Npp1 fusion proteins

Abstract

The present invention provides a novel fusion polypeptide containing a catalytic domain of NPP1 fused to a targeting moiety, nucleic acids encoding the fusion polypeptide, a vector containing the nucleic acid integrated thereinto, a host cell transformed with the vector and Pharmaceutical compositions comprising the fusion polypeptide.

1 . An isolated polypeptide comprising an ectonucleotide pyrophosphatase/phosphodiesterase 1 (NPP1) component, wherein the NPP1 component is a portion of NPP1 that comprises the cysteine-rich region and the C-terminus catalytic domain of NPP1, but having the N-terminal cytosolic and the transmembrane domains removed, said isolated polypeptide having NPP1 enzymatic activity. 
     
     
         2 . (canceled) 
     
     
         3 . The polypeptide of  claim 1 , wherein said polypeptide is a recombinant protein. 
     
     
         4 - 7 . (canceled) 
     
     
         8 . The polypeptide of  claim 1 , wherein said catalytic domain of NPP1 comprises P99 through D925 of SEQ ID NO: 1. 
     
     
         9 - 18 . (canceled) 
     
     
         19 . The polypeptide of  claim 1 , wherein said polypeptide further comprises an Fc region of an immunoglobulin. 
     
     
         20 . The polypeptide of  claim 1 , wherein said polypeptide further comprises a signal peptide. 
     
     
         21 . The polypeptide of  claim 1 , wherein said polypeptide forms a homodimer. 
     
     
         22 . The polypeptide of  claim 1 , wherein said polypeptide protein is a monomer. 
     
     
         23 . An isolated nucleic acid encoding the polypeptide of  claim 1 . 
     
     
         24 . A replication or expression vector carrying the isolated nucleic acid of  claim 23 . 
     
     
         25 . A host cell transformed with the replication or expression vector according to  claim 23 . 
     
     
         26 . The host cell of  claim 25 , wherein said host cell is selected from the group consisting of CHO cell, HEK293 cell, or COS cell. 
     
     
         27 . The host cell of  claim 20 , wherein said host cell is an avian tumor cell. 
     
     
         28 . A transgenic animal producing the polypeptide of  claim 1 . 
     
     
         29 . The transgenic animal of  claim 28 , wherein said transgenic animal is mammalian or avian. 
     
     
         30 . A method producing said polypeptide of  claim 1  by an avian cell comprising culturing an avian cell transfected with at least one expression vector comprising a transcription unit having a nucleotide sequence encoding said polypeptide of  claim 1  operably linked to a promoter and a transcriptional terminator, and wherein the cultured avian cell produces said polypeptide. 
     
     
         31 . (canceled) 
     
     
         32 . A method of treating a disorder in a subject in need thereof comprising administering an effective amount of a pharmaceutical composition comprising the polypeptide of  claim 1 . 
     
     
         33 - 34 . (canceled) 
     
     
         35 . The method of  claim 32 , wherein said disorder is arterial calcification, insulin resistance, hypophosphatemic rickets, or ossification of posterior longitudinal ligament of spine. 
     
     
         36 . The method of  claim 35 , wherein said disorder is arterial calcification. 
     
     
         37 . (canceled) 
     
     
         38 . The method of  claim 36 , wherein the arterial calcification is generalized arterial calcification of infancy. 
     
     
         39 . An isolated polypeptide comprising a targeting moiety and a C-terminus catalytic domain of NPP1.