US20130323302A1 - Treatment of amd using aav sflt-1 - Google Patents

Treatment of amd using aav sflt-1 Download PDF

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Publication number
US20130323302A1
US20130323302A1 US13/889,275 US201313889275A US2013323302A1 US 20130323302 A1 US20130323302 A1 US 20130323302A1 US 201313889275 A US201313889275 A US 201313889275A US 2013323302 A1 US2013323302 A1 US 2013323302A1
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United States
Prior art keywords
seq
sflt
sequence
pharmaceutical composition
aspects
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
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US13/889,275
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English (en)
Inventor
Ian J. Constable
Elizabeth P. Rakoczy
Chooi-May Lai
Thomas W. Chalberg, JR.
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Avalanche Australia Pty Ltd
Original Assignee
Avalanche Australia Pty Ltd
Lions Eye Institute of Western Australia Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=49584435&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=US20130323302(A1) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Priority to US13/889,275 priority Critical patent/US20130323302A1/en
Application filed by Avalanche Australia Pty Ltd, Lions Eye Institute of Western Australia Inc filed Critical Avalanche Australia Pty Ltd
Assigned to AVALANCHE AUSTRALIA PTY LTD. reassignment AVALANCHE AUSTRALIA PTY LTD. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: CHALBERG, THOMAS W., JR.
Assigned to LIONS EYE INSTITUTE LIMITED reassignment LIONS EYE INSTITUTE LIMITED ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: LAI, Chooi-may, CONSTABLE, IAN J., RAKOCZY, ELIZABETH P.
Publication of US20130323302A1 publication Critical patent/US20130323302A1/en
Priority to US14/281,783 priority patent/US20140371438A1/en
Priority to US14/281,749 priority patent/US9943573B2/en
Priority to US14/281,765 priority patent/US20140341977A1/en
Assigned to AVALANCHE AUSTRALIA PTY LTD. reassignment AVALANCHE AUSTRALIA PTY LTD. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: LIONS EYE INSTITUTE LIMITED
Assigned to LIONS EYE INSTITUTE LIMITED reassignment LIONS EYE INSTITUTE LIMITED CORRECTIVE ASSIGNMENT TO CORRECT THE THE LISTING OF ASSIGNOR NAME. PREVIOUSLY RECORDED ON REEL 030457 FRAME 0046. ASSIGNOR(S) HEREBY CONFIRMS THE ASSIGNOR'S NAME. Assignors: RAKOCZY, P. ELIZABETH, LAI, Chooi-may, CONSTABLE, IAN J.
Priority to US15/851,650 priority patent/US10004788B2/en
Priority to US15/961,654 priority patent/US20180311319A1/en
Priority to US17/328,705 priority patent/US20220111015A1/en
Abandoned legal-status Critical Current

