Compositions and methods for inhibition of vegf

Abstract

Disclosed herein are siRNA compositions and methods useful for inhibiting expression of vascular endothelial growth factor (VEGF) isoforms. 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 small interfering RNAs as disclosed.

1 . An isolated siRNA comprising a sense RNA strand and an antisense RNA strand, wherein the sense and the antisense RNA strands form an RNA duplex, and wherein the sense RNA strand comprises a nucleotide sequence identical to a target sequence of about 19 to about 25 contiguous nucleotides in human VEGF mRNA, wherein the sense RNA strand comprises a nucleotide sequence that consists of SEQ ID NO: 119, and the antisense strand comprises a nucleotide sequence that consists of SEQ ID NO: 120. 
     
     
         2 . The siRNA of  claim 1 , wherein the first and second RNA strands forming the RNA duplex are covalently linked by a single-stranded hairpin. 
     
     
         3 . The siRNA of  claim 1 , wherein the siRNA further comprises non-nucleotide material. 
     
     
         4 . The siRNA of  claim 1 , wherein the first and second RNA strands are stabilized against nuclease degradation. 
     
     
         5 . A pharmaceutical composition comprising an effective amount of an isolated siRNA comprising a sense RNA strand and an antisense RNA strand, wherein the sense and the antisense RNA strands form an RNA duplex, and wherein the sense RNA strand comprises a nucleotide sequence that consists of SEQ ID NO: 119, and the antisense strand comprises a nucleotide sequence that consists of SEQ ID NO: 120. 
     
     
         6 . The pharmaceutical composition of  claim 5 , wherein the first and second RNA strands are stabilized against nuclease degradation. 
     
     
         7 . A method of treating an angiogenic disease in a subject comprising: administering to a subject an effective amount of a short interfering ribonucleic acid (siRNA) comprising a sense RNA strand and an antisense RNA strand, wherein the sense and the antisense RNA strands form an RNA duplex, and wherein the sense RNA strand comprises a nucleotide sequence identical to a target sequence of about 19 to about 25 contiguous nucleotides in human vascular endothelial growth factor (VEGF) mRNA, and wherein the sense RNA strand comprises a nucleotide sequence that consists of SEQ ID NO: 119, and the antisense strand comprises a nucleotide sequence that consists of SEQ ID NO: 120, such that angiogenesis associated with the angiogenic disease is inhibited. 
     
     
         8 . The method of  claim 7 , wherein the angiogenic disease comprises a tumor associated with a cancer. 
     
     
         9 . The method of  claim 8 , wherein the cancer is selected from the group consisting of breast cancer, lung cancer, head and neck cancer, brain cancer, abdominal cancer, colon cancer, colorectal cancer, esophagus cancer, gastrointestinal cancer, glioma, liver cancer, tongue cancer, neuroblastoma, osteosarcoma, ovarian cancer, pancreatic cancer, prostate cancer, retinoblastoma, Wilm's tumor, multiple myeloma, skin cancer, lymphoma, and blood cancer. 
     
     
         10 . The method of  claim 7 , wherein the angiogenic disease is selected from the group consisting of diabetic retinopathy, age-related macular degeneration, and inflammatory diseases. 
     
     
         11 . The method of  claim 10 , wherein the inflammatory disease is psoriasis or rheumatoid arthritis. 
     
     
         12 . The method of  claim 7 , wherein the angiogenic disease is age-related macular degeneration. 
     
     
         13 . The method of  claim 7 , wherein the pharmaceutical composition is administered in combination with a pharmaceutical agent for treating the angiogenic disease, which pharmaceutical agent is different from the short interfering ribonucleic acid (siRNA). 
     
     
         14 . The method of  claim 7 , wherein the angiogenic disease is cancer, and the pharmaceutical agent comprises a chemotherapeutic agent. 
     
     
         15 . The method of  claim 14 , wherein the chemotherapeutic agent is selected from the group consisting of cisplatin, carboplatin, cyclophosphamide, 5-fluorouracil, adriamycin, daunorubicin, and tamoxifen. 
     
     
         16 . The method of  claim 7 , wherein the pharmaceutical composition is administered to a subject in combination with another therapeutic method designed to treat the angiogenic disease. 
     
