometry.

[0460] As shown in Figures 28A and 28B, the anti-Aβ antibody inhibited Aβ in a concentration-dependent manner. 1-42 These results show that the antibodies of the present invention inhibit Aβ in brain tissue. 1-42 This suggests that it may be possible to drive the clearance of

[0461] Example 14. Distribution of total Aβ and pyroglutamic acid-modified Aβ in brain tissue from patients with advanced AD The ex vivo IHC method described above and herein was performed on AD brain tissue and Aβ 1-XX (detected with N-terminal anti-Aβ antibody) and anti-Aβ pE3-42 The distribution of was determined.

[0462] Aβ 1-XX and Aβ pE3-42 Evaluation of Aβ confirmed the widespread distribution of both species in tissues from patients with advanced AD. pE3-42 Aβ compared to 1-XX The distribution pattern (Figures 29A(1) and 29A(2) (and enlarged views 29B(1) and 29B(2) respectively)) and quantification (Figure 29C) of the area percent covered by Aβ pE3-42 This was consistent with previous studies suggesting that N-terminal Aβ antibodies target a relatively small pool of modified Aβ mixed with unmodified Aβ. pE3-42 are shown in Figures 29A(2) and 29B(2), and intact N-terminal Aβ is shown in Figures 29A(1) and 29B(1). pE3-42 The antibody is Aβ 1-42 did not cross-react with (data not shown).

[0463] The boxes in Figure 29A(1) and Figure 29B(1) show intact N-terminal Aβ and modified Aβ proximal to blood vessels. pE3-42 Table 15 below reports the quantification of staining in the plaques in Figures 29B(1) and 29B(2), which is presented graphically in Figure 29C. The difference between the means is statistically significant (p=0.007, paired two-tailed t-test). [Table 15]

[0464] Example 15. Anti-Aβ antibody h2731 co-localizes with AβpE3-42 in AD brains h2731 immunostaining and Aβ pE3-42 Colocalization of Aβ was assessed by immunofluorescence microscopy. An N-terminal anti-Aβ antibody (in this case, h2731) was pre-conjugated to a Cy3-secondary anti-human antibody (Jackson Laboratories) and then applied to the tissue. pE3-42 , mouse anti-Aβ pE3-42 Antibodies were detected using a 488-AlexaFluor-conjugated anti-mouse secondary antibody. Slides were imaged using a Metamorph-assisted IX81 Olympus microscope connected to a Hamamatsu camera (C10600-10B).

[0465] FIG. 30 (Panel A) shows the localization of h2731 to Aβ plaques, and FIG. 30 (Panel B) shows the localization of anti-Aβ pE3-42 Showing the localization of antibody signals, Figure 30 (Panel C) shows the localization of h2731 and anti-Aβ to Aβ plaques. pE3-42 Colocalization of antibody signals is shown, with overlapping signals more prominent in the dense core region of the plaque.

[0466] Example 16. The anti-Aβ antibodies of the present invention inhibit Aβ ex vivo from AD brain tissue in a dose-dependent manner with greater efficacy than aducanumab. pE3-42 Promotes clearance. Using the methods described above and elsewhere herein, Aβ was isolated from AD brain tissue. pE3-42 The ability of aducanumab and antibodies of the invention (e.g., h2731) to clear proteins was evaluated.

[0467] A physiologically relevant dose-response series of h2731 (3 ng / ml, 10 ng / ml, 30 ng / ml, and 100 ng / ml) was incubated with brain tissue slices from AD patients and primary mouse microglia for 72 hours. pE3-42The results are presented in Table 16 below and Figure 31A. [Table 16]

[0468] h2731 inhibited the microglial phagocytosis of Aβ from brain tissue slices of AD patients in a concentration-dependent manner and within a relatively short incubation period (72 hours). pE3-42 Therefore, the antibody of the present invention potently promotes the clearance of Aβ from the brains of AD patients at a concentration range expected to be achieved by subcutaneous administration. pE3-42 promotes ex vivo clearance of

[0469] Another series of experiments was performed comparing h2731 at 25 ng / ml and 75 ng / ml with aducanumab at 25 ng / ml and 225 ng / ml. The results are presented in Table 17 and Figure 31B. [Table 17]

[0470] h2731 demonstrated superior Aβ inhibition compared to aducanumab, even at a 9-fold lower concentration pE3-42 It showed clearance activity.

[0471] Another physiologically relevant dose-response series of h2731 and aducanumab (3 ng / ml, 25 ng / ml, and 225 ng / ml) was incubated with brain tissue slices from AD patients and primary mouse microglia for 72 hours (both compared to an IgG1 isotype control). Both h2731 and aducanumab concentration-dependently downregulated Aβ pE3-42 h2731 promoted the clearance of β-amyloid, but again h2731 promoted it significantly more potently, with a p-value of <0.0001 at a concentration 9-fold lower than the concentration required for aducanumab to reach a p-value of 0.0005. The results are presented in Table 18 below and Figure 32A. [Table 18]

[0472] To verify that h2731-mediated ex vivo phagocytic activity is microglia-dependent, we performed a + / - microglia experiment. Microglia alone phagocytose Aβ from AD patient tissue sections. pE3-42 Although h2731 drives some clearance, clearance is significantly more robust in combination with h2731 and microglia. h2731 appears to require the presence of microglia for clearance activity, as h2731 alone shows no activity without microglia. The results are presented in Table 19 and Figure 32B. [Table 19]

[0473] Antibody concentrations tested were based on estimated CNS ranges at 0.1% of steady-state plasma minimum and maximum concentrations from modeled pharmacokinetics following monthly administration of 3 mg / kg subcutaneous h2731 (25-75 ng / ml) or 10 mg / kg intravenous aducanumab (25-225 ng / ml) in humans (Figure 33).

[0474] The antibodies of the present invention can inhibit Aβ from the brains of AD patients with a concentration range expected to be achieved by subcutaneous administration and with higher biological activity than aducanumab. pE3-42 promotes ex vivo clearance of

[0475] Antibody h2731 inhibits Aβ in AD brains pE3-42 Figure 34 shows the Aβ pE3-42 (Staining indicated by white arrows) was observed in plaques (white triangles) and associated with blood vessels (circles in Figures 34A and 34C) in AD brains treated with a human IgG isotype control antibody (Figures 34A and 34B). Treatment with h2731 reduced microglia-mediated Aβ production, as evidenced by the reduction in plaques. pE3-42 The antibodies of the present invention, as exemplified by h2731, enhanced the reduction of Aβ levels in tissues (Figures 34C and 34D). pE3-42Reduces plaque containing

[0476] Example 17. Target engagement of h2731 Female APP×PS1 mice expressing mutant human amyloid precursor protein (hAPP[V717I]) and mutant human presenilin 1 (hPS1[A246E]) were used to evaluate the ability of h2731 and aducanumab to cross the blood-brain barrier and bind to amyloid beta (Aβ) plaques in the brain after peripheral administration. The average age of the animals at the start of the study was 6.7 months. The day before drug administration, all animals received an injection of anti-CD4 antibody (20 mg / kg, intravenous) to prevent the formation of anti-drug antibodies in mice treated with h2731 or aducanumab, both fully humanized antibodies. h2731 (3 or 10 mg / kg, subcutaneous, SC) or aducanumab (10 mg / kg, intravenous) was administered weekly for 3 weeks, after which the animals were euthanized 1 week later. After transcardial perfusion with ice-cold saline, brains were removed from mice, snap-frozen in 2-methylbutane on dry ice, and stored at −80°C.

