Chemically modified G-CSF

Abstract

The present invention provides a chemically-modified protein prepared by binding polyethylene glycol to a polypeptide characterized by being the product of expression by a host cell of an exogenous DNA sequence and substantially having the following amino acid sequence: (Met) n Thr Pro Leu Gly Pro Ala Ser Ser Leu Pro Gln Ser Phe Leu Leu Lys Cys Leu Glu Gln Val Arg Lys Ile Gln Gly Asp Gly Ala Ala Leu Gln Glu Lys Leu Cys Ala Thr Tyr Lys Leu Cys His Pro Glu Glu Leu Val Leu Leu Gly His Ser Leu Gly Ile Pro Trp Ala Pro Leu Ser Ser Cys Pro Ser Gln Ala Leu Gln Leu Ala Gly Cys Leu Ser Gln Leu His Ser Gly Leu Phe Leu Tyr Gln Gly Leu Leu Gln Ala Leu Glu Gly Ile Ser Pro Glu Leu Gly Pro Thr Leu Asp Thr Leu Gln Leu Asp Val Ala Asp Phe Ala Thr Thr Ile Trp Gln Gln Met Glu Glu Leu Gly Met Ala Pro Ala Leu Gln Pro Thr Gln Gly Ala Met Pro Ala Phe Ala Ser Ala Phe Gln Arg Arg Ala Gly Gly Val Leu Val Ala Ser His Leu Gln Ser Phe Leu Glu Val Ser Tyr Arg Val Leu Arg His Leu Ala Gln Pro (n = 0 (SEQ ID NO: 1) or 1 (SEQ ID NO: 2)) The chemically-modified protein according to the present invention has a neutrophils-increasing activity much more lasted than that of the intact human G-CSF, enabling fewer numbers of administration with a lower dose.

1 . A chemically-modified protein prepared by binding polyethylene glycol to a polypeptide characterized by being the product of expression by a host cell of an exogenous DNA sequence and substantially having the following amino acid sequence:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                                                        (Met) n   
                     
                 
                     
                   Thr Pro Leu Gly Pro Ala Ser Ser Leu Pro Gln 
                 
                     
                     
                 
                     
                   Ser Phe Leu Leu Lys Cys Leu Glu Gln Val Arg 
                 
                     
                     
                 
                     
                   Lys Ile Gln Gly Asp Gly Ala Ala Leu Gln Glu 
                 
                     
                     
                 
                     
                   Lys Leu Cys Ala Thr Tyr Lys Leu Cys His Pro 
                 
                     
                     
                 
                     
                   Glu Glu Leu Val Leu Leu Gly His Ser Leu Gly 
                 
                     
                     
                 
                     
                   Ile Pro Trp Ala Pro Leu Ser Ser Cys Pro Ser 
                 
                     
                     
                 
                     
                   Gln Ala Leu Gln Leu Ala Gly Cys Leu Ser Gln 
                 
                     
                     
                 
                     
                   Leu His Ser Gly Leu Phe Leu Tyr Gln Gly Leu 
                 
                     
                     
                 
                     
                   Leu Gln Ala Leu Glu Gly Ile Ser Pro Glu Leu 
                 
                     
                     
                 
                     
                   Gly Pro Thr Leu Asp Thr Leu Gln Leu Asp Val 
                 
                     
                     
                 
                     
                   Ala Asp Phe Ala Thr Thr Ile Trp Gln Gln Met 
                 
                     
                     
                 
                     
                   Glu Glu Leu Gly Met Ala Pro Ala Leu Gln Pro 
                 
                     
                     
                 
                     
                   Thr Gln Gly Ala Met Pro Ala Phe Ala Ser Ala 
                 
                     
                     
                 
                     
                   Phe Gln Arg Arg Ala Gly Gly Val Leu Val Ala 
                 
                     
                     
                 
                     
                   Ser His Leu Gln Ser Phe Leu Glu Val Ser Tyr 
                 
                     
                     
                 
                     
                   Arg Val Leu Arg His Leu Ala Gln Pro 
                 
                     
                   (n = 0 (SEQ ID NO:1) or 1 (SEQ ID NO:2)) 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         2 . The chemically-modified protein according to  claim 1  wherein polyethylene glycol is bound through an amino group of the amino acid(s) of the polypeptide.  
     
     
         3 . The chemically-modified protein according to  claim 1  wherein polyethylene glycol is bound through a carboxyl group of the amino acid(s) of the polypeptide.