Natriuretic compounds, conjugates, and uses thereof

Abstract

Modified natriuretic compounds and conjugates thereof are disclosed in the present invention. In particular, conjugated forms of hBNP are provided that include at least one modifying moiety attached thereto. The modified natriuretic compound conjugates retain activity for stimulating cGMP production, binding to NPR-A receptor, and in some embodiments an improved half-life in circulation as compared to unmodified counterpart natriuretic compounds. Oral, parenteral, enteral, subcutaneous, pulmonary, and intravenous forms of the compounds and conjugates may be prepared as treatments and/or therapies for heart conditions particularly congestive heart failure. Modifying moieties comprising oligomeric structures having a variety of lengths and configurations are also disclosed. Analogs of the natriuretic compound are also disclosed, having an amino acid sequence that is other than the native sequence.

1 . A natriuretic compound conjugate comprising: 
 a biologically active natriuretic compound comprising:    a natriuretic molecule NPR-A binding site; and    at least one modifying moiety conjugation site; and    at least one modifying moiety attached to said modifying moiety conjugation site;    wherein said natriuretic compound conjugate exhibits one or more advantages selected from the group consisting of increased resistance to enzymatic degradation relative to a corresponding unconjugated natriuretic compound, increased circulating half life, increased bioavailability, and prolonged duration of effect.    
     
     
         2 . The natriuretic compound conjugate of  claim 1  wherein the natriuretic compound comprises a peptide or a biologically active peptide segment of brain natriuretic peptide, atrial natriuretic peptide, C-type natriuretic peptide, or dendroaspis natriuretic peptide.  
     
     
         3 . The natriuretic compound conjugate of  claim 1  wherein the natriuretic compound comprises a native hBNP sequence having one or more mutations selected from the group consisting of Lys3Arg, Lys14Arg, Arg30Lys, Lys27Arg, and Arg31Lys.  
     
     
         4 . The natriuretic compound conjugate of  claim 1  wherein the natriuretic compound comprises a native hBNP sequence, having one or more insertions, deletions or extensions.  
     
     
         5 . The natriuretic compound conjugate of  claim 1  wherein the modifying moiety has a formula:  
       
         
           
           
               
               
           
         
         wherein  
         each C is independently selected and is an alkyl moiety having m carbons and m is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; and  
         each PAG is independently selected and is a polyalkylene glycol moiety having n subunits and n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25; and  
         each X is independently selected and is a linking moiety.  
       
     
     
         6 . The natriuretic compound conjugate of  claim 5 , wherein the modifying moiety has a formula:  
       
         
           
           
               
               
           
         
       
     
     
         7 . The natriuretic compound conjugate of  claim 5 , wherein the modifying moiety has a formula:  
       
         
           
           
               
               
           
         
       
     
     
         8 . A method of treating a condition characterized by an excessive level of extracellular fluid, the method comprising administering to a subject in need thereof a pharmaceutically acceptable amount of a natriuretic compound conjugate of  claim 5 .  
     
     
         9 . The natriuretic compound conjugate of  claim 1 , wherein the modifying moiety has a formula:  
       
         
           
           
               
               
           
         
       
     
     
         10 . The natriuretic compound conjugate of  claim 5 , wherein the modifying moiety has a formula:  
       
         
           
           
               
               
           
         
       
     
     
         11 . The natriuretic compound conjugate of  claim 1 , wherein the modifying moiety has a formula:  
       
         
           
           
               
               
           
         
       
     
     
         12 . The natriuretic compound conjugate of  claim 5 , wherein the modifying moiety has a formula:  
       
         
           
           
               
               
           
         
       
     
     
         13 . The natriuretic compound conjugate of  claim 5 , wherein the modifying moiety has a formula:  
       
         
           
           
               
               
           
         
         and the natriuretic compound is hBNP, and the modifying moiety is coupled at Lys3, and the natriuretic compound conjugate is a monoconjugate.  
       
     
     
         14 . The natriuretic compound conjugate of  claim 9  wherein the natriuretic compound is hBNP.  
     
     
         15 . The natriuretic compound conjugate of  claim 10  wherein the natriuretic compound is hBNP.  
     
     
         16 . The natriuretic compound conjugate of  claim 11  wherein the natriuretic compound is hBNP.  
     
     
         17 . The natriuretic compound conjugate of  claim 12  wherein the natriuretic compound is hBNP.  
     
     
         18 . The natriuretic compound conjugate of  claim 9  wherein the natriuretic compound is hBNP, and the modifying moiety is coupled at Lys3, and the natriuretic compound conjugate is a monoconjugate.  
     
     
         19 . The natriuretic compound conjugate of  claim 11  wherein the natriuretic compound is hBNP, and the modifying moiety is coupled at Lys3, and the natriuretic compound conjugate is a monoconjugate.  
     
