Monoclonal Antibodies Against Pathological Alpha-Synuclein, and Methods Using Same

Abstract

The present disclosure provides monoclonal antibodies that bind α-Synuclein. In certain aspects, the antibodies preferentially bind to α-Synuclein fibrils over α-Synuclein monomer. In other aspects, the invention comprises a method of treating α-Synucleopathic disease in a subject, comprising administering any of the antibodies of the invention to the subject. In yet other aspects, the invention comprises methods of detecting α-Synuclein fibrils using any of the antibodies of the invention.

What is claimed is: 
     
         1 . A method of reducing phosphorylated α-Synuclein (p-α-Syn) in a neuron, the method comprising contacting the neuron with an antibody or an antigen binding fragment thereof,
 wherein the antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), 
 wherein the VL comprises: 
 a CDR1 region comprising the amino acid sequence of SEQ ID NOs: 1, 11, or 21; 
 a CDR2 region comprising the amino acid sequence of SEQ ID NOs: 2, 12, or 22; and 
 a CDR3 region comprising the amino acid sequence of SEQ ID NOs: 3, 13, or 23, and 
 wherein the VH comprises: 
 a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6; 
 a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and 
 a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8. 
 
     
     
         2 . The method of  claim 1 ,
 wherein the VL comprises:   a CDR1 region comprising the amino acid sequence of SEQ ID NOs: 1 or 11;   a CDR2 region comprising the amino acid sequence of SEQ ID NOs: 2 or 12; and   a CDR3 region comprising the amino acid sequence of SEQ ID NOs: 3 or 13, and   wherein the VH comprises:   a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6;   a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and   a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8.   
     
     
         3 . The method of  claim 1 , wherein one of the following applies:
 (a)   the VL comprises:   a CDR1 region comprising the amino acid sequence of SEQ ID NO: 1;   a CDR2 region comprising the amino acid sequence of SEQ ID NO: 2; and   a CDR3 region comprising the amino acid sequence of SEQ ID NO: 3; and   the VH comprises:   a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6;   a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and   a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8;   (b)   the VL comprises:   a CDR1 region comprising the amino acid sequence of SEQ ID NO: 11;   a CDR2 region comprising the amino acid sequence of SEQ ID NO: 12; and   a CDR3 region comprising the amino acid sequence of SEQ ID NO: 13, and   the VH comprises:   a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6;   a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and   a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8;   (c)   the VL comprises:   a CDR1 region comprising the amino acid sequence of SEQ ID NO: 21;   a CDR2 region comprising the amino acid sequence of SEQ ID NO: 22; and   a CDR3 region comprising the amino acid sequence of SEQ ID NO: 23, and   the VH comprises:   a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6;   a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and   a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8.   
     
     
         4 . The method of  claim 1 , wherein the VL comprises the amino acid sequence of SEQ ID NOs: 4 or 14, and wherein the VH comprises the amino acid sequence of SEQ ID NO: 9. 
     
     
         5 . The method of  claim 1 , wherein the VL comprises the amino acid sequence of SEQ ID NOs: 4, and wherein the VH comprises the amino acid sequence of SEQ ID NO: 9. 
     
     
         6 . The method of  claim 1 , wherein the VL comprises the amino acid sequence of SEQ ID NO: 14, and the VH comprises SEQ ID NO: 9. 
     
     
         7 . The method of  claim 1 , wherein the antibody is a monoclonal antibody. 
     
     
         8 . The method of  claim 1 , wherein the antibody or antigen binding fragment thereof is humanized or labeled. 
     
     
         9 . The method of  claim 1 , wherein the antibody or antigen binding fragment thereof is a component of a pharmaceutical composition further comprising at least one pharmaceutical excipient. 
     
     
         10 . The method of  claim 1 , further comprising providing the antibody or antigen binding fragment thereof,
 wherein providing the antibody or antigen binding fragment thereof comprises expressing the antibody or antigen binding fragment thereof using an expression vector comprising a polynucleotide comprising:   (a) the nucleic acid sequences of SEQ ID NOs: 5 and 10;   (b) the nucleic acid sequences of SEQ ID NOs: 15 and 20; or   (c) the nucleic acid sequences of SEQ ID NOs: 25 and 30.   
     
     
         11 . The method of  claim 1 , wherein the neuron is in the brain of a subject. 
     
     
         12 . The method of  claim 11 , wherein the p-α-Syn causes a synucleopathic disease in the subject. 
     
     
         13 . The method of  claim 12 , wherein the synucleopathic disease is at least one from the group consisting of Parkinson's disease, Parkinson's disease with dementia, dementia with Lewy bodies, Alzheimer's disease, Down's syndrome, multiple-system atrophy, prion diseases, and other α-Syn related neurodegenerative disorders. 
     
     
         14 . The method of  claim 11 , wherein the subject is administered a therapeutic effective amount of the antibody or antigen binding fragment thereof to treat or ameliorate the synucleopathic disease. 
     
     
         15 . The method of  claim 14 , wherein the antibody or antigen binding fragment thereof is administered to the subject parenterally. 
     
     
         16 . The method of  claim 11 , wherein the subject is a human. 
     
     
         17 . A method of detecting α-Syn fibrils in a subject, the method comprising:
 administering to the subject a labeled antibody or a labeled antigen binding fragment of the antibody; and 
 detecting presence or absence of a complex of the labeled antibody or antigen binding fragment thereof with any α-Syn fibrils present in the subject, 
 wherein the antibody comprise a light chain variable region (VL) and a heavy chain variable region (VH),
 wherein the VL comprises: 
 a CDR1 region comprising the amino acid sequence of SEQ ID NOs: 1, 11, or 21; 
 a CDR2 region comprising the amino acid sequence of SEQ ID NOs: 2, 12, or 22; and 
 a CDR3 region comprising the amino acid sequence of SEQ ID NOs: 3, 13, or 23, and 
 wherein the VH comprises: 
 a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6; 
 a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and
 a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8, and 
 
 wherein, if presence of the complex of the labeled antibody or antigen binding fragment thereof with α-Syn fibrils is detected, α-Syn fibrils are present in the subject. 
 
 
     
     
         18 . The method of  claim 17 , wherein the antibody is a monoclonal antibody. 
     
     
         19 . The method of  claim 17 , wherein the antibody or antigen binding fragment thereof is humanized. 
     
     
         20 . The method of  claim 17 , wherein the subject is a human.