NPP1 fusion proteins
View Patent ↗The present invention provides a novel fusion polypeptide containing a catalytic portion 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. A recombinant nucleic acid that encodes a NPP1 fusion protein, wherein the NPP1 fusion protein comprises an NPP1 component, a targeting moiety, and an Fc region of an immunoglobulin, wherein a) the NPP1 component comprises the cysteine-rich region and the C-terminus catalytic domain of NPP1, b) the targeting moiety and the Fc region are each located C-terminally to the NPP1 component, and c) the NPP1 fusion protein has NPP1 enzymatic activity.
2. The recombinant nucleic acid of claim 1 , wherein said NPP1 enzymatic activity is pyrophosphatase activity, phosphodiesterase activity or pyrophosphatase and phosphodiesterase activity.
3. The recombinant nucleic acid of claim 1 , wherein said NPP1 component consists of P99 through D925 of SEQ ID NO: 1.
4. The recombinant nucleic acid of claim 3 , wherein said targeting moiety is a peptide comprising at least four negatively charged amino acid residues.
5. The recombinant nucleic acid of claim 3 , wherein said targeting moiety is a peptide comprising between about five and about fifteen negatively charged amino acid residues.
6. The recombinant nucleic acid of claim 5 , wherein said negatively charged amino acid residues comprise about eight consecutive aspartic acid or glutamic acid residues.
7. The recombinant nucleic acid of claim 5 , wherein said negatively charged amino acid residues comprise at least four aspartic acid residues.
8. The recombinant nucleic acid of claim 5 , wherein said negatively charged amino acid residues comprise at least four glutamic acid residues.
9. The recombinant nucleic acid of claim 5 , wherein said negatively charged amino acid residues comprise eight consecutive aspartic acid residues.
10. The recombinant nucleic acid of claim 3 , wherein said targeting moiety is fused to the C-terminus of said catalytic domain of NPP1.
11. The recombinant nucleic acid of claim 3 , wherein said Fc region is fused to the C-terminus of said catalytic domain of NPP1.
12. The recombinant nucleic acid of claim 1 , wherein said fusion protein further comprises a polypeptide linker between said targeting moiety and said catalytic domain of NPP1.
13. The recombinant nucleic acid of claim 1 , wherein said recombinant nucleic acid further encodes a signal peptide at the N-terminus of the fusion protein.
14. The recombinant nucleic acid of claim 1 , wherein said fusion protein forms a homodimer.
15. The recombinant nucleic acid of claim 1 , wherein said fusion protein is a monomer.
16. The recombinant nucleic acid of claim 1 , wherein the recombinant nucleic acid is an expression vector.