Factor IX polypeptide mutant, its uses and a method for its production

Abstract

Disclosed are a modified FIX (factor IX) polypeptide comprising a leucine, cysteine, aspartic acid, glutamic acid, histidine, lysine, asparagine, glutamine or tyrosine in position 338; pharmaceutical preparations containing said modified FIX polypeptide; a nucleotide sequence coding for the modified FIX polypeptide; and a method for producing the modified FIX polypeptide.

The invention claimed is: 
     
       1. A pharmaceutical composition comprising:
 a modified recombinant FIX (factor IX) polypeptide comprising at least 70% identity to SEQ ID NO: 2 and a leucine in position 338 of SEQ ID NO: 2; the modified recombinant FIX polypeptide in an amount suitable to provide a daily dosage comprised between 0.1 μg/kg and 400 μg/kg body weight, and   at least one pharmaceutically acceptable excipient.   
     
     
       2. The pharmaceutical composition according to  claim 1 , wherein the polypetide comprises at least 90% identity to SEQ ID NO: 2. 
     
     
       3. The pharmaceutical composition according to  claim 1  formulated for oral, parenteral or rectal administration or suited to administration by inhalation or insufflation. 
     
     
       4. A method of treatment, the method comprising administering to an individual in need thereof the pharmaceutical composition according to  claim 1 . 
     
     
       5. The method according to  claim 4  wherein the administering is performed to treat at least one coagulopathy in the individual. 
     
     
       6. A sequence of nucleotides encoding a modified FIX (Factor IX) polypeptide comprising at least 70% identity to SEQ ID NO: 2 and a leucine at position 338 of SEQ ID NO: 2. 
     
     
       7. The sequence of nucleotides according to  claim 6 , wherein the nucleotide sequence has at least 90% homology with the sequence having accession number K02402 (GenBank). 
     
     
       8. A nucleotide sequence encoding a modified FIX (Factor IV) polypeptide comprising 100% identity to SEQ ID NO: 2 except for an alanine at position 148 and a leucine at position 338 of SEQ ID NO: 2 in combination with positions corresponding to positions 34098, 34099 and 34100 of a sequence having accession number K02402 (GenBank) having a triplet selected from the group consisting of: TTA, UUA, TTG, UUG, CTT, CUU, CTC, CUC, CTA, CUA, CTG, and CUG, GAT, GAU and GAC. 
     
     
       9. A nucleic acid comprising the nucleotide sequence according to  claim 6 . 
     
     
       10. The nucleic acid according to  claim 9 , the nucleic acid comprising a promoter in operational linkage with said nucleotide sequence. 
     
     
       11. A vector comprising the nucleic acid according to  claim 9 . 
     
     
       12. A pharmaceutical composition comprising the sequence of nucleotide according to  claim 6 . 
     
     
       13. A method for producing a modified FIX (Factor IX) polypeptide, the method comprising
 expressing the modified FIX polypeptide by means of a nucleic acid according to  claim 9 .   
     
     
       14. A method to perform gene therapy, the method comprising
 administering to an individual in need thereof the nucleotide sequence according to  claim 6  via a vector configured for gene therapy.   
     
     
       15. The method according to  claim 14 , wherein the administering is performed to treat at least one coagulopathy in the individual. 
     
     
       16. The nucleotide sequence of  claim 8 , wherein the triplet is selected from the group consisting of CTC and CTG.  
     
     
       17. The nucleotide sequence of  claim 8 , wherein the triplet is selected from the group consisting of CUC and CUG.  
     
     
       18. The nucleotide sequence of  claim 8 , wherein the triplet is CTC.  
     
     
       19. The nucleotide sequence of  claim 8 , wherein the triplet is CTG.  
     
     
       20. The nucleotide sequence of  claim 8 , wherein the triplet is CUC.  
     
     
       21. The nucleotide sequence of  claim 8 , wherein the triplet is CUG.