     
         17 . The method of  claim 7 , wherein the angiogenic disease is cancer, and the pharmaceutical composition is administered in combination with radiation therapy, chemotherapy or surgery. 
     
     
         18 . A method of inhibiting expression of human vascular endothelial growth factor (VEGF) comprising: administering to a subject an effective amount of a short interfering ribonucleic acid (siRNA) comprising a sense RNA strand and an antisense RNA strand, wherein the sense and the antisense RNA strands form an RNA duplex, and wherein the sense RNA strand comprises a nucleotide sequence identical to a target sequence of about 19 to about 25 contiguous nucleotides in human vascular endothelial growth factor (VEGF) mRNA and wherein the sense RNA strand comprises a nucleotide sequence that consists of SEQ ID NO: 119, and the antisense strand comprises a nucleotide sequence that consists of SEQ ID NO: 120. 
     
     
         19 . The method of  claim 18 , wherein the effective amount comprises from about 1 nM to about 100 nM of the short interfering ribonucleic acid (siRNA). 
     
     
         20 . The method of  claim 18 , wherein the pharmaceutical composition further comprises a delivery reagent. 
     
     
         21 . The method of  claim 18 , wherein the delivery agent is selected from the group consisting of lipofectin, lipofectamine, cellfectin, polycations, and liposomes. 
     
     
         22 . The method of  claim 21 , wherein the delivery agent is a liposome. 
     
     
         23 . The method of  claim 22 , wherein the liposome comprises a ligand which targets the liposome to cells at or near the site of angiogenesis. 
     
     
         24 . The method of  claim 23 , wherein the ligand binds to receptors on tumor cells or vascular endothelial cells. 
     
     
         25 . The method of  claim 23 , wherein the ligand comprises a monoclonal antibody. 
     
     
         26 . The method of  claim 22 , wherein the liposome is modified with an opsonization-inhibition moiety. 
     
     
         27 . The method of  claim 26 , wherein the opsonization-inhibiting moiety comprises a PEG, PPG, or derivatives thereof. 
     
     
         28 . The method of  claim 18 , wherein the short interfering ribonucleic acid (siRNA) is expressed from a recombinant plasmid. 
     
     
         29 . The method of  claim 18 , wherein the short interfering ribonucleic acid (siRNA) is expressed from a recombinant viral vector. 
     
     
         30 . The method of  claim 29 , wherein the recombinant viral vector comprises an adenoviral vector, an adeno-associated viral vector, a lentiviral vector, a retroviral vector, or a herpes virus vector. 
     
     
         31 . The method of  claim 30 , wherein the recombinant viral vector is pseudotyped with surface proteins from vesicular stomatitis virus, rabies virus, Ebola virus, or Mokola virus. 
     
     
         32 . The method of  claim 29 , wherein the recombinant viral vector comprises an adeno-associated viral vector. 
     
     
         33 . The method of  claim 18 , wherein the pharmaceutical composition is administered by an enteral administration route. 
     
     
         34 . The method of  claim 33 , wherein the enteral administration route is selected from the group consisting of oral, rectal, and intranasal. 
     
     
         35 . The method of  claim 18 , wherein the pharmaceutical composition is administered by a parenteral administration route. 
     
     
         36 . The method of  claim 35 , wherein the parenteral administration route is selected from the group consisting of intravascular administration, peri- and intra-tissue injection, subcutaneous injection or deposition, subcutaneous infusion, and direct application at or near the site of neovascularization. 
     
     
         37 . The method of  claim 35 , wherein the intravascular administration is selected from the group consisting of intravenous bolus injection, intravenous infusion, intra-arterial bolus injection, intra-arterial infusion and catheter instillation into the vasculature. 
     
     
         38 . A method of degrading human vascular endothelial growth factor (VEGF) mRNA comprising: administering to a subject an effective amount of a short interfering ribonucleic acid (siRNA) comprising a sense RNA strand and an antisense RNA strand, wherein the sense and the antisense RNA strands form an RNA duplex, and wherein the sense RNA strand comprises a nucleotide sequence identical to a target sequence of about 19 to about 25 contiguous nucleotides in human vascular endothelial growth factor (VEGF) mRNA and wherein the sense RNA strand comprises a nucleotide sequence that consists of SEQ ID NO: 119, and the antisense strand comprises a nucleotide sequence that consists of SEQ ID NO: 120.