Compositions and methods of treating Huntington's disease

Patent metadata

Field	Value
Publication number	US-11752181-B2
Application number	US-201816611041-A
Country	US
Kind code	B2
Filing date	May 4, 2018
Priority date	May 5, 2017
Publication date	Sep 12, 2023
Grant date	Sep 12, 2023

Abstract

The present invention relates to adeno-associated viral (AAV) particles encoding siRNA molecules and methods for treating Huntington's Disease (HD).

First claim

We claim:

1. An adeno-associated viral (AAV) genome comprising a first inverted terminal repeat (ITR), a promoter operably linked to a nucleic acid comprising a modulatory polynucleotide encoding a sense strand sequence and an antisense strand sequence, and a second ITR; wherein the encoded antisense strand sequence comprises the nucleotide sequence of SEQ ID NO: 918 and the sense strand sequence comprises the nucleotide sequence of SEQ ID NO: 1079; and wherein the AAV viral genome comprises one or more of:
(i) a first ITR sequence comprising the nucleotide sequence of any one of SEQ ID NOs: 1380 or 1381-1383;
(ii) a promoter comprising a chicken β-actin (CBA) promoter, a cytomegalovirus (CMV) promoter, a phosphoglycerate kinase 1 (PGK) promoter, an H1 promoter, a T7 promoter, a ubiquitin c (UBC) promoter, a β-glucuronidase (GUSB) promoter, a neuron-specific enolase (NSE) promoter, a synapsin promoter, a methyl-CpG binding protein 2 (MeCP2) promoter, or a glial fibrillary acidic protein (GFAP) promoter;
(iii) a promoter comprising the nucleotide sequence of any one of SEQ ID NOs: 1410, 1411-1414; or
(iv) a second ITR sequence comprising the nucleotide sequence of any one of SEQ ID NOs: 1382, 1380, 1381, or 1383.

2. The AAV viral genome of claim 1 , wherein the nucleotide sequence encoding the antisense strand sequence comprises the nucleotide sequence of SEQ ID NO: 1335, and the nucleotide sequence encoding the sense strand sequence comprises the nucleotide sequence of SEQ ID NO: 1331.

3. The AAV viral genome of claim 1 , wherein:
(i) the encoded sense strand sequence and the encoded antisense strand sequence comprises a 3′ overhang of at least 1 or 2 nucleotide; and/or
(ii) the encoded antisense strand sequence and/or encoded sense strand sequence independently comprises at least 21, 22, 23 or 24 nucleotides in length.

4. The AAV viral genome of claim 1 , wherein the modulatory polynucleotide comprises in 5′ to 3′ order:
(i) a 5′ flanking region, the sense strand sequence, a loop region, the antisense strand sequence, and a 3′ flanking region; or
(ii) a 5′ flanking region, the antisense strand sequence, a loop region, the sense strand sequence, and a 3′ flanking region.

5. The AAV viral genome of claim 4 , wherein:
(i) the 5′ flanking region comprises the nucleotide sequence of any one of SEQ ID NOs: 1163, 1167, 1161, 1162, or 1164-1166;
(ii) the loop region comprises the nucleotide sequence of any one of SEQ ID NOs: 1172, 1175, 1169, 1168, 1170, 1171, 1173, or 1174; and/or
(iii) the 3′ flanking region comprises the nucleotide sequence of any one of SEQ ID NOs: 1178, 1182, or 1179-1181.

6. The AAV viral genome of claim 1 , wherein the modulatory polynucleotide comprises the nucleotide sequence of SEQ ID NO: 1262.

7. The AAV viral genome of claim 1 , wherein:
(i) the first ITR sequence comprises the nucleotide sequence of any one of SEQ ID NOs: 1380 or 1381-1383;
(ii) the promoter comprises the nucleotide sequence of any one of SEQ ID NOs: 1410, 1411-1414; and
(iii) the second ITR sequence comprises the nucleotide sequence of any one of SEQ ID NOs: 1382, 1380, 1381, or 1383.

8. The AAV viral genome of claim 1 , which further comprises:
(i) an enhancer;
(ii) at least 1 or at least 2 multiple cloning site (MCS);
(iii) an intron;
(iv) an exon;
(v) a filler sequence;
(vi) a microRNA (miRNA) binding site; and/or
(vii) a polyadenylation (polyA) signal sequence region.

