Compositions and methods for inhibiting expression of the PCSK9 gene
View Patent ↗The invention relates to a double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of the PCSK9 gene (PCSK9 gene), comprising an antisense strand having a nucleotide sequence which is less that 30 nucleotides in length, generally 19-25 nucleotides in length, and which is substantially complementary to at least a part of the PCSK9 gene. The invention also relates to a pharmaceutical composition comprising the dsRNA together with a pharmaceutically acceptable carrier and method for treating diseases caused by PCSK9 gene expression.
1. A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of a human proprotein convertase subtilisin kexin 9 (PCSK9) gene in a cell, wherein the dsRNA comprises a sense strand and an antisense strand complementary to at least 15 contiguous nucleotides of nucleotides 1-19 of SEQ ID NO:453 and comprises a duplex structure between 15 and 30 base pairs in length.
2. The dsRNA of claim 1 comprising a duplex structure between 19 and 21 base pairs in length.
3. The dsRNA of claim 1 wherein the sense strand comprises the nucleotide sequence of nucleotides 1-19 of SEQ ID NO:453 and the antisense strand comprises the nucleotide sequence of nucleotides 1-19 of SEQ ID NO:454.
4. The dsRNA of claim 1 , the sense strand consisting of SEQ ID NO:453 and the antisense strand consisting of SEQ ID NO:454.
5. The dsRNA of claim 4 , comprising at least one modified nucleotide.
6. The dsRNA of claim 1 , wherein the dsRNA comprises at least one modified nucleotide.
7. The dsRNA of claim 1 , wherein the dsRNA comprises at least one 2′-O-methyl modified nucleotide and at least one nucleotide comprising a 5′-phosphorothioate group.
8. The dsRNA claim 1 , wherein the dsRNA comprises at least one modified nucleotide, wherein the modified nucleotide is chosen from the group of: a 2′-O-methyl modified nucleotide, a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, 2′-amino-modified nucleotide, 2′-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, a nucleotide comprising a 5′-phosphorothioate group, a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group, and a non-natural base comprising nucleotide.
9. A cell comprising the dsRNA of claim 1 .
10. A pharmaceutical composition comprising the dsRNA of claim 1 and a pharmaceutically acceptable carrier.
11. A composition comprising the dsRNA of claim 1 and a lipid formulation.
12. A composition comprising the dsRNA of claim 1 and a lipid formulation, wherein the lipid formulation comprises a cationic lipid comprising ND-98.
13. A vector comprising a regulatory sequence operably linked to a nucleotide sequence that encodes at least one strand of the dsRNA of claim 1 .
14. A cell comprising the vector of claim 13 .
15. The dsRNA of claim 1 , wherein contacting a cell in vitro with 30 nM or less of the dsRNA and maintaining the cell for a time sufficient to obtain degradation of a mRNA transcript of a PCSK9 gene, inhibits expression of the PCSK9 gene in the cell.
16. The dsRNA of claim 1 , wherein contacting HepG2 cells expressing the PCSK9 gene in vitro with the dsRNA and maintaining the cells for a time sufficient to obtain degradation of a mRNA transcript of a PCSK9 gene, inhibits expression of the PCSK9 gene in the cell by at least 20%.
17. The dsRNA of claim 1 , wherein administering the dsRNA to an animal decreases total serum cholesterol in the animal.
18. A method for inhibiting expression of a proprotein convertase subtilisin kexin 9 (PCSK9) gene in a cell comprising contacting the cell with the dsRNA of claim 1 and maintaining the cell for a time sufficient to obtain degradation of a mRNA transcript of a PCSK9 gene, thereby inhibiting expression of the PCSK9 gene in the cell.
19. A method of treating or managing pathological processes which can be mediated by down regulating expression of a proprotein convertase subtilisin kexin 9 (PCSK9) gene comprising administering to a patient in need of such treatment or management a therapeutically effective amount of the dsRNA of claim 1 .
20. A method of treating a proprotein convertase subtilisin kexin 9 (PCSK9) gene-associated disorder comprising administering to a patient in need of such treatment, a therapeutically effective amount of the dsRNA of claim 1 .
21. A cell comprising the dsRNA of claim 4 .
22. A pharmaceutical composition comprising the dsRNA of claim 4 and a pharmaceutically acceptable carrier.
23. A composition comprising the dsRNA of claim 4 and a lipid formulation.
24. A composition comprising the dsRNA of claim 4 and a lipid formulation, wherein the lipid formulation comprises a cationic lipid comprising ND-98.
25. A method for inhibiting expression of a proprotein convertase subtilisin kexin 9 (PCSK9) gene in a cell comprising contacting the cell with the dsRNA of claim 4 and maintaining the cell for a time sufficient to obtain degradation of a mRNA transcript of a PCSK9 gene, thereby inhibiting expression of the PCSK9 gene in the cell.
26. A method of treating or managing pathological processes which can be mediated by down regulating expression of a proprotein convertase subtilisin kexin 9 (PCSK9) gene comprising administering to a patient in need of such treatment or management a therapeutically effective amount of the dsRNA of claim 4 .
27. A method of treating a proprotein convertase subtilisin kexin 9 (PCSK9) gene-associated disorder comprising administering to a patient in need of such treatment, a therapeutically effective amount of the dsRNA of claim 4 .