Dual targeting siRNA agents
The invention relates to dual targeting siRNA agents targeting a PCSK9 gene and a second gene, and methods of using dual targeting siRNA agents to inhibit expression of PCSK.9 and to treat PCSK.9 related disorders, e.g., hyperlipidemia.
1. A dual targeting siRNA agent comprising a first dsRNA targeting a PCSK9 gene, the first dsRNA comprising a first sense strand comprising the nucleotide sequence of SEQ ID NO:4148 and a first antisense strand comprising the nucleotide sequence of SEQ ID NO:4150, and a second dsRNA targeting a XBP-1 gene, the second dsRNA comprising a second sense strand comprising the nucleotide sequence of SEQ ID NO:4154 and a second strand comprising the nucleotide sequence of SEQ ID NO:4156 wherein the first dsRNA and the second dsRNA are linked with a covalent linker.
2. The dual targeting siRNA agent of claim 1 , the first dsRNA consisting of AD-10792, the second dsRNA consisting of AD-18038, the covalent linker consisting of a disulfide Q51 linker and linking the first sense strand to the second sense strand.
3. The dual targeting siRNA agent of claim 1 , wherein the first dsRNA comprises AD-10792.
4. The dual targeting siRNA agent of claim 1 , wherein the second dsRNA comprises AD-18038.
5. The dual targeting siRNA agent of claim 1 , wherein the first and second dsRNA each comprises at least one modified nucleotide.
6. The dual targeting siRNA agent of claim 5 , wherein 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.
7. The dual targeting siRNA agent of claim 5 , wherein 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, a 2′-amino-modified nucleotide, a 2′-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide.
8. The dual targeting siRNA agent of claim 1 , wherein each strand of each dsRNA is 19-23 bases in length.
9. The dual targeting siRNA agent of claim 1 , wherein the first and second dsRNAs are linked with a disulfide linker.
10. The dual targeting siRNA agent of claim 1 , wherein the covalent linker links the sense strand of the first dsRNA to the sense strand of the second dsRNA.
11. The dual targeting siRNA agent of claim 1 , wherein the covalent linker links the antisense strand of the first dsRNA to the antisense strand of the second dsRNA.
12. The dual targeting siRNA agent of claim 1 , further comprising a ligand.
13. The dual targeting siRNA agent of claim 1 , wherein administration of the dual targeting siRNA agent to a cell inhibits expression of the PCSK9 gene and the XBP-1 gene at a level equivalent to inhibition of expression of both genes obtained by the administration of each siRNA individually.
14. A pharmaceutical composition comprising the dual targeting siRNA agent of claim 1 and a pharmaceutical carrier.
15. The pharmaceutical composition of claim 14 , wherein the pharmaceutical carrier is a lipid formulation.
16. A method of inhibiting expression of a PCSK9 gene and a XBP-1 gene in a cell, the method comprising (a) introducing into the cell the dual targeting siRNA agent of claim 1 ; and (b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of the PCSK9 gene and the XBP-1 gene, thereby inhibiting expression of the PCSK9 gene and the XBP-1 gene in the cell.
17. The dual targeting siRNA agent of claim 2 , further comprising a ligand.
18. A pharmaceutical composition comprising the dual targeting siRNA agent of claim 2 and a pharmaceutical carrier.
19. The pharmaceutical composition of claim 18 , wherein the pharmaceutical carrier is a lipid formulation.
20. A method of inhibiting expression of a PCSK 9 gene and a XBP-1 gene in a cell, the method comprising (a) introducing into the cell the dual targeting siRNA agent of claim 2 ; and (b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of the PCSK9 gene and the XBP-1 gene, thereby inhibiting expression of the PCSK9 gene and the XBP-1 gene in the cell.