iRNA Agents Targeting VEGF

Abstract

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. 
     
     
         4 . The method of  claim 2 , wherein the subject is diagnosed as having macular degeneration. 
     
     
         5 . The method of  claim 2 , wherein the subject is diagnosed as having diabetic retinopathy. 
     
     
         6 . The method of  claim 2 , wherein the subject is diagnosed as having cancer. 
     
     
         7 . The method of  claim 1 , wherein the method does not trigger an interferon response. 
     
     
         8 . The method of  claim 1 , wherein the iRNA agent is formulated with a pharmaceutically acceptable carrier or diluent. 
     
     
         9 . The method of  claim 1 , wherein the iRNA agent comprises a non-nucleotide moiety. 
     
     
         10 . The method of  claim 9 , wherein the sense and the antisense strands are stabilized against nucleolytic degradation. 
     
     
         11 . 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. 
     
     
         12 . 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. 
     
     
         13 . 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. 
     
     
         14 . The method of  claim 1 , wherein at least one nucleotide of at least one strand of the iRNA agent is a 2′-modified nucleotide. 
     
     
         15 . The method of  claim 14 , 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). 
     
     
         16 . The method of  claim 1 , wherein the iRNA agent comprises a cholesterol moiety. 
     
     
         17 . The method of  claim 16 , wherein the cholesterol moiety is conjugated to the 3′-end of the sense strand of the iRNA agent. 
     
     
         18 . The method of  claim 16 , wherein the cholesterol moiety is conjugated to the 3′-end of the antisense strand of the iRNA agent. 
     
     
         19 . The method of  claim 16 , wherein the cholesterol moiety is conjugated to the 3′-end of both the sense strand and the antisense strand of the iRNA agent. 
     
     
         20 . The method of  claim 1 , wherein contacting the cell with the iRNA agent results in at least 80% inhibition of expression of an endogenous human VEGF121 gene in HeLA cells measured by ELISA assay. 
     
     
         21 . The method of  claim 1 , wherein contacting the cells with the iRNA agent results in at least 95% inhibition of expression of an endogenous human VEGF121 gene in HeLA cells under hypoxic conditions as measured by ELISA assay. 
     
     
         22 . The method of  claim 1 , wherein contacting the cells with the iRNA agent results in greater than 90% inhibition of expression of an endogenous human VEGF121 gene in HeLA cells as measured by ELISA assay, and wherein the iRNA agent is modified with a phosphorothioate linkage, a 2′-O-methyl nucleotide, or a 2′-fluoro-modified nucleotide. 
     
     
         23 . The method of  claim 1 , wherein the iRNA agent is more stable in rat retina lysates than in rat whole eye extracts. 
     
     
         24 . 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. 
     
     
         25 . 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. 
     
     
         26 . The method of  claim 25 , wherein administration is at or near the site of VEGF expression. 
     
     
         27 . The method of  claim 25 , wherein administration is to the eye by injection. 
     
     
         28 . The method of  claim 25 , wherein administration is to a cell or cells in a choroid region of the eye. 
     
     
         29 . The method of  claim 25 , wherein administration 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. 
     
     
         30 . The method of  claim 25 , comprising multiple administrations. 
     
     
         31 . The method of  claim 25 , wherein the method is prophylactic. 
     
     
         32 . 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. 
     
     
         33 . An isolated double stranded iRNA agent, consisting of SEQ ID NO: 609 and SEQ ID NO: 608. 
     
     
         34 . An isolated double stranded iRNA agent, consisting of SEQ ID NO:671 and SEQ ID NO:939.