iRNA agents targeting VEGF
The features of the present invention relate to compounds, compositions and methods useful for modulating the expression of vascular endothelial growth factor (VEGF), such as by the mechanism of RNA interference (RNAi). The compounds and compositions include iRNA agents that can be unmodified or chemically-modified.
1. A method of inhibiting VEGF expression in a cell comprising contacting the cell with a double stranded iRNA agent, wherein the iRNA agent comprises a sense strand and an antisense strand, wherein the sense strand nucleotide sequence consists of SEQ ID NO:608 and the antisense strand nucleotide sequence consists of SEQ ID NO:609.
2. The method of claim 1 , wherein the cell is a human cell of a human subject.
3. The method of claim 2 , wherein the subject is diagnosed as having an ophthalmic disorder or macular degeneration or diabetic retinopathy of cancer.
4. The method of claim 1 , wherein the method does not trigger an interferon response.
5. The method of claim 1 , wherein the iRNA agent is formulated with a pharmaceutically acceptable carrier or diluent.
6. The method of claim 1 , wherein the iRNA agent comprises a non-nucleotide moiety.
7. The method of claim 1 , wherein the iRNA agent further comprises a phosphorothioate at the first internucleotide linkage at the 5′ end of the antisense and sense strands.
8. The method of claim 1 , wherein the iRNA agent further comprises a phosphorothioate at the first internucleotide linkage at the 3′ end of the antisense and sense strands.
9. The method of claim 1 , wherein the iRNA agent further comprises a phosphorothioate at the first internucleotide linkage at the 5′ end of the antisense and sense strands and at the first internucleotide linkage at the 3′ end of the antisense and sense strands.
10. The method of claim 1 , wherein at least one nucleotide of at least one strand of the iRNA agent is a 2′-modified nucleotide.
11. The method of claim 10 , wherein the 2′-modified nucleotide comprises a modification selected from the group consisting of: 2′-deoxy, 2′-deoxy-2′-fluoro, 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), 2′-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), and 2′-O—N-methylacetamido (2′-O-NMA).
12. The method of claim 1 , wherein the iRNA agent comprises a cholesterol moiety.
13. The method of claim 12 , wherein the cholesterol moiety is conjugated to the 3′-end of the sense strand of the iRNA agent.
14. The method of claim 12 , wherein the cholesterol moiety is conjugated to the 3′-end of the antisense strand of the iRNA agent.
15. The method of claim 12 , wherein the cholesterol moiety is conjugated to the 3′-end of both the sense strand and the antisense strand of the iRNA agent.
16. The method of claim 1 , wherein contacting the cell with the iRNA agent results in at least 80% or at least 95% inhibition of expression of an endogenous human VEGF121 gene in HeLA cells measured by ELISA assay.
17. A method of inhibiting VEGF expression in a cell comprising the step of contacting the cell with a double stranded iRNA agent, wherein the iRNA agent comprises a sense strand and an antisense strand, wherein the sense strand consists of SEQ ID NO:671 and the antisense strand consists of SEQ ID NO:939.
18. A method of treating a disease or condition associated with VEGF expression in a subject comprising administration of an effective amount of a double stranded iRNA agent to the subject, wherein the iRNA agent comprises a sense strand and an antisense strand, wherein the sense strand nucleotide sequence consists of SEQ ID NO:608 and the antisense strand nucleotide sequence consists of SEQ ID NO:609.
19. The method of claim 18 , wherein administration is at or near the site of VEGF expression.
20. The method of claim 18 , wherein administration is to a cell or cells in a choroid region of the eye and is performed at a unit dose selected from a group consisting of: about 0.00001 mg to about 3 mg per eye; about 0.0001-0.001 mg per eye; about 0.03-3.0 mg per eye; about 0.1-3.0 mg per eye; and about 0.3-3.0 mg per eye.
21. A method of treating a disease or condition associated with VEGF expression in a subject comprising administration of an effective amount of a double stranded iRNA agent to the subject, wherein the iRNA agent comprises a sense strand and an antisense strand, wherein the sense strand consists of SEQ ID NO:671 and the antisense strand consists of SEQ ID NO:939.
22. The method of claim 18 or 21 , wherein the disease or condition is cancer.
23. The method of claim 18 or 21 , wherein the disease or condition is colon cancer or breast cancer or renal cancer.
24. The method of claim 18 or 21 , wherein the disease or condition is presence of a tumor.
25. The method of claim 1 , 17 , 18 , and 21 , wherein the iRNA is complexed to a delivery agent comprising a lipid.