[0477] Serial sagittal 10-μm-thick cryosections were prepared using a Leica 3050S cryostat. Sections were thaw-mounted directly onto positively charged glass slides and stored at -20°C until use. Prior to IHC, slides were immersed in 10% neutral buffered formalin solution for 10 minutes at 4°C, rinsed with PBS, and then incubated in glucose oxidase solution (20 mM beta-D(+) glucose, 2 mM sodium azide, and 2 units / mL glucose oxidase in 1x PBS) at 37°C for 1 hour. Slides were rinsed three times for 5 minutes in PBS before being transferred to a staining rack for processing on an automated stainer. Biotin SP-conjugated goat anti-human IgG (H+L) (Jackson ImmunoResearch Laboratories #109-065-088) was used to detect h2731 or aducanumab in APP×PS1 brain tissue. Staining was performed using a Bond Research kit (DS980, Leica Biosystems) in an automated Leica Bond Rx stainer (Leica Biosystems). Nuclear hematoxylin counterstain was then applied to the sections, which were then dehydrated in an ascending alcohol series, cleared in xylene, cover-slipped, and air-dried. All sections were imaged using a NanoZoomer 2.0HT slide scanner (Hamamatsu Corporation, Japan). Morphometric analysis of the digitized images was performed using Halo software (V2.1.1537). After delineating the cerebral cortex as the region of interest, the percentage of stained tissue area was determined. The data are presented in Table 20. [Table 20]

[0478] A reduction in the number or size of Aβ plaques in Alzheimer's disease may correlate with a slowing or reversal of disease progression. The ability of the anti-Aβ antibodies of the present invention to bind to and clear Aβ in vivo after peripheral administration supports the potential utility of these antibodies as therapeutic agents.

[0479] Thus, the antibodies of the present invention inhibit microglia-mediated Aβ agonism in brain tissue from AD patients. 1-42 Although the antibodies of the invention may not directly target pyroglutamate modifications, they inhibit Aβ with greater potency and greater bioactivity than aducanumab at concentrations predicted to be clinically relevant, as exemplified by h2731. pE3-42 These antibodies can effectively remove pyroglutamic acid species. Clearance of pyroglutamic acid species by these antibodies may be due to the ability of microglia to recognize opsonized plaques and engulf large particles with diverse content. Therefore, the antibodies of the present invention may remove other neurotoxic elements co-deposited in plaques by this same mechanism.

[0480] Example 18. Reduction of amyloid plaques in patients To reduce amyloid plaques, which are associated with inhibiting, reducing, and / or reversing symptoms of Alzheimer's disease, a pharmaceutically effective amount of an anti-Aβ antibody (or antigen-binding fragment thereof) is administered to the patient (e.g., one or more of the antibodies described in the Examples above).

[0481] Patients: Individuals suspected of or diagnosed as having an amyloid plaque-associated disease, such as Alzheimer's disease, are selected for treatment with anti-Aβ antibodies.

[0482] Treatment: Patients receive 70 mg of the anti-Aβ antibody h2931 subcutaneously approximately once every four weeks. Patients receive 200 mg of the anti-Aβ antibody h2931 subcutaneously approximately once every four weeks.

[0483] Plaque reduction and amelioration of Alzheimer's disease symptoms can be measured as described herein.

[0484] Patients with Alzheimer's disease treated with 70 mg or 200 mg of the anti-Aβ antibodies h2731, h2726, h2831, or h2931 administered subcutaneously approximately once every four weeks are treated with a reduction in amyloid plaque burden as measured by PET imaging.

[0485] Example 19. Phase 1 Single Ascending Dose Study to Evaluate the Safety, Tolerability, Immunogenicity, and Pharmacokinetics of h2731 This example describes a Phase 1, randomized, double-blind, placebo-controlled, single ascending dose (SAD) study to evaluate the safety, tolerability, and immunogenicity of h2731 at doses of 70 mg and 200 mg. The purpose of this study was to characterize the plasma pharmacokinetic (PK) profile of h2731 in healthy volunteers (HVs) and patients with AD, specifically in AD patients with parenchymal amyloid burden confirmed by molecular imaging. AD subjects were required to meet the National Institute on Aging and Alzheimer's Association (NIA-AA) study criteria and guidelines for AD (McKhann, 2011) or mild cognitive impairment (MCI) due to AD (Albert, 2011).

[0486] Rationale for the test Preclinical studies in transgenic mice that produce excess Aβ have shown that antibodies targeting the N-terminus of Aβ can enter the brain and reduce amyloid deposition in brain tissue and the cerebral vasculature (Bard, 2000). Clinical evidence indicates that monoclonal antibodies directed against N-terminal amyloid can clear and reduce Aβ aggregate deposition from the brain and alleviate cognitive decline (Sevigny, 2016; Swanson, 2021; Aduhelm USPI, 2021).

[0487] As shown in the example above, preclinical studies have shown that h2731 rapidly and robustly removes Aβ plaques by enhancing microglia-mediated clearance mechanisms. These data suggest that h2731 may slow clinical decline in patients with AD. The 70 mg and 200 mg doses for the Phase 1 clinical trial are based on CNS fraction occupancy modeled from predicted clinical exposure at these doses.

[0488] Test Purpose The primary objectives of this study are to evaluate the safety and tolerability of h2731 when administered as a single dose, including (1) general safety, tolerability, and immunogenicity in all subjects, and (2) target-related safety and tolerability in subjects with confirmed parenchymal amyloid.

[0489] Secondary objectives of this study are to characterize the PK profile of h2731 following SC administration as a single dose and the cerebrospinal fluid (CSF) PK profile of h2731.

[0490] Study design Figure 35 is a schematic diagram of the study design. The study includes at least two dose cohorts of subjects with biologically confirmed AD. The study also includes two healthy volunteer ("HV") cohorts. These four cohorts include: AD Cohort 1 (70 mg), AD Cohort 2 (200 mg), HV Cohort 1 (70 mg), and AD Cohort 2 (200 mg). Each of these will be administered subcutaneously ("SC") at their respective doses.

[0491] After initial site observation, subjects will have four follow-up visits over approximately 12 weeks, during which time safety assessments and PK collection will be completed. Selected cohorts may undergo additional CSF collection by lumbar puncture on days 3 and 29.

[0492] Evaluation items Primary endpoints include: Adverse event ("AE") reporting (incidence of AEs, SAEs, and h2731-related AEs), safety and tolerability based on ECG, clinical laboratory tests, vital signs, and physical examination Immunogenicity measured by confirming the presence of ADA in plasma Amyloid-related imaging abnormalities (ARIA-H and ARIA-E) and other urgent radiological findings

[0493] Secondary endpoints include: Plasma PK of h2731 ·CSF PK in h2731 C of h2731 at each sampling time obs

[0494] Selection Criteria Each cohort will contain approximately 8 subjects with a body mass index (BMI) between 18.0 and 32.0 kg / m2.

