RNAi modulation of APOB and uses thereof
View Patent ↗The invention relates to compositions and methods for modulating the expression of apolipoprotein B, and more particularly to the downregulation of apolipoprotein B by chemically modified oligonucleotides.
1. An iRNA agent comprising a sense strand and an antisense strand, wherein said sense strand comprises a first sequence and said antisense strand comprises a second sequence, wherein said first sequence is complementary to said second sequence, and wherein said second sequence comprises at least 15 contiguous nucleotides of SED ID NO: 98, and wherein each strand of said iRNA agent is between 15 and 30 base pairs in length.
2. The iRNA agent of claim 1 , comprising a non-nucleotide moiety.
3. The iRNA agent of claim 1 , comprising a phosphorothioate linkage.
4. The iRNA agent of claim 3 , wherein said iRNA agent comprises a phosphorothioate at the first and second internucleotide linkage at the 3′ end of the antisense strand.
5. The iRNA agent of claim 3 , wherein said iRNA agent comprises a phosphorothioate at the first internucleotide linkage at the 3′ end of the sense strand.
6. The iRNA agent of claim 1 , comprising a 2′-modified nucleotide.
7. The iRNA agent of claim 6 , wherein the 2′-modification is chosen from the group of: 2′-deoxy, 2′-deoxy-2′-fluoro, 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-0-aminopropyl (2′-O-AP), 2′-0-dimethylaminoethyl (2′-O-DMAOE), 2′-dimethylaminopropyl (2′-O-DMAP), 2′-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), and 2′-O—N-methylacetamido (2′-O-NMA).
8. The iRNA agent of claim 6 , comprising: at least one 5′-uridine-adenine-3′ (5′-UA-3′) dinucleotide wherein the uridine is a 2′-modified nucleotide; at least one 5′-uridine-guanine-3′ (5′-UG-3′) dinucleotide, wherein the 5′-uridine is a 2′-modified nucleotide; at least one 5′-cytidine-adenine-3′ (5′-CA-3′) dinucleotide, wherein the 5′-cytidine is a 2′-modified nucleotide; or at least one 5′-uridine-uridine-3′ (5′-UU-3′) dinucleotide, wherein the 5′-uridine is a 2′-modified nucleotide.
9. The iRNA agent of claim 8 , wherein said antisense strand consists of SEQ ID NO:254.
10. The iRNA agent of claim 8 , wherein the 5′-most pyrimidines in all occurrences of sequence motif 5′-UA-3′,5′-CA-3′,5′-UU-3′, and 5′-UG-3′ on the antisense strand are 2′-modified nucleotides.
11. The iRNA agent of claim 10 , wherein said antisense strand consists of SEQ ID NO:256.
12. The iRNA agent of claim 1 , wherein a 5′-end of said iRNA agent comprises an adenosine, cytidine, guanosine, thymidine, or uridine.
13. The iRNA agent of claim 1 , comprising a nucleotide overhang having 1 to 4 nucleotides.
14. The iRNA agent of claim 13 , wherein the nucleotide overhang has 2 or 3 unpaired nucleotides.
15. The iRNA agent of claim 13 , wherein the nucleotide overhang is at the 3′-end of the antisense strand of the iRNA agent.
16. The mRNA agent of claim 1 , comprising a cholesterol moiety.
17. The iRNA agent of claim 16 , wherein the cholesterol moiety is conjugated to the 3′-end of the sense strand of the iRNA agent.
18. The iRNA agent of claim 9 or 11 , wherein said sense strand is selected from the group consisting of: SEQ ID NO:253, SEQ ID NO:255, and SEQ ID NO: 261.
19. The iRNA agent of claim 9 , wherein said sense strand consists of SEQ ID NO:253.
20. The iRNA agent of claim 1 , wherein said sense strand consists of SEQ ID NO:97 and said antisense strand consists of SEQ ID NO:98.
21. The iRNA agent of claim 1 , wherein said sense strand consists of SEQ ID NO:275 and said antisense strand consists of SEQ ID NO:276.
22. The iRNA agent of claim 1 , wherein said antisense strand consists of SEQ ID NO:98.
23. The iRNA agent of claim 1 , wherein said iRNA agent reduces the amount of ApoB mRNA present in cultured mouse cells of hepatic origin.
24. The mRNA agent of claim 1 , wherein the agent reduces the amount of ApoB mRNA in cultured human HepG2 cells after incubation with the agent by more than 50% compared to cells that have not been incubated with the agent, or reduces the amount of ApoB protein secreted into cell culture supernatant by more than 50%.
25. A method for reducing the expression levels of ApoB in a subject, comprising the step of administering an iRNA agent to said subject, wherein said sense strand comprises a first sequence and said antisense strand comprises a second sequence, wherein said first sequence is complementary to said second sequence, and wherein said second sequence comprises at least 15 contiguous nucleotides of SED ID NO: 98, and wherein each strand of said iRNA agent is between 15 and 30 base pairs in length.