Specific murine and humanized monoclonal antibodies detecting pathology associated secondary structure changes in proteins and peptides

Abstract

The present invention relates to antibodies and binding fragments thereof that bind the β-sheet secondary structure of a pathological monomeric or oligomeric non-fibrillar proteins without binding to the non-toxic, non-pathological forms of these proteins or peptides. These antibodies and binding fragments thereof are suitable for the diagnosis, prevention, and treatment of protein conformational disorders including all amyloid diseases.

1 . An antibody or binding fragment thereof comprising a heavy chain variable region, wherein said heavy chain variable region comprises:
 a complementarity-determining region 1 (H-CDR1) comprising an amino acid sequence of any one of SEQ ID NOs: 23-26, and 50, or a modified amino acid sequence of any one of SEQ ID NOs: 23-26, and 50, said modified sequence containing 1, 2, or 3 amino acid residue modifications as compared to any one of SEQ ID NOs: 23-26 and 50;   a complementarity-determining region 2 (H-CDR2) comprising an amino acid sequence of any one of SEQ ID NOs: 27-30, and 51, or a modified amino acid sequence of any one of SEQ ID NOs: 27-30, and 51, said modified sequences containing 1, 2, 3, or 4 amino acid residue modifications as compared to any one of SEQ ID NOs: 27-30, and 51; and   a complementarity-determining region 3 (H-CDR3) comprising an amino acid sequence of any one of SEQ ID NOs: 31-34, and 52, or a modified amino acid sequence of any one of SEQ ID NO: 31-34, and 52, said modified sequence containing 1, 2, or 3 amino acid residue modifications as compared to any one of SEQ ID NOs: 31-34 and 52.   
     
     
         2 .- 4 . (canceled) 
     
     
         5 . The antibody or binding fragment thereof of  claim 1 , wherein said antibody or binding fragment thereof is a monoclonal antibody or binding fragment thereof. 
     
     
         6 . The antibody or binding fragment thereof of  claim 1 , wherein said heavy chain variable region comprises:
 the H-CDR1 comprising the amino acid sequence of SEQ ID NO: 23, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1 or 2 amino acid residue modifications as compared to SEQ ID NO: 23;   the H-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1, 2, 3, or 4 amino acid residue modifications as compared to SEQ ID NO: 27; and   the H-CDR3 comprising the amino acid sequence of SEQ ID NO: 31, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1 or 2 amino acid modifications as compared to SEQ ID NO: 31.   
     
     
         7 . The antibody or binding fragment thereof of  claim 1 , wherein said heavy chain variable region comprises:
 the H-CDR1 comprising the amino acid sequence of SEQ ID NO: 24, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1 or 2 amino acid residue modifications as compared to SEQ ID NO: 24;   the H-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1, 2, 3, or 4 amino acid residue modifications as compared to SEQ ID NO: 28; and   the H-CDR3 comprising the amino acid sequence of SEQ ID NO: 32, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1, 2, or 3 amino acid modifications as compared to SEQ ID NO: 32.   
     
     
         8 . The antibody or binding fragment thereof of  claim 1 , wherein said heavy chain variable region comprises:
 the H-CDR1 comprising the amino acid sequence of SEQ ID NO: 25, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1 or 2 amino acid residue modifications as compared to SEQ ID NO: 25;   the H-CDR2 comprising the amino acid sequence of SEQ ID NO: 29, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1, 2, 3, or 4 amino acid residue modifications as compared to SEQ ID NO: 29; and   the H-CDR3 comprising the amino acid sequence of SEQ ID NO: 33, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1 or 2 amino acid residue modifications as compared to SEQ ID NO: 33.   
     
     
         9 . The antibody or binding fragment thereof of  claim 1 , wherein said heavy chain variable region comprises:
 the H-CDR1 comprising the amino acid sequence of SEQ ID NO: 26, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1 or 2 amino acid residue modifications as compared to SEQ ID NO: 26;   the H-CDR2 comprising the amino acid sequence of SEQ ID NO: 30, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1, 2, 3, or 4 amino acid residue modifications as compared to SEQ ID NO: 30; and   the H-CDR3 comprising the amino acid sequence of SEQ ID NO: 34, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1 or 2 amino acid residue modifications as compared to SEQ ID NO: 34.   
     
     
         10 . The antibody or binding fragment thereof of  claim 1 , wherein said heavy chain variable region comprises:
 the H-CDR1 comprising the amino acid sequence of SEQ ID NO: 50, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1, 2, or 3 amino acid residue modifications as compared to SEQ ID NO: 50;
 the H-CDR2 comprising the amino acid sequence of SEQ ID NO: 51, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1, 2, 3, or 4 amino acid residue modifications as compared to SEQ ID NO: 51; and 
 the H-CDR3 comprising the amino acid sequence of SEQ ID NO: 52, or a modified amino acid sequence thereof, said modified amino acid sequence containing 1, 2, or 3 amino acid residue modifications as compared to SEQ ID NO: 52. 
   