[0495] AD subjects will be selected according to inclusion criteria, including: (a) have a definite or probable diagnosis of AD based on either demonstrable AD with evidence of AD pathophysiological process according to the National Institute on Aging and Alzheimer's Disease Association (NIA-AA) criteria (McKhann et al., Alzheimers Dement., 7(3):264-9, 2011) or a high probability of AD according to the NIA-AA criteria (Albert et al., Alzheimers Dement., 7(3):270-79, 2011); (b) have a gradual and progressive change in memory function over 6 months as reported by the subject or study partner; (c) Mini-Mental State Examination (MMSE) score ≥ 18 at screening, and (d) Evidence of AD pathological process confirmed by amyloid PET scan.

[0496] Exclusion criteria Subjects will be selected based on the exclusion criteria, including the following: Subjects must not meet any of the exclusion criteria, including the following criteria: (a) coagulation disorders (prothrombin time 1.2 × ULN) or other blood clotting disorders; (b) severe and clinically significant (persistent neurological deficit or structural brain injury) central nervous system (CNS) trauma (e.g., cerebral contusion), history of epilepsy; (c) Contraindications to MRI examination, including claustrophobia, the presence of contraindicated metal (ferromagnetic) implants, or cardiac pacemakers. (d) Anticoagulants within 3 months of screening (not scheduled to be initiated before randomization) or a prolonged history of bleeding after minor trauma. Note: Low-dose aspirin is permitted (maximum 162 mg / day). (e) History or presence of posterior reversible encephalopathy syndrome (PRES) (Fugate, et al., Posterior reversible encephalopathy syndrome: clinical and radiological manifestations, pathophysiology, and outstanding questions. Lancet Neurol. 2015;14(9):914-25.)

[0497] Clinical laboratory evaluation Perform laboratory analyses of hematology, clinical chemistry, coagulation, urinalysis plasma, biomarkers, and CSF.Perform central and local pregnancy testing.

[0498] Brain MRIs will be locally interpreted and scans will be submitted to a centralized MRI vendor to finalize MRI eligibility and provide central evaluation of baseline ARIA findings. MRIs must be performed using a 1.5- or 3.0-T scanner, and the same scanner will be used for each subject throughout the study. The first MRI will be performed during the screening period as a baseline measurement to confirm eligibility criteria based on structural brain imaging. MRI scans will include, but are not limited to, the following sequences: T2-weighted FLAIR, two-dimensional (2D) T2*-weighted gradient echo (GRE), or susceptibility-weighted imaging (SWI): diffusion-weighted, three-dimensional (3D) T1-weighted GRE. MRI scans will be assessed and interpreted by a central MRI reader, who will provide diagnostic interpretation and evaluation of MRI outcome measures. MRI data (Day 29 MRI data from all subjects and any other available MRI and safety data) will be available to the DSMB to confirm the h2731 dose.

[0499] If CSF analysis is performed, the cohort will undergo two LPs: one on Day 3 (48 hours after study drug administration) and a second on Day 29. CSF will be analyzed to determine h2731 levels. CSF samples with clear evidence of blood contamination should not be used for PK evaluation. CSF analysis will also include standard analyses including, but not limited to, blood pressure, color, glucose, protein, lactate, red blood cells, and white blood cells.

[0500] Amyloid PET imaging is used to biologically confirm the diagnosis of AD by providing evidence of the presence of pathological features of neurofibrillary tangles comprising beta-amyloid. Isotopically labeled compounds with high affinity for aggregated forms of beta-amyloid (tracers) can provide in vivo evidence of beta-amyloid. Three radioligands are being used for screening purposes: [18F]florbetapir / AV45 (Amyvid), [18F]flutemetamol (Vizamyl), and [18F]florbetaben (Neuraceq). Subjects in cohorts 1-4 will undergo amyloid PET imaging to biologically confirm the diagnosis of AD. Positive PET scans using [18F]florbetapir / AV45, [18F]flutemetamol, or [18F]florbetaben obtained outside the study protocol within 18 months prior to the first screening visit will be acceptable for confirming patient inclusion criteria with central reader review.

[0501] APOE4 status (e.g., APOE4 / APOE4, APOE4 / APOE3, APOE3 / APOE3, APOE4 / APOE2, APOE3 / APOE2) will be determined by central assessment for subjects enrolled in cohorts 1–4.

[0502] Additional evaluation The Cogstate CBB (Maruff, 2013) will be administered to subjects at the first screening visit (days -72 to -8). The CBB is a brief (approximately 15-minute) computer-based cognitive test battery designed to measure memory, working memory, psychomotor function, and attention. The CBB has been shown to be a sensitive tool for detecting AD-related cognitive decline in healthy older adults and adults with amnestic mild cognitive impairment (MCI) (Darby, 2002; Lim, 2013), as well as for cognitive improvement resulting from treatment with cognition-enhancing medications (Davison, 2011; Jaeger, 2011; Nathan, 2013).

[0503] The MMSE (Folstein, 1975) will be administered to subjects at the first screening visit (days -72 to -8) prior to the Cogstate CBB assessment to determine whether the subject meets the inclusion criteria for cognitive impairment.

[0504] Subjects will undergo plasma sampling for biomarkers of AD pathology, including but not limited to Aβ42 / 40, and biomarkers associated with tau pathology, including but not limited to total tau, p181-tau, and p217-tau.

[0505] Subjects will undergo PK sampling for plasma and CSF h2731.

[0506] Plasma anti-h2731 antibody levels are measured (antibodies are detected by electrochemiluminescence assay (ECLIA)).

[0507] ARIA Assessment: A screening MRI scan will be used to exclude subjects with pre-existing vasogenic edema (ARIA-E), more than four microhemorrhages, or more than one superficial hemosiderosis (ARIA-H). In addition to scheduled MRIs, unscheduled MRIs may be performed at the investigator's discretion if ARIA is suspected based on symptomatic features. MRIs will be scheduled for all subjects prior to study drug administration (baseline) and for AD subjects at the Day 29 and Day 85 visits (28 and 84 days post-dose, respectively) to assess, classify, and document radiographic evidence of ARIA.

[0508] Example 20: Phase 1 Multiple Ascending Dose Study to Evaluate the Safety, Tolerability, Immunogenicity, Pharmacokinetics, and Pharmacodynamics of h2721 in Subjects with Alzheimer's Disease This example describes a Phase 1, randomized, double-blind, placebo-controlled, multiple ascending dose (MAD) study to evaluate the safety, tolerability, and immunogenicity, PK, and pharmacodynamic (PD) effects of h2731 in patients with AD. Figure 36 is a schematic diagram of the study design.

[0509] Test Purpose As discussed in more detail below, the primary objective of this study is to evaluate the safety, tolerability, and immunogenicity of h2731 after multiple SC administration. Secondary objectives of this study are to characterize the PK profile of h2731 after multiple SC administration, characterize the plasma and CSF PK profiles of h2731 after multiple SC administration, and evaluate the PD effect of h2731 on cerebral amyloid plaque deposition after multiple SC administration. Exploratory objectives of this study are to evaluate the PD effect of h2731 on blood and CSF biomarkers after multiple SC administration and to evaluate ARIA findings by apolipoprotein E4 (APOE4) status.

