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

Patent metadata

Field	Value
Publication number	US-2016376325-A1
Application number	US-201615261599-A
Country	US
Kind code	A1
Filing date	Sep 9, 2016
Priority date	Mar 11, 2014
Publication date	Dec 29, 2016
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

What is claimed is:

1 . A viral vector comprising:
(a) a polynucleotide that comprises a first isolated nucleic acid segment that encodes a modified, virally-derived, M013 polypeptide; and (b) a second isolated nucleic acid segment encoding a secretion signal peptide; wherein each nucleic acid segment is operably linked to a promoter sequence that expresses the encoded polypeptides in one or more selected mammalian host cells transformed with a viral particle comprising the viral vector.

2 . The viral vector of claim 1 , wherein the vector is a retroviral vector, a lentiviral vector, an adenoviral vector, a Herpes viral vector, a Hepatitis viral vector, an SV40 vector, an EBV vector, or an adeno-associated virus (AAV) vector.

3 . The viral vector of claim 2 , wherein the vector is packaged in an viral particle that comprises a viral capsid protein, in which at least one surface-exposed amino acid is substituted by a non-native residue, wherein the viral particle has an increased transduction efficiency, an altered cellular specificity, or a combination thereof, as compared to a viral particle comprising the wild-type capsid protein, when the particle is introduced into a selected mammalian host cell.

4 . The viral vector of claim 1 , wherein the vector is a self-complementary rAAV (scAAV) vector.

5 . The viral vector of claim 1 , a) 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) packaged within a virion particle comprising 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 AAV0 capsid protein, an AAV10 capsid protein, an AAV11 capsid protein, or an AAV12 capsid protein.

6 . The viral vector 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 viral vector 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 viral vector 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.

9 . The viral vector of claim 1 , 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.

10 . The viral vector 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 one or more mammalian cells comprising the vector.

11 . The viral vector of claim 1 , further comprising a cell-penetrating peptide selected from any one of the amino acid sequences set forth in SEQ ID NO:13 to SEQ ID NO:111.

12 . The viral vector of claim 1 , wherein the one or more protein transduction domains comprise an N-terminal mammalian secretion signal peptide.

13 . The viral vector of claim 1 , 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 NO:1 to 12, and combinations thereof.

14 . The viral vector of claim 1 , further comprising an rAAV packaging component, such as a) a host cell that expresses at least one rep gene and/or at least one cap gene, b) a helper virus gene product; or c) a helper virus selected from adenovirus or herpes virus that facilitates expression of the rAAV viral vector in a mammalian host cell.

15 . The viral vector of claim 1 , comprised within a viral particle or an infectious virion.

16 . A recombinant adeno-associated virus (rAAV) particle, comprising a rAAV nucleic acid vector comprising a 5′ inverted terminal repeat (ITR), a transgene, and a 3′ ITR.

17 . The rAAV particle of claim 16 , wherein the transgene is operatively linked to a sequence that regulates expression of the transgene in a cell.

18 . The rAAV particle of claim 17 , wherein the sequence that regulates expression of the transgene in a cell is a promoter.

19 . The rAAV particle of claim 16 , wherein the transgene is about 2- to 5-kb in length.

20 . The rAAV particle of claim 16 , wherein the nucleic acid vector is a self-complementary (sc) vector.

21 . The rAAV particle of claim 16 , wherein the transgene is a reporter transgene.

22 . A plurality of virions or infectious viral particles comprising the viral vector of claim 1 .

23 . An isolated, non-human, mammalian host cell comprising the viral vector of claim 1 .

24 . A composition comprising: a) the viral vector of claim 1 , the plurality of virions or infectious viral particles of claim 16 ; or the rAAV particle of claim 17 ; and b) a pharmaceutically-acceptable buffer, diluent, or excipient.

25 . The composition of claim 24 , comprised within a kit for diagnosing, preventing, treating or ameliorating one or more symptoms of a disease, defect, disorder, dysfunction, injury, or trauma in a mammal.

26 . A kit comprising:
(1) a component selected from the group consisting of:
(a) the viral vector of claim 1 ;
(b) the plurality of virions or infectious viral particles of claim 16 ; or
(c) the rAAV particle of claim 17 ; and
(2) instructions for using the component in the diagnosis, prevention, treatment, or amelioration of one or more symptoms of inflammation or disease in a mammal.

27 . The kit of claim 26 , further comprising an rAAV packaging component, such as a) a host cell that expresses at least one rep gene and/or at least one cap gene, b) a helper virus gene product; or c) a helper virus selected from adenovirus or herpes virus that facilitates expression of the rAAV viral vector in a mammalian host cell.

28 . A method for the treatment or the amelioration of one or more symptoms of a disease or an abnormal condition that is associated with, or that results from, oxidative stress or inflammation in one or more cells of a mammalian subject, comprising

Assignees

• Univ Florida

Inventors

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

Classifications

• A61P37/00

  Drugs for immunological or allergic disorders · CPC title

• A61P29/00

  Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID] · CPC title

• A61K9/0048

  Eye, e.g. artificial tears · CPC title

• C12N2750/14143

  viral genome or elements thereof as genetic vector · CPC title

• C12N15/86

  Viral vectors · CPC title