Novel ifn beta protein analogs

Abstract

The present invention provides a composition comprising an interferon-beta (IFN-beta) protein of which at least 80% is deamidated, a deamidated IFN-beta 1a protein, methods of producing deamidated proteins, and therapeutic uses of such compositions and deamidated IFN-beta 1a proteins.

1 - 21 . (canceled) 
     
     
         22 . A composition comprising interferon-beta (IFN-beta) protein at least 80% of which is deamidated at the asparagine residue located at an amino acid position corresponding to amino acid position 25 of an interferon-beta 1a protein according to SEQ ID NO:1. 
     
     
         23 . The composition of  claim 22 , wherein said IFN-beta protein is IFN-beta 1a. 
     
     
         24 . The composition of  claim 22 , wherein said asparagine is replaced by an aspartate residue, an iso-aspartate residue or a cyclic imid residue. 
     
     
         25 . The composition of  claim 24 , wherein about 65% to about 70% of the asparagine residue is replaced by an aspartate residue. 
     
     
         26 . The composition of  claim 22 , said composition exhibiting an increased immunomodulatory activity. 
     
     
         27 . The composition of  claim 22 , wherein said IFN-beta protein is glycosylated. 
     
     
         28 . The composition of  claim 27 , wherein said IFN-beta protein has the glycosylation pattern of an IFN-beta protein produced in CHO cells. 
     
     
         29 . The composition of  claim 27 , wherein, in said IFN-beta protein all glycan chains are sialylated. 
     
     
         30 . A modified IFN-beta 1a protein wherein the amino acid asparagine at an amino acid position corresponding to amino acid position 25 of an interferon-beta 1a protein according to SEQ ID NO:1 is deamidated. 
     
     
         31 . A method of treating a subject comprising administering a modified IFN-beta 1a protein according to  claim 30 , or a composition thereof, to a subject in need of treatment. 
     
     
         32 . The method of  claim 31 , wherein said method modulates the immune system of said subject. 
     
     
         33 . The method of  claim 31 , wherein said method comprises administering a vaccine or cancer immunotherapy to said subject in combination with said modified IFN-beta 1a protein or a composition thereof. 
     
     
         34 . The method of  claim 31 , wherein said composition further comprises an anti-oxidant. 
     
     
         35 . The method of  claim 31 , wherein said subject is in need of treatment of a condition selected from the group consisting of viral infections, cancer, and neuronal disorders. 
     
     
         36 . The method of  claim 35 , wherein said subject is treated for a cancer and said composition is administered in combination with chemoimmunotherapy, chemotherapy, radiotherapy, a cytokine or a chemotherapeutic antibody. 
     
     
         37 . A method of deamidating a protein, comprising:
 (a) incubating the protein to be deamidated under alkaline conditions for about 16 to about 24 hours; and   (b) purifying the deamidated protein.   
     
     
         38 . The method according to  claim 37 , wherein said protein is IFN-beta. 
     
     
         39 . The method according to  claim 37 , wherein said incubation is conducted at a pH of about 8.9 to about 9.5. 
     
     
         40 . The method according to  claim 37 , wherein said incubation is at a temperature of about 20° C. to about 25° C. 
     
     
         41 . The method according to  claim 37 , wherein said purification comprises ultra-filtration.