Polynucleotide agents targeting hydroxyacid oxidase (glycolate oxidase, HAO1) and methods of use thereof
The invention relates to polynucleotide agents targeting an hydroxyacid oxidase (HAO1) gene, and methods of using such polynucleotide agents to inhibit expression of HAO1 and to treat subjects having an HAO1-associated disease, e.g., hyperoxaluria.
1. A single stranded antisense polynucleotide agent for inhibiting hydroxyacid oxidase (HAO1) expression, comprising at least 15 contiguous nucleotides of the nucleotide sequence 5′-GCACAGUGUCUCUUUGUCAA-3′ (SEQ ID NO:272), wherein the agent comprises
a gap segment consisting of linked deoxynucleotides;
a 5′-wing segment consisting of linked nucleotides;
a 3′-wing segment consisting of linked nucleotides;
wherein the gap segment is positioned between the 5′-wing segment and the 3′-wing segment and wherein each nucleotide of each wing segment comprises a modified sugar, and
wherein the agent is about 18 to about 24 nucleotides in length.
2. The single stranded antisense polynucleotide agent of claim 1 , wherein substantially all of the nucleotides are modified nucleotides.
3. The single stranded antisense polynucleotide agent of claim 1 , which is 20 nucleotides in length.
4. The single stranded antisense polynucleotide agent of claim 1 , wherein the gap segment is 5 to 14 2′-deoxynucleotides in length and each of the wing segments is 1 to 6 nucleotides in length.
5. The single stranded antisense polynucleotide agent of claim 1 , wherein the agent further comprises a ligand at the 3′-terminus.
6. The single stranded antisense polynucleotide agent of claim 5 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
7. A pharmaceutical composition for inhibiting expression of an hydroxyacid oxidase (HAO1) gene comprising the single stranded antisense polynucleotide agent of claim 1 .
8. A pharmaceutical composition comprising the single stranded antisense polynucleotide agent of claim 1 , and a lipid formulation.
9. A method of inhibiting hydroxyacid oxidase (HAO1) expression in a cell, the method comprising:
(a) contacting the cell with the single stranded antisense polynucleotide agent of claim 1 or a pharmaceutical composition of claim 7 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain antisense inhibition of an HAO1 gene, thereby inhibiting expression of the HAO1 gene in the cell.
10. The method of claim 9 , wherein the cell is within a subject.
11. The method of claim 10 , wherein the subject is a human.
12. A method of treating a subject having a disease or disorder that would benefit from reduction in hydroxyacid oxidase (HAO1) expression, the method comprising administering to the subject a therapeutically effective amount of the single stranded antisense polynucleotide agent of claim 1 or a pharmaceutical composition of claim 7 , thereby treating the subject.
13. A method of preventing at least one symptom in a subject having a disease or disorder that would benefit from reduction in hydroxyacid oxidase (HAO1) expression, the method comprising administering to the subject a prophylactically effective amount of the single stranded antisense polynucleotide agent of claim 1 or a pharmaceutical composition of claim 7 , thereby preventing at least one symptom in the subject having a disorder that would benefit from reduction in HAO1 expression.
14. The method of claim 12 or 13 , wherein the disorder is an hydroxyacid oxidase (HAO1)-associated disease.
15. The method of claim 14 , wherein the HAO1-associated disease is selected from the group consisting of primary hyperoxaluria and secondary hyperoxaluria.
16. The single stranded antisense polynucleotide agent of claim 1 , wherein the modified sugar moiety is selected from the group consisting of a 2′-O-methoxyethyl modified sugar moiety, a 2′-methoxy modified sugar moiety, a 2′-O-alkyl modified sugar moiety, and a bicyclic sugar moiety.
17. The single stranded antisense polynucleotide agent of claim 1 , further comprising a modified internucleoside linkage.
18. The single stranded antisense polynucleotide agent of claim 4 , wherein the 5′-wing segment is 4 to 6 nucleotides in length, the 3′-wing segment is 4 to 6 nucleotides in length, and the gap segment is 9 to 13 nucleotides in length.
19. The pharmaceutical composition of claim 7 , wherein the single stranded antisense polynucleotide agent is present in an unbuffered solution.
20. The method of claim 12 or 13 , wherein the single stranded antisense polynucleotide agent is administered to the subject at a dose of about 0.01 mg/kg to about 10 mg/kg or about 0.5 mg/kg to about 50 mg/kg.
21. The method of claim 20 , wherein the single stranded antisense polynucleotide agent is administered to the subject once a week; twice a week; or twice a month.
22. The method of claim 12 or 13 , wherein the single stranded antisense polynucleotide agent is administered to the subject subcutaneously.
23. The single stranded antisense polynucleotide agent of claim 1 , wherein all of the nucleotides are modified nucleotides.