Treatments for ocular neovascularization
Provided herein are compositions and methods for the treatment of ocular neovascular diseases such as wet age-related macular degeneration.
1 . A synthetic ribonucleic acid (RNA) molecule, wherein the synthetic RNA molecule comprises a sense strand comprising a nucleotide sequence set forth as SEQ ID NO:19 and an antisense strand comprising a nucleotide sequence set forth as SEQ ID NO:20.
2 . The synthetic RNA molecule according to claim 1 , wherein the RNA is a small hairpin RNA (shRNA) or an artificial microRNA (miRNA).
3 . A nucleic acid comprising a nucleotide sequence to be transcribed to an RNA, wherein the RNA comprises a synthetic RNA molecule according to claim 1 .
4 . The nucleic acid of claim 3 wherein the nucleic acid comprises (i) a nucleotide sequence to be transcribed to an artificial miRNA comprising a sense sequence comprising the nucleotide sequence set forth as SEQ ID No: 19 and an antisense sequence comprising the nucleotide sequence set forth as SEQ ID No: 20 and further comprises (ii) a nucleotide sequence encoding a first anti-angiogenic polypeptide that inhibits the activity of VEGF-A, wherein the nucleotide sequence of (i) and (ii) are operably linked to an expression control sequence.
5 . The nucleic acid according to claim 4 , wherein the first anti-angiogenic polypeptide is aflibercept.
6 . The nucleic acid according to claim 5 , wherein the nucleotide sequence encoding aflibercept comprises the nucleotide sequence of SEQ ID NO:50 or a sequence at least 90% identical thereto.
7 . The nucleic acid according to claim 6 , wherein the nucleotide sequence encoding the first anti-angiogenic polypeptide and the nucleotide sequence encoding said artificial miRNA are under the control of the same ubiquitous promoter.
8 . The nucleic acid according to claim 7 , wherein the promoter is a CAG or CBA promoter.
9 . The nucleic acid according to claim 8 , wherein the promoter is a CAG promoter and wherein the nucleotide sequence encoding the artificial miRNA is located within the hybrid chicken β-actin and rabbit β-globin intron of the CAG promoter.
10 . The nucleic acid according to claim 9 , wherein the nucleic acid comprises the nucleotide sequence set forth in SEQ ID No: 69 or a sequence at least 90% identical thereto.
11 . The nucleic acid according to claim 10 , wherein the nucleic acid comprises the nucleotide sequence set forth as SEQ ID NO:69.
12 . A vector comprising the nucleic acid according to claim 5 .
13 . A vector comprising the nucleic acid according to claim 11 .
14 . The vector according to claim 13 , wherein the vector is a recombinant adeno-associated virus (rAAV) vector.
15 . The vector according to claim 14 , wherein the rAAV comprises a capsid protein comprising an amino acid sequence at least 90% identical to SEQ ID NO:48.
16 . The vector according to claim 15 , wherein the rAAV comprises a capsid comprising a capsid protein comprising an amino acid sequence at least 95% identical to SEQ ID NO:48 and a heterologous nucleic acid comprising the sequence set forth as SEQ ID No: 69 or a sequence at least 95% identical thereto.
17 . The vector according to claim 16 , wherein the rAAV comprises a capsid protein comprising the amino acid sequence set forth as SEQ ID NO:48 and a heterologous nucleic acid sequence comprising the sequence set forth as SEQ ID NO:69.
18 . A host cell comprising the vector according to claim 17 .
19 . A pharmaceutical composition comprising the vector according to claim 17 and a pharmaceutically acceptable carrier.
20 . A method for the treatment of an ocular disease associated with VEGF-A in a subject in need of such treatment, the method comprising intraocularly administering to the subject a pharmaceutical composition according to claim 19 .
21 . The method according to claim 20 , wherein the pharmaceutical composition is administered by intravitreal injection.
22 . The method according to claim 21 , wherein the pharmaceutical composition comprises 1× 10 8 to 1× 10 15 vector particles or 1× 10 10 to 5× 10 13 vector particles.
23 . The method according to claim 22 , wherein the disease associated with VEGF-A is selected from the group consisting of wet age-related macular degeneration; macular edema following retinal vein occlusion; retinal neovascularization resulting from retinal vein occlusion; diabetic macular edema, diabetic retinopathy, myopic macular degeneration, branch retinal vein occlusion, hemi-retinal vein occlusion, and central retinal vein occlusion; retinopathy of prematurity; idiopathic choroidal neovascularization; myopia macular degeneration and secondary retinal and choroidal neovascularization; retinal telangiectasia; neovascular glaucoma; vitreous hemorrhage; retinal and choroidal neovascularization secondary to retinal diseases, including but not limited to uveitis, trauma, retinal degenerative disorders, genetic retinal and/or choroidal disease, tumors of the eye, corneal and iris neovascularization.
24 . The method according to claim 23 , wherein the disease associated with VEGF-A is selected from wet age-related macular degeneration; diabetic macular edema; macular edema following retinal vein occlusion; diabetic retinopathy; and myopic choroidal neovascularization.