FIELD OF THE INVENTION

This invention relates to the field of antibodies directed to amyloid-beta (Aβ) peptides and therapeutic methods using the antibodies. In particular, antibodies can be used for identifying and treating amyloid-related disorders.

REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

This application contains a sequence listing, which is submitted electronically via EFS-Web as an ASCII formatted sequence listing with a file name “JAB7013USPSP Sequence Listing” and a creation date of Mar. 11, 2019, and having a size of 76 kb. The sequence listing submitted via EFS-Web is part of the specification and is herein incorporated by reference in its entirety.

BACKGROUND

Alzheimer's disease (AD) is a degenerative brain disorder characterized clinically by progressive loss of memory, cognition, reasoning, judgment and emotional stability that gradually leads to profound mental deterioration and ultimately death. Alzheimer's disease is a common cause of progressive mental failure (dementia) in the elderly. Alzheimer's disease has been observed worldwide and represents a major public health issue. The disease is currently estimated to affect more than five million individuals in the United States alone. At present it is incurable, and no treatment effectively prevents AD or reverses its symptoms or course.

The brains of individuals with AD exhibit characteristic lesions termed amyloid plaques, amyloid angiopathy (amyloid deposits in blood vessels) and neurofibrillary tangles. Large numbers of these lesions, particularly amyloid plaques and neurofibrillary tangles, are generally found in several areas of the brain important for memory and cognitive function. Amyloid plaques and amyloid angiopathy also characterize the brains of individuals with Trisomy 21 (Down's Syndrome), diffuse Lewy body disease and hereditary cerebral hemorrhage with amyloidosis of the Dutch-type (HCHWA-D).

A major constituent of amyloid plaques is a variety of amyloid-beta (Aβ) peptides that are produced by cleavage of the β-amyloid precursor protein (APP). Deposition of Aβ peptides in brain is hypothesized to be an early and necessary step in the disease cascade leading to AD. The identification of mutations in the amyloid precursor protein and presenillin genes resulting in altered Aβ production and causing familial early onset AD provides strong evidence that altered amyloid metabolism is a central event in the pathogenic process underlying the disease.

Amyloid-β peptides having pyroglutamate at the third residue (3pE Aβ) are a major species deposited in the brain of AD patients. 3pE Aβ is present in almost all diffuse and mature plaques in AD, is metabolically stable, and can play a role in both plaque seeding and stabilization (Cynis et al., Molecular Neurodegeneration, 2016; 11:48). Detectable amounts of 3pE Aβ have not been reported in CSF or plasma, thus suggesting that the target peptide is pathology specific (DeMattos et al., Neuron, 2012; 76:1-13). Antibodies that selectively bind to 3pE Aβ can be useful for immunotherapy.

SUMMARY OF THE INVENTION

As embodied and fully described, the invention relates to antibodies and antigen binding fragments thereof that bind to amyloid-β having pyroglutamate at the third residue (3pE Aβ), methods of producing antibodies or antigen binding fragments thereof that bind to 3pE Aβ, assay methods using such antibodies or antigen binding fragments thereof, and use of the antibodies or antigen binding fragments thereof of the invention for the manufacture of a medicament, for treating, delaying the onset of or reversing at least one pathology or symptom of Alzheimer's disease and other β-amyloid-related diseases. Antibodies of the invention preferentially bind Aβ peptide containing 3pE over Aβ peptide that does not contain 3pE.

In particular, described herein is an isolated monoclonal antibody or antigen-binding fragment thereof comprising a heavy complementarity determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3, having the polypeptide sequences of:a. SEQ ID NOs: 1, 2, 3, 4, 5, and 6, respectively;b. SEQ ID NOs: 1, 7, 3, 4, 5, and 6, respectively;c. SEQ ID NOs: 1, 7, 3, 8, 5, and 6, respectively;d. SEQ ID NOs: 1, 2, 3, 8, 5, and 6, respectively;e. SEQ ID NOs: 56, 57, 3, 8, 5, and 6, respectively;f. SEQ ID NOs: 56, 57, 3, 4, 5, and 6, respectively;g. SEQ ID NOs: 56, 58, 3, 4, 5, and 6, respectively;h. SEQ ID NOs: 56, 7, 3, 8, 5, and 6, respectively;i. SEQ ID NOs: 1, 57, 3, 8, 5, and 6, respectively;j. SEQ ID NOs: 56, 7, 3, 4, 5, and 6, respectively;k. SEQ ID NOs: 1, 57, 3, 4, 5, and 6, respectively;l. SEQ ID NOs: 1, 58, 3, 4, 5, and 6, respectively; orm. SEQ ID NOs: 56, 2, 3, 4, 5, and 6, respectively;
wherein the antibody or antigen-binding fragment thereof specifically binds 3pE Aβ, preferably human 3pE Aβ.

