Use of aav-expressed m013 protein as an anti-inflammatory therapeutic

Patent metadata

Field	Value
Publication number	US-2024101607-A1
Application number	US-202318314773-A
Country	US
Kind code	A1
Filing date	May 9, 2023
Priority date	Mar 11, 2014
Publication date	Mar 28, 2024
Grant date	—

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

1 . A composition comprising an adeno-associated virus (AAV) particle, wherein the AAV particle comprises an AAV capsid protein and a polynucleotide that comprises:
(a) a first nucleic acid segment that encodes a modified, virally-derived, M013 polypeptide; and (b) a second nucleic acid segment-encoding a cell penetrating peptide and a secretion signal peptide; wherein the first and second nucleic acid segments are operably linked to a promoter sequence that is active in one or more mammalian cells comprising the polynucleotide.

2 . (canceled)

3 . The composition of claim 1 , wherein the AAV capsid protein comprises at least one surface-exposed amino acid which is substituted by a non-native residue, wherein the AAV particle has an increased transduction efficiency, an altered cellular specificity, or a combination thereof, as compared to an AAV particle comprising a wild-type AAV capsid protein, when the AAV particle is introduced into the one or more mammalian cells.

4 . The composition of claim 1 , wherein the polynucleotide is a self-complementary recombinant AAV vector.

5 . The composition of claim 1 , wherein a) the polynucleotide is characterized as an AAV serotype 1 vector (AAV1), an AAV serotype 2 vector (AAV2), an AAV serotype 3 vector (AAV3), an AAV serotype 4 vector (AAV4), an AAV serotype 5 vector (AAV5), an AAV serotype 6 vector (AAV6), an AAV serotype 7 vector (AAV7), an AAV serotype 9 vector (AAV9), an AAV serotype 10 vector (AAV10), an AAV serotype 11 vector (AAV11), or an AAV serotype 12 vector; or b) the AAV capsid protein is an AAV1 capsid protein, an AAV2 capsid protein, an AAV3 capsid protein, an AAV4 capsid protein, an AAV5 capsid protein, an AAV6 capsid protein, an AAV7 capsid protein, an AAV8 capsid protein, an AAV9 capsid protein, an AAV10 capsid protein, an AAV11 capsid protein, or an AAV12 capsid protein.

6 . The composition 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.

7 . The composition of claim 1 , wherein the promoter is a homologous promoter, a heterologous promoter, 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.

8 . The composition of claim 1 , wherein the polynucleotide further comprises a sequence that expresses or encodes a therapeutic molecule, a diagnostic molecule, or a combination thereof.

9 . The composition of claim 8 , wherein the therapeutic molecule or the 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.

10 . The composition of claim 1 , wherein the modified, virally-derived M013 polypeptide comprises a TatM013 peptide or an M013 peptide, fused to one or more protein transduction domains that facilitate or aid in transport of the resulting fusion protein across a mammalian cell membrane, or facilitates secretion of the resulting fusion protein from the one or more mammalian cells comprising the polynucleotide.

11 . The composition of claim 1 , wherein the cell-penetrating peptide comprises a sequence selected from any one of the amino acid sequences set forth in SEQ ID NOs:13-111.

12 . The composition of claim 10 , wherein the one or more protein transduction domains comprise an N-terminal mammalian secretion signal peptide.

13 . The composition of claim 10 , wherein the one or more protein transduction domains comprise an N-terminal mammalian secretion signal peptide selected from the group consisting of an IgK peptide, a glucagon-like peptide, a CNTF 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: 1-12, and any combination thereof.

14 .- 15 . (canceled)

16 . The composition of claim 1 , wherein the AAV particle is a recombinant adeno-associated virus (rAAV) particle, comprising a vector comprising a 5′ AAV inverted terminal repeat (ITR), the polynucleotide, and a 3′ AAV ITR.

17 .- 21 . (canceled)

22 . The composition of claim 16 , wherein the composition comprises a plurality of the rAAV particle.

23 .- 42 . (canceled)

43 . A polynucleotide comprising:
(a) a first nucleic acid segment that encodes a modified, virally-derived, M013 polypeptide; and (b) a second nucleic acid segment-encoding a cell penetrating peptide and a secretion signal peptide; wherein the first and second nucleic acid segments are operably linked to a promoter sequence that is active in one or more mammalian cells comprising the polynucleotide.

44 . The polynucleotide of claim 43 , wherein the polynucleotide further comprises a sequence that expresses or encodes a therapeutic molecule, a diagnostic molecule, or a combination thereof.

45 . The polynucleotide of claim 43 , wherein the modified, virally-derived M013 polypeptide comprises a TatM013 peptide or an M013 peptide, fused to one or more protein transduction domains that facilitate or aid in transport of the resulting fusion protein across a mammalian cell membrane, or facilitates secretion of the resulting fusion protein from the one or more mammalian cells comprising the polynucleotide.

46 . A vector comprising a 5′ adeno-associated virus (AAV) inverted terminal repeat (ITR), the polynucleotide of claim 43 , and a 3′ AAV ITR.

47 . A cell comprising the polynucleotide of claim 43 .

48 . A recombinant adeno-associated virus (rAAV) comprising at least one AAV capsid protein and the vector of claim 46 .

Assignees

• Univ Florida

Inventors

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

Classifications

• C07K14/005 Primary

  from viruses · CPC title

• A61K9/0048

  Eye, e.g. artificial tears · CPC title

• A61K38/162

  from virus · CPC title

• A61K48/005 Primary

  characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered · CPC title

• C12N7/00

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title