Use of AAV-expressed M013 protein as an anti-inflammatory therapeutic

Patent metadata

Field	Value
Publication number	US-11685767-B2
Application number	US-201615261599-A
Country	US
Kind code	B2
Filing date	Sep 9, 2016
Priority date	Mar 11, 2014
Publication date	Jun 27, 2023
Grant date	Jun 27, 2023

Abstract

Disclosed are methods and compositions for preventing, treating, and/or ameliorating one or more symptoms of inflammation in a mammal. In particular, viral vectors and medicaments containing them are disclosed, which are useful in the prophylaxis, therapy, or amelioration of symptoms of one or more inflammatory-mediated mammalian diseases, such as age-related macular degeneration (AMD), arthritis, Bechet's disease, Best macular dystrophy, corneal inflammation, diabetic retinopathy, drusen formation, dry AMD, dry eye, geographic atrophy, glaucomaocular neovascularization, Lupus erythematosus, macular degeneration, Mallatia Leventinese and Doyne honeycomb retinal dystrophy, nephritis, ocular hypertension, ocular inflammation, recurrent uveitis, Sorsby fundus dystrophy, vasculitis, vitreoretinopathy, wet AMD, or related disorders. In exemplary methods, administration of a pharmaceutical composition comprising a recombinant viral vector that delivers a secretable and cell-penetrating M013 protein or peptide to a subject in need thereof facilitated treatment of particular human disorders such as AMD, ocular neovascularization, uveitis, and related inflammatory ocular disease.

First claim

What is claimed is:

1. A method of reducing or inhibiting secretion of proinflammatory cytokines from retinal microglia during conditions of inflammation, comprising contacting a population of retinal cells in one or both eyes of a mammalian subject with a composition that comprises an effective amount of a viral particle comprising an AAV vector comprising a polynucleotide that comprises
(a) a first nucleic acid segment that encodes a virally-derived M013 polypeptide; and
(b) a second nucleic acid segment encoding a polypeptide comprising a cell penetrating peptide comprising the sequence set forth in SEQ ID NO: 23 and a secretion signal peptide;
wherein the first and second nucleic acid segments are operably linked to a promoter sequence that expresses the encoded polypeptides in one or more retinal cells transduced with the viral particle;
wherein the viral particle has a serotype that is AAV2 or AA2-based, and the viral particle comprises an AAV2(quadY-F+T−V) capsid protein; wherein the composition is contacted to the one or both eyes by intravitreal injection; wherein the population comprises one or more retinal microglia; and wherein the contacting provides a reduction or inhibition in secretion of proinflammatory cytokines from the retinal microglia.

2. A method for treating or ameliorating one or more symptoms of inflammation in a mammal, the method comprising administering to the mammal a composition that comprises a viral particle comprising an AAV vector comprising a polynucleotide that comprises
(a) a first nucleic acid segment that encodes a virally-derived M013 polypeptide; and
(b) a second nucleic acid segment encoding a polypeptide comprising a cell penetrating peptide comprising the sequence set forth in SEQ ID NO: 23 and a secretion signal peptide;
wherein the first and second nucleic acid segments are operably linked to a promoter sequence that expresses the encoded polypeptides in one or more retinal cells transduced with the viral particle; in an amount and for a time sufficient to treat or ameliorate the one or more symptoms of inflammation in the mammal, wherein the AAV vector is administered to one or both eyes of the mammal by intravitreal injection;
wherein the viral particle has a serotype that is AAV2 or AA2-based, and the viral particle comprises an AAV2(quadY-F+T−V) capsid protein;
wherein the one or both eyes comprises one or more retinal microglia; and wherein the administering provides a reduction or inhibition in secretion of proinflammatory cytokines from the retinal microglia.

3. The method of claim 2 , wherein the mammal is a human that has been diagnosed with macular degeneration, age-related macular degeneration (AMD), geographic atrophy, wet AMD, dry AMD, drusen formation, dry eye, diabetic retinopathy, vitreoretinopathy, Best macular dystrophy, Sorsby fundus dystrophy, Mallatia Leventinese and Doyne honeycomb retinal dystrophy, corneal inflammation, uveitis, ocular hypertension, glaucoma, or any combination thereof.

4. The method of claim 2 , wherein expression of the encoded polypeptides in the retinal cells: a) treats or ameliorates one or more symptoms of inflammation, b) preserves one or more photoreceptor cells, or c) a combination thereof.

