Anti-angiogenic methods and compositions

• US 8,012,946 B1
• Filed: 01/09/2008
• Issued: 09/06/2011
• Est. Priority Date: 02/28/2005
• Status: Expired due to Fees

• 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:
  US12/008,245
• Time in Patent Office: 1,336 Days
• Field of Search: 514/44.R, 514/7.6, 514/8.2, 514/19.3
• US Class Current: 514/44.00R
• 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 a malignancy in a human or a mouse animal, wherein the method comprises introducing a fusion protein comprising a zinc finger protein and an activation domain into the malignancy of the human or a 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 activates PEDF expression in the human or a mouse animal such the malignancy 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:

F1;

RSDALSR (SEQ ID NO;

14)

F2;

QSGDLTR (SEQ ID NO;

15)

F3;

QSGDLTR (SEQ ID NO;

15)

F4;

TSGHLSR (SEQ ID NO;

16)

F5;

RSDHLSN (SEQ ID NO;

17)

F6;

QSATRIT (SEQ ID NO;

18).

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 fusion protein comprises two p65 activation domains.

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

6. The method of claim 5, wherein the cytokine is granulocyte-macrophage colony-stimulating factor (GM-CSF).

7. The method of claim 6, wherein expression of the endogenous GM-CSF gene is activated by binding of a fusion protein comprising an activation domain and a zinc finger protein that binds to a target site in the endogenous GM-CSF gene.

8. The method of claim 7, wherein the animal is a human and the GM-CSF-binding 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:

F1;

RSDALSE (SEQ ID NO;

65)

F2;

DSSHRTR (SEQ ID NO;

60)

F3;

RSDHLSA (SEQ ID NO;

61)

F4;

ANSNRIK (SEQ ID NO;

62)

F5;

QSSDLSR (SEQ ID NO;

58)

F6;

RSDALAR (SEQ ID NO;

32).

9. The method of claim 7, wherein the activation domain is selected from the group consisting of the p65 activation domain, the VP64 activation domain and the VP16 activation domain.

10. The method of claim 1, wherein the malignancy occurs in a disease selected from the group consisting of head and neck cancer, glioblastoma, prostate cancer and pancreatic cancer.