     
         20 . The natriuretic compound conjugate of  claim 12  wherein the natriuretic compound is hBNP, and the modifying moiety is coupled at Lys3, and the natriuretic compound conjugate is a monoconjugate.  
     
     
         21 . The natriuretic compound conjugate of  claim 9  wherein the natriuretic compound is hBNP, and the modifying moiety is coupled at Lys3, and the natriuretic compound conjugate is a monoconjugate.  
     
     
         22 . A pharmaceutical composition comprising the natriuretic compound conjugate of  claim 9  and a pharmaceutically acceptable carrier.  
     
     
         23 . A pharmaceutical composition comprising the natriuretic compound conjugate of  claim 10  and a pharmaceutically acceptable carrier.  
     
     
         24 . A pharmaceutical composition comprising the natriuretic compound conjugate of  claim 11  and a pharmaceutically acceptable carrier.  
     
     
         25 . A pharmaceutical composition comprising the natriuretic compound conjugate of  claim 12  and a pharmaceutically acceptable carrier.  
     
     
         26 . A pharmaceutical composition comprising the natriuretic compound conjugate of  claim 14  and a pharmaceutically acceptable carrier.  
     
     
         27 . A pharmaceutical composition comprising the natriuretic compound conjugate of  claim 15  and a pharmaceutically acceptable carrier.  
     
     
         28 . A pharmaceutical composition comprising the natriuretic compound conjugate of  claim 16  and a pharmaceutically acceptable carrier.  
     
     
         29 . A pharmaceutical composition comprising the natriuretic compound conjugate of  claim 17  and a pharmaceutically acceptable carrier.  
     
     
         30 . A pharmaceutical composition comprising the natriuretic compound conjugate of  claim 18  and a pharmaceutically acceptable carrier.  
     
     
         31 . A pharmaceutical composition comprising the natriuretic compound conjugate of  claim 19  and a pharmaceutically acceptable carrier.  
     
     
         32 . A pharmaceutical composition comprising the natriuretic compound conjugate of  claim 20  and a pharmaceutically acceptable carrier.  
     
     
         33 . A pharmaceutical composition comprising the natriuretic compound conjugate of  claim 21  and a pharmaceutically acceptable carrier.  
     
     
         34 . A method of making the natriuretic compound conjugate of  claim 9 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.  
     
     
         35 . A method of making the natriuretic compound conjugate of  claim 10 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.  
     
     
         36 . A method of making the natriuretic compound conjugate of  claim 11 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.  
     
     
         37 . A method of making the natriuretic compound conjugate of  claim 12 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.  
     
     
         38 . A method of making the natriuretic compound conjugate of  claim 14 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.  
     
     
         39 . A method of making the natriuretic compound conjugate of  claim 15 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.  
     
     
         40 . A method of making the natriuretic compound conjugate of  claim 16 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.  
     
     
         41 . A method of making the natriuretic compound conjugate of  claim 17 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.  
     
     
         42 . A method of making the natriuretic compound conjugate of  claim 18 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.  
     
     
         43 . A method of making the natriuretic compound conjugate of  claim 19 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.  
     
     
         44 . A method of making the natriuretic compound conjugate of  claim 20 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.  
     
     
         45 . A method of making the natriuretic compound conjugate of  claim 21 , the method comprising chemically coupling the modifying moiety to the natriuretic compound.  
     
     
         46 . A pharmaceutically pure composition of the natriuretic compound conjugate of  claim 9 .  
     
     
         47 . A pharmaceutically pure composition of the natriuretic compound conjugate of  claim 10 .  
     
     
         48 . A pharmaceutically pure composition of the natriuretic compound conjugate of  claim 11 .  
     
     
         49 . A pharmaceutically pure composition of the natriuretic compound conjugate of  claim 12 .  
     
     
         50 . A pharmaceutically pure composition of the natriuretic compound conjugate of  claim 14 .  
     
     
         51 . A pharmaceutically pure composition of the natriuretic compound conjugate of  claim 15 .  
     
     
         52 . A pharmaceutically pure composition of the natriuretic compound conjugate of  claim 16 .  
     
     
         53 . A pharmaceutically pure composition of the natriuretic compound conjugate of  claim 17 .  
     
     
         54 . A pharmaceutically pure composition of the natriuretic compound conjugate of  claim 18 .  
     
     
         55 . A pharmaceutically pure composition of the natriuretic compound conjugate of  claim 19 .  
     
     
         56 . A pharmaceutically pure composition of the natriuretic compound conjugate of  claim 20 .  
     
     
         57 . A pharmaceutically pure composition of the natriuretic compound conjugate of  claim 21 .  
     
     
         58 . The natriuretic compound conjugate of  claim 5  wherein m is 1, X is —O—, n is 4, and C is —CH 3 .