Methods and compositions to treat and detect misfolded-sod1 mediated diseases

Abstract

The invention provides a method for treating a medical condition, disease, or disorder mediated by a misfolded form of superoxide dismutase (SOD) in a subject in need of treatment. The method optionally comprises administering to the subject a composition comprising a pharmaceutically acceptable vehicle and an agent selected from (1) an exogenous antibody or fragment thereof that binds selectively to the misfolded form of SOD, and/or (2) an immunogen that elicits production of an endogenous antibody that binds selectively to the misfolded form of SOD, and/or (3) a nucleic acid sequence encoding (1) or (2). In certain embodiments, the invention provides methods of treating diseases such as Alzheimer's Disease, Parkinson's Disease or amyotrophic lateral sclerosis using amyotrophic disease-specific epitopes, and compositions including these epitopes. The invention also provides antibodies that bind to monomeric or misfolded SOD1, and not on the molecular surface of native homodimeric SOD1. In addition, the invention includes methods of diagnosing Alzheimer's Disease, Parkinson's Disease or amyotrophic lateral sclerosis in a subject. Also, the invention provides methods of identifying substances for the treatment or prevention of Alzheimer's Disease, Parkinson's Disease or amyotrophic lateral sclerosis and kits using the binding proteins of the invention.

We claim: 
     
         1 - 92 . (canceled) 
     
     
         93 . An isolated antibody that specifically binds at least 5 contiguous residues of any one of SEQ ID NO: 2, 3 or 6, optionally where the isolated antibody specifically binds a sequence selected from: 
       
         
           
                 
                 
               
                     
                   (amino acid (aa) 1-8 of SEQ ID NO: 2) 
                 
                     
                   DLGKGGNE; 
                 
                     
                     
                 
                     
                   (aa 12-18 of SEQ ID NO: 2) 
                 
                     
                   KTGNAGS; 
                 
                     
                     
                 
                     
                   (aa 8-14 of SEQ ID NO: 3) 
                 
                     
                   GGPKDEE; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   (aa 3-9 of SEQ ID NO: 6) 
                 
                     
                   IIGRTLV. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         94 . The isolated antibody according to  claim 93 , wherein the antibody is monoclonal, polyclonal, chimeric or humanized. 
     
     
         95 . The isolated antibody according to  claim 93 , wherein the antibody is an antibody fragment. 
     
     
         96 . The isolated antibody according to  claim 95 , wherein the antibody fragment is a Fab, Fab′, F(ab′) 2 , scFv, dsFv, ds-scFv, dimers, minibodies, diabodies, or multimers thereof or bispecific antibody fragments. 
     
     
         97 . A method of making an antibody according to  claim 93 , wherein the antibody specifically binds an isolated peptide comprising the sequence DLGKGGNE (aa 1-8 of SEQ ID NO:2), KTGNAGS (aa 12-18 of SE ID NO:2), GGPKDEE (aa 8-14 of SEQ ID NO:3) or IIGRTLV (aa 3-9 of SEQ ID NO:6), by:
 i) immunizing an animal with an immunogenic form of the isolated peptide;   ii) screening an expression library; or   iii) using phage display.   
     
     
         98 . The method according to  claim 97 , further comprising forming antibody producing hybridomas. 
     
     
         99 . An immunogen comprising a polypeptide of up to 50 SOD1 residues coupled to an immune enhancing agent, the polypeptide comprising: a) a peptide of at least 5 contiguous residues of any one of SEQ ID NO: 2, 3, or 6, or b) a peptide as in a) comprising one or more oxidized or nitrated amino acids, optionally wherein the peptide of at least 5 contiguous residues is selected from DLGKGGNES (aa 1-8 of SEQ ID NO:2), KTGNAGS (aa 12-18 of SEQ ID NO:2), GGPKDEE (aa 8-14 of SEQ ID NO:3) and IIGRTLV (aa 3-9 of SEQ ID NO:6). 
     
     
         100 . A method of detecting or diagnosing a medical condition, disease, or disorder mediated by a misfolded form of superoxide dismutase (SOD1) in a subject comprising the steps of:
 (a) contacting a test sample of said subject with an antibody according to  claim 93 ;   (b) to produce an antibody-antigen complex;   (c) measuring the amount of the antibody-antigen complex in the test sample; and   (d) comparing the amount of antibody-antigen complex in the test sample to a control;   wherein a difference in the amount of antibody-antigen complex in the test sample as compared to the control is indicative of a medical condition, disease, or disorder mediated by a misfolded form of SOD1 in a subject.   
     
     
         101 . The method according to  claim 100 , wherein the medical condition, disease or disorder is a neurodegenerative condition, disease or disorder. 
     
     
         102 . The method according to  claim 100 , wherein the neurodegenerative condition, disease or disorder comprises Alzheimer's Disease, Parkinson's Disease, Lewy Body Disease or ALS. 
     
