Antibodies for the treatment of synucleinopathies and neuroinflammation

Abstract

Anti-alpha-synuclein antibodies and antigen-binding fragments thereof (e.g., scFvs) are provided. Also provided are methods of using the same in therapy

What is claimed is: 
     
         1 - 62 . (canceled) 
     
     
         63 . An isolated polynucleotide comprising a nucleic acid sequence encoding an antibody or antigen-binding fragment thereof capable of binding to soluble monomers, oligomers, and/or preformed fibrils of alpha-synuclein, wherein the antibody or antigen-binding fragment thereof comprises:
 i) a complementary determining region (CDR) H1 comprising the amino acid sequence encoded by the nucleic acid sequences set forth in SEQ ID NO:7, nucleotides 142 to 171 of SEQ ID NO:19, nucleotides 142 to 165 of SEQ ID NO:23, nucleotides 142 to 165 of SEQ ID NO:27, nucleotides 142 to 165 of SEQ ID NO:31, or nucleotides 142 to 171 of SEQ ID NO: 35;   ii) a CDR H2 comprising the amino acid sequence encoded by the nucleic acid sequences set forth in SEQ ID NO:8, nucleotides 223 to 243 of SEQ ID NO:19, nucleotides 217 to 240 of SEQ ID NO:23, nucleotides 217 to 240 of SEQ ID NO:27, nucleotides 217 to 240 of SEQ ID NO:31, or nucleotides 223 to 243 of SEQ ID NO: 35 ;   iii) a CDR H3 comprising the amino acid sequence encoded by the nucleic acid sequences set forth in SEQ ID NO:9, nucleotides 358 to 402 of SEQ ID NO: 19, nucleotides 355 to 378 of SEQ ID NO:23, nucleotides 355 to 381 of SEQ ID NO:27, nucleotides 355 to 390 of SEQ ID NO:31, or nucleotides 358 to 390 of SEQ ID NO: 35;   iv) a CDR L1 comprising the amino acid sequence encoded by the nucleic acid sequences set forth in SEQ ID NO: 10, nucleotides 133 to 156 of SEQ ID NO:20, nucleotides 136 to 159 of SEQ ID NO:24, nucleotides 136 to 153 of SEQ ID NO:28, nucleotides 133 to 159 of SEQ ID NO:32, or nucleotides 133 to 156 of SEQ ID NO: 36;   v) a CDR2 L2 comprising the amino acid sequence encoded by the nucleic acid sequences set forth in SEQ ID NO:11, nucleotides 208 to 216 of SEQ ID NO:20, nucleotides 211 to 219 of SEQ ID NO:24, nucleotides 205 to 213 of SEQ ID NO:28, nucleotides 211 to 219 of SEQ ID NO:32, or nucleotides 208 to 216 of SEQ ID NO: 36;   vi) and a CDR3 L3 comprising the amino acid sequence encoded by the nucleic acid sequences set forth in SEQ ID NO:12, nucleotides 325 to 357 of SEQ ID NO:20, nucleotides 328 to 360 of SEQ ID NO:24, nucleotides 322 to 354 of SEQ ID NO:28, nucleotides 328 to 357 of SEQ ID NO:32, or nucleotides 325 to 357 of SEQ ID NO: 36.   
     
     
         64 . The isolated polynucleotide of  claim 63 , wherein the antibody or antigen binding fragment thereof comprises a VH and a VL,
 wherein the VH comprises the amino acid sequence encoded by the nucleic acid sequences set forth in SEQ ID NOs:19, 23, 27, 31, or 35;   wherein the VL comprises the amino acid sequence encoded by the nucleic acid sequences set forth in SEQ ID NOs:20, 24, 28, 32, or 36;   wherein the VH and VL comprise the amino acid sequences encoded by the nucleic acid sequences set forth in SEQ ID NOs:19 and 20, respectively, SEQ ID NOs:23 and 24, respectively, SEQ ID NOs:27 and 28, respectively, SEQ ID NOs: 31 and 32, respectively, or SEQ ID NOs:35 and 36, respectively.   
     
