Composition and methods of RNAi therapeutics for treatment of cancer and other neovascularization diseases

Abstract

Compositions and methods are provided for treatment of diseases involving unwanted neovascularization (NV). The invention provides treatments that control NV through selective inhibition of pro-angiogenic biochemical pathways, including inhibition of the VEGF pathway gene expression and inhibition localized at pathological NV tissues. Tissue targeted nanoparticle compositions comprising polymer conjugates and nucleic acid molecules that induce RNA interference (RNAi) are provided. The nanoparticle compositions of the invention can be used alone or in combination with other therapeutic agents such as VEGF pathway antagonists. The compositions and methods can be used for the treatment of NV diseases such as cancer, ocular disease, arthritis, and inflammatory diseases.

1 - 38 . (canceled) 
     
     
         39 . A composition selected from the group consisting of:
 a composition comprising a siRNA identified by SEQ ID NO. 2, a siRNA identified by SEQ ID NO. 9 a siRNA identified by SEQ ID NO. 14 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 2, a siRNA identified by SEQ ID NO. 9, a siRNA identified by SEQ ID NO. 16 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 2, a siRNA identified by SEQ ID NO. 9, a siRNA identified by SEQ ID NO. 18 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 2, a siRNA identified by SEQ ID NO. 11, a siRNA identified by SEQ ID NO. 14 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 2, a siRNA identified by SEQ ID NO. 11, a siRNA identified by SEQ ID NO. 16 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 2, a siRNA identified by SEQ ID NO. 11, a siRNA identified by SEQ ID NO. 18 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 2, a siRNA with the sequence of SEQ ID NO. 21, a siRNA identified by SEQ ID NO. 14 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 2, a siRNA with the sequence of SEQ ID NO. 21, a siRNA identified by SEQ ID NO. 16 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 2, a siRNA with the sequence of SEQ ID NO. 21, a siRNA identified by SEQ ID NO. 18 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 4, a siRNA identified by SEQ ID NO. 9, a siRNA identified by SEQ ID NO. 14 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 4, a siRNA identified by SEQ ID NO. 9, a siRNA identified by SEQ ID NO. 16 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 4, a siRNA identified by SEQ ID NO. 9, a siRNA identified by SEQ ID NO. 18 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 4, a siRNA identified by SEQ ID NO. 11, a siRNA identified by SEQ ID NO. 14 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 4, a siRNA identified by SEQ ID NO. 11, a siRNA identified by SEQ ID NO. 16 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 4, a siRNA identified by SEQ ID NO. 11, a siRNA identified by SEQ ID NO. 18 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 4, a siRNA with the sequence of SEQ ID NO. 21, a siRNA identified by SEQ ID NO. 14 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 4, a siRNA with the sequence of SEQ ID NO. 21, a siRNA identified by SEQ ID NO. 16 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 4, a siRNA with the sequence of SEQ ID NO. 21, a siRNA identified by SEQ ID NO. 18 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 6, a siRNA identified by SEQ ID NO. 9, a siRNA identified by SEQ ID NO. 14 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 6, a siRNA identified by SEQ ID NO.9, a siRNA identified by SEQ ID NO. 16 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 6, a siRNA identified by SEQ ID NO. 9, a siRNA identified by SEQ ID NO. 18 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 6, a siRNA identified by SEQ ID NO. 11, a siRNA identified by SEQ ID NO. 14 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 6, a siRNA identified by SEQ ID NO. 11, a siRNA identified by SEQ ID NO. 16 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 6, a siRNA identified by SEQ ID NO. 11, a siRNA identified by SEQ ID NO. 18 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 6, a siRNA with the sequence of SEQ ID NO. 21, a siRNA identified by SEQ ID NO. 14 and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 6, a siRNA with the sequence of SEQ ID NO. 21, a siRNA identified by SEQ ID NO. 16 and a pharmaceutically acceptable carrier; and   a composition comprising a siRNA identified by SEQ ID NO. 6, a siRNA with the sequence of SEQ ID NO. 21, a siRNA identified by SEQ ID NO. 18 and a pharmaceutically acceptable carrier.   
     
     
         40 . A composition selected from the group consisting of:
 a composition comprising a siRNA identified by SEQ ID NO. 2, a siRNA identified by SEQ ID NO. 9, and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 2, a siRNA identified by SEQ ID NO. 11, and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 2, a siRNA with the sequence of SEQ ID NO. 21, and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 4, a siRNA identified by SEQ ID NO. 9, and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 4, a siRNA identified by SEQ ID NO. 11, and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 4, a siRNA with the sequence of SEQ ID NO. 21, and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 6, a siRNA identified by SEQ ID NO. 9, and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 6, a siRNA identified by SEQ ID NO. 11, and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 6, a siRNA with the sequence of SEQ ID NO. 21, and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 2, a siRNA identified by SEQ ID NO. 14, and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 2, a siRNA identified by SEQ ID NO. 16, and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 4, a siRNA identified by SEQ ID NO. 14, and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 4, a siRNA identified by SEQ ID NO. 16, and a pharmaceutically acceptable carrier;   a composition comprising a siRNA identified by SEQ ID NO. 6, a siRNA identified by SEQ ID NO. 14, and a pharmaceutically acceptable carrier; and   a composition comprising a siRNA identified by SEQ ID NO. 6, a siRNA identified by SEQ ID NO. 16, and a pharmaceutically acceptable carrier.   
     