[0510] Study population The study consists of two parts, each testing a different group of subjects: Group A, AD subjects who are heterozygous or non-carriers of the apolipoprotein E4 (APOE4) allele (referred to as the non-homozygous population), and Group B, AD subjects who are homozygous for APOE4 (referred to as the homozygous population). The dose levels of h2731 evaluated are the same in the non-homozygous (Group A) and homozygous (Group B) populations.

[0511] Apolipoprotein E (APOE) genotype status has been demonstrated to influence not only the development of AD (Corder, 1993; van Duijn, 1994) but also the rate of ARIA after treatment with anti-Aβ antibodies (Arrighi, 2016; Ketter, 2017; Muralidharan, 2022). The onset of ARIA is dose-dependent, with most events occurring within the first few months of anti-Aβ treatment (Muralidharan, 2022). AD patients with one copy of the APOE4 allele may be at higher risk for anti-Aβ antibody-mediated ARIA compared with APOE4 non-carriers, although the risk demonstrated across clinical trials has been somewhat inconsistent. However, APOE4 homozygous (having two alleles) patients consistently demonstrate a higher incidence of ARIA compared with both APOE4 heterozygous and non-carrier patients.

[0512] Rationale for dose selection The proposed 45 mg, 70 mg, and 200 mg doses are based on an analysis of estimated target engagement (fraction occupancy [fOcc]) levels resulting from simulating clinical drug exposure levels at these doses, balancing the prediction of effective exposure with the potential to induce ARIA. These doses are further supported by the results of completed nonclinical toxicology studies with repeated dose administration for up to 3 months and confirmed based on certain information from the single ascending dose (SAD) study in Example 19. · All safety and tolerability data through Day 15 for selected cohorts dosed at 70 mg and / or 200 mg; PK data through Day 29 for selected cohorts receiving 70 mg and / or 200 mg (approximately 8 subjects per cohort; 6 received h2731 and 2 received placebo); and Day 29 MRI results from selected cohorts (approximately 12 subjects; 9 received h2731 and 3 received placebo) who received 70 mg and / or 200 mg.

[0513] Study design This Phase 1, randomized, double-blind, placebo-controlled, multiple ascending dose study will be conducted in two dose cohorts in subjects with biologically confirmed AD to evaluate the safety, tolerability, immunogenicity, PK, and PD of h2731.

[0514] The study consists of two parts, each evaluating a different group of subjects: Group A, AD subjects who are heterozygous or non-carriers of the APOE4 allele, and Group B, AD subjects who are homozygous for APOE4. The three dose cohorts for each group are listed in Table 21. [Table 21]

[0515] AD subjects who are APOE4 heterozygous or non-carriers of the APOE4 allele are assigned to group A, and subjects who are homozygous for APOE4 are assigned to group B.

[0516] For each cohort in Group A, approximately 32 subjects will be randomly assigned to h2731 or placebo in a 3:1 ratio: approximately 24 subjects will receive h2731 and approximately 8 subjects will receive placebo. Randomization will be stratified by APOE4 carrier status (APOE4 heterozygous or APOE4 non-carrier).

[0517] For each cohort in Arm B, approximately 12 subjects will be randomly assigned to h2731 or placebo in a 3:1 ratio: approximately 9 subjects will receive h2731 and approximately 3 subjects will receive placebo.

[0518] Treatment duration Study drug (45 mg, 70 mg, or 200 mg) will be administered every 4 weeks, starting on Day 1, for a total of up to 6 doses. Subjects will receive the first dose of study drug (h2731 or placebo) subcutaneously on Day 1. Subjects will undergo safety assessments, including adverse event (AE) monitoring, clinical laboratory tests, vital signs, physical examination, and electrocardiogram (ECG), as well as blood collection for PK, anti-drug antibody (ADA), and biomarker (BM) analyses. Subjects will be discharged from the study site 8 hours after dosing after completing scheduled post-dose assessments.

[0519] Dosing will continue every 4 weeks. Safety assessments and blood sampling for PK, ADA, and BM analyses will be completed. Centrally reviewed MRI findings must be reviewed prior to dosing to assess for the presence of ARIA.

[0520] After the final dose of study drug, subjects will return to the study site for a Week 24 (Day 169) visit and complete an End of Treatment (EOT) visit, which includes safety assessments, blood collection for PK, ADA, and BM analyses, and amyloid PET imaging assessment.

[0521] Subjects participating in optional CSF collection will be scheduled for CSF collection within 1-5 days after the imaging visit (MRI and PET) at Week 24 (Day 169). Subjects may be discharged after a 4-hour observation period.

[0522] Dose escalation / dose titration The dose level for each cohort will be determined by a limited number of unblinded sponsor representatives based on review and interpretation of all available safety, tolerability, PD, and PK information for h2731.

[0523] Safety and tolerability data will be evaluated on an ongoing and periodic basis throughout this study and the SAD study in Example 19. Safety and tolerability data will be evaluated and recommendations will be provided regarding the decision to enroll in a cohort if minimum data requirements are met.

[0524] Subject-level administration interruptions Dosage will depend on the presence and severity of amyloid-related imaging findings related to underlying vasogenic edema (ARIA-E) or hemorrhage (ARIA-H) observed before each dose of h2731 or placebo, based on MRI findings assessed by a central MRI reader, and potential symptoms of ARIA reported by the subject or observed by the investigator.

[0525] MRI visits will be scheduled at least 7 days before each dosing visit. The severity classification of ARIA based on radiographic findings is summarized in Table 22. MRI results should be reviewed prior to dosing. [Table 22]

[0526] Any intracerebral hemorrhage larger than 1 cm is considered radiologically severe.

[0527] Based on either ARIA-E clinical severity or ARIA-E radiological severity, administration can be interrupted based on moderate and / or severe ARIA-E findings. With the exception of asymptomatic mild ARIA-H, any new findings of ARIA-H (microhemorrhages or superficial hemosiderosis) will lead to interruption of administration of h2731 to the subject.

[0528] If dosing is interrupted, the scheduled dose should be omitted entirely, and subsequent visits should continue as scheduled. If the subject is able to resume treatment (Section 7.11.3), dosing of h2731 or placebo should be resumed at the same dose at the next scheduled dose. Do not replace a skipped dose.

[0529] Evaluation items Primary endpoints include: Safety and tolerability based on AE reporting (incidence of AEs, SAEs, and H2731-related AEs), vital signs, physical and neurological examinations, 12-lead ECG, and laboratory tests Injection site reactions as assessed by the investigator The nature, frequency, severity, and timing of MRI findings of ARIA, including the incidence of symptomatic ARIA-E and / or ARIA-H; the incidence of isolated ARIA-E (ARIA-E only, no ARIA-H) and isolated ARIA-H (ARIA-H only...

Claims

1. A method of treating Alzheimer's disease in a subject, comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks.

2. A method for reducing amyloid plaques in a subject, comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks.

3. A method for converting a subject from amyloid-positive to amyloid-negative, comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks.

