Compositions and methods for sirna inhibition of angiogenesis

Abstract

RNA interference using small interfering RNAs which are specific for the vascular endothelial growth factor (VEGF) gene and the VEGF receptor genes Flt-1 and Flk-1/KDR inhibit expression of these genes. Diseases which involve angiogenesis stimulated by overexpression of VEGF, such as diabetic retinopathy, age related macular degeneration and many types of cancer, can be treated by administering the small interfering RNAs.

1 . An isolated siRNA comprising of a duplex of a first RNA strand and a second RNA strand, said first RNA strand comprising a nucleotide sequence identical to a target sequence of about 19 to about 25 contiguous nucleotides in said human VEGF mRNA. 
     
     
         2 . The siRNA of  claim 1 , wherein said human VEGF mRNA is selected from VEGF 121  mRNA (SEQ ID NO: 2); VEGF 165  mRNA (SEQ ID NO: 3); VEGF 189  mRNA (SEQ ID NO: 4) and VEGF 206  mRNA (SEQ ID NO: 5). 
     
     
         3 . The siRNA of  claim 1 , wherein the first and second RNA strands forming the RNA duplex are covalently linked by a single-stranded hairpin. 
     
     
         4 . The siRNA of  claim 1 , wherein the siRNA further comprises non-nucleotide material. 
     
     
         5 . The siRNA of  claim 1 , wherein the first and second RNA strands are stabilized against nuclease degradation. 
     
     
         6 . The siRNA of  claim 1 , further comprising a 3′ overhang. 
     
     
         7 . The siRNA of  claim 6 , wherein the 3′ overhang comprises from 1 to about 6 nucleotides. 
     
     
         8 . The siRNA of  claim 6 , wherein the 3′ overhang comprises about 2 nucleotides. 
     
     
         9 . The siRNA of  claim 1 , wherein the sense RNA strand comprises a first 3′ overhang, and the antisense RNA strand comprises a second 3′ overhang. 
     
     
         10 . The siRNA of  claim 9 , wherein the first and second 3′ overhangs each comprise from 1 to about 6 nucleotides. 
     
     
         11 . The siRNA of  claim 9 , wherein the first 3′ overhang comprises a dinucleotide and the second 3′ overhang comprises a dinucleotide. 
     
     
         12 . The siRNA of  claim 11 , where the dinucleotide comprising the first and second 3′ overhangs is dithymidylic acid (TT) or diuridylic acid (uu). 
     
     
         13 . The siRNA of  claim 6 , wherein the 3′ overhang is stabilized against nuclease degradation. 
     
     
         14 . A pharmaceutical composition comprising a siRNA and a pharmaceutically acceptable carrier, said siRNA comprised of a duplex of a first RNA strand and a second RNA strand, said first RNA strand comprising a nucleotide sequence identical to a target sequence of about 19 to about 25 contiguous nucleotides in said human VEGF mRNA. 
     
     
         15 . The pharmaceutical composition of  claim 14 , wherein said human VEGF mRNA is selected from VEGF 121  mRNA (SEQ ID NO: 2); VEGF 165  mRNA (SEQ ID NO: 3); VEGF 189  mRNA (SEQ ID NO: 4) and VEGF 206  mRNA (SEQ ID NO: 5). 
     
     
         16 . The pharmaceutical composition of  claim 14 , wherein the first and second RNA strands are stabilized against nuclease degradation. 
     
     
         17 . The pharmaceutical composition of  claim 14 , further comprising at least one 3′ overhang. 
     
     
         18 . The pharmaceutical composition of  claim 17 , wherein the at least one 3′ overhang comprises about 2 nucleotides. 
     
     
         19 . The pharmaceutical composition of  claim 17 , where the at least one 3′ overhang comprises a dithymidylic acid (TT) or diuridylic acid (uu). 
     
     
         20 . The pharmaceutical composition of  claim 14 , wherein the sense RNA strand comprises a first 3′ overhang, and the antisense RNA strand comprises a second 3′ overhang.