Regulation of angiogenesis with zinc finger proteins

• US 7,067,317 B2
• Filed: 04/30/2001
• Issued: 06/27/2006
• Est. Priority Date: 12/07/2000
• Status: Expired due to Fees

• Current Assignee: Sangamo Biosciences, Inc.
• Original Assignee: Sangamo Biosciences, Inc.
• Inventors: Jamieson, Andrew, Liu, Qiang, Rebar, Edward, Eisenberg, Stephen P., Wolffe, Alan, Jarvis, Eric, Liu, Pei-Qi
• Primary Examiner(s): Andres, Janet L.
• Assistant Examiner(s): Seharaseyon, Jegatheesan
• Application Number:
  US09/846,033
• Publication Number:
  US 20030044404A1
• Time in Patent Office: 1,884 Days
• Field of Search: 514/2, 514/12, 530/12, 530/351, 435/69.1, 435/6, 435/7.1, 435/455, 435/320.1, 435/325, 536/23.1, 536/23.5
• US Class Current: 435/455
• 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: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• 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: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C07K 14/52 Cytokines; Lymphokines; Interferons
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 2799/021 for the expression of a heterologous nucleic acid

Abstract

Provided herein are a variety of methods and compositions for regulating angiogenesis, such methods and compositions being useful in a variety of applications where modulation of vascular formation is useful, including, but not limited to, treatments for ischemia and wound healing. Certain of the methods and compositions accomplish this by using various zinc finger proteins that bind to particular target sites in one or more VEGF genes. Nucleic acids encoding the zinc finger proteins are also disclosed. Methods for modulating the expression of one or more VEGF genes with the zinc finger proteins and nucleic acids are also disclosed. Such methods can also be utilized in a variety of therapeutic applications that involve the regulation of endothelial cell growth. Pharmaceutical compositions including the zinc finger proteins or nucleic acids encoding them are also provided.

Claims

1. A method for modulating angiogenesis comprising introducing a zinc finger protein into an animal having a genome comprising a target site within a vascular endothelial growth factor (VEGF) gene, whereby the zinc finger protein binds to the target site and thereby modulates angiogenesis in the animal, wherein the zinc finger protein binds to a target site specified in Table 3 (SEQ ID NOS:

1–

29 and

     244) or Table 4 (SEQ ID NOS;

117–

119).

2. The method according to claim 1, wherein positions −

1 to +6 in each of three zinc fingers are occupied by first (SEQ ID NOS;

30–

58), second (SEQ ID NOS;

59–

87, 112, and 245–

252) and third segments (SEQ ID NOS;

42, 68 and 88–

116) of seven contiguous amino acids as specified in a row of Table 3.

3. The method according to claim 1, wherein the zinc finger protein comprises six zinc fingers, and positions −

1 to +6 in each of the six zinc fingers are occupied by first (SEQ ID NOS;

120–

122), second (SEQ ID NOS;

123–

125), third (SEQ ID NOS;

126–

128), fourth (SEQ ID NOS;

129–

131), fifth (SEQ ID NOS;

132–

134) and sixth (SEQ ID NOS;

135–

137) segments of seven contiguous amino acids as specified in a row of Table 4.

4. The method according to claim 1, wherein the target site is present in a plurality of VEGF genes, whereby the zinc finger protein binds to the target site in the plurality of genes, thereby modulating expression of the plurality of VEGF genes.

5. A method for treating a wound comprising introducing a zinc finger protein into an animal having a genome comprising a target site within a VEGF gene, whereby the zinc finger protein binds to the target site, such binding accelerating healing of the wound, and wherein the zinc finger protein binds to a target site specified in Table 3 (SEQ ID NOS:

1–

29 and

     244) or Table 4 (SEQ ID NOS;

117–

119).

6. The method according to claim 5, wherein positions −

1 to +6 in each of three zinc fingers are occupied by first (SEQ ID NOS;

30–

58), second (SEQ ID NOS;

59–

87, 112, and 245–

252) and third segments (SEQ ID NOS;

42, 68 and 88–

116) of seven contiguous amino acids as specified in a row of Table 3.

7. The method according to claim 5, wherein the zinc finger protein comprises six zinc fingers, and positions −

1 to +6 in each of the six zinc fingers are occupied by first (SEQ ID NOS;

120–

122), second (SEQ ID NOS;

123–

125), third (SEQ ID NOS;

126–

128), fourth (SEQ ID NOS;

129–

131), fifth (SEQ ID NOS;

132–

134) and sixth (SEQ ID NOS;

135–

137) segments of seven contiguous amino acids as specified in a row of Table 4.

8. The method of claim 5, wherein the zinc finger protein is applied topically.

10. The method according to claim 5, wherein the zinc finger protein comprises at least three fingers of the C2H2 class of zinc fingers.

9. A method of treating ischemia, comprising administering a zinc finger protein that binds to a target site within a VEGF gene into an animal having ischemia, wherein the animal has a genome comprising a VEGF gene comprising the target site and the zinc finger protein binds to the target site, and wherein the zinc finger protein is administered in an amount effective to treat ischemia, wherein the zinc finger protein binds to a target site specified in Table 3 (SEQ ID NOS:

1–

29 and

     244) or Table 4 (SEQ ID NOS;

117–

119).

11. The method according to claim 9, wherein positions −

1 to +6 in each of three zinc fingers are occupied by first (SEQ ID NOS;

30–

58), second (SEQ ID NOS;

59–

87, 112, and 245–

252) and third segments (SEQ ID NOS;

42, 68 and 88–

116) of seven contiguous amino acids as specified in a row of Table 3.

12. The method according to claim 9, wherein the zinc finger protein comprises six zinc fingers, and positions −

1 to +6 in each of the six zinc fingers are occupied by first (SEQ ID NOS;

120–

122), second (SEQ ID NOS;

123–

125), third (SEQ ID NOS;

126–

128), fourth (SEQ ID NOS;

129–

131), fifth (SEQ ID NOS;

132–

134) and sixth (SEQ ID NOS;

135–

137) segments of seven contiguous amino acids as specified in a row of Table 4.

13. The method of claim 9, wherein the zinc finger protein is applied to a specific tissue of the animal.