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 . An isolated double stranded iRNA agent, wherein the iRNA agent comprises a sense strand and an antisense strand, wherein the antisense strand comprises a sequence complementary to 19- to 23 nucleotides of a VEGF nucleotide sequence and each strand is 15-30 nucleotides in length. 
     
     
         2 . The iRNA agent of  claim 1 , wherein the sense sequence comprises a sequence that differs by no more than 1, 2, or 3 nucleotides from a sequence selected from the group consisting of SEQ ID NOs:2-401 and SEQ ID NO:456, SEQ ID NO:546, SEQ ID NO:548, SEQ ID NO:550, SEQ ID NO:552, SEQ ID NO:590, SEQ ID NO:592, SEQ ID NO:594, SEQ ID NO:596, SEQ ID NO:608, SEQ ID NO:610, SEQ ID NO:612, SEQ ID NO:614, SEQ ID NO:634, SEQ ID NO:636, SEQ ID NO:638, SEQ ID NO:640, SEQ ID NO:646, SEQ ID NO: 648, SEQ ID NO:650 and SEQ ID NO:671. 
     
     
         3 . The iRNA agent of  claim 1 , wherein each strand is 19-23 nucleotides in length. 
     
     
         4 . The iRNA agent of  claim 1 , wherein at least one strand comprises at least one nucleotide overhang having 1 to 4 nucleotides. 
     
     
         5 . The iRNA agent of  claim 1 , wherein the iRNA agent comprises at least one non-nucleotide moiety. 
     
     
         6 . The iRNA agent of  claim 1 , wherein at least one nucleotide of at least one strand of the iRNA agent is a 2′-modified nucleotide comprising 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′-β-DMAEOE), and 2′-O—N-methylacetamido (2′-O-NMA). 
     
     
         7 . The iRNA agent of  claim 1 , wherein the iRNA agent comprises a cholesterol moiety. 
     
     
         8 . The iRNA agent of  claim 1 , wherein the iRNA agent comprises a cholesterol moiety conjugated to the 3′-end of the sense strand of the iRNA agent and/or to the 3′-end of the antisense strand of the iRNA agent. 
     
     
         9 . A cell comprising the iRNA agent of  claim 1 . 
     
     
         10 . A pharmaceutical composition comprising the iRNA agent of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         11 . A method of inhibiting VEGF expression in a cell comprising contacting the cell with the iRNA agent of  claim 1  and maintain the cell for a time sufficient to obtain degradation of the mRNA transcript of a VEGF gene, thereby inhibiting expression of VEGF in the cell. 
     
     
         12 . The method of  claim 11 , wherein the cell is a human cell of a human subject. 
     
     
         13 . The method of  claim 12 , wherein the subject is diagnosed as having an ophthalmic disorder or macular degeneration or diabetic retinopathy or cancer. 
     
     
         14 . The method of  claim 11 , 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 or 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 or 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. 
     
     
         15 . A method of treating a disease or condition associated with VEGF expression in a subject comprising administration of an effective amount of the iRNA agent of  claim 1  to the subject. 
     
     
         16 . The method of  claim 15 , wherein disease or condition comprises adult onset macular degeneration, diabetic retinopathy, neovascular glaucoma, colon cancer, breast cancer, renal cancer, pulmonary disease, rheumatoid arthritis, or psoriasis. 
     
     
         17 . The method of  claim 15 , wherein administration is at or near the site of VEGF expression. 
     
     
         18 . The method of  claim 15 , wherein administration is to a cell or cells in a choroid region of the eye by injection. 
     
     
         19 . The method of  claim 18 , 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. 
     
     
         20 . The method of  claim 15 , comprising multiple administrations.