Antibodies for guanylyl cyclase receptors

Abstract

Monoclonal antibodies that act as potentiators, stimulators and agonists of guanylyl cyclase receptors are disclosed.

1 . An isolated antibody or antigen-binding portion thereof that selectively binds to an epitope located in the extracellular domain of natriuretic peptide receptor A (NPRA) wherein said epitope comprises amino acids residues within 
       
         
           
                 
               
                   a) 7-28  
                 
                   (SEQ ID NO: 30) 
                 
                   (NLTVAWLPLANTSYPWSWARV); 
                 
                     
                 
                   b) 28-87  
                 
                   (SEQ ID NO: 35) 
                 
                   (VGPAVELALAQVKARPDLLPGWTVRTVLGSSENALGVCSDTAAP 
                 
                   LAAVDLKWEHNPAVFL); 
                 
                     
                 
                   c) 96-113  
                 
                   (SEQ ID NO: 36) 
                 
                   (APVGRFTAHWRVPLLTAG); 
                 
                     
                 
                   d) 121-129  
                 
                   (SEQ ID NO: 31) 
                 
                   (VKDEYALTT); 
                 
                     
                 
                   e) 293-301  
                 
                   (SEQ ID NO: 37) 
                 
                   (PEYLEFLKQ); 
                 
                     
                 
                   f) 310-312  
                 
                   (FNF); 
                 
                     
                 
                   g) 334-335  
                 
                   (IQ); 
                 
                     
                 
                   h) 313-320  
                 
                   (SEQ ID NO: 32) 
                 
                   (TMEDGLVN); 
                 
                     
                 
                   i) 327-333  
                 
                   (SEQ ID NO: 33) 
                 
                   (HDGLLLY); 
                 
                     
                 
                   j) 347-351  
                 
                   (SEQ ID NO: 34) 
                 
                   (VTDGE);  
                 
                   or 
                 
                     
                 
                   k) 352-362  
                 
                   (SEQ ID NO: 38) 
                 
                   (NITQRMWNRSF). 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         2 . The isolated antibody or antigen-binding portion thereof according to  claim 1 , wherein the mammalian natriuretic peptide receptor A (NPRA) is a human natriuretic peptide receptor A (NPRA). 
     
     
         3 - 6 . (canceled) 
     
     
         7 . The isolated antibody or antigen-binding portion thereof according to  claim 2 , wherein a ligand of NPRA is ANP or BNP. 
     
     
         8 - 10 . (canceled) 
     
     
         11 . The isolated antibody or antigen-binding portion thereof of  claim 1 , wherein the EC 50  of ligand induced intracellular cGMP production is at least 2-fold lower than it would be if the antibody or antigen-binding portion thereof was not present. 
     
     
         12 - 14 . (canceled) 
     
     
         15 . The isolated antibody or antigen-binding portion thereof according to  claim 1 , wherein the antibody or antigen-binding portion thereof is a monoclonal antibody. 
     
     
         16 . The isolated antibody or antigen-binding portion thereof according to  claim 1 , wherein the antibody is an IgG antibody. 
     
     
         17 . The isolated antibody or antigen-binding portion thereof according to  claim 1 , wherein the antibody is an IgG4 or an IgG1 antibody. 
     
     
         18 . The isolated antibody or antigen-binding portion thereof according to  claim 1 , wherein the antibody or antigen-binding portion thereof is selected from the group consisting of: a human antibody, a humanized antibody, a murine antibody, a chimeric antibody, a peptibody, a single chain antibody, a single domain antibody, a Fab fragment, a F(ab′) 2  fragment, a Fv fragment, scF v  fragment and fusion proteins. 
     
     
         19 . An isolated antibody or antigen-binding portion thereof that selectively binds to an epitope located in an extracellular domain of natriuretic peptide receptor A (NPRA) wherein said epitope forms when said NPRA is bound to atrial natriuretic peptide (ANP) or brain natriuretic peptide (ANP) a and wherein said antibody is not reactive with the atrial natriuretic peptide (ANP) or brain natriuretic peptide (ANP) binding site of NPRA. 
     
     
         20 . The antibody of  claim 19 , wherein said antibody potentiates the activity of said ligand or activating protein through said receptor. 
     
