JPWO2020069461A5 - - Google Patents

Claims (33)

An adeno-associated virus (AAV) vector genome comprising an engineered promoter and a payload region encoding a frataxin (FXN) protein , wherein said engineered promoter is at least 90% identical to SEQ ID NO: 1742 an adeno-associated virus (AAV) vector genome that contains a sequence having a unique property and is 50-300 nucleotides in length . 2. The AAV vector genome of claim 1, wherein said engineered promoter comprises a sequence having at least 95% sequence identity to SEQ ID NO:1742. 3. The AAV vector genome of claim 1 or 2 , wherein said engineered promoter comprises a sequence having at least 99% sequence identity to SEQ ID NO:1742. The AAV vector genome of any one of claims 1-3, wherein said engineered promoter comprises the sequence of SEQ ID NO:1742. Claim 1 , wherein said frataxin protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity to SEQ ID NO: 1725, 1726 or 1727. 5. The AAV vector genome of any one of items 1-4 . The AAV vector genome of any one of claims 1-5 , wherein said frataxin protein comprises the amino acid sequence of SEQ ID NO:1725. a nucleotide sequence wherein said frataxin protein has at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity to SEQ ID NO: 1728, 1729 or 1730, or a functional fragment thereof The AAV vector genome of any one of claims 1-6 , encoded by 8. The AAV vector genome of claim 7 , wherein the nucleotide sequence encoding the frataxin protein comprises the nucleotide sequence of SEQ ID NO: 1728 or a functional fragment thereof. 9. The AAV vector genome of claim 7 or 8 , wherein the nucleotide sequence encoding the frataxin protein comprises or consists of nucleotides 221-853 of SEQ ID NO:1728. Claims 1-6 , wherein the nucleotide sequence encoding the frataxin protein has at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity to SEQ ID NO: 1823 or 1824. The AAV vector genome of any one of Claims 1 to 3. 11. The AAV vector genome of claim 10 , wherein the nucleotide sequence encoding the frataxin protein comprises the nucleotide sequence of SEQ ID NO: 1823 or 1824. The AAV vector genome of any one of claims 1-11, wherein said AAV vector genome further comprises a 5' terminal inverted sequence (ITR). 13. The AAV vector genome of claim 12 , wherein said 5' ITR is the AAV2 ITR. 14. The AAV vector genome of claim 12 or 13, wherein said AAV vector genome further comprises a 3'ITR. 15. The AAV vector genome of claim 14 , wherein said 3' ITR is the AAV2 ITR. 16. Any one of claims 1 to 15 , wherein the AAV vector genome comprises one or more of the following components: introns, microRNA (miR) binding sites, polyadenylation (polyA) sequences or filler sequences. AAV vector genomes as described. 17. Any of claims 1-16, wherein the AAV vector genome comprises an intron, a microRNA (miR) binding site, and a polyadenylation (polyA) sequence, the intron comprising or consisting of an enhancer sequence. AAV vector genome according to paragraph 1. wherein the AAV vector genome contains an intron;
18. The AAV vector genome of claim 16 or 17 , wherein said intron comprises a sequence selected from the group consisting of SEQ ID NOS: 1815-1821. wherein the AAV vector genome comprises a microRNA (miR) binding site;
19. The AAV vector genome of any one of claims 16-18 , wherein said m i R binding site comprises a sequence having at least 95% sequence identity to SEQ ID NO:1827. 20. The AAV vector genome of Claim 19 , wherein said miR binding site comprises the sequence of SEQ ID NO:1827. 21. The AAV vector genome of claim 19 or 20 , wherein said AAV vector genome comprises: 3 copies of said miR binding site . 22. The AAV vector genome of any one of claims 1-21, wherein said AAV vector genome comprises a sequence having at least 90% sequence identity to SEQ ID NO:1797. 23. The AAV vector genome of any one of claims 1-22, wherein said AAV vector genome comprises a sequence having at least 95% sequence identity to SEQ ID NO:1797. 24. The AAV vector genome of any one of claims 1-23, wherein said AAV vector genome comprises a sequence having at least 99% sequence identity to SEQ ID NO:1797. 25. The AAV vector genome of any one of claims 1-24, wherein said AAV vector genome comprises the sequence of SEQ ID NO:1797. An AAV particle comprising an AAV vector genome and an AAV capsid according to any one of claims 1-25. 27. The AAV particle of claim 26, wherein said AAV capsid is an AAV5 variant capsid or an AAV9 variant capsid. 28. A pharmaceutical composition comprising the AAV particles of claim 26 or 27 . 29. A pharmaceutical composition according to claim 28 for the treatment of a neurological or neuromuscular disorder in a subject . 30. The pharmaceutical composition of claim 29 , wherein the neurological or neuromuscular disorder is a disorder associated with decreased frataxin protein levels or Friedreich's ataxia. administration of said pharmaceutical composition to said subject compared to frataxin expression in equivalent target cells in subjects without disorders associated with decreased frataxin protein levels and Friedreich's ataxia 31. The pharmaceutical composition of claim 30 , which results in a 0.5x to 3.0x increase in frataxin protein expression in target cells. 28. Use of AAV particles according to claims 26 or 27 for the manufacture of a therapeutic agent for a neurological or neuromuscular disorder in a subject. 33. Use according to claim 32, wherein the neurological or neuromuscular disorder is a disorder associated with decreased frataxin protein levels or Friedreich's ataxia.