Factor ix gene therapy

Abstract

The invention relates to a new, more potent, coagulation Factor IX (FIX) expression cassette for gene therapy of Haemophilia B (HB). Disclosed is a vector for expressing factor IX protein, the vector comprising a promoter, a nucleotide sequence encoding for a functional factor IX protein, and an intron sequence, wherein the intron sequence is positioned between exon 1 and exon 2 of the nucleotide sequence encoding for a functional factor IX protein, and wherein the intron sequence has at least 80% identity to the sequence of SEQ ID NO. 1 as disclosed herein.

1 .- 46 . (canceled) 
     
     
         47 . A vector for expressing factor IX protein, the vector comprising a promoter, a nucleotide sequence encoding for a functional factor IX protein, and an intron sequence, wherein the intron sequence is positioned between exon 1 and exon 2 of the nucleotide sequence encoding for the functional factor IX protein, and wherein:
 the intron sequence has at least 80% identity to the sequence of SEQ ID NO. 1,   the intron sequence has at least 95% identity to the sequence of SEQ ID NO. 1, or   the intron sequence has the sequence of SEQ ID NO. 1.   
     
     
         48 . The vector of  claim 47 , wherein:
 the promoter has a nucleotide sequence which has at least 80% identity to the sequence of SEQ ID NO. 4,   the promoter has a nucleotide sequence which has at least 95% identity to the sequence of SEQ ID NO. 4, or   the promoter has the nucleotide sequence of SEQ ID NO. 4.   
     
     
         49 . The vector of  claim 47 , wherein:
 the nucleotide sequence encoding for the functional protein has at least 80% identity to the sequence of SEQ ID NO. 2,   the nucleotide sequence encoding for the functional FIX protein has at least 95% identity to the sequence of SEQ ID NO. 2, or   the nucleotide sequence encoding for the functional FIX protein has the sequence of SEQ ID NO. 2.   
     
     
         50 . The vector of  claim 47 , wherein:
 the nucleotide sequence encoding the functional FIX protein has at least 80% identity to the sequence of SEQ ID NO. 6,   the nucleotide sequence encoding the functional FIX protein has at least 95% identity to the sequence of SEQ ID NO. 6, or   the nucleotide sequence encoding the functional FIX protein has the sequence of SEQ ID NO. 6.   
     
     
         51 . The vector of  claim 47 , wherein:
 the nucleotide sequence encoding for the functional FIX protein, including the intron sequence between exon 1 and 2, has 80% identity to the sequence of SEQ ID NO. 3,   the nucleotide sequence encoding for the functional FIX protein, including the intron sequence between exon 1 and 2, has 95% identity to the sequence of SEQ ID NO. 3, or   the nucleotide sequence encoding for the functional FIX protein, including the intron sequence between exon 1 and 2, has the sequence of SEQ ID NO. 3.   
     
     
         52 . The vector of  claim 47 , wherein the nucleotide sequence encoding for the functional factor IX protein comprises:
 a nucleotide sequence that has 80% identity, 95% identity, or 100% identity to nucleotides 1-88 (exon 1) of Genbank accession number J00137.1; or   a nucleotide sequence that has 80% identity, 95% identity; or 100% identity to nucleotides 89-197 (exon 2, partial) of Genbank accession number J00137.1.   
     
     
         53 . The vector of  claim 52 , wherein the nucleotide sequence encoding for the functional factor IX protein is codon optimized. 
     
     
         54 . The vector of  claim 53 , wherein the nucleotide sequence encoding for the functional factor IX protein comprises a nucleotide sequence that has 80% identity to Genbank accession number J00137.1 
     
     
         55 . The vector of  claim 47 ; wherein:
 the vector comprises a nucleotide sequence which has 80% identity to the sequence of SEQ ID NO. 5,   the vector comprises a nucleotide sequence which has 95% identity to the sequence of SEQ ID NO. 5, or   the vector comprises a nucleotide sequence which has the sequence of SEQ ID NO. 5.   
     
     
         56 . The vector of  claim 47 , wherein:
 the intron sequence has at least 95% identity to the sequence of SEQ ID NO. 1,   the nucleotide sequence encoding for the functional FIX protein has at least 95% identity to the sequence of SEQ ID NO. 2, and   the promoter has a nucleotide sequence which has at least 95% identity to the sequence of SEQ ID NO. 4, and   wherein the vector is a single stranded vector.   
     
     
         57 . The vector of  claim 47 , wherein the vector:
 is an AAV vector,   is a single stranded vector, or   further comprises a bovine growth hormone poly A tail.   
     
     
         58 . A pharmaceutical composition comprising the vector of  claim 47 , and one or more pharmaceutically acceptable excipients. 
     
     
         59 . A method of treating haemophilia B comprising administering a therapeutically effective amount of the vector of  claim 47  to a patient suffering from haemophilia B.