Anti-angiogenic methods and compositions

• US 7,893,022 B2
• Filed: 10/31/2007
• Issued: 02/22/2011
• Est. Priority Date: 02/28/2005
• Status: Active Grant

• Current Assignee: Sangamo Biosciences, Inc.
• Original Assignee: Sangamo Biosciences, Inc.
• Inventors: Rebar, Edward J., Zhang, H. Steve, Gregory, Philip D.
• Primary Examiner(s): WEHBE, ANNE MARIE SABRINA
• Application Number:
  US11/981,386
• Publication Number:
  US 20080118469A1
• Time in Patent Office: 1,210 Days
• Field of Search: 514/2, 514/44.R
• US Class Current: 514/13.3
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 14/4705 stimulating, promoting or activating activity
• CPC Class Codes: C07K 14/811 Serine protease (E.C. 3.4.21) inhibitors
• CPC Class Codes: C07K 2319/71 containing domain for transcriptional activaation, e.g. VP16
• CPC Class Codes: C07K 2319/81 containing a Zn-finger domain for DNA binding
• CPC Class Codes: C12N 2799/022 where the vector is derived from an adenovirus
• CPC Class Codes: C12N 2799/025 where the vector is derived from a parvovirus
• CPC Class Codes: C12N 2799/028 where the vector is derived from a herpesvirus

Abstract

Disclosed herein are methods and compositions for treatment of conditions requiring inhibition of angiogenesis. Such conditions include those characterized by neovascularization, such as retinopathies, macular degeneration and various malignancies.

Claims

1. A method for the treatment of ocular neovascularization in a human or a mouse animal, the method comprising intra-ocularly introducing a fusion protein comprising a zinc finger protein and an activation domain into the human or mouse animal having a genome, the genome comprising a target site within a pigment epithelium derived factor (PEDF) gene, whereby the zinc finger protein binds to the target site and thereby modulates PEDF expression in the human or mouse animal such that ocular neovascularization is treated, and further wherein the zinc finger protein binds to a target site specified in Table 1 (SEQ ID NOs:

8-13).

2. The method of claim 1, wherein the animal is a human and the zinc finger protein comprises six zinc fingers and the amino acid sequence of the recognition region of each of the zinc fingers is as follows:

3. The method of claim 1, wherein the activation domain is selected from the group consisting of the VP16 activation domain, the VP64 activation domain and the p65 activation domain.

4. The method of claim 3, wherein the protein comprises two p65 activation domains.

5. The method of claim 1, wherein the method further comprises inhibiting the expression of an endogenous gene encoding an angiogenic factor in one or more cells of the human or mouse animal.

6. The method of claim 5, wherein the angiogenic factor is a vascular endothelial growth factor (VEGF).

7. The method of claim 6, wherein the VEGF is vascular endothelial growth factor A (VEGF-A).

8. The method of claim 7, wherein expression of the endogenous VEGF-A gene is inhibited by binding of a zinc finger protein to a target site in the endogenous VEGF-A gene, wherein the zinc finger protein further comprises a repression domain.

9. The method of claim 8, wherein the animal is a human and the VEGF-A binding zinc finger protein comprises three zinc fingers and the amino acid sequence of the recognition region of each of the zinc fingers is as follows:

10. The method of claim 9, wherein the repression domain is selected from the group consisting of the v-erbA repression domain and the KOX repression domain.

11. The method of claim 1, wherein the ocular neovascularization occurs in a disease selected from the group consisting of age-related macular degeneration (AMD), diabetic retinopathy (DR) and retinopathy of prematurity.

12. The method of claim 1, wherein the fusion protein is introduced intra-ocularly as a polynucleotide expression vector into one or more cells of the human or mouse animal.

13. The method of claim 9, wherein the fusion protein and the VEGF-binding zinc finger proteins are introduced intra-ocularly as a polynucleotide expression vector into one or more cells of the human.

14. The method of claim 12, in which the polynucleotide is introduced into one or more retinal epithelial cells.

15. The method of claim 12, in which the polynucleotide is encapsidated in a viral vector selected from the group consisting of adeno-associated virus (AAV), Adenovirus and lentivirus.

16. The method of claim 15, in which the viral vector is an adeno-associated virus (AAV).

17. The method of claim 16, in which the viral vector is AAV Type 2 or AAV Type 4.