Modified Human Four Helical Bundle Polypeptides and Their Uses

Abstract

Modified human four helical bundle (4HB) polypeptides and uses thereof are provided.

1 - 5 . (canceled) 
     
     
         6 . A four helical bundle (4HB polypeptide wherein said 4HB polypeptide is linked to a second polypeptide wherein said second polypeptide comprises a non-naturally encoded amino acid. 
     
     
         7 . (canceled) 
     
     
         8 . The 4HB polypeptide of  claim 6 , wherein the second polypeptide is linked to a water soluble polymer wherein said water soluble polymer comprises a poly(alkylene glycol) moiety. 
     
     
         9 . (canceled) 
     
     
         10 . The 4HB polypeptide of  claim 6 , selected from the group consisting of G-CSF, erythropoietin, interferon, and growth hormone. 
     
     
         11 - 15 . (canceled) 
     
     
         16 . The 4HB polypeptide of  claim 6 , wherein the non-naturally encoded amino acid comprises a carbonyl group, an aminooxy group, a hydrazine group, a hydrazide group, a semicarbazide group, an azide group, or an alkyne group. 
     
     
         17 . The 4HB polypeptide of  claim 16 , wherein the non-naturally encoded amino acid comprises a carbonyl group. 
     
     
         18 . The 4HB polypeptide of  claim 17 , wherein the non-naturally encoded amino acid has the structure: 
       
         
           
           
               
               
           
         
         wherein n is 0-10; R 1  is an alkyl, aryl, substituted alkyl, or substituted aryl; R 2  is H, an alkyl, aryl, substituted alkyl, and substituted aryl; and R 3  is H, an amino acid, a polypeptide, or an amino terminus modification group, and R 4  is H, an amino acid, a polypeptide, or a carboxy terminus modification group. 
       
     
     
         19 . The 4HB polypeptide of  claim 16 , wherein the non-naturally encoded amino acid comprises an aminooxy group. 
     
     
         20 . The 4HB polypeptide of  claim 16 , wherein the non-naturally encoded amino acid comprises a hydrazide group. 
     
     
         21 . The 4HB polypeptide of  claim 16 , wherein the non-naturally encoded amino acid comprises a hydrazine group. 
     
     
         22 . The 4HB polypeptide of  claim 16 , wherein the non-naturally encoded amino acid residue comprises a semicarbazide group. 
     
     
         23 . The 4HB polypeptide of  claim 16 , wherein the non-naturally encoded amino acid residue comprises an azide group. 
     
     
         24 . The 4HB polypeptide of  claim 23 , wherein the non-naturally encoded amino acid has the structure: 
       
         
           
           
               
               
           
         
         wherein n is 0-10; R 1  is an alkyl, aryl, substituted alkyl, substituted aryl or not present; X is O, N, S or not present; m is 0-10; R 2  is H, an amino acid, a polypeptide, or an amino terminus modification group, and R 3  is H, an amino acid, a polypeptide, or a carboxy terminus modification group. 
       
     
     
         25 . The 4HB polypeptide of  claim 16 , wherein the non-naturally encoded amino acid comprises an alkyne group. 
     
     
         26 . The 4HB polypeptide of  claim 25 , wherein the non-naturally encoded amino acid has the structure: 
       
         
           
           
               
               
           
         
         wherein n is 0-10; R 1  is an alkyl, aryl, substituted alkyl, or substituted aryl; X is O, N, S or not present; m is 0-10, R 2  is H, an amino acid, a polypeptide, or an amino terminus modification group, and R 3  is H, an amino acid, a polypeptide, or a carboxy terminus modification group. 
       
     
     
         27 . The 4HB polypeptide of  claim 8 , wherein the water soluble polymer has a molecular weight of between about 0.1 kDa and about 100 kDa. 
     
     
         28 . The 4HB polypeptide of  claim 27 , wherein the water soluble polymer has a molecular weight of between about 0.1 kDa and about 50 kDa. 
     
     
         29 - 83 . (canceled)