Methods of treatment using fusion proteins

Abstract

The invention relates to method of treatment using fusion proteins. The fusion proteins may be RAGE fusion proteins produce by methods for controlling the glycosylation of the fusion protein. The fusion protein may comprise a RAGE polypeptide linked to an immunoglobulin polypeptide.

1 . A method of treating a condition selected from the group consisting of: diabetic nephropathy, diabetic retinopathy, a diabetic foot ulcer, a cardiovascular complication, diabetic neuropathy, amyloidosis, Alzheimer's disease, kidney failure, inflammation, osteoporosis, kidney failure, transplant rejection, and inflammation or rejection associated with transplantation, comprising administering a composition comprising an amount of a fusion protein, wherein the fusion protein comprises a RAGE polypeptide linked to an immunoglobulin polypeptide,
 a) wherein the RAGE polypeptide comprises a fragment of human RAGE (SEQ ID NO: 3) wherein the fragment of human RAGE comprises a ligand binding site and at least one amino acid residue that may be glycosylated,   b) wherein the immunoglobulin polypeptide comprises a C H 2 domain or a portion of a C H 2 domain of an immunoglobulin and a C H 3 domain of an immunoglobulin, and   c) wherein the N-terminal residue of the immunoglobulin polypeptide is linked to the C-term inal residue of the RAGE polypeptide; and   wherein at least 0.5% of the amount of the fusion protein is aglycosylated and wherein no more than 53.2% of the amount of the fusion protein is aglycosylated.   
     
     
         2 . The method of  claim 1 , wherein the condition is inflammation, and wherein the inflammation is associated with inflammatory bowel disease, rheumatoid arthritis, psoriasis, multiple sclerosis, hypoxia, stroke, heart attack, hemorrhagic shock, sepsis, organ transplantation, impaired wound healing, or autoimmunity. 
     
     
         3 . The method of  claim 1 , wherein at least 30% of the total amount of the fusion protein is aglycosylated. 
     
     
         4 . The method of  claim 1 , wherein the percentage of the amount of the fusion protein in the fully glycosylated form is less than the percentage of the amount of the fusion protein in all of the non-fully glycosylated forms. 
     
     
         5 . The method of  claim 1 , wherein the fusion protein comprises at least three amino acid residues that may be glycosylated, wherein a first potential site of glycosylation is an amino acid residue of the RAGE ligand binding site, a second potential site of glycosylation is an amino acid residue of the RAGE polypeptide, and a third potential site of glycosylation is an amino acid residue of the immunoglobulin polypeptide. 
     
     
         6 . The method of  claim 1 , wherein the RAGE polypeptide comprises a sequence selected from the group consisting of SEQ ID NO:13, SEQ ID NO:14, and SEQ ID NO:15. 
     
     
         7 . The method of  claim 1 , wherein the RAGE polypeptide comprises a sequence selected from the group consisting of SEQ ID NO:16, SEQ ID NO:17, and SEQ ID NO:18. 
     
     
         8 . A method of treating a condition selected from the group consisting of: diabetic nephropathy, diabetic retinopathy, a diabetic foot ulcer, a cardiovascular complication, diabetic neuropathy, amyloidosis, Alzheimer's disease, kidney failure, inflammation, osteoporosis, kidney failure, transplant rejection, and inflammation or rejection associated with transplantation, comprising administering a composition comprising an amount of a fusion protein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO:1 without the signal sequence which comprises from amino acid residue 1 through amino acid residue number 23 or from amino acid residue 1 through amino acid residue number 22, wherein said fusion protein lacks the terminal lysine residue (Lys438), and further wherein at least 0.5% of the amount of the fusion protein is aglycosylated and wherein no more than 53.2% of the amount of the fusion protein is aglycosylated. 
     
     
         9 . The method of  claim 8 , wherein the condition is inflammation, and wherein the inflammation is associated with inflammatory bowel disease, rheumatoid arthritis, psoriasis, multiple sclerosis, hypoxia, stroke, heart attack, hemorrhagic shock, sepsis, organ transplantation, impaired wound healing, or autoimmunity.