     
         21 - 32 . (canceled) 
     
     
         33 . An isolated antibody or antigen-binding portion thereof that selectively binds to an epitope located in an extracellular domain of natriuretic peptide receptor A (NPRA) and competes with a monoclonal antibody comprising a VH and VL chain, each VH and VL chain comprising hypervariable regions CDR1, CDR2 and CDR3 separated by framework amino acid sequences, the hypervariable regions having amino acid sequences in each VH and VL wherein
 VH CDR1  of said antibody has a sequence of SEQ ID NO:3;   VH CDR2  of said antibody has a sequence selected from the group consisting of SEQ ID NO:4 and 10,   VH CDR3  of said antibody has a sequence of SEQ ID NO:13,   VL CDR1  of said antibody has a sequence of SEQ ID NO:14,   VL CDR2  of said antibody has a sequence of SEQ ID NO:15; and   
       VL CDR3  of said antibody has a sequence selected from the group consisting of SEQ ID NO:16, 17 and 18. 
     
     
         34 . An isolated antibody or antigen-binding portion thereof according to  claim 33  that binds to the same epitope bound by an antibody comprising a VH and VL chain, each VH and VL chain comprising hypervariable regions CDR1, CDR2 and CDR3 separated by framework amino acid sequences, the hypervariable regions having amino acid sequences in each VH and VL wherein
 VH CDR1  of said antibody has a sequence of SEQ ID NO:3; 
 VH CDR2  of said antibody has a sequence selected from the group consisting of SEQ ID NO:4 and 10, 
 VH CDR3  of said antibody has a sequence of SEQ ID NO: 13, 
 VL CDR1  of said antibody has a sequence of SEQ ID NO:14, 
 VL CDR2  of said antibody has a sequence of SEQ ID NO:15; and 
 VL CDR3  of said antibody has a sequence selected from the group consisting of SEQ ID NO:16, 17 and 18. 
 
     
     
         35 . An isolated antibody or antigen-binding portion thereof according to  claim 34  comprising a heavy chain variable region (V H ) and a light chain variable region (V L ), each V H  and V L  comprising hypervariable regions CDR1, CDR2 and CDR3 separated by framework amino acid sequences, the hypervariable regions having amino acid sequences in each VH and VL chain of each VH and VL chains of:
 VH CDR1  having a sequence of SEQ ID NO:3; 
 VH CDR2  having a sequence selected from the group consisting of SEQ ID NO:4 and 10, 
 VH CDR3  having a sequence of SEQ ID NO:13, 
 VL CDR1  having a sequence of SEQ ID NO:14, 
 VL CDR2  having a sequence of SEQ ID NO:15; and 
 VL CDR3  having a sequence selected from the group consisting of SEQ ID NO: 16, 17 and 18. 
 
     
     
         36 - 38 . (canceled) 
     
     
         39 . An isolated nucleic acid molecule comprising a sequence encoding an antibody or antigen-binding portion thereof according to  claim 33 . 
     
     
         40 - 45 . (canceled) 
     
     
         46 . A vector comprising and capable of expressing the nucleic acid molecule according to  claim 39 . 
     
     
         47 . A host cell transformed with the vector of  claim 46 . 
     
     
         48 - 49 . (canceled) 
     
     
         50 . A pharmaceutical composition comprising the purified antibody or antigen-binding portion thereof according to  claim 1  and a pharmaceutically acceptable carrier or excipient thereof. 
     
     
         51 . (canceled) 
     
     
         52 . A method of producing an isolated antibody or antigen-binding portion thereof comprising the steps of culturing the host cell according to  claim 47  under suitable conditions and recovering the antibody or antigen-binding portion thereof. 
     
     
         53 . (canceled) 
     
     
         54 . A method for potentiating an apparent affinity of a ligand or an activating protein to an extracellular domain of natriuretic peptide receptor A (NPRA), comprising contacting NPRA with the purified antibody or antigen-binding portion thereof according to  claim 1  in the presence of the ligand or activating protein. 
     
     
         55 - 72 . (canceled) 
     
     
         73 . A method for treating a disorder or condition associated with a decreased level of catalytic activity of natriuretic peptide receptor A (NPRA) in a subject comprising administering to the subject in need thereof an effective amount of a purified antibody or antigen-binding portion thereof according to  claim 1 . 
     
     
         74 . A method for treating a disorder or condition associated with a decreased level of catalytic activity of natriuretic peptide receptor A (NPRA) in a subject comprising administering to the subject in need thereof an effective amount of the pharmaceutical composition according to  claim 50 . 
     
     
         75 - 94 . (canceled)