ELASTIN-LIKE POLYMER DELIVERY VEHICLES

• US 20070265197A1
• Filed: 05/11/2007
• Published: 11/15/2007
• Est. Priority Date: 05/12/2006
• Status: Active Grant

• Current Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Original Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Inventors: Bae, Younsoo, Furgeson, Darin
• Granted Patent: US 8,367,626 B2
• Time in Patent Office: Days
• US Class Current: 514/1.200
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 47/6435 the peptide or protein in the drug conjugate being a connective tissue peptide, e.g. collagen, fibronectin or gelatin
• CPC Class Codes: A61K 48/0041 the non-active part being polymeric
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00

Abstract

In invention concerns elastin-like polymer (ELP) delivery compositions and methods for the use thereof. In some aspects ELP compositions may be used to deliver therapeutic nucleic acids, polypeptides of small molecules. In some aspects, in vivo delivery with ELP compositions can directed to specific target sites by the application of local hyperthermia therapy. Compositions and methods for ELP gene therapy are provided.

Claims

1. A bioplex composition comprising a therapeutic nucleic acid in complex with a fusion protein wherein the fusion protein comprises an elastin-like repeat and a nucleotide binding sequence.

2. The bioplex of claim 1, wherein the fusion protein further comprises a spacer region between the elastin-like repeat and a nucleotide binding sequence.

3. The bioplex of claim 1, wherein the elastin-like repeat comprises 10 to 500 repeats of the sequence VPGXG (SEQ ID NO:

1) wherein X is any amino acid except proline.

6. The bioplex of claim 3, wherein X is valine, alanine or glycine.

7. The bioplex of claim 1, wherein 25% or more of the amino acids in the nucleic acid binding sequence are positively charged at neutral pH.

8. The bioplex of claim 7, wherein 25% or more of the amino acids in the nucleic acid binding sequence are lysine residues.

9. The bioplex of claim 8, wherein the nucleic acid binding sequence comprises 4 to 100 repeats of the sequence VKG.

10. The bioplex of claim 8, wherein the nucleic acid binding sequence comprises 4 to 100 repeats of the sequence VK.

11. The bioplex of claims 2, wherein the spacer region comprises 3 or more histidine residues.

12. The bioplex of claims 1, wherein the fusion protein further comprises a cell targeting sequence, a cell penetrating sequence or a localization signal.

22. The bioplex of claim 2, wherein the spacer comprises a cleavable linker.

23. The bioplex of claims 22, wherein the cleavable linker is cleaved at low pH.

24. The bioplex of claims 22, wherein the cleavable linker is cleaved by an intracellular enzyme.

25. The bioplex of claim 1, wherein the bioplex has a median thermal transition temperature of less than about 48°

C.

26. The bioplex of claim 1, wherein thermal transition of the bioplex occurs over a range of less than about 5°

C.

27. The bioplex of claim 8, wherein the bioplex has an amino acid nitrogen to nucleic acid phosphate (N/P) ratio of less than about 50 to 1.

31. The bioplex of claim 1, wherein bioplex is less than about 1 μ

m in diameter.

32. The bioplex of claim 31, wherein bioplex is less than about 500 nm in diameter.

33. The bioplex of claim 32, wherein bioplex is less than about 200 nm in diameter.

34. The bioplex of claim 1, wherein the therapeutic nucleic acid is a DNA.

35. The bioplex of claim 34, wherein the DNA is a DNA expression vector.

36. The bioplex of claim 1, wherein the therapeutic nucleic acid is a RNA.

37. The bioplex of claim 36, wherein the RNA is a mRNA, a siRNA or a miRNA.

4-5. -5. (canceled)

13-21. -21. (canceled)

28-30. -30. (canceled)

38. A method for delivery of a therapeutic nucleic acid to a cell comprising:

i) mixing the therapeutic nucleic acid with a fusion protein comprising an elastin-like repeat and a nucleotide binding sequence to form a bioplex; and

ii) contacting the cell with the bioplex.

75. The method of claim 38, the method further comprising:

i) mixing a therapeutic nucleic acid with a fusion protein comprising an elastin-like repeat and a nucleotide binding sequence to form a bioplex;

ii) contacting the cell with the bioplex; and

iii) increasing the temperature around the bioplex.

76. The method of claim 75, wherein increasing the temperature around the bioplex decreases the aqueous solubility of the bioplex.

77. The method of claim 38, wherein the cell is in an animal.

78. The method of claim 75, wherein the animal is a human.

79. The method of claim 77, wherein the nanbioplex is administered intravenously, intradermally, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostaticaly, intrapleurally, intratracheally, intranