Chimeric clotting factors

Abstract

The invention provides chimeric clotting factors comprising an activatable clotting factor and an enhancer moiety. The activatable clotting factor allows the chimeric clotting factor to be activated at the site of coagulation. The enhancer moiety can additionally improve procoagulation activities of the chimeric clotting factors. The chimeric clotting factors can further be improved by fusion to a half-life extender, which improves a pharmacokinetics property of the chimeric clotting factor. The invention also includes methods of making and methods of using these chimeric clotting factors.

1 - 79 . (canceled) 
     
     
         80 . A nucleic acid molecule encoding a chimeric protein or a complement thereof, wherein the chimeric protein comprises (i) an activatable clotting factor (Ac), (ii) an enhancer moiety (Em), and (iii) an optional linker moiety (L or L1) between the activatable clotting factor and an enhancer moiety. 
     
     
         81 . (canceled) 
     
     
         82 . A vector comprising the nucleic acid molecule of  claim 80 . 
     
     
         83 . (canceled) 
     
     
         84 . The vector according to  claim 82 , further comprising a nucleotide sequence encoding an intracellular processing enzyme which processes at least one of the intracellular processing sites in the chimeric protein or a complement thereof. 
     
     
         85 . A host cell comprising the vector according to  claim 82 . 
     
     
         86 . The host cell of  claim 85 , which is a HEK293 cell, a BHK cell, or a CHO cell. 
     
     
         87 . A method for producing a chimeric protein comprising culturing the host cell according to  claim 85  and recovering the chimeric protein from the culture medium. 
     
     
         88 - 100 . (canceled) 
     
     
         101 . A chimeric protein comprising a linear structure from amino terminus to carboxy terminus represented by the formula Ac-L1-Em, wherein Ac is a protease activatable FVII clotting factor, Em is soluble tissue factor, and L1 is a linker moiety comprising a gly/ser peptide connecting the carboxy terminus of Ac and the amino terminus of Em; wherein the protease activatable FVII clotting factor Ac comprises a light chain (LC) and a heavy chain (HC), and a thrombin-cleavage site inserted between the carboxy terminus of the LC and the amino terminus of the HC. 
     
     
         102 - 106 . (canceled) 
     
     
         107 . A host cell comprising the vector according to  claim 84 . 
     
     
         108 . The host cell of  claim 107 , which is a HEK293 cell, a BHK cell, or a CHO cell. 
     
     
         109 . A method for producing a chimeric protein comprising culturing the host cell according to  claim 107  and recovering the chimeric protein from the culture medium. 
     
     
         110 . A method for treating, ameliorating, or preventing a bleeding disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of a nucleic acid molecule encoding a chimeric protein, wherein the chimeric protein comprises (i) an activatable clotting factor (Ac), (ii) an enhancer moiety (Em), and (iii) an optional linker moiety (L or L1) between the activatable clotting factor and an enhancer moiety. 
     
     
         111 . The method according to  claim 110 , wherein the bleeding disease or disorder is hemophilia A or hemophilia B. 
     
     
         112 . The method according to  claim 110 , wherein the subject is a human subject.