4. 4. The method of claim 1, wherein the anti-amyloid beta antibody or antigen-binding fragment thereof binds to an epitope located within the N-terminus of the Aβ peptide, and the epitope comprises at least one amino acid selected from amino acids 1 to 10 of the Aβ peptide.

5. The method of claim 4, wherein the anti-amyloid β antibody or antigen-binding fragment thereof binds to an epitope comprising at least one amino acid selected from amino acids 1 to 7 of the Aβ peptide.

6. The anti-amyloid beta antibody or antigen-binding fragment thereof binds to amyloid beta with an apparent KD of about 5 nM or less. １－４２ The method of any one of claims 1 to 5, wherein the method binds to protofibrils.

7. The anti-amyloid beta antibody or antigen-binding fragment thereof binds to amyloid beta with an apparent KD of about 1 nM or less. １－４２ The method of any one of claims 1 to 6, wherein the method binds to protofibrils.

8. The anti-amyloid beta antibody or antigen-binding fragment thereof binds to amyloid beta with an apparent KD of about 10 nM or less. １－２８ The method according to any one of claims 1 to 7, wherein the monomer is bound to the polymer.

9. The method of any one of claims 1 to 8, comprising administering about 20 mg to about 100 mg of the anti-amyloid β antibody or antigen-binding fragment thereof.

10. The method of any one of claims 1 to 8, comprising administering about 100 mg to about 200 mg of the anti-amyloid β antibody or antigen-binding fragment thereof.

11. The method of any one of claims 1 to 8, comprising administering about 45 mg of the anti-amyloid beta antibody or antigen-binding fragment thereof.

12. The method of any one of claims 1 to 8, comprising administering about 70 mg of the anti-amyloid beta antibody or antigen-binding fragment thereof.

13. The method of any one of claims 1 to 8, comprising administering about 200 mg of the anti-amyloid beta antibody or antigen-binding fragment thereof.

14. The method of any one of claims 1 to 13, wherein the anti-amyloid beta antibody or antigen-binding fragment thereof is administered as a pharmaceutical composition comprising the anti-amyloid beta antibody or antigen-binding fragment thereof and a pharmaceutically acceptable diluent.

15. The method of any one of claims 1 to 14, wherein the anti-amyloid beta antibody is administered approximately once every four weeks.

16. The method of any one of claims 1 to 15, wherein the administration is intravenous or subcutaneous.

17. 17. The method of claim 16, wherein the administration is subcutaneous.

18. the anti-amyloid beta antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3, and a light chain variable region comprising light chain CDR1, CDR2, and CDR3; heavy chain CDR1 comprises the amino acid sequence of one of SEQ ID NOs: 16, 19, or 20; the heavy chain CDR2 comprises the amino acid sequence of one of SEQ ID NOs: 20, 21, 22, or 23; the heavy chain CDR3 comprises the amino acid sequence of one of SEQ ID NOs: 18, 24, or 25; the light chain CDR1 comprises the amino acid sequence of one of SEQ ID NOs: 26, 29, 31, or 32; the light chain CDR2 comprises the amino acid sequence of one of SEQ ID NOs: 33, 34, 35, or 36; The method of any one of claims 1 to 17, wherein the light chain CDR3 comprises the amino acid sequence of one of SEQ ID NOs: 28, 38, or 39.

19. the anti-amyloid β antibody or antigen-binding fragment thereof a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 16; a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 20; a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 18; a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 29; a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 34; and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 38; 20. The method of claim 18, comprising:

20. 20. The method of claim 19, wherein the heavy chain variable region, excluding the CDRs, is at least 95% identical to the amino acid sequence of SEQ ID NO:3, and the light chain variable region, excluding the CDRs, is at least 95% identical to the amino acid sequence of SEQ ID NO:

9.

21. 20. The method of claim 19, wherein the heavy chain variable region, excluding the CDRs, is at least 98% identical to the amino acid sequence of SEQ ID NO:3, and the light chain variable region, excluding the CDRs, is at least 98% identical to the amino acid sequence of SEQ ID NO:

9.

22. 21. The method of claim 20, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 3 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:

9.

23. 23. The method of claim 22, wherein the heavy chain variable region consists of the amino acid sequence of SEQ ID NO: 3 and the light chain variable region consists of the amino acid sequence of SEQ ID NO:

9.

24. The method according to any one of claims 1 to 23, wherein the anti-amyloid β antibody is a humanized IgG1.

25. The method of any one of claims 1 to 24, wherein the anti-amyloid β antibody is a complete antibody, a chimeric antibody, a CDR-grafted antibody, or a recombinant antibody.

26. The method of any one of claims 1 to 25, wherein the anti-amyloid beta antibody or antigen-binding fragment thereof further comprises a heavy chain constant region comprising an amino acid sequence that is at least 95% identical to SEQ ID NO: 40 and / or a light chain constant region comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:

41.

27. The method of any one of claims 1 to 26, wherein the anti-amyloid beta antibody or antigen-binding fragment thereof further comprises a heavy chain constant region comprising an amino acid sequence that is at least 98% identical to SEQ ID NO: 40 and / or a light chain constant region comprising an amino acid sequence that is at least 98% identical to SEQ ID NO:

41.

28. The method of any one of claims 1 to 27, wherein the anti-amyloid beta antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 40, with or without the C-terminal lysine, and a light chain constant region comprising the amino acid sequence of SEQ ID NO:

41.

29. The method of any one of claims 1 to 28, wherein the anti-amyloid beta antibody comprises a heavy chain constant region consisting essentially of the amino acid sequence of SEQ ID NO: 40, with or without the C-terminal lysine, and a light chain constant region consisting essentially of the amino acid sequence of SEQ ID NO:

41.

30. 30. The method of any one of claims 1 to 29, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without the C-terminal lysine, and a light chain of SEQ ID NO:

102.

31. The method according to any one of claims 1 to 30, wherein the anti-amyloid beta antibody is h2731.

32. 1. A method of treating Alzheimer's disease in a subject, the method comprising subcutaneously administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody about once every four weeks, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

33. A method of treating Alzheimer's disease in a subject, the method comprising subcutaneously administering to the subject about 45 mg of an anti-amyloid beta antibody about once every four weeks, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

34. A method of treating Alzheimer's disease in a subject, the method comprising subcutaneously administering to the subject about 70 mg of an anti-amyloid beta antibody about once every four weeks, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

35. A method of treating Alzheimer's disease in a subject, the method comprising subcutaneously administering to the subject about 200 mg of an anti-amyloid beta antibody about once every four weeks, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

36. 1. A method for reducing amyloid plaques in a subject, the method comprising subcutaneously administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody about once every four weeks, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

37. A method for reducing amyloid plaques in a subject, the method comprising subcutaneously administering to the subject about 45 mg of an anti-amyloid beta antibody approximately once every four weeks, the anti-amyloid beta antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

38. A method for reducing amyloid plaques in a subject, the method comprising subcutaneously administering to the subject about 70 mg of an anti-amyloid beta antibody approximately once every four weeks, the anti-amyloid beta antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

39. A method for reducing amyloid plaques in a subject, the method comprising subcutaneously administering to the subject about 200 mg of an anti-amyloid beta antibody about once every four weeks, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

40. 1. A method of treating Alzheimer's disease in a subject, comprising administering to the subject about 20 μg / mL to about 40 μg / mL of C ａｖｅ the method comprising subcutaneously administering a dose of an anti-Aβ antibody sufficient to obtain a C-terminal lysine level, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

41. 1. A method of treating Alzheimer's disease in a subject, comprising administering to the subject an AUC of about 15,000 hr*ug / mL to about 30,000 hr*ug / mL. ０－ｔａｕ the method comprising subcutaneously administering a dose of an anti-Aβ antibody sufficient to obtain a C-terminal lysine level, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

42. The maximum concentration (C) of the anti-amyloid beta antibody or antigen-binding fragment thereof in the subject over an administration interval. ｍａｘ 42. The method of any one of claims 1 to 41, wherein the concentration of erythritol in the blood is about 30 μg / mL to about 60 μg / mL.

