MODIFIED COAGULATION FACTOR VIIa WITH EXTENDED HALF-LIFE

Abstract

The present invention relates to the fields of Factor VII (FVII) and Factor VIIa (FVIIa) albumin linked polypeptides. More specifically, the invention relates to cDNA sequences coding for human Factor VII and Factor VIIa and derivatives genetically fused to a cDNA coding for human serum albumin which may be linked by oligonucleotides which code for intervening peptidic linkers such encoded derivatives exhibiting improved stability and extended functional plasma half-life, recombinant expression vectors containing such cDNA sequences, host cells transformed with such recombinant expression vectors, recombinant polypeptides and derivatives which do have biological activities of the unmodified wild type protein but having improved stability and prolonged shelf-life and processes for the manufacture of such recombinant proteins and their derivatives. The invention also covers a transfer vector for use in human gene therapy, which comprises such modified DNA sequences.

1 - 29 . (canceled) 
     
     
         30 . A method of treating a bleeding disorder, comprising administering an effective amount of an albumin fusion polypeptide to an individual in need thereof, wherein the albumin fusion polypeptide comprises:
 (a) Factor VII or Factor VIIa,   (b) albumin, and   (c) a peptide linker that joins the Factor VII or Factor VIIa to the N-terminus of the albumin,   wherein the peptide linker is at least 25 amino acids in length and comprises Ser-Ser-(Gly-Gly-Ser) n -Gly-Ser, wherein n is an integer greater than or equal to 7.   
     
     
         31 . The method of  claim 30 , wherein n is an integer greater than or equal to 9, and wherein the peptide linker is at least 31 amino acids in length. 
     
     
         32 . The method of  claim 30 , wherein the linker comprises Ser-Ser-(Gly-Gly-Ser) n -Gly-Ser, and n is 9. 
     
     
         33 . The method of  claim 30 , wherein the linker comprises Ser-Ser-(Gly-Gly-Ser) n -Gly-Ser, n is 9, and the peptide linker is 31 amino acids in length. 
     
     
         34 . The method of  claim 30 , wherein the linker comprises at least one N-glycosylation site of the structure Asn-X-Ser/Thr, wherein X denotes any amino acid except proline. 
     
     
         35 . The method of  claim 30 , wherein the albumin fusion polypeptide has a Factor VII/VIIa molar specific activity that is increased by at least 100% as compared to a Factor VII- or Factor VIIa-albumin fusion polypeptide without a linker. 
     
     
         36 . The method of  claim 30 , wherein the albumin fusion polypeptide has an increased functional plasma half-life in vivo as compared to an unfused Factor VII or Factor VIIa. 
     
     
         37 . The method of  claim 36 , wherein the functional plasma half-life of the albumin fusion polypeptide is increased by at least 100% as compared to the unfused Factor VII or Factor VIIa. 
     
     
         38 . The method of  claim 30 , wherein the peptide linker contains a protease cleavage site. 
     
     
         39 . The method of  claim 38 , wherein the cleavage site can be cleaved by one or more of Factor IIa, Factor IXa, Factor Xa, Factor XIa, Factor XIIa, activated protein C, elastase, and kallikrein. 
     
     
         40 . The method of  claim 30 , wherein the albumin fusion polypeptide is modified to comprise an activation peptide from a vitamin K-dependent polypeptide other than Factor VII or Factor VIIa. 
     
     
         41 . The method of  claim 30 , wherein the Factor VII or Factor VIIa has procoagulant activity. 
     
     
         42 . The method of  claim 30 , wherein the albumin fusion polypeptide is in a pharmaceutical composition that further comprises a pharmaceutically acceptable carrier or excipient. 
     
     
         43 . The method of  claim 30 , wherein the bleeding disorder is a blood coagulation disorder. 
     
     
         44 . The method of  claim 30 , wherein the bleeding disorder is hemophilia. 
     
     
         45 . The method of  claim 30 , wherein the bleeding disorder is hemophilia A. 
     
     
         46 . A nucleic acid molecule encoding an albumin fusion polypeptide, wherein the albumin fusion polypeptide comprises:
 (a) Factor VII or Factor VIIa,   (b) albumin, and   (c) a peptide linker that joins the Factor VII or Factor VIIa to the N-terminus of the albumin,   wherein the peptide linker is at least 25 amino acids in length and comprises Ser-Ser-(Gly-Gly-Ser) n -Gly-Ser, wherein n is an integer greater than or equal to 7.   
     
     
         47 . A plasmid or vector comprising the nucleic acid molecule of  claim 46 . 
     
     
         48 . The plasmid or vector of  claim 47 , wherein the plasmid or vector is an expression vector operable for use in human gene therapy. 
     
     
         49 . A host cell comprising the nucleic acid molecule of  claim 46 .