     
         103 . A kit for diagnosing a medical condition, disease, or disorder mediated by a misfolded form of superoxide dismutase (SOD1) in a subject according to the method of  claim 100  comprising an antibody specific for a sequence selected from the group consisting of: 
       
         
           
                 
                 
                 
               
                     
                   DLGKGGNE; 
                   (aa 1-8 of SEQ ID NO: 2) 
                 
                     
                     
                 
                     
                   KTGNAGS; 
                   (aa 12-18 of SEQ ID NO: 2) 
                 
                     
                     
                 
                     
                   GGPKDEE; 
                   (aa 8-14 of SEQ ID NO: 3) 
                 
                     
                   and 
                     
                 
                     
                     
                 
                     
                   IIGRTLV. 
                   (aa 3-9 of SEQ ID NO: 6) 
                 
             
                
                
                
                
                
                
                
                
               
            
           
         
         and instructions for the use thereof. 
       
     
     
         104 . The kit according to  claim 103 , further comprising a recombinant peptide standard consisting of a sequence selected from the group consisting of: 
       
         
           
                 
                 
                 
               
                     
                   DLGKGGNE; 
                   (aa 1-8 of SEQ ID NO: 2) 
                 
                     
                     
                 
                     
                   KTGNAGS; 
                   (aa 12-18 of SEQ ID NO: 2) 
                 
                     
                     
                 
                     
                   GGPKDEE; 
                   (aa 8-14 of SEQ ID NO: 3) 
                 
                     
                   and 
                     
                 
                     
                     
                 
                     
                   IIGRTLV. 
                   (aa 3-9 of SEQ ID NO: 6) 
                 
             
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         105 . A diagnostic agent comprising (1) an antibody of  claim 93  attached to (2) a label that produces a detectable signal, directly or indirectly. 
     
     
         106 . The diagnostic agent of  claim 105 , wherein the label comprises a radioisotope, a fluorescent compound, a chemiluminescent compound, an enzyme, an imaging agent or a metal ion. 
     
     
         107 . A method for treating a medical condition, disease, or disorder mediated by a misfolded form of superoxide dismutase (SOD1) in a subject in need of treatment, the method comprising administering to the subject a composition comprising a pharmaceutically acceptable vehicle and an agent selected from (1) an exogenous antibody or fragment thereof that binds selectively and at least 2 fold more efficiently to a misfolded form of wild type human SOD1 compared to a natively folded form of wild type human SOD1, (2) an immunogen that elicits production of an endogenous antibody that binds selectively to the misfolded form of SOD1, and/or (3) a nucleic acid sequence encoding (1) or (2),
 wherein the antibody or fragment thereof binds selectively to at least 5 contiguous amino acids of any one of SEQ ID NO: 2, 3 or 6, optionally wherein the antibody or fragment thereof binds selectively to DLGKGGNE (aa 1-8 of SEQ ID NO:2), KTGNAGS (aa 12-18 of SEQ ID NO:2), GGPKDEE (aa 8-14 of SEQ ID NO:3), or IIGRTLV (aa 3-9 of SEQ ID NO:6).   
     
     
         108 . The method according to  claim 107 , wherein the medical condition, disease or disorder is a neurodegenerative condition, disease or disorder selected from ALS, Alzheimer's disease or Parkinson's disease. 
     
     
         109 . The method according to  claim 107 , wherein the agent of (2) is conjugated to a molecule that enhances immunogenicity of the agent. 
     
     
         110 . The method according to  claim 107 , wherein the misfolded SOD1 has wild type sequence. 
     
     
         111 . The method according to  claim 107 , wherein the antibody or fragment thereof binds any one of SEQ ID NOs: 46-56, 57, 58, 61, and/or 62. 
     
     
         112 . A pharmaceutical composition useful in the treatment of a subject having a medical condition, disease, or disorder mediated by a misfolded form of superoxide dismutase 1 (SOD1), comprising a pharmaceutically acceptable vehicle and an agent selected from (1) an exogenous antibody or fragment thereof that binds selectively to a misfolded or aggregated form of SOD1 of  claim 93 , and/or (2) an immunogen comprising a peptide consisting of at least 5 contiguous residues selected from any one of DLGKGGNE (aa 1-8 of SEQ ID NO:2), KTGNAGS (aa 12-18 of SEQ ID NO:2), GGPKDEE (aa 8-14 of SEQ ID NO:3), or IIGRTLV (aa 3-9 of SEQ ID NO:6) that elicits production by said subject of endogenous antibody that binds selectively to a misfolded or aggregated form of SOD1,
 wherein the isolated antibody of fragment thereof binds selectively to at least 5 contiguous residues of any of DLGKGGNE (aa 1-8 of SEQ ID NO:2), KTGNAGS (aa 12-18 of SEQ ID NO:2), GGPKDEE (aa 8-14 of SEQ ID NO:3), and IIGRTLV (aa 3-9 of SEQ ID NO:6).   
     
     
         113 . A method for alleviating a symptom associated with neurodegenerative condition, disease, or disorder mediated by a misfolded or aggregated form of superoxide dismutase 1 (SOD1) in a subject in need of such alleviation, the method comprising administering to the subject a composition comprising a pharmaceutically acceptable vehicle and the immunogen of  claim 99  wherein the immunogen elicits production of an endogenous antibody that binds selectively and at least 2 fold more efficiently to an epitope on misfolded form of SOD1 compared to natively folded SOD1. 
     
     
         114 . The method according to  claim 113 , wherein the neurodegenerative condition, disease or disorder is ALS, Alzheimer's disease or Parkinson's disease.