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 pharmaceutical composition comprising a nucleic acid sequence encoding an anti-VEGF protein operatively linked to a promoter sequence.
43 . The pharmaceutical composition according to claim 42 , wherein the anti-VEGF protein comprises a functional fragment of human sFLT1.
44 . The pharmaceutical composition according to claim 42 , wherein the functional fragment has a sequence identity of 90% or more to SEQ ID NO:109.
45 . The pharmaceutical composition according to claim 44 , wherein the functional fragment is a ligand binding domain of sFLT1.
46 . The pharmaceutical composition according to claim 44 , wherein the functional fragment is a dimerization domain of sFLT1.
47 . The pharmaceutical composition according to claim 42 , wherein the promoter sequence and the nucleic acid sequence encoding the anti-VEGF protein are separated by a sequence that is 300 base pairs or more.
48 . The pharmaceutical composition according to claim 42 , wherein the promoter sequence and the nucleic acid sequence encoding the anti-VEGF protein are separated by a UTR sequence.
49 . The pharmaceutical composition according to claim 42 , wherein the pharmaceutical composition does not comprise a prokaryotic regulatory sequence.
50 . The pharmaceutical composition according to claim 42 , wherein the pharmaceutical composition does not comprise an antibiotic resistance sequence.
51 . The pharmaceutical composition according to claim 42 , wherein the nucleic acid sequence encoding an anti-VEGF protein operatively linked to a promoter sequence is comprised by a recombinant virus.
52 . The pharmaceutical composition according to claim 51 , wherein the virus is selected from the group consisting of adeno-associated virus (AAV), adenovirus, helper-dependent adenovirus, retrovirus, herpes simplex virus, lentivirus, poxvirus, hemagglutinatin virus of Japan-liposome (HVJ) complex, Moloney murine leukemia virus, and HIV-based virus.
53 . The pharmaceutical composition according to claim 52 , wherein the virus is AAV.
54 . The pharmaceutical composition according to claim 51 , wherein the viral genome of the recombinant virus comprises nucleic acid elements in the following order:
a) a first ITR sequence;
b) the promoter sequence;
c) an intron sequence;
d) a first UTR sequence;
e) the sequence encoding the anti-VEGF protein;
f) a second UTR sequence;
g) a poly A sequence; and
h) a second ITR sequence.
55 . The pharmaceutical composition according to claim 51 , wherein the composition comprises at most 1×10 13 vector genomes of recombinant virus.
56 . The pharmaceutical composition according to claim 51 , wherein the composition comprises at most 1×10 11 vector genomes of recombinant virus.
57 . The pharmaceutical composition according to claim 51 , wherein the composition comprises at least 1×10 8 vector genomes of recombinant virus.
58 . The pharmaceutical composition according to claim 51 , wherein the vector genomes are formulated in a volume of 0.1 to 0.5 ml.
59 . The pharmaceutical composition according to claim 51 , wherein the vector genomes are formulated in a volume of 100 μl.
60 . A unit dose of a pharmaceutical composition, the unit dose comprising at most 1×10 13 vector genomes of recombinant virus comprising a nucleic acid that encodes for an anti-VEGF protein.
61 . The unit dose of a pharmaceutical composition according to claim 60 , wherein the unit dose comprises at most 1×10 11 vector genomes of the recombinant virus.
62 . The unit dose of a pharmaceutical composition according to claim 60 , wherein the unit dose comprises at least 1×10 8 vector genomes of the recombinant virus.
63 . The unit dose pharmaceutical composition according to claim 60 , wherein the unit dose is a volume of 0.1 to 0.5 ml.
64 . The unit dose of pharmaceutical composition according to claim 63 , wherein the unit dose is a volume of 100 μl volume.