     
         41 . The composition of  claim 39  or  40 , further comprising one or more additional nucleic acid molecules that induce RNA interference and decrease the expression of a gene of interest. 
     
     
         42 . The composition of  claim 41 , wherein at least one of the one or more additional nucleic acid molecules decreases the expression of a gene that promotes angiogenesis. 
     
     
         43 . The composition of  claim 41 , wherein at least one of the one or more additional nucleic acid molecules decreases the expression of a gene selected from the group consisting of; VEGFR1, VEGFR2, VEGFR3, PDGF, PDGFR-α, PDGFR-β, EGF, EGFR, RAF-a, RAF-c, AKT, RAS, NFKB, HIF, bFGF, bFGFR, Her-2, c-Met, c-Myc and HGF. 
     
     
         44 . The composition of  claim 39  or  40 , wherein the carrier is a nucleic acid delivery vehicle. 
     
     
         45 . The composition of  claim 44 , wherein the nucleic acid delivery vehicle is synthetic. 
     
     
         46 . The composition of  claim 45  wherein the synthetic nucleic acid delivery vehicle comprises a core region comprising a cationic polymer and the nucleic acid. 
     
     
         47 . The composition of  claim 46 , wherein the cationic polymer is polyethyleneimine. 
     
     
         48 . The composition of  claim 46 , wherein the cationic polymer is a histidine-lysine co-polymer. 
     
     
         49 . The composition of  claim 46 , wherein the synthetic nucleic acid delivery vehicle further comprises a protective hydrophilic layer. 
     
     
         50 . The composition of  claim 49 , wherein the protective hydrophilic layer comprises a component selected from the group consisting of: polyethylene glycol, a polyacetal and a polyoxazoline. 
     
     
         51 . The composition of  claim 46 , wherein the synthetic nucleic acid vehicle further comprises a targeting moiety. 
     
     
         52 . The composition of  claim 49 , wherein the synthetic nucleic acid vehicle further comprises a targeting moiety. 
     
     
         53 . The composition of  claim 51 , wherein the targeting moiety binds a tumor specific molecule or an angiogenesis-specific molecule. 
     
     
         54 . The composition of  claim 51 , wherein the targeting moiety binds endothelial cells. 
     
     
         55 . The composition of  claim 54 , wherein the targeting moiety binds an integrin on vascular endothelial cells. 
     
     
         56 . The composition of  claim 55 , wherein the targeting moiety is a peptide comprising the amino acid sequence RGD. 
     
     
         57 . The composition of  claim 56 , wherein the peptide comprising RGD is a cyclic peptide. 
     
     
         58 . The composition of  claim 55 , wherein the synthetic nucleic acid delivery vehicle comprises:
 (a) a core region comprising PEI;   (b) a protective hydrophilic layer comprising PEG; and   (c) a cyclic RGD-containing peptide targeting moiety.   
     
     
         59 . The composition of  claim 39  or  40  comprising an additional therapeutic agent selected from the group consisting of: an anti-cancer agent, an anti-inflammatory agent and an anti-infective agent. 
     
     
         60 . The composition of  claim 39  or  40  comprising an anti-angiogenic agent. 
     
     
         61 . The composition of  claim 60 , wherein the anti-angiogenic agent is an inhibitor selected from the group consisting of: an inhibitor of human VEGF, an inhibitor of human VEGFR1 and an inhibitor of human VEGFR2. 
     
     
         62 . A method for reducing neovascularization in a subject in need thereof comprising the step of administering to the subject the composition of  claim 39  or  40 . 
     
     
         63 . A method for reducing neovascularization in a subject in need thereof comprising the step of administering to the subject the composition of  claim 58 . 
     
     
         64 . A method of reducing tumor growth in a subject in need thereof, comprising the step of administering to the subject the composition of  claim 39  or  40 . 
     
     
         65 . A method of reducing tumor growth in a subject in need thereof, comprising the step of administering to the subject the composition of  claim 58 . 
     
     
         66 . A method for decreasing one or more of VEGF, VEGFR1 or VEGFR2 protein levels in a cell comprising introducing into the cell the nucleic acid molecules of  claim 39 . 
     
     
         67 . A method for decreasing the protein level of VEGF or VEGFR1, or of both VEGF and VEGFR1, in a cell comprising introducing into the cell the nucleic acid molecules of  claim 40 . 
     
     
         68 . A method for decreasing the protein level of VEGF or VEGFR2, or of both VEGF and VEGFR2, in a cell comprising introducing into the cell the nucleic acid molecules of  claim 40 .