43. C of the anti-amyloid β antibody or antigen-binding fragment thereof in the subject ｍａｘ 43. The method of any one of claims 1 to 42, wherein the value is from about 50 μg / mL to about 60 μg / mL.

44. C of the anti-amyloid β antibody or antigen-binding fragment thereof in the subject ｍａｘ 44. The method of any one of claims 1 to 43, wherein the value does not exceed about 60 μg / mL.

45. Said C ｍａｘ The value is serum C ｍａｘ The method according to any one of claims 42 to 44, wherein the value is

46. Said C ｍａｘ The value is plasma C ｍａｘ The method according to any one of claims 42 to 44, wherein the value is

47. The average concentration (C) of the anti-amyloid beta antibody or antigen-binding fragment thereof in the subject over the administration interval. ａｖｅ 47. The method of any one of claims 1 to 46, wherein the IL-10 value is from about 20 μg / mL to about 40 μg / mL.

48. C of the anti-amyloid β antibody or antigen-binding fragment thereof in the subject ａｖｅ 48. The method of any one of claims 1 to 47, wherein the value is from about 30 μg / mL to about 40 μg / mL.

49. C of the anti-amyloid β antibody or antigen-binding fragment thereof in the subject ａｖｅ 48. The method of any one of claims 1 to 47, wherein the value does not exceed about 40 μg / mL.

50. Said C ａｖｅ The value is serum C ａｖｅ 50. The method according to any one of claims 40 and 47 to 49, wherein the value is

51. Said C ａｖｅ The value is plasma C ａｖｅ 50. The method according to any one of claims 40 and 47 to 49, wherein the value is

52. The area under the concentration-time curve (AUC) for the administration interval of the anti-amyloid β antibody or antigen-binding fragment thereof in the subject ０－ｔａｕ 52. The method of any one of claims 1 to 51, wherein the HR value is from about 15,000 hr*ug / mL to about 30,000 hr*ug / mL.

53. AUC of the anti-amyloid beta antibody or antigen-binding fragment thereof in the subject ０－ｔａｕ 53. The method of any one of claims 1 to 52, wherein the value is from about 20,000 hr*ug / mL to about 30,000 hr*ug / mL.

54. AUC of the anti-amyloid beta antibody or antigen-binding fragment thereof in the subject ０－ｔａｕ 54. The method of any one of claims 1 to 53, wherein the value is a dose not exceeding about 30,000 hr*ug / mL.

55. The AUC ０－ｔａｕ The value is the serum AUC ０－ｔａｕ 55. The method according to any one of claims 41 and 52 to 54, wherein the value is

56. The AUC ０－ｔａｕ The value is the plasma AUC ０－ｔａｕ 55. The method according to any one of claims 41 and 52 to 54, wherein the value is

57. 57. The method of any one of claims 1 to 56, wherein amyloid plaques are reduced in the subject.

58. 58. The method of any one of claims 2, 3, and 57, wherein the reduction in cerebral amyloid beta plaques comprises a reduction of at least about 30 centiloids to about 70 centiloids.

59. 58. The method of any one of claims 3, 3, and 57, wherein the reduction in cerebral amyloid beta plaques comprises a reduction of about 45 centiloids to about 80 centiloids.

60. 58. The method of any one of claims 2, 3, and 57, wherein the reduction in cerebral amyloid beta plaques comprises a reduction of about 50 centiloids to about 85 centiloids.

61. 58. The method of any one of claims 2, 3, and 57, wherein the reduction in cerebral amyloid beta plaques comprises a reduction of at least about 40% to about 90%.

62. 58. The method of any one of claims 2, 3, and 57, wherein the reduction in cerebral amyloid beta plaques comprises a reduction of about 60% to about 100%.

63. 58. The method of any one of claims 2, 3, and 57, wherein the reduction in cerebral amyloid beta plaques comprises a reduction of about 65% to about 100%.

64. 64. The method of any one of claims 2, 3, and 57-63, wherein the reduction in cerebral amyloid plaques comprises a reduction compared to baseline.

65. 65. The method of any one of claims 2, 3, and 57-64, wherein the reduction in cerebral amyloid beta plaques comprises a reduction compared to the subject before administration of the anti-amyloid beta antibody.

66. 66. The method of any one of claims 2, 3, and 57-65, wherein reduction in cerebral amyloid beta plaques is achieved after 6 months of treatment.

67. 66. The method of any one of claims 2, 3, and 57-65, wherein reduction in cerebral amyloid beta plaques is achieved after about 12 months of treatment.

68. 66. The method of any one of claims 2, 3, and 57-65, wherein reduction in cerebral amyloid beta plaques is achieved after about 18 months of treatment.

69. 69. The method of any one of claims 2, 3, and 57-68, wherein the reduction of cerebral amyloid beta plaques is assessed by positron emission tomography (PET).

70. 70. The method of any one of claims 1 to 69, wherein the subject is converted from amyloid positive to amyloid negative.

71. 71. The method of any one of claims 1-70, wherein treating comprises increasing the likelihood of converting the subject from amyloid positive to amyloid negative.

72. 72. The method of any one of claims 1-71, wherein treating comprises about a 10% to about 40% chance of converting the subject from amyloid positive to amyloid negative.

73. 72. The method of any one of claims 1-71, wherein treating comprises about a 30% to about 60% chance of converting the subject from amyloid positive to amyloid negative.

74. 72. The method of any one of claims 1-71, wherein treating comprises about a 40% to about 80% chance of converting the subject from amyloid positive to amyloid negative.

75. 75. The method of any one of claims 72-74, wherein the likelihood of converting the subject from amyloid positive to amyloid negative is after about 6 months of treatment.

76. 75. The method of any one of claims 72-74, wherein the likelihood of converting the subject from amyloid positive to amyloid negative is after about 12 months of treatment.

77. 75. The method of any one of claims 72-74, wherein the likelihood of converting the subject from amyloid positive to amyloid negative is about 18 months after treatment.

78. 78. The method of any one of claims 1-77, wherein treating comprises slowing, stopping, or reversing cognitive decline.

79. 79. The method of claim 78, wherein treating comprises slowing cognitive decline.

80. 81. The method of claim 78 or 80, wherein cognitive function is measured by at least one of the following: CRD-SB, ADAS-Cog14, ADCOMS, and ADCS MCI-ADL.

