Claims for Patent: 8,222,222

Summary for Patent: 8,222,222

Title:	Compositions and methods for inhibiting expression of the PCSK9 gene
Abstract:	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.
Inventor(s):	Tan; Pamela (Kulmbach, DE), Bramlage; Birgit (Kulmbach, DE), Frank-Kamenetsky; Maria (Brookline, MA), Fitzgerald; Kevin (Brookline, MA), Akinc; Akin (Needham, MA), Kotelianski; Victor E. (Boston, MA)
Assignee:	Alnylam Pharmaceuticals, Inc. (Cambridge, MA)
Application Number:	12/554,231
Patent Claims:	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.