In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment comprises a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:9, 11, 13, 15, 16, 17, 19, 20, or 21, or a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:10, 12, 14, 18, 22, 53, or 55.

In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof comprises:a. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:21, and a light chain variable region having the polypeptide sequence of SEQ ID NO:22;b. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:9, and a light chain variable region having the polypeptide sequence of SEQ ID NO:10;c. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:11, and a light chain variable region having the polypeptide sequence of SEQ ID NO:12;d. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:13, and a light chain variable region having the polypeptide sequence of SEQ ID NO:14;e. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:15, and a light chain variable region having the polypeptide sequence of SEQ ID NO:14;f. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:16, and a light chain variable region having the polypeptide sequence of SEQ ID NO:14;g. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:20, and a light chain variable region having the polypeptide sequence of SEQ ID NO:14;h. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:17, and a light chain variable region having the polypeptide sequence of SEQ ID NO:18;i. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:19, and a light chain variable region having the polypeptide sequence of SEQ ID NO:18;j. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:21, and a light chain variable region having the polypeptide sequence of SEQ ID NO:53; ork. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:21, and a light chain variable region having the polypeptide sequence of SEQ ID NO:55.

In certain embodiments, the monoclonal antibody or antigen-binding fragment thereof is chimeric. In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof is human or humanized.

In certain embodiments, the isolated monoclonal antibody comprises:a. a heavy chain amino acid sequence comprising SEQ ID NO:37 and a light chain amino acid sequence comprising SEQ ID NO:38;b. a heavy chain amino acid sequence comprising SEQ ID NO:39 and a light chain amino acid sequence comprising SEQ ID NO:38c. a heavy chain amino acid sequence comprising SEQ ID NO:37 and a light chain amino acid sequence comprising SEQ ID NO:52; ord. a heavy chain amino acid sequence comprising SEQ ID NO:39 and a light chain amino acid sequence comprising SEQ ID NO:54.

In certain embodiments, the antigen binding fragment is selected from the group of fragments consisting of Fv, F(ab′), F(ab′)2 and scFv. The antibody or antigen binding fragment thereof selectively binds to 3pE Aβ peptide (e.g., Aβ3pE-40 and Aβ3pE-42), with little or no cross-reactivity to other Aβ peptides or β-amyloid precursor protein (APP).

Also provided are isolated nucleic acids encoding the monoclonal antibodies or antigen-binding fragments thereof of the invention disclosed herein.

Also provided are vectors comprising the isolated nucleic acids encoding the monoclonal antibodies or antigen-binding fragments thereof of the invention.

Also provided are host cells comprising the vectors comprising the isolated nucleic acids encoding the monoclonal antibodies or antigen-binding fragments thereof of the invention. Also provided are hybridomas that produce the isolated monoclonal antibody or antigen-binding fragment thereof of the invention.

In certain embodiments, provided is a pharmaceutical composition comprising an isolated monoclonal antibody or antigen-binding fragment thereof of the invention and a pharmaceutically acceptable carrier.

Also provided are methods of treating a condition associated the formation of plaques containing beta-amyloid protein in a subject in need thereof. The methods comprise administering a monoclonal antibody or antigen binding fragment thereof of the invention or the pharmaceutical composition of the invention to the subject in need thereof. In certain embodiments, the condition is Alzheimer's disease. In certain embodiments, the condition is selected form the group consisting of dementia associated with Trisomy 21 (Down's Syndrome), diffuse Lewy body disease, inclusion body myositis, cerebral amyloid angiopathy and hereditary cerebral hemorrhage with amyloidosis of the Dutch-type (HCHWA-D).

Also provided are methods of reducing plaques associated with Alzheimer's disease in a subject in need thereof. The methods comprise administering a monoclonal antibody or antigen-binding fragment thereof of the invention or the pharmaceutical composition of the invention to the subject in need thereof.

Also provided are methods of preventing seeding activity of 3pE Aβ in a subject in need thereof. The methods comprise administering a monoclonal antibody or antigen-binding fragment thereof of the invention or the pharmaceutical composition of the invention to the subject in need thereof.

Also provided are methods of producing the monoclonal antibody or antigen-binding fragment thereof of the invention, the methods comprise culturing a cell comprising a nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof under conditions to produce the monoclonal antibody or antigen-binding fragment thereof and recovering the antibody or antigen-binding fragment thereof.

Also provided are methods of producing a pharmaceutical composition of the invention. The methods comprise combining the monoclonal antibody or antigen-binding fragment thereof of the invention with a pharmaceutically acceptable carrier to obtain the pharmaceutical composition.

An embodiment includes kits and devices comprising the antibody or antigen binding fragment thereof described above.

Further objects, features, and advantages of the present invention will be apparent to those skilled in the art from detailed consideration of the preferred embodiments that follow.