81. 81. The method of claim 80, wherein cognitive function is measured by ADCOMS.

82. 1. A method of modulating a biomarker in a subject, comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody about once every 3 to 5 weeks, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

83. 1. A method for increasing the Aβ42 / 40 ratio in a subject, comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody about once every 3 to 5 weeks, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

84. 1. A method for reducing phospho-tau levels in a subject, comprising administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody thereof about once every 3 to 5 weeks, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

85. 85. The method of any one of claims 82-84, wherein the administration comprises subcutaneous injection.

86. 86. The method of any one of claims 1 to 85, wherein a biomarker in the subject is modulated.

87. 87. The method of claim 86, wherein the biomarker in the subject is modulated compared to baseline.

88. 88. The method of any one of claims 1 to 87, further comprising detecting a biomarker in a sample taken from the subject.

89. 89. The method of any one of claims 1 to 88, further comprising quantifying biomarkers in a sample taken from the subject.

90. 90. The method of any one of claims 82 and 86-89, wherein the biomarker comprises the ratio of Aβ42 / 40 in the subject.

91. The method of any one of claims 83 and 90, wherein the ratio of Aβ42 / 40 in the subject.

92. 91. The method of any one of claims 83 and 90, wherein the ratio of Aβ42 / 40 in the subject is increased by at least 25%.

93. 91. The method of any one of claims 83 and 90, wherein the ratio of Aβ42 / 40 in the subject is increased by at least 50%.

94. 91. The method of any one of claims 83 and 90, wherein the ratio of Aβ42 / 40 in the subject is increased by about 25% to about 100%.

95. 91. The method of any one of claims 83 and 90, wherein the ratio of Aβ42 / 40 in the subject is increased by about 50% to about 100%.

96. 90. The method of any one of claims 82 and 86 to 89, wherein the biomarker is phospho-tau levels.

97. 97. The method of any one of claims 84 and 96, wherein the phospho-tau value comprises at least one of the following: a p181-tau value, a p212-tau value, a p217-tau value, a p231-tau value, and a p235-tau value.

98. 97. The method of any one of claims 84 and 96, wherein the phospho-tau level comprises a p181-tau level.

99. 97. The method of any one of claims 84 and 96, wherein the phospho-tau level comprises a p212-tau level.

100. 97. The method of any one of claims 84 and 96, wherein the phospho-tau level comprises a p217-tau level.

101. 97. The method of any one of claims 84 and 96, wherein the phospho-tau level comprises a p231-tau level.

102. 97. The method of any one of claims 84 and 96, wherein the phospho-tau level comprises a p235-tau level.

103. 103. The method of any one of claims 84 and 96-102, wherein the phospho-tau level is decreased.

104. 104. The method of claim 103, wherein the phospho-tau level is reduced by at least about 10%.

105. 104. The method of any one of claims 84 and 103, wherein the phospho-tau level is reduced by about 10% to about 30%.

106. 104. The method of any one of claims 84 and 103, wherein the phospho-tau level is reduced by about 20% to about 30%.

107. 91. The method of any one of claims 89 and 90, wherein the sample comprises blood or a portion thereof taken from the subject.

108. 91. The method of any one of claims 89 and 90, wherein the sample comprises plasma collected from the subject.

109. 91. The method of any one of claims 89 and 90, wherein the sample comprises serum taken from the subject.

110. 91. The method of any one of claims 89 and 90, wherein the sample comprises cerebrospinal fluid ("CSF") taken from the subject.

111. 111. The method of any one of claims 1-110, comprising a risk of ARIA-E that is less than about 45%.

112. 111. The method of any one of claims 1-110, comprising a risk of ARIA-E of about 25% to about 45%.

113. 111. The method of any one of claims 1-110, comprising a risk of ARIA-E of less than about 75%.

114. 111. The method of any one of claims 1-110, comprising a risk of ARIA-E of about 50% to about 75%.

115. 115. The method of any one of claims 1-114, comprising a risk of symptomatic ARIA-E that is less than about 15%.

116. 115. The method of any one of claims 1-114, comprising a risk of symptomatic ARIA-E that is less than about 30%.

117. The method of any one of claims 111 to 116, wherein the risk of ARIA-E is a risk of severe ARIA-E.

118. 118. The method of any one of claims 111-117, wherein the risk of ARIA-E is after about 6 months of treatment.

119. 118. The method of any one of claims 111-117, wherein the risk of ARIA-E is after about 12 months of treatment.

120. 118. The method of any one of claims 111-117, wherein the risk of ARIA-E is after about 18 months of treatment.

121. 118. The method of any one of claims 1-117, wherein the subject does not experience symptomatic ARIA-E during treatment.

122. 122. The method of any one of claims 1-121, comprising a risk of ARIA-H that is less than about 35%.

123. 122. The method of any one of claims 1-121, comprising a risk of ARIA-H of about 10% to about 35%.

124. The method of claim 112 or 123, wherein the risk of ARIA-H is a risk of severe ARIA-H.

125. 125. The method of any one of claims 123-124, wherein the risk of ARIA-H is after about 6 months of treatment.

126. 126. The method of any one of claims 1-125, wherein the subject does not experience symptomatic ARIA-H during treatment.

127. 127. The method of any one of claims 111-126, wherein ARIA is assessed by magnetic resonance imaging ("MRI").

128. The method of any one of claims 1 to 127, wherein the subject is an APOE4 homozygous subject.

129. The method of any one of claims 1 to 127, wherein the subject is an APOE4 heterozygous subject or an APOE4 non-carrier.

130. 130. The method of any one of claims 1 to 129, further comprising determining the APOE4 status of the subject prior to administration.

131. 131. The method of any one of claims 1 to 130, wherein the duration of the treatment is at least 6 months.

132. 131. The method of any one of claims 1 to 130, wherein the duration of the treatment is at least 12 months.

133. 131. The method of any one of claims 1 to 130, wherein the duration of the treatment is at least 18 months.

134. 134. The method of any one of claims 1 to 133, wherein the administering is performed using a syringe.

135. 134. The method of any one of claims 1 to 133, wherein the administration is performed using an autoinjector.

136. The method of any one of claims 1 to 135, wherein the subject is a mammal.

137. The method of any one of claims 1 to 136, wherein the subject is a human.

138. A pharmaceutical composition comprising an anti-amyloid beta antibody or antigen-binding fragment thereof for treating Alzheimer's disease in a subject by administering to the subject about 20 mg to about 200 mg of the antibody or antigen-binding fragment thereof about once every 3 to 5 weeks.

139. A pharmaceutical composition comprising an anti-amyloid beta antibody or an antigen-binding fragment thereof for reducing amyloid plaques in a subject by administering to the subject about 20 mg to about 200 mg of the anti-amyloid beta antibody or an antigen-binding fragment thereof about once every 3 to 5 weeks.

140. A pharmaceutical composition comprising an anti-amyloid β antibody or an antigen-binding fragment thereof for converting a subject from amyloid-positive to amyloid-negative by administering about 20 mg to about 200 mg of the anti-amyloid β antibody or an antigen-binding fragment thereof to the subject about once every 3 to 5 weeks.