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Definitions

  • VEGF vascular endothelial growth factor
  • VEGFR VEGF receptor
  • PDGF platelet-derived growth factor
  • HIF hypoxia inducible factor
  • Ang angiopoietin
  • MAPK mitogen-activated protein kinases
  • the disclosure provides for a nucleic acid encoding the sFLT-1 which is operatively linked to a promoter selected from the group consisting of: cytomegalovirus (CMV) promoter, Rous sarcoma virus (RSV) promoter, MMT promoter, EF-1 alpha promoter, UB6 promoter, chicken beta-actin promoter, CAG promoter, RPE65 promoter and opsin promoter.
  • a promoter selected from the group consisting of: cytomegalovirus (CMV) promoter, Rous sarcoma virus (RSV) promoter, MMT promoter, EF-1 alpha promoter, UB6 promoter, chicken beta-actin promoter, CAG promoter, RPE65 promoter and opsin promoter.
  • the disclosure provides for the human subject as resistant to treatment with VEGF inhibitors.
  • the disclosure provides for administering of the pharmaceutical composition at a frequency less than 3 times a year in the human subject.
  • the disclosure provides for a pharmaceutical composition followed by gas/fluid exchange in the vitreous chamber.
  • the disclosure provides for a unit dose of a pharmaceutical composition
  • a pharmaceutical composition comprising recombinant viruses of 1 ⁇ 10 6 to 1 ⁇ 10 15 vector genomes, wherein the recombinant viruses comprise a nucleic acid encoding sFLT-1 operatively linked to a promoter.
  • the unit dose of the pharmaceutical composition comprises 1 ⁇ 10 10 to 3 ⁇ 10 12 vector genomes.
  • the disclosure provides for the nucleic acid for use, wherein the pharmaceutical composition is capable of elevating levels of sFLT-1 protein in the vitreous of the human subject after at least 72 hours after administration of said pharmaceutical composition to said human subject, compared to levels of sFLT-1 protein in the vitreous of said human prior to said administration.
  • VEGF has been implicated in eye diseases, including but not limited to ischemic retinopathy, intraocular neovascularization, age-related macular degeneration (AMD), wet-AMD, dry-AMD, retinal neovascularization, diabetic macular edema, diabetic retina ischemia, diabetic retinal edema, proliferative diabetic retinopathy, retinal vein occlusion, central retinal vein occlusion, branched retinal vein occlusion.
  • AMD age-related macular degeneration
  • AMD age-related macular degeneration
  • AMD age-related macular degeneration
  • AMD age-AMD
  • dry-AMD dry-AMD
  • retinal neovascularization diabetic macular edema
  • diabetic retina ischemia diabetic retinal edema
  • proliferative diabetic retinopathy retinal vein occlusion
  • central retinal vein occlusion central retinal vein occlusion
  • recombinant viruses of the disclosure are at most about 1 ⁇ 10 1 , 1 ⁇ 10 2 , 1 ⁇ 10 3 , 1 ⁇ 10 4 , 1 ⁇ 10 5 , 1 ⁇ 10 6 , 1 ⁇ 10 7 , 1 ⁇ 10 8 , 1 ⁇ 10 10 , 1 ⁇ 10 11 , 1 ⁇ 10 12 , 1 ⁇ 10 13 , 1 ⁇ 10 14 , 1 ⁇ 10 15 , 1 ⁇ 10 16 , 1 ⁇ 10 17 , and 1 ⁇ 10 18 vector genomes.
  • the nucleic acid may be delivered without the use of a virus (i.e. with a non-viral vector), and may be measured as the quantity of nucleic acid.
  • a virus i.e. with a non-viral vector
  • any suitable amount of nucleic acid may be used with the compositions and methods of this disclosure.
  • a polyA or termination sequence may include but is not limited to sequences such as 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.
  • 5′ UTRs sequences may comprise various sequence elements.
  • the present disclosure provides for 5′ UTR sequences that may include but are not limited to sequence elements such as one or more ribosome binding sites (RBS), linker sequences, spacer sequences, regulatory sequences, regulatory response elements, riboswitches, sequences that promote or inhibit translation initiation, regulatory sequences for mRNA transport or variants and/or combinations thereof.
  • RBS ribosome binding sites
  • linker sequences spacer sequences
  • regulatory sequences regulatory response elements
  • riboswitches sequences that promote or inhibit translation initiation, regulatory sequences for mRNA transport or variants and/or combinations thereof.
