FVII or FVIIa Gla domain variants

Abstract

Gla domain variants of human Factor VII or human Factor VIIa, comprising 1-15 amino acid modifications relative to human Factor VII or human Factor VIIa, wherein a hydrophobic amino acid residue has been introduced by substitution in position 34; or having an amino acid substitution in position 36; and use of the variants for the treatment of intracerebral haemorrhage (ICH) or trauma.

1 . A Factor VII (FVII) or Factor VIa (FVIIa) polypeptide variant having an amino acid sequence comprising 1-15 amino acid modifications relative to human Factor VII (hFVII) or human Factor VIIa (hFVIIa) with the amino acid sequence shown in SEQ ID NO:1, the variant comprising an amino acid substitution in position 36.  
     
     
         2 . The variant of  claim 1 , wherein a negatively charged amino acid residue has been introduced by substitution in position 36.  
     
     
         3 . The variant of  claim 2 , wherein said substitution is R36D.  
     
     
         4 . The variant of  claim 2 , wherein said substitution is R36E.  
     
     
         5 . The variant of  claim 1 , further comprising an amino acid substitution in position 34.  
     
     
         6 . The variant of  claim 5 , wherein a negatively charged amino acid residue has been introduced by substitution in position 34.  
     
     
         7 . The variant of  claim 6 , wherein said substitution is A34E.  
     
     
         8 . The variant of  claim 7  comprising the substitutions A34E+R36E.  
     
     
         9 . The variant of  claim 1 , further comprising an amino acid substitution in position 10 and/or 32.  
     
     
         10 . The variant of  claim 9 , comprising the substitution K32E.  
     
     
         11 . The variant of  claim 9 , comprising the substitution P10Q.  
     
     
         12 . The variant of  claim 9 , comprising the substitutions P10Q+K32E.  
     
     
         13 . The variant of  claim 12 , comprising the substitutions P10Q+K32E+A34E+R36E.  
     
     
         14 . The variant of  claim 1 , wherein at least one amino acid residue comprising an attachment group for a non-polypeptide moiety has been introduced in a position located outside the Gla domain.  
     
     
         15 . The variant of  claim 14 , wherein said attachment group is an in vivo N-glycosylation site introduced by substitution.  
     
     
         16 . The variant of  claim 15 , wherein said in vivo N-glycosylation site is introduced by a substitution selected from the group consisting of A51N, G58N, T106N, K109N, G124N, K143N+N145T, A175T, I205S, I205T, V253N, T267N, T267N+S269T, S314N+K316S, S314N+K316T, R315N+V317S, R315N+V317T, K316N+G318S, K316N+G318T, G318N, D334N and combinations thereof.  
     
     
         17 . The variant of  claim 16 , comprising at least one substitution selected from the group consisting of T106N, I205T and V253N.  
     
     
         18 . The variant of  claim 17 , comprising at least two substitutions selected from the group consisting of T106N, I205T and V253N.  
     
     
         19 . The variant of  claim 17 , comprising the substitutions P10Q+K32E+A34E+R36E+T106N+V253N.  
     
     
         20 . The variant of  claim 1 , wherein said variant is in its activated form.  
     
     
         21 . A nucleotide sequence encoding a variant as defined in  claim 1 .  
     
     
         22 . An expression vector comprising the nucleotide sequence of  claim 21 .  
     
     
         23 . A host cell comprising the nucleotide sequence of  claim 21  or an expression vector comprising said nucleotide sequence.  
     
     
         24 . A method for producing the FVII or FVIIa polypeptide variant of  claim 1 , comprising culturing a eukaryotic host cell capable of in vivo N-glycosylation under conditions conducive for the expression of the polypeptide variant, and recovering the polypeptide variant.  
     
     
         25 . A composition comprising the variant of  claim 1  and at least one pharmaceutical acceptable carrier or excipient.  
     
     
         26 . A method for treating a mammal having a disease or a disorder wherein clot formation is desirable, comprising administering to a mammal in need thereof an effective amount of the FVII or FVIIa polypeptide variant of  claim 1 .  
     
     
         27 . The method of  claim 26 , wherein said disease or disorder is selected from the group consisting of hemorrhages, including brain hemorrhages, severe uncontrolled bleedings, such as trauma, bleedings in patients undergoing transplantations or resection, variceal bleeding, and hemophilia.  
     
     
         28 . The method of  claim 27 , wherein said disease or disorder is trauma.  
     
     
         29 . The method of  claim 27 , wherein said disease or disorder is hemophilia.  
     
     
         30 . A method for treating intracerebral haemorrhage or traumatic brain injury, comprising administering to a patient in need thereof an effective amount of the FVII or FVIIa polypeptide variant of  claim 1.