9. The AAV viral genome of claim 8 , wherein:
(i) the enhancer comprises a CMV enhancer;
(ii) the enhancer comprises SEQ ID NO: 1408 or 1409;
(iii) the MCS comprises the nucleotide sequence any one of SEQ ID NOs: 1384-1387 or 1389 or the nucleotide sequence of TCGAG;
(iv) the intron comprises an SV40 intron or a beta-globin intron,
(v) the intron comprises the nucleotide sequence of any one of SEQ ID NOs: 1417-1419;
(vi) the exon comprises a beta-globin exon;
(vii) the exon comprises the nucleotide sequence of SEQ ID NO: 1415 or 1416;
(viii) the polyA signal sequence region comprises a rabbit globin polyadenylation signal sequence region; and/or
(ix) the polyA signal sequence region comprises the nucleotide sequence of any one of SEQ ID NOs: 1420-1423.

10. The AAV viral genome of claim 1 , comprising in order:
(i) a 5′ inverted terminal repeat (ITR) sequence region, wherein the 5′ ITR sequence region comprises the nucleotide sequence of SEQ ID NO: 1380 or 1381;
(ii) an enhancer sequence region, wherein the enhancer sequence region comprises the nucleotide sequence of SEQ ID NO: 1408 or 1409;
(iii) a promoter sequence region, wherein the promoter sequence region comprises the nucleotide sequence of any one of SEQ ID NOs: 1410-1414;
(iv) a first intron sequence region, wherein the first intron sequence comprises the nucleotide sequence of any one of SEQ ID NOs: 1417-1419;
(v) a modulatory polynucleotide sequence region, wherein the modulatory polynucleotide sequence region comprises the nucleotide sequence of SEQ ID NO: 1262;
(vi) a polyadenylation (polyA) signal sequence region, wherein the polyA signal sequence region comprises the nucleotide sequence of any one of SEQ ID NOs: 1420-1423; and
(vii) a 3′ ITR sequence region, wherein the 3′ ITR sequence region comprises the nucleotide sequence of SEQ ID NO: 1382 or 1383.

11. The AAV viral genome of claim 1 , comprising, from 5′ to 3′:
(i) a 5′ inverted terminal repeat (ITR) sequence region comprising the nucleotide sequence of SEQ ID NO: 1380;
(ii) a first multiple cloning site (MCS) sequence region comprising the nucleotide sequence of SEQ ID NO: 1384;
(ii) an enhancer sequence region comprising the nucleotide sequence of SEQ ID NO: 1408;
(iii) a promoter sequence region comprising the nucleotide sequence of SEQ ID NO: 1410;
(iv) an intron sequence region comprising the nucleotide sequence of SEQ ID NO: 1417;
(v) a modulatory polynucleotide sequence region comprising the nucleotide sequence of SEQ ID NO: 1262;
(vi) a second MCS sequence region comprising the nucleotide sequence of TCGAG;
(vii) a polyA signal sequence region comprising the nucleotide sequence of SEQ ID NO: 1420; and
(viii) a 3′ ITR sequence region comprising the nucleotide sequence of SEQ ID NO: 1382.

12. An AAV viral particle comprising the AAV viral genome of claim 1 , and an AAV capsid protein, wherein the AAV capsid protein comprises an AAV1 capsid protein, an AAV5 capsid protein, or an AAV9 capsid protein.

13. A vector comprising the AAV viral genome claim 1 .

14. A cell comprising the AAV viral genome of claim 1 , wherein the cell is a mammalian cell, a medium spiny neuron, a cortical neuron, or an astrocyte.

15. A pharmaceutical composition comprising an AAV particle comprising the AAV viral genome of claim 1 , and a pharmaceutically acceptable excipient.

16. A method of inhibiting expression of a Huntingtin (HTT) gene, mRNA, and/or protein in a cell, comprising contacting the cell with the AAV viral genome of claim 1 .

17. The method of claim 16 , wherein the cell is in a subject, and the subject has or has been diagnosed with having Huntington's Disease (HD).

18. A method of treating Huntington's Disease (HD) in a subject, comprising administering to the subject an effective amount of an AAV viral particle comprising the AAV viral genome of claim 1 , thereby treating HD in the subject.

19. The method of claim 18 , wherein the HD is:
(i) a juvenile form HD in a subject of 2 to 20 years of age;
(ii) an early stage HD;
(iii) a late stage HD;
(iv) a fully penetrant HD, wherein the

Assignees

• Voyager Therapeutics Inc

Inventors

• Sah Dinah Wen-Yee
• Chen Fen
• Zhou Pengcheng
• Wang Xin
• Shu Yanqun
• Hou Jinzhao
• Deckert Jochen
• Hossbach Markus

Classifications

• C12N2750/14143

  viral genome or elements thereof as genetic vector · CPC title

• C12N2750/14123

  Virus like particles [VLP] · CPC title

• C12N2750/14122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• A61P25/14

  for treating abnormal movements, e.g. chorea, dyskinesia · CPC title

• A61K48/005

  characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered · CPC title