141. 1. Use of an anti-amyloid beta antibody or antigen-binding fragment for the manufacture of a medicament for treating Alzheimer's disease in a subject, wherein the medicament is for administering to the subject about 20 mg to about 200 mg of the anti-amyloid beta antibody or antigen-binding fragment thereof about once every 3 to 5 weeks.

142. Use of an anti-amyloid beta antibody or antigen-binding fragment thereof for the manufacture of a medicament for reducing amyloid plaques in a subject by administering about 20 mg to about 200 mg of the anti-amyloid beta antibody or antigen-binding fragment to the subject about once every 3 to 5 weeks.

143. Use of an anti-amyloid beta antibody or an antigen-binding fragment thereof for the manufacture of a medicament for converting a subject from amyloid-positive to amyloid-negative by administering about 20 mg to about 200 mg of the anti-amyloid beta antibody or antigen-binding fragment thereof to the subject approximately once every 3 to 5 weeks.

144. the anti-amyloid beta antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3, and a light chain variable region comprising light chain CDR1, CDR2, and CDR3; heavy chain CDR1 comprises the amino acid sequence of one of SEQ ID NOs: 16, 19, or 20; the heavy chain CDR2 comprises the amino acid sequence of one of SEQ ID NOs: 20, 21, 22, or 23; the heavy chain CDR3 comprises the amino acid sequence of one of SEQ ID NOs: 18, 24, or 25; the light chain CDR1 comprises the amino acid sequence of one of SEQ ID NOs: 26, 29, 31, or 32; the light chain CDR2 comprises the amino acid sequence of one of SEQ ID NOs: 33, 34, 35, or 36; The pharmaceutical composition of any of claims 138 to 140 or the use of any of claims 141 to 143, wherein the light chain CDR3 comprises the amino acid sequence of one of SEQ ID NOs: 28, 38, or 39.

145. the anti-amyloid β antibody or antigen-binding fragment thereof a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 16; a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 20; a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 18; a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 29; a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 34; and Light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 38 A pharmaceutical composition according to any one of claims 138 to 140 or a use according to any one of claims 141 to 143, comprising:

146. 144. The pharmaceutical composition of any of claims 138 to 140 or the use of any of claims 141 to 143, wherein the heavy chain variable region, excluding the CDRs, is at least 95% identical to the amino acid sequence of SEQ ID NO: 3 and the light chain variable region, excluding the CDRs, is at least 95% identical to the amino acid sequence of SEQ ID NO:

9.

147. 144. The pharmaceutical composition of any of claims 138 to 140 or the use of any of claims 141 to 143, wherein the heavy chain variable region, excluding the CDRs, is at least 98% identical to the amino acid sequence of SEQ ID NO: 3 and the light chain variable region, excluding the CDRs, is at least 98% identical to the amino acid sequence of SEQ ID NO:

9.

148. 144. The pharmaceutical composition of any of claims 138 to 140 or the use of any of claims 141 to 143, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 3 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:

9.

149. The pharmaceutical composition of any of claims 138 to 140 or the use of any of claims 141 to 143, wherein the heavy chain variable region consists of the amino acid sequence of SEQ ID NO: 3 and the light chain variable region consists of the amino acid sequence of SEQ ID NO:

9.

150. The pharmaceutical composition of any one of claims 138 to 140 or the use of any one of claims 141 to 143, wherein the anti-amyloid beta antibody is a humanized IgG1.

151. The pharmaceutical composition of any of claims 138 to 140 or the use of any of claims 141 to 143, wherein the anti-amyloid beta antibody or antigen-binding fragment thereof further comprises a heavy chain constant region comprising an amino acid sequence that is at least 98% identical to SEQ ID NO: 40 and / or a light chain constant region comprising an amino acid sequence that is at least 98% identical to SEQ ID NO:

41.

152. The pharmaceutical composition of any of claims 138 to 140 or the use of any of claims 141 to 143, wherein the anti-amyloid beta antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 40, with or without the C-terminal lysine, and a light chain constant region comprising the amino acid sequence of SEQ ID NO:

41.

153. A pharmaceutical composition according to any one of claims 138 to 140 or a use according to any one of claims 141 to 143, wherein the anti-amyloid beta antibody comprises a heavy chain constant region consisting essentially of the amino acid sequence of SEQ ID NO: 40, with or without the C-terminal lysine, and a light chain constant region consisting essentially of the amino acid sequence of SEQ ID NO:

41.

154. A pharmaceutical composition according to any one of claims 138 to 140 or a use according to any one of claims 141 to 143, wherein the anti-amyloid beta antibody comprises a heavy chain of SEQ ID NO: 101, with or without the C-terminal lysine, and a light chain of SEQ ID NO:

102.

155. The pharmaceutical composition of any one of claims 138 to 140 or the use of any one of claims 141 to 143, wherein the anti-amyloid beta antibody is h2731.

156. The pharmaceutical composition of any of claims 138 to 140 or the use of any of claims 141 to 143, wherein the administering comprises subcutaneously administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody about once every four weeks, the anti-amyloid beta antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

157. A pharmaceutical composition described in any of claims 138 to 140 or a use described in any of claims 141 to 143, wherein the administering comprises subcutaneously administering to the subject about 45 mg of an anti-amyloid beta antibody approximately once every four weeks, the anti-amyloid beta antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

158. The pharmaceutical composition of any of claims 138 to 140 or the use of any of claims 141 to 143, wherein the administering comprises subcutaneously administering to the subject about 70 mg of an anti-amyloid beta antibody about once every four weeks, the anti-amyloid beta antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

159. The pharmaceutical composition of any of claims 138 to 140 or the use of any of claims 141 to 143, wherein the administering comprises subcutaneously administering to the subject about 200 mg of an anti-amyloid beta antibody about once every four weeks, the anti-amyloid beta antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

160. The pharmaceutical composition of any of claims 138 to 140 or the use of any of claims 141 to 143, wherein the administering comprises subcutaneously administering to the subject about 20 mg to about 200 mg of an anti-amyloid beta antibody about once every four weeks, the anti-amyloid beta antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

161. A pharmaceutical composition described in any of claims 138 to 140 or a use described in any of claims 141 to 143, wherein the administering comprises subcutaneously administering to the subject about 45 mg of an anti-amyloid beta antibody approximately once every four weeks, the anti-amyloid beta antibody comprising a heavy chain of SEQ ID NO: 101, with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

162. The pharmaceutical composition of any of claims 138 to 140 or the use of any of claims 141 to 143, wherein the administering comprises subcutaneously administering to the subject about 70 mg of an anti-amyloid beta antibody about once every four weeks, the anti-amyloid beta antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine, and a light chain of SEQ ID NO:

102.

163. The pharmaceutical composition of any of claims 138 to 140 or the use of any of claims 141 to 143, wherein the administering comprises subcutaneously administering to the subject about 200 mg of an anti-amyloid beta antibody about once every four weeks, the anti-amyloid beta antibody comprising a heavy chain of SEQ ID NO: 101 with or without a C-terminal lysine, and a light chain of SEQ ID NO: 102.

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