     
         65 . An isolated antibody or antigen binding fragment thereof comprising a VH and a VL, wherein the VH and VL comprise the mature forms of SEQ ID NOs: 21 and 22, respectively, SEQ ID NOs: 25 and 26, respectively, SEQ ID NOs: 29 and 30, respectively, SEQ ID NOs: 33 and 34, respectively, SEQ ID NOs: 37 and 38, respectively, or wherein the VH comprises the same amino acid sequence as the VH encoded by the plasmid of ATCC Deposit Designation PTA-125004 and the VL comprises the same amino acid sequence as the VL encoded by the plasmid of ATCC Deposit Designation PTA-125004. 
     
     
         66 . The isolated antibody or antigen binding fragment thereof of  claim 65 , wherein said fragment is selected from the group consisting of Fab, F(ab′)2, Fv, scFv, scFv-Fc, dsFv and a single domain molecule. 
     
     
         67 . The isolated antibody or antigen binding fragment thereof of  claim 66 , wherein said fragment is a scFv-Fc. 
     
     
         68 . The isolated antibody or antigen binding fragment thereof of  claim 66 , wherein said fragment is a scFv. 
     
     
         69 . A method of treating a synucleinopathic disease in a subject, the method comprising administering to the subject a therapeutically effective amount of the antibody or antigen-binding fragment thereof of  claim 65 , thereby treating the synucleinopathic disease. 
     
     
         70 . The method of  claim 69 , wherein the synucleinopathic disease is selected from the group consisting of Parkinson’s Disease, Parkinson’s Disease Dementia, dementia with Lewy bodies, juvenile-onset generalized neuroaxonal dystrophy (Hallervorden-Spatz disease) pure autonomic failure, multiple system atrophy, and neurodegeneration with brain iron accumulation type-I. 
     
     
         71 . The method of  claim 70 , wherein the synucleinopathic disease is Parkinson’s Disease. 
     
     
         72 . The method of  claim 70 , wherein the synucleinopathic disease is Parkinson’s Disease Dementia. 
     
     
         73 . The method of  claim 70 , wherein the synucleinopathic disease dementia with Lewy bodies. 
     
     
         74 . A method of treating neuroinflammation in a subject, the method comprising administering to the subject a therapeutically effective amount of the antibody or antigen-binding fragment thereof of  claim 65 , thereby treating the neuroinflammation. 
     
     
         75 . A method of diagnosing a synucleinopathic disease in a subject, comprising:
 (a) assessing the level, localization, conformation or a combination thereof of misfolded alpha-synuclein in a subject to be diagnosed with the antibody or antigen-binding fragment thereof of  claim 65 ;   (b) comparing the level, localization, conformation or combination thereof of misfolded alpha-synuclein in the subject to one or more reference standards derived from one or more control samples,   wherein a difference or similarity between the level, localization, conformation or combination thereof of alpha-synuclein in the subject and the reference standard indicates whether the subject has a synucleinopathic disease.   
     
     
         76 . The method of  claim 75 , wherein the synucleinopathic disease is selected from the group consisting of Parkinson’s Disease, Parkinson’s Disease Dementia, dementia with Lewy bodies, juvenile-onset generalized neuroaxonal dystrophy (Hallervorden-Spatz disease) pure autonomic failure, multiple system atrophy, and neurodegeneration with brain iron accumulation type-I. 
     
     
         77 . A method of detecting misfolded alpha-synuclein, the method comprising:
 (a) contacting a biological sample with the antibody or antigen-binding fragment thereof of  claim 65  under conditions which allow formation of immunocomplexes; and   (b) determining the presence and/or level of said immunocomplexes.   
     
     
         79 . The isolated antibody or antigen binding fragment thereof of  claim 68 , wherein the scFv comprises the amino acid sequence encoded by the plasmid of ATCC Deposit Designation PTA-125004. 
     
     
         80 . A method of treating a synucleinopathic disease in a subject, the method comprising administering to the subject a therapeutically effective amount of the antibody or antigen-binding fragment thereof of  claim 79 , thereby treating the synucleinopathic disease. 
     
     
         81 . A method of treating neuroinflammation in a subject, the method comprising administering to the subject a therapeutically effective amount of the antibody or antigen-binding fragment thereof of  claim 79 , thereby treating the neuroinflammation. 
     
     
         82 . A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof of  claim 79 .