5. The method of claim 2 , wherein expression of the encoded polypeptides persists in the retinal cells for a period of at least three months following intravitreal injection of the AAV vector to the one or both eyes of the mammal.

6. The method of claim 2 , wherein the intravitreal injection occurs as a single injection, or wherein the intravitreal injection includes sequential injection of the AAV vector to the one or both eyes over a pre-determined treatment period of several weeks to several months.

7. The method of claim 2 , wherein expression of the encoded polypeptides persists in the retinal cells for a period of at least six months following intravitreal injection of the AAV vector to the one or both eyes of the mammal.

8. The method of claim 2 , wherein the cell penetrating peptide undergoes proteolytic cleavage from the encoded polypeptide expressed by the remainder of the second nucleic acid segment.

9. The method of claim 3 , wherein the vector is intravitreally injected in an amount and for a time sufficient to treat or ameliorate symptoms of dry AMD.

10. The method of claim 1 , wherein the AAV vector is a self-complementary rAAV (scAAV) vector.

11. The method of claim 2 , wherein the AAV vector is a self-complementary rAAV (scAAV) vector.

12. The method of claim 1 , wherein the polynucleotide further comprises an enhancer, a post-transcriptional regulatory sequence, a polyadenylation signal, an inverted terminal repeat, a multiple cloning site, or any combination thereof, operably linked to the first and the second nucleic acid segments.

13. The method of claim 2 , wherein the polynucleotide further comprises an enhancer, a post-transcriptional regulatory sequence, a polyadenylation signal, an inverted terminal repeat, a multiple cloning site, or any combination thereof, operably linked to the first and the second nucleic acid segments.

14. The method of claim 1 , wherein the promoter is an endogenous promoter, an exogenous promoter, a viral-derived promoter, a mammalian-specific promoter, a tissue-specific promoter, a cell-specific promoter, a constitutive promoter, an inducible promoter, a human cell-specific promoter, or any combination thereof.

15. The method of claim 2 , wherein the promoter is an endogenous promoter, an exogenous promoter, a viral-derived promoter, a mammalian-specific promoter, a tissue-specific promoter, a cell-specific promoter, a constitutive promoter, an inducible promoter, a human cell-specific promoter, or any combination thereof.

16. The method of claim 1 , wherein the polynucleotide further comprises a sequence region that expresses or encodes a first therapeutic molecule, a first diagnostic molecule, or a combination thereof.

17. The method of claim 16 , wherein the first therapeutic molecule or the first diagnostic molecule is selected from the group consisting of a polypeptide, a peptide, a ribozyme, a peptide nucleic acid, an siRNA, an RNAi, an antisense oligonucleotide, an antisense polynucleotide, an antibody, an antigen binding fragment, and any combination thereof.

18. The method of claim 2 , wherein the polynucleotide further comprises a sequence region that expresses or encodes a first therapeutic molecule, a first diagnostic molecule, or a combination thereof.

19. The method of claim 18 , wherein the first therapeutic molecule or the first diagnostic molecule is selected from the group consisting of a polypeptide, a peptide, a ribozyme, a peptide nucleic acid, an siRNA, an RNAi, an antisense oligonucleotide, an antisense polynucleotide, an antibody, an antigen binding fragment, and any combination thereof.

20. The method of claim 1 , wherein the secretion signal peptide is selected from the group consisting of an N-terminal mammalian secretion signal peptide selected from the group consisting of an IgK peptide, a glucagon-like peptide, a PEDF peptide, a FGF10 peptide, a PDGF-A peptide, a Gas6 peptide, a CFH peptide, a GDNF peptide, an IL-8 peptide, an MCP-1 peptide, a TIMP3 peptide, a synthetic peptide, a peptide signal sequence as set forth in any one of SEQ ID NOs: 1 to 12, and any combination thereof.

21. The method of claim 2 , wherein the secretion signal peptide is selected from the group consisting of an N-terminal mammalian secretion signal peptide selected

Assignees

• Univ Florida

Inventors

• Mcfadden Douglas Grant
• Lewin Alfred S
• Lucas Alexandra Rose
• Ildefonso Cristhian J
• Rahman Mohammed Masmudur

Classifications

• A61K9/0048

  Eye, e.g. artificial tears · CPC title

• A61K48/00

  Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy · CPC title

• C12N15/86

  Viral vectors · CPC title

• C12N2710/24022

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• A61K38/162

  from virus · CPC title