Methods and compositions for the prevention and treatment of anemia

Abstract

Methods for increasing and maintaining hematocrit in a mammal comprising administering a hyperglycosylated analog of erythropoietin are disclosed. An analog may be administered less frequently than an equivalent molar amount of recombinant human erythropoietin to obtain a comparable target hematocrit and treat anemia. Alternatively, a lower molar amount of a hyperglycosylated analog may be administered to obtain a comparable target hematocrit and treat anemia. Also disclosed are new hyperglycosylated erythopoietin analogs, methods of production of the analogs, and compositions comprising the analogs.

1 - 44 . (canceled) 
     
     
         45 . An analog of human erythropoietin having the in vivo activity of raising and maintaining hematocrit wherein the analog comprises one or more amino acid changes within residues 1-165 as shown in SEQ ID NO: 1 which provide for one or more additional N-linked glycosylation site(s) as compared to human erythropoietin, wherein one additional N-linked glycosylation site is introduced at any one of positions 52, 53, 55, 86 or 114 and an N-linked carbohydrate chain is attached at said additional site. 
     
     
         46 . The analog of  claim 45  selected from: 
       
         
           
                 
                 
               
                     
                   Asn 52  Thr 54  Epo; 
                 
                     
                     
                 
                     
                   Asn 53  Thr 55  Epo; 
                 
                     
                     
                 
                     
                   Asn 114  Thr 116  Epo; 
                 
                     
                     
                 
                     
                   Asn 30  Thr 32  Asn 53  Thr 55  Val 87  Asn 88  Thr 90  Epo; 
                 
                     
                     
                 
                     
                   Asn 55  Thr 57  Epo; 
                 
                     
                     
                 
                     
                   Asn 86  Val 87  Thr 88  Epo; 
                 
                     
                     
                 
                     
                   Asn 30  Thr 32  Asn 53  Thr 55  Epo; 
                 
                     
                     
                 
                     
                   Asn 30  Thr 32  Asn 114  Thr 116  Epo; 
                 
                     
                     
                 
                     
                   Asn 30  Thr 32  Asn 53  Thr 55  Val 87  Asn 88  Thr 90  Asn 114   
                 
                     
                     
                 
                     
                   Thr 116  Epo; 
                 
                     
                     
                 
                     
                   Asn 30  Thr 32  Asn 55  Thr 57  Epo; 
                 
                     
                     
                 
                     
                   Asn 30  Thr 32  Asn 55  Thr 57  Val 87  Asn 88  Thr 90  Epo; 
                 
                     
                     
                 
                     
                   Asn 30  Thr 32  Asn 55  Thr 57  Asn 114  Thr 116  Epo; 
                 
                     
                     
                 
                     
                   Asn 30  Thr 32  Val 87  Asn 88  Thr 90  Asn 114  Thr 116  Epo; 
                 
                     
                     
                 
                     
                   Asn 30  Thr 32  Asn 55  Thr 57  Val 87  Asn 88  Thr 90  Asn 114   
                 
                     
                     
                 
                     
                   Thr 116  Epo; 
                 
                     
                     
                 
                     
                   Asn 30  Thr 32  Asn 55  Thr 57  Val 87  Asn 88  Thr 90  Pro 124   
                 
                     
                     
                 
                     
                   Thr 125  Thr 126  Epo; 
                 
                     
                     
                 
                     
                   Asn 30  Thr 32  Asn 55  Thr 57  Asn 114  Thr 116  Pro 124  Thr 125   
                 
                     
                     
                 
                     
                   Thr 126  Epo; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   Asn 30  Thr 32  Asn 55  Thr 57  Val 87  Asn 88  Thr 90  Asn 114   
                 
                     
                     
                 
                     
                   Thr 116  Pro 124  Thr 125  Thr 126  Epo, 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         47 . The analog of  claim 45  or  46  which comprises conservative amino acid substitutions within residues 1-165 of SEQ ID NO: 1. 
     
     
         48 . The analog of  claim 45  or  46  which comprises deletions or additions of 5 to 10 amino acid residues within residues 1-165 of SEQ ID NO: 1. 
     
     
         49 . The analog of  claim 45  or  46  which comprises deletions or additions of 2 to amino acid residues within residues 1-165 of SEQ ID NO: 1. 
     
     
         50 . The analog of  claim 45  or  46  which comprises deletions or additions of 1 to 2 amino acid residues within residues 1-165 of SEQ ID NO: 1. 
     
     
         51 . The analog of  claim 45  or  46  which is the product of expression of an exogenous DNA sequence. 
     
     
         52 . The analog of  claim 45  or  46  which is produced in a mammalian host cell. 
     
     
         53 . A composition comprising the analog of  claim 45  or  46  and a pharmaceutically acceptable diluent, carrier, solubilizer, emulsifier, preservative, anti-oxidant and/or adjuvant.