Polymer conjugates of interferon Beta-1a and uses

Abstract

An interferon beta polypeptide comprising interferon-beta 1a coupled to a polymer containing a polyalkylene glycol moiety wherein the interferon-beta-1a and the polyalkylene glycol moiety are arranged such that the interferon-beta-1a has an enhanced activity relative to another therapeutic form of interferon beta (interferon-beta-1b) and exhibits no decrease in activity as compared to non-conjugated interferon-beta-1a. The conjugates of the invention are usefully employed in therapeutic as well as non-therapeutic, e.g., diagnostic, applications.

1 - 40 . (canceled) 
     
     
         41 . A composition comprising a glycosylated interferon-beta-1a comprising the amino acid sequence set forth in any one of SEQ ID NOs: 27-39 and 42-56 coupled to a non-naturally-occurring polymer at an N-terminal end of said glycosylated interferon-beta-1a, said polymer comprising a polyalkylene glycol moiety. 
     
     
         42 . The composition of  claim 41 , wherein the polyalkylene moiety is coupled to the interferon-beta by way of a group selected from an aldehyde group, a maleimide group, a vinylsulfone group, a haloacetate group, plurality of histidine residues, a hydrazine group and an aminothiol group. 
     
     
         43 . The composition of  claim 41 , wherein the interferon-beta-1a of any one of SEQ ID NOs: 27-39 and 42-56 is an interferon-beta-1a fusion protein. 
     
     
         44 . The composition of  claim 43 , wherein the interferon-beta-1a fusion protein comprises a portion of an immunoglobulin molecule. 
     
     
         45 . A physiologically active interferon-beta composition comprising a physiologically active interferon-beta-1a comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 27-39 and 42-56, coupled to a polymer comprising a polyalkylene glycol moiety, wherein the interferon-beta-1a is coupled to the polymer at a site on the interferon-beta-1a that is an N-terminal end, wherein the physiologically active interferon-beta-1a and the polyalkylene glycol moiety are arranged such that the physiologically active interferon-beta-1a in the physiologically active interferon-beta composition has an activity at least 2-fold greater relative to physiologically active interferon-beta-1b, when measured by an antiviral assay. 
     
     
         46 . The composition of  claim 45 , wherein the interferon-beta-1a is coupled to the polymer at a site by way of a glycan moiety of the interferon-beta-1a. 
     
     
         47 . The composition of  claim 45 , wherein the interferon-beta-1a is an interferon-beta-1a fusion protein. 
     
     
         48 . The composition of  claim 47 , wherein the interferon-beta-1a fusion protein comprises a portion of an immunoglobulin molecule. 
     
     
         49 . A physiologically active interferon-beta composition comprising a physiologically active glycosylated interferon-beta-1a comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 27-39 and 42-56, N-terminally coupled to a polymer comprising a polyalkylene glycol moiety, wherein the physiologically active interferon-beta-1a and the polyalkylene glycol moiety are arranged such that the physiologically active interferon-beta 1a in the physiologically active interferon-beta composition has equal activity relative to physiologically active interferon-beta lacking said moiety, when measured by an antiviral assay. 
     
     
         50 . The composition of  claim 49 , wherein the interferon-beta is coupled to the polymer at a site by way of a glycan moiety on the interferon-beta. 
     
     
         51 . The composition of  claim 49 , wherein the interferon-beta-1a is an interferon beta fusion protein. 
     
     
         52 . The composition of  claim 51 , wherein the interferon beta fusion protein comprises a portion of an immunoglobulin molecule. 
     
     
         53 . A stable, aqueously soluble, conjugated interferon-beta-1a complex comprising a interferon-beta-1a comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 27-40 and 42-56 N-terminally coupled to a polyethylene glycol moiety, wherein the interferon-beta-1a is coupled to the polyethylene glycol moiety by a labile bond, wherein the labile bond is cleavable by biochemical hydrolysis and/or protcolysis. 
     
     
         54 . An interferon-beta composition according to  claim 41 , wherein the polymer has a molecular weight of from about 5 to 40 kilodaltons. 
     
     
         55 . An interferon-beta composition according to  claim 49 , wherein the polymer has a molecular weight of from about 5 to 40 kilodaltons. 
     
     
         56 . A interferon-beta composition according to  claim 53 , wherein the polymer has a molecular weight of from about 5 to 40 kilodaltons. 
     
     
         57 . A pharmaceutical composition comprising the interferon-beta composition of  claim 54 . 
     
     
         58 . A protein comprising the amino acid sequence set forth in any one of SEQ ID NOs: 25-56, coupled to a non-naturally-occurring polymer at the C-terminal end of said protein, said polymer comprising a polyalkylene glycol moiety. 
     
     
         59 . A protein comprising the amino acid sequence set forth in any one of SEQ ID NOs: 25-56, coupled to a non-naturally-occurring polymer, said polymer comprising a polyalkylene glycol moiety, and said polymer is attached to an amino, carboxylic, hydroxyl, guanidyl, or glycan moiety of said protein. 
     
     
         60 . A protein comprising the amino acid sequence set forth in any one of SEQ ID NOs: 25-56, coupled to a non-naturally-occurring polymer at the N-terminal end of said protein, said polymer comprising a polyalkylene glycol moiety. 
     
     
         61 . A method of treating multiple sclerosis in a subject comprising administering to a subject in need thereof a therapeutically effect amount of a protein comprising the amino acid sequence set forth in any one of SEQ ID NOs: 25-56 coupled to a non-naturally-occurring polymer, said polymer comprising a polyalkylene glycol moiety. 
     
     
         62 . A method of preparing the protein of  claim 60 , comprising reacting a protein with a non-naturally-occurring polymer under reductive alkylation conditions, said protein comprising the amino acid sequence set forth in any one of SEQ ID NOs: 25-56, and said polymer comprising a polyalkylene glycol moiety and a terminal aldehyde moiety.