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 proprotein convertase subtilisin kexin 9 (PCSK9) gene, comprising an antisense strand having a nucleotide sequence which is less than 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 methods for treating diseases caused by PCSK9 gene expression by using the pharmaceutical composition.
1. A double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of a human proprotein convertase subtilisin kexin 9 (PCSK9) gene in a cell, wherein said dsRNA comprises at least two sequences that are complementary to each other and wherein a sense strand comprises a first sequence and an antisense strand comprises a second sequence comprising at least 15 contiguous nucleotides of SEQ ID NO:1230.
2. The dsRNA of claim 1 , wherein said first sequence comprises SEQ ID NO:1229 and said second sequence comprises SEQ ID NO:1230.
3. The dsRNA of claim 1 , wherein said dsRNA comprises at least one modified nucleotide.
4. The dsRNA of claim 2 , wherein said dsRNA comprises at least one modified nucleotide.
5. The dsRNA of claim 3 , wherein said modified nucleotide is chosen from the group of: a 2′-O-methyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, and a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group.
6. The dsRNA of claim 3 , wherein said modified nucleotide is chosen from the group of: 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, morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide.
7. The dsRNA of claim 1 , wherein said sense strand consists of SEQ ID NO:1229 and said antisense strand consists of SEQ ID NO:1230.
8. The dsRNA of claim 6 , wherein said first sequence comprises SEQ ID NO:1229 and said second sequence comprises SEQ ID NO:1230.
9. A cell comprising the dsRNA of claim 1 .
10. A pharmaceutical composition for inhibiting the expression of the proprotein convertase subtilisin kexin 9 (PCSK9) gene in an organism, comprising a dsRNA and a pharmaceutically acceptable carrier, wherein the dsRNA comprises at least two sequences that are complementary to each other and wherein a sense strand comprises a first sequence and an antisense strand comprises a second sequence comprising at least 15 contiguous nucleotides of SEQ ID NO:1230 and wherein said dsRNA is capable of causing a decrease in serum lipid levels.
11. The pharmaceutical composition of claim 10 , wherein said first sequence of said dsRNA comprises SEQ ID NO:1229, and said second sequence of said dsRNA comprises SEQ ID NO:1230.
12. A vector for inhibiting the expression of a proprotein convertase subtilisin kexin 9 (PCSK9) gene in a cell, said vector comprising a regulatory sequence operably linked to a nucleotide sequence that encodes at least one strand of the dsRNA of claim 1 .
13. A cell comprising the vector of claim 12 .
14. The dsRNA of claim 1 , wherein said dsRNA, upon contact with a cell expressing said PCSK9 gene, inhibits expression of said PCSK9 gene.
15. The dsRNA of claim 14 , wherein said contact is performed in vitro at 30 nM or less.
16. The dsRNA of claim 1 , wherein said dsRNA, upon contact with HepG2 cells expressing PCSK9, inhibits expression of said PCSK9 gene by at least 20%.
17. The pharmaceutical composition of claim 10 , wherein said dsRNA, upon contact with a cell expressing said PCSK9, inhibits expression of said PCSK9 gene.
18. The dsRNA of claim 1 , wherein administration of said dsRNA to an animal results in a decrease in total serum cholesterol.
19. The pharmaceutical composition of claim 10 , wherein administration of said pharmaceutical composition to an animal results in a decrease in total serum cholesterol.
20. The dsRNA of pharmaceutical composition of claim 10 , wherein said sense strand consist of SEQ ID NO:1229 and said antisense strand consist of SEQ ID NO:1230.