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; methods for treating diseases caused by PCSK9 gene expression and the expression of the PCSK9 gene using the pharmaceutical composition; and methods for inhibiting expression of a PCSK9 gene in a cell.
1. A method for inhibiting the expression of a proprotein convertase subtilisin kexin 9 (PCSK9) gene in a cell, the method comprising:
(a) contacting the cell with a double-stranded ribonucleic acid (dsRNA), wherein said dsRNA comprises a sense strand and an antisense strand and wherein the sense strand comprises a first sequence and the antisense strand comprises a second sequence comprising at least 15 contiguous nucleotides of SEQ ID NO:1228; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of the PCSK9 gene, thereby inhibiting expression of the PCSK9 gene in the cell.
2. The method of claim 1 , wherein the method is performed in vitro.
3. The method of claim 1 , wherein said dsRNA, upon contact with HepG2 cells expressing PCSK9, inhibits expression of said PCSK9 gene by at least 20%.
4. The method of claim 1 , wherein said method is performed in vivo.
5. The method of claim 1 , wherein administration of the dsRNA to an animal results in a decrease in total serum cholesterol.
6. The method of claim 1 , wherein said first sequence comprises SEQ ID NO:1227 and said second sequence comprises SEQ ID NO:1228.
7. The method of claim 1 , wherein said sense strand comprises SEQ ID NO:1227 and said antisense strand comprises SEQ ID NO:1228.
8. The method of claim 1 , wherein said sense strand consists of SEQ ID NO:1227 and said antisense strand consists of SEQ ID NO:1228.
9. The method of claim 1 , 6 , 7 , or 8 wherein said dsRNA comprises at least one modified nucleotide.
10. The method of claim 1 , 6 , 7 , or 8 wherein the dsRNA comprises at least one modified nucleotide and the 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.
11. The method of claim 1 , 6 , 7 , or 8 wherein the dsRNA comprises at least one modified nucleotide and the 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.
12. The method of claim 1 , 6 , 7 , or 8 wherein the dsRNA comprises at least one 2′-O-methyl modified nucleotide and at least one nucleotide comprising a 5′-phosphorothioate group.
13. The method of claim 1 , 6 , 7 , or 8 wherein the dsRNA is lipid formulated.
14. The method of claim 1 , 6 , 7 , or 8 wherein the dsRNA is LNP-01 lipid formulated.
15. 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 a dsRNA, wherein said dsRNA comprises a sense strand and an antisense strand and wherein the sense strand comprises a first sequence and the antisense strand comprises a second sequence comprising at least 15 contiguous nucleotides of SEQ ID NO:1228.
16. 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 a dsRNA, wherein said dsRNA comprises a sense strand and an antisense strand and wherein the sense strand comprises a first sequence and the antisense strand comprises a second sequence comprising at least 15 contiguous nucleotides of SEQ ID NO:1228.
17. The method of claim 15 , wherein said sense strand comprises SEQ ID NO:1227 and said antisense strand comprises SEQ ID NO:1228.
18. The method of claim 15 , wherein said sense strand consists of SEQ ID NO:1227 and said antisense strand consists of SEQ ID NO:1228.
19. The method of claim 16 , wherein said sense strand comprises SEQ ID NO:1227 and said antisense strand comprises SEQ ID NO:1228.
20. The method of claim 16 , wherein said sense strand consists of SEQ ID NO:1227 and said antisense strand consists of SEQ ID NO:1228.
21. The method of claim 1 , wherein the sense strand consists of the nucleotide sequence of SEQ ID NO:1227 and the antisense strand consists of the nucleotide sequence of SEQ ID NO:1228 and each strand is modified as follows to include a 2′-O-methyl ribonucleotide as indicated by a lower case letter “c” or “u” and a phosphorothioate as indicated by a lower case letter “s” and the sense strand consists of SEQ ID NO:1229 (5′-uucuAGAccuGuuuuGcuuTsT-3′) and the antisense strand consists of SEQ ID NO: 1230 (5′-AAGcAAAAcAGGUCuAGAATsT-3′).
22. The method of claim 15 , wherein the sense strand consists of the nucleotide sequence of SEQ ID NO:1227 and the antisense strand consists of the nucleotide sequence of SEQ ID NO:1228 and each strand is modified as follows to include a 2′-O-methyl ribonucleotide as indicated by a lower case letter “c” or “u” and a phosphorothioate as indicated by a lower case letter “s” and the sense strand consists of SEQ ID NO:1229 (5′-uucuAGAccuGuuuuGcuuTsT-3′) and the antisense strand consists of SEQ ID NO:1230 (5′-AAGcAAAAcAGGUCuAGAATsT-3′).
23. The method of claim 16 , wherein the sense strand consists of the nucleotide sequence of SEQ ID NO:1227 and the antisense strand consists of the nucleotide sequence of SEQ ID NO:1228 and each strand is modified as follows to include a 2′-O-methyl ribonucleotide as indicated by a lower case letter “c” or “u” and a phosphorothioate as indicated by a lower case letter “s” and the sense strand consists of SEQ ID NO:1229 (5′-uucuAGAccuGuuuuGcuuTsT-3′) and the antisense strand consists of SEQ ID NO:1230 (5′-AAGcAAAAcAGGUCuAGAATsT-3′).
24. The method of claim 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , or 23 , wherein the dsRNA is lipid formulated.
25. The method of claim 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , or 23 , wherein the dsRNA is LNP-01 lipid formulated.
26. The method of claim 9 , wherein the dsRNA is lipid formulated.
27. The method of claim 9 , wherein the dsRNA is LNP-01 lipid formulated.