Delivery of Nrf2 as therapy for protection against reactive oxygen species

Patent metadata

Field	Value
Publication number	US-11053291-B2
Application number	US-201515120318-A
Country	US
Kind code	B2
Filing date	Feb 19, 2015
Priority date	Feb 19, 2014
Publication date	Jul 6, 2021
Grant date	Jul 6, 2021

Abstract

The present invention provides methods and compositions for treating and/or preventing age related macular degeneration and other conditions involving macular degeneration, ocular neovascularization, or inflammation, including ocular inflammation. In some embodiments, the methods comprise administering an expression vector that delivers a secretable and cell penetrating Nrf2 to a subject in need thereof.

First claim

What is claimed is:

1. An AAV vector comprising an expression construct comprising a nucleotide sequence encoding a recombinant polypeptide,
wherein the recombinant polypeptide comprises a cell penetrating peptide comprising a Tat peptide, an Nrf2 peptide consisting of a 14-mer peptide having the amino acid sequence LQLDEETGEFLPIQ (SEQ ID NO: 1), and a secretion signal, and
wherein the AAV vector is encapsidated in a viral particle comprising a modified AAV2 capsid protein having non-tyrosine residues corresponding to the positions Y272, Y444, Y500, and Y730 of a wild-type AAV2 capsid.

2. The AAV vector of claim 1 , wherein the recombinant polypeptide further comprises a carrier protein.

3. The AAV vector of claim 2 , wherein the secretion signal comprises a sequence selected from SEQ ID NOs: 2-14, the cell penetrating peptide comprises a sequence comprising SEQ ID NO: 25, and the carrier protein is selected from opticin, human serum albumin, dihidrofolate reductase (DHFR) destabilization domain, and FK506 binding protein (FKBP) destabilization domain.

4. The AAV vector of claim 2 , wherein the carrier protein and cell penetrating peptide are linked to one another by an enzyme cleavage site.

5. The AAV vector of claim 4 , wherein the enzyme cleavage site is a furin cleavage site.

6. The AAV vector of claim 3 , wherein the secretion signal is an Igκ signal.

7. A composition comprising a pharmaceutically acceptable carrier and the AAV vector of claim 1 .

8. A method for the amelioration or treatment of a disease or condition associated with oxidative stress or inflammation in a subject, the method comprising administration of a therapeutically effective amount of the AAV vector of claim 1 to the eye of the subject.

9. The method of claim 8 , wherein the recombinant polypeptide of the AAV vector further comprises a carrier protein.

10. The method of claim 9 , wherein the secretion signal comprises a sequence selected from SEQ ID NOs: 2-14, the cell penetrating peptide comprises a sequence comprising SEQ ID NO: 25, and the carrier protein is selected from opticin, human serum albumin, dihidrofolate reductase (DHFR) destabilization domain, and FK506 binding protein (FKBP) destabilization domain.

11. The method of claim 9 , wherein the carrier protein and cell penetrating peptide are linked to one another by an enzyme cleavage site.

12. The method of claim 11 , wherein the enzyme cleavage site is a furin cleavage site.

13. The method of claim 8 , wherein said administration results in secretion of the recombinant polypeptide from cells transfected with the AAV vector.

Assignees

• Univ Florida

Inventors

• Ildefonso Cristhian J
• Lewin Alfred S
• Li Qiuhong

Classifications

• A61K48/0075

  characterised by an aspect of the delivery route, e.g. oral, subcutaneous · CPC title

• A61K38/1709

  from mammals · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title

• C07K2319/02

  containing a signal sequence · CPC title

• C07K2319/50

  containing protease site · CPC title