TREATMENT OF AMD USING AAV SFLT-1
The present disclosure provides compositions and methods for the prevention or treatment of ocular neovascularization, such as AMD, in a human subject, by administering subretinally a pharmaceutical composition comprising a pharmaceutically effective amount of a vector comprising a nucleic acid encoding soluble Fms-related tyrosine kinase-1 (sFlt-1) protein to the human subject.
1 - 41 . (canceled)
42 . A method for the treatment or prophylaxis of ocular neovascularization in a human subject having or at risk for developing ocular neovascularization, the method comprising: administering to the retina of the human subject a pharmaceutically effective amount of a pharmaceutical composition comprising a nucleic acid encoding sFLT-1.
43 . The method of claim 42 , wherein the ocular neovascularization is associated with a condition selected from the group consisting of: age-related macular degeneration (AMD), retinal neovascularization, choroidal neovascularization, diabetic retinopathy, retinal vein occlusion, diabetic macular edema, diabetic retinal ischemia, ischemic retinopathy and diabetic retinal edema.
44 . The method of claim 43 , wherein the condition is AMD.
45 . The method of claim 42 , wherein the pharmaceutical composition comprises a recombinant virus, the virus selected from the group consisting of: adeno-associated virus (AAV), adenovirus, helper-dependent adenovirus, retrovirus, herpes simplex virus, lentivirus, poxvirus, hemagglutination virus of Japan-liposome (HVJ) complex, Moloney murine leukemia virus, and HIV-based virus.
46 . The method of claim 45 , wherein the virus is AAV.
47 . The method of claim 1 , wherein the sFLT-1 nucleic acid encodes at least 1 dimerization domain.
48 . The method of claim 42 , wherein a reduction in neovascularization, as observed by a Fluorscein Angiography (FA), follows the administering of the pharmaceutical composition.
49 . The method of claim 42 , wherein no superficial, anterior segment or vitreous inflammatory signs are present in the human subject at least 1 week after injection.
50 . The method of claim 42 , wherein the human subject has received one or more treatments with a VEGF inhibitor prior to the administering of the pharmaceutical composition.
51 . The method of claim 42 , wherein the human subject has not previously received treatment with a VEGF inhibitor before the administering of the pharmaceutical composition.
52 . The method of claim 42 , wherein the human subject does not require treatment with a VEGF inhibitor for at least 30 days after the administering of the pharmaceutical composition.
53 . The method of claim 42 , wherein the administering of the pharmaceutical composition is performed at a frequency less than 3 times a year in the human subject.
54 . The method of claim 42 , further comprising removing vitreous gel prior to or within one week of the administering of the pharmaceutical composition.
55 . The method of claim 42 , wherein said removing comprises the use of a vitrectomy system comprising a cannula having a 20-27 gauge bore size.
56 . The method of claim 42 , wherein said pharmaceutical composition is administered using a cannula having a 27-45 gauge bore size.
57 . A pharmaceutical composition comprising recombinant viruses or plasmids comprising a nucleic acid comprising at least 1 promoter sequence operatively linked to an sFLT-1 transgene sequence, wherein the promoter sequence and the sFLT-1 transgene sequence are separated by a UTR sequence.
58 . The pharmaceutical composition of claim 57 , wherein the promoter sequence and the sFLT-1 transgene sequence are separated by a sequence greater than 300 base pairs.
59 . The pharmaceutical composition of claim 57 , wherein the nucleic acid sequence comprises at least 3 linker sequences each comprising at least 50 base pairs.
60 . A pharmaceutical composition comprising nucleic acid elements in the following order: a. a promoter sequence selected from the group consisting of SEQ ID No. 1, SEQ ID No. 2, SEQ ID No. 3, SEQ ID No. 4, SEQ ID No. 5, SEQ ID No. 6, SEQ ID No. 7, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10, SEQ ID No. 11, SEQ ID No. 12, SEQ ID No. 13, SEQ ID No. 14, SEQ ID No. 15, SEQ ID No. 16, SEQ ID No. 17, SEQ ID No. 18, SEQ ID No. 19, SEQ ID No. 20, SEQ ID No. 21, SEQ ID No. 22, SEQ ID No. 23, SEQ ID No. 24, SEQ ID No. 25, SEQ ID No. 26, SEQ ID No. 27, SEQ ID No. 28, SEQ ID No. 29, SEQ ID No. 30, SEQ ID No. 31 and SEQ ID No. 32; b. a sequence encoding a VEGF inhibitor selected from the group consisting of SEQ ID No. 102, SEQ ID No. 103, SEQ ID No. 104, SEQ ID No. 105, SEQ ID No. 106, SEQ ID No. 107, SEQ ID No. 108 and SEQ ID No. 122; c. an intron sequence consisting of SEQ ID No. 48, SEQ ID No. 115, SEQ ID No. 116, SEQ ID No. 117, SEQ ID No. 118, SEQ ID No. 119 and SEQ ID No. 120; d. a UTR sequence selected from the group consisting of SEQ ID No. 91, SEQ ID No. 2, SEQ ID No. 92, SEQ ID No. 93, SEQ ID No. 94, SEQ ID No. 95, SEQ ID No. 96, SEQ ID No. 97, SEQ ID No. 98, SEQ ID No. 99, SEQ ID No. 100, and SEQ ID No. 101; and e. a termination sequence selected from the group consisting of SEQ ID No. 49, SEQ ID No. 50, SEQ ID No. 51, SEQ ID No. 52, SEQ ID No. 53, SEQ ID No. 54, and SEQ ID No. 55.
61 . A unit dose of a pharmaceutical composition comprising recombinant viruses of 1×10 6 to about 1×10 15 vector genomes, wherein the recombinant viruses comprise a nucleic acid encoding sFLT-1 operatively linked to a promoter.