Chemokine-immunoglobulin fusion polypeptides, compositions, method of making and use thereof

Abstract

This application is directed to chemokine-immunoglobulin fusion polypeptides and chemokine-polymer conjugates. The fusion polypeptides and conjugates can be used for treating chemokine receptor-mediated disorders and modulating inflammation, inflammatory cell motility, cancer cell motility, or cancer cell survival.

1 . An isolated chemokine-immunoglobulin fusion polypeptide, comprising
 a chemokine moiety selected from the group consisting of human and functional variants thereof; and   an immunoglobulin moiety selected from the group consisting of the constant region of human IgG1 (IgG1Fc), the constant region of human IgG2 (I gG1Fc), the constant region of human IgG3 (IgG1Fc), the constant region of human IgG4 (IgG1Fc), and functional variants thereof.   
     
     
         2 . The isolated eheinokine-immunoglobulin fusion polypeptide  claim 1 , wherein said fusion polypeptide is selected from the group consisting of CCL7-IgG1Fc, var-CCL7-IgG1Fc, CCL7-IgG2Fc, var-CCL7-IgG2Fc, CCL7-IgG3Fc, var-CCL7-IgG3Fc, CCL7-IgG4Fc, var-CCL7-IgG4Fc, CCL8-IgG1Fc, var-CCL8-IgG1Fc, CCL8-IgG2Fc, var-CL8-IgG2Fc, CCL8-IgG3Fc, var-CCL8-IgG3Fe, CCL8-IgG4Fc and var-CCL8-IgG4Fc. 
     
     
         3 . The isolated chemokine-immunoglobuliri fusion polypeptide of  claim 1 , wherein said fusion polypeptide is selected from the group consisting of CCL7-IgG1Fc, CCL7(5-76)-IgG1Fc, CCL7(5-76/H→A)-IgG1Fc, CCL7-IgG2Fc, CCL7(5-76)-IgG2Fc, CCL7(5-76K/H→A)-IgG2Fc, CCL7-IgG3Fc, CCL7(5-76)-IgG3Fc, CCL7(5-76K/H→A)-IgG3Fc, CCL7-IgG4Fe, CCL7(5-76)-IgG4Fc, CCL7(5-76K/H→A)-IgG4Fc, CCL8-IgG1Fc, CCL8(5-76)-IgG1Fc, CCL8(5-76K/H→A)-IgG1Fc, CCL8-IgG2Fc, CCL8(5-76)-IgG2Fc, CCL8(5-76K/H→A)-IgG2Fc, CCL8-IgG3Fc, CCL8(5-76)-IgG3Fc, CCL8(5.-76K/H→A)-IgG3Fc, CCL8-IgG4Fc, CCL8(5-76)-IgG4Fc and CCL8(5-76K/H→A)-IgG4Fc 
     
     
         4 . The isolated chemokine-immunoglobulin fusion polypeptide of  claim 1 , wherein said fusion polypeptide is a pegylated fusion polypeptide. 
     
     
         5 . The isolated chemokine-immunoglobulin fusion polypeptide of  claim 4 , wherein said pegylated fusion polypeptide has a molecular weight of at least about 500,000 dalton. 
     
     
         6 . The isolated chemokine-immunoglobulin fusion polypeptide of  claim 4 , wherein said pegylated fusion polypeptide has a PEG-to-fusion polypeptide molar ratio of no more than about 10:1. 
     
     
         7 . The isolated chemokine-immunoglobulin fusion polypeptide of  claim 4 , wherein said pegylated fusion polypeptide has a PEG-to-fusion polypeptide molar ratio of no more than about 2:1. 
     
     
         8 . The isolated chemokine mmunoglobulin fusion polypeptide of  claim 1 , wherein said functional variants of chemokines comprise an N-terminal truncation. 
     
     
         9 . An isolated polynucleotide, encoding the chemokine-immunoglobulin fusion polypeptide of  claim 1 . 
     
     
         10 . An isolated polynucleotide, encoding the chemokine-immunoglobulin fusion polypeptide of  claim 3 . 
     
     
         11 . An expression vector, comprising:
 a regulatory element; and   polynucleotide operably linked to said regulatory element,   wherein said polynucleotide encodes the chemokine -immunoglobulin fusion polypeptide of  claim 1 .   
     
     
         12 . The expression vector of  claim 11 , wherein said vector is a plasmid-based expression vector or a virus-based expression vector. 
     
     
         13 . A pharmaceutical composition, comprising:
 the chemokine-immunoglobulin fusion polypeptide of  claim 1 ; and   a pharmaceutically acceptable carrier.   
     
     
         14 . A pharmaceutical composition, comprising:
 the expression vector of  claim 11 ; and   a pharmaceutically acceptable carrier.   
     
     
         15 . A method for treating a chemokine receptor-mediated disorder in a subject, comprising:
 administering to said subject an effective amount of the pharmaceutical composition of  claim 13 .   
     
     
         16 . The method of  claim 15 , wherein said chemokine receptor-mediated disorder is selected from the group consisting of leukemia, carcinoma, melanoma, sarcoma and lymphoma. 
     
     
         17 . A method for modulating inflammation in a subject, comprising:
 administering to said subject an effective amount of the pharmaceutical composition of  claim 1 .   
     
     
         18 . The method of  claim 17 , wherein said inflammation is selected from the group consisting of inflammation of the central or peripheral nervous system; inflammation of the urogenital system; inflammation of the digestive system; inflammation of the respiratory system; inflammation of the skin, integument and musculoskeletal system; inflammation of the cardiovascular system; autoimmune disorders; cat scratch disease; infections of the eye; Lyme disease; lymphadenopathy; lymphatic inflammation; radiation-induced inflammation; sarcoidosis; Sjogren's syndrome; systemic lupus erythematosus; and inflammation resulting from microbial infections and inflammatory molecules. 
     
     
         19 . A method for treating a chemokine receptor-mediated disorder in a subject, comprising:
 administering to said subject an effective amount of a pegylated chemokine or a pagylated chemokine-immunoglobulin fusion polypeptide, wherein the chemokine is selected from the group consisting of human and functional variants thereof.   
     
     
         20 . The method of  claim 19 , wherein said pegylated chemokine is selected from the group consisting of CCL7-PEG, var-CCL7-PEG, CCL8-PEG and var-CCL8-PEG.