     
     
         11 .- 18 . (canceled) 
     
     
         19 . The antibody or binding fragment thereof of  claim 1 , wherein said heavy chain variable region further comprises human or a humanized immunoglobulin heavy chain framework regions. 
     
     
         20 . The antibody or binding fragment thereof of  claim 19 , wherein said heavy chain variable region comprises an amino acid sequence that is at least 60% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 6, 10, 14, and 20. 
     
     
         21 .- 25 . (canceled) 
     
     
         26 . The antibody or binding fragment thereof of  claim 1 , wherein said antibody or binding fragment thereof is a single domain antibody. 
     
     
         27 . (canceled) 
     
     
         28 . An isolated polynucleotide encoding the antibody or binding fragment thereof of  claim 1 . 
     
     
         29 . A vector comprising the isolated polynucleotide of  claim 28 . 
     
     
         30 . A host cell comprising the vector of  claim 29 . 
     
     
         31 . A pharmaceutical composition comprising:
 the antibody or binding fragment thereof of  claim 1 , and   a pharmaceutical carrier.   
     
     
         32 . (canceled) 
     
     
         33 . A method of treating a condition mediated by an amyloidogenic protein or peptide in a subject, said method comprising:
 administering to the subject the pharmaceutical composition of  claim 31 , wherein said composition is administered in an amount effective to treat the condition or one or more symptoms mediated by the amyloidogenic protein or peptide in the subject.   
     
     
         34 . (canceled) 
     
     
         35 . The method of  claim 33 , wherein the condition is selected from the group consisting of Alzheimer's disease (AD) and all its variations, preclinical AD, Rapid Progressive dementia, Down syndrome (DS), fronto-temporal dementia (FTD), Lewy Body Dementia (LBD), Parkinson's disease (PD), hereditary cerebral hemorrhage with amyloidosis (HCHWA), kuru, Creutzfeldt-Jakob disease (CJD, including familial, sporadic or new Variant (nV) forms), chronic wasting disease (CWD) and its adapted forms in other mammals, Gerstmann-Straussler-Scheinker disease (GSS), bovine spongiform encephalopathy (BSE) and its adapted forms in other mammals, ovine Scrapie (Sc) and its adapted forms in other mammals, Huntington's disease (HD) and all its glutamine expansion repeats, fatal familial insomnia, British familial dementia and all its variations, Danish familial dementia and all its variations, frontotemporal lobar degeneration associated with protein tau (FTLD-tau), frontotemporal lobar degeneration associated with protein FUS (FTLD-FUS), FTD-TDP-43, Amyotrophic lateral sclerosis (ALS), FTD and ALS with all repeat expansions due to mutations on C9orf72, Mild Cognitive Impairment (MCI), familial corneal amyloidosis, Familial corneal dystrophies, medullary thyroid carcinoma, insulinoma, type 2 diabetes, isolated atrial amyloidosis, pituitary amyloidosis, aortic amyloidosis, plasma cell disorders, familial amyloidosis, senile cardiac amyloidosis, inflammation-associated amyloidosis, familial Mediterranean fever (FMF), dialysis-associated amyloidosis, systemic amyloidosis, familial systemic amyloidosis, motor neuron disease, traumatic brain injury (TBI), ad chronic traumatic encephalopathy. 
     
     
         36 . A method of treating a subject having or at risk of having a condition mediated by a pathological protein having a β-sheet secondary structure, said method comprising:
 administering to the subject the pharmaceutical composition of  claim 31 , wherein said composition is administered in an amount effective to treat, inhibit, or slow the progression of the condition or one or more symptoms associated with the conditions mediated by the pathological protein having a β-sheet secondary structure. 
 
     
     
         37 . (canceled) 
     
     
         38 . A method of diagnosing an amyloid disease in a subject, said method comprising:
 detecting, in the subject, the presence of an amyloidogenic protein or peptide using a diagnostic reagent, wherein the diagnostic reagent comprises the antibody or binding fragment thereof of  claim 1 , and   diagnosing the amyloid disease in the subject based on said detecting.   
     
     
         39 .- 40 . (canceled) 
     
     
         41 . A method of identifying a subject's risk for developing a condition mediated by an amyloidogenic protein or peptide, said method comprising:
 detecting, in the subject, the presence of an amyloidogenic protein or peptide using a diagnostic reagent, wherein the diagnostic reagent comprises the antibody or binding fragment thereof of  claim 1 , and   identifying the subject's risk of developing the condition mediated by the amyloidogenic protein or peptide based on said detecting.   
     
     
         42 .- 43 . (canceled) 
     
     
         44 . A diagnostic kit comprising:
 the antibody or binding fragment thereof of  claim 1  and   a detectable label.