  • the recombinant virus and/or plasmid used to generate recombinant virus comprises nucleic acid elements in the following order: a) a first ITR sequence; b) a first linker sequence; c) a promoter sequence; d) a second linker sequence; e) an intron sequence; f) a third linker sequence; g) a first UTR sequence; h) a sequence encoding a VEGF inhibitor; i) a second UTR sequence; j) a fourth linker sequence; k) a poly A sequence; l) a fifth linker sequence; and m) a second ITR sequence.
  • salts formed with cations such as sodium, potassium, ammonium, magnesium, calcium, polyamides such as spermine and spermidine, and the like
  • inorganic acids for example hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid and the like
  • salts formed with organic acids such as, for example, acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, napthalenesulfonic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, polygalacturonic
  • the nucleic acid may be delivered without the use of a virus (i.e. with a non-viral vector), and may be measured as the quantity of nucleic acid.
  • a virus i.e. with a non-viral vector
  • any suitable amount of nucleic acid may be used with the compositions and methods of this disclosure.
  • CNV choroidal neovascularization
  • One or multiple (e.g. 2, 3, or more) blebs can be created.
  • the total volume of bleb or blebs created by the methods and systems of the disclosure cannot exceed the fluid volume of the eye, for example about 4 ml in a typical human subject.
  • the total volume of each individual bleb is preferably at about 0.1-0.2 ml.
  • a patient's best corrected visual acuity improves by 1, 2 3, 4, 5 or more lines.
  • sFLT-1 was first identified in human umbilical vein endothelial cells (HUVEC), but it has since been found to occur naturally in the placenta and circulating systematically in pregnant women.
  • the sFLT-1 used in generating rAAV.sFlt-1 contains an open reading frame encoding only the first six extracellular immunoglobulin-like domains of the full length membrane-spanning FLT-1, followed by a unique 31-amino acid long C-terminal extension, representing the alternatively splices, secreted soluble FLT-1 isoform described earlier.
  • Frag001m was formed from the following sequential nucleic acid elements which were chemically synthesized by Blue Heron Biotech, LLC (Bothell, Wash.) and cloned into a BHKan backbone: an ITR (AAV serotype 2), CMV-IE promoter, chimeric intron, 5′ untranslated region (UTR), sFlt-1 coding sequence, SV40 polyA region, ITR (AAV serotype 2).
  • the plasmid V109-pFB-AAV-CMV-SV40pA-Kan was obtained from Virovek, Inc. (Hayward, Calif.).
  • results of the studies for rAAV.sFlt-1 and rAAV(bv).sFlt-1 are presented in FIG. 25A and FIG. 25B .
  • the concentration of sFlt-1 protein expressed by HEK293 cells 72 hours after transduction with rAAV.sFlt-1 and rAAV(bv).sFlt-1 ranged from 100-1,000 pg/mL at an MOI of 1 ⁇ 10 4 , 100-10,000 pg/mL at an MOI of 1 ⁇ 10 5 and 1,000-10,000 pg/mL at an MOI of 1 ⁇ 10 6 .
  • mice from either the rAAV.sFlt-1 or control treatment groups were then assessed at one week and one month post-injection for the presence of immune cells (leucocytes, macrophages and B- and T-lymphocytes).
  • Flow cytometric examination of the posterior eye cup showed that at one week post-injection there was a statistically significant increase in CD45+ leucocytes (6.6-fold increase compared to control; P ⁇ 0.05) and CD11b+ macrophages (5.7-fold increase compared to control; P ⁇ 0.036).
  • CD19+, CD8+ and CD4+ B- and T-lymphocytes
  • Plasma hsFLT-1 levels in the monkeys did not show any trend at the different sampling times ( FIG. 3B ). This suggests that the injection of rAAV.sFlt-1 did not have an obvious effect on the plasma hsFLT-1 level. The fluctuating levels did not have any effect on the well-being of the monkeys.

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US14/281,749 US9943573B2 (en) 2012-05-15 2014-05-19 Treatment of ocular neovascularization using anti-VEGF proteins
US14/281,765 US20140341977A1 (en) 2012-05-15 2014-05-19 Treatment of amd using aav sflt-1
US15/851,650 US10004788B2 (en) 2012-05-15 2017-12-21 Treatment of ocular neovascularization using anti-VEGF proteins
US15/961,654 US20180311319A1 (en) 2012-05-15 2018-04-24 Treatment of ocular neovascularization using anti-vegf proteins
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