sRAGE mimetibody, compositions, methods and uses

Abstract

Mammalian sRAGE mimetibody polypeptides and nucleic acids are disclosed. Methods of utilizing the polypeptides to reduce or inhibit the binding of RAGE and its ligands and to treat RAGE-related diseases are also disclosed.

1 . A polypeptide according to formula (II):  
         (Rg-Lk-V2—Hg—C H 2—C H 3) (t)  (II)  
       where Rg is a mammalian sRAGE sequence, Lk is a polypeptide or chemical linkage, V2 is a portion of a C-terminus of an immunoglobulin variable region, Hg is at least a portion of an immunoglobulin variable hinge region, C H 2 is an immunoglobulin heavy chain C H 2 constant region and C H 3 is an immunoglobulin heavy chain C H 3 constant region and t is independently an integer from 1 to 10.  
     
     
         2 . The polypeptide of  claim 1  wherein Rg is a biologically active fragment of or a polypeptide substantially homologous to SEQ ID NO: 1.  
     
     
         3 . The polypeptide of  claim 1  wherein Rg has the amino acid sequence shown in residues 31 to 106, 31 to 215 or 31 to 308 of SEQ ID NO: 1.  
     
     
         4 . The polypeptide of  claim 1  wherein Hg, C H 2 and C H 3 are of the IgG1 subclass.  
     
     
         5 . The polypeptide of  claim 4  wherein Cys220 of Hg is substituted with Ala, and Leu234 and Leu235 of C H 2—C H 3 are mutated to Ala234 and Ala235.  
     
     
         6 . The polypeptide of  claim 1  wherein Hg is of the IgG4 subclass, and C H 2 and C H 3 are of the IgG1 subclass.  
     
     
         7 . The sRAGE mimetibody polypeptide of  claim 1  wherein Hg, C H 2 and C H 3 are of the IgG4 subclass.  
     
     
         8 . The polypeptide of  claim 1  wherein Ser228 of Hg is substituted with Pro and Phe234 and Leu235 of C H 2—C H 3 are mutated to Ala234 and Ala235.  
     
     
         9 . The polypeptide of  claim 1  wherein the polypeptide binds to at least one RAGE ligand.  
     
     
         10 . The polypeptide of  claim 9  wherein the ligand is at least one of advanced glycated endproducts (AGE), amphoterin, S100/calgranulin and β-amyloid.  
     
     
         11 . A polypeptide comprising a polypeptide having the sequence shown in SEQ ID NO: 3, 5, 7, 9, 11 or 13.  
     
     
         12 . A polynucleotide encoding a polypeptide according to  claim 1 .  
     
     
         13 . A polynucleotide comprising a polynucleotide having the sequence shown in SEQ ID NO: 4, 6, 8, 10, 12 or 14 or a complementary sequence.  
     
     
         14 . A polynucleotide comprising a polynucleotide encoding the amino acid sequence shown in SEQ ID NO: 3, 5, 7, 9, 11 or 13.  
     
     
         15 . A vector comprising the polynucleotide of  claim 13  or  14 .  
     
     
         16 . A cell line expressing a polypeptide according to  claim 1 .  
     
     
         17 . A cell line comprising the vector of  claim 15 .  
     
     
         18 . The cell line of  claim 17  wherein the cell line is HEK293, NSO, SP2/0 or CHO cells.  
     
     
         19 . A pharmaceutical composition comprising an effective amount of at least one polypeptide according to  claim 1  and a pharmaceutically acceptable carrier or diluent.  
     
     
         20 . A method of modifying the biological activity of RAGE in a mammal comprising administering the pharmaceutical composition of  claim 19  to the mammal.  
     
     
         21 . A method of reducing the symptoms of, or treating at least one RAGE-related condition or disorder, comprising administering the pharmaceutical composition of  claim 19  to a patient in need thereof.  
     
     
         22 . The method of  claim 21  wherein the RAGE-related condition or disorder is an immune disorder, a cardiovascular disorder, a metabolic disease, a malignant disorder, or a neurological disorder.  
     
     
         23 . The method of  claim 21  wherein the RAGE-related condition or disorder is diabetes, Alzheimer's disease, atherosclerosis, tumor growth and metastasis, colitis, delayed type hypersensitivity, multiple sclerosis or oxidant stress.  
     
     
         24 . A method to produce a polypeptide comprising the steps of culturing the cell line of  claim 16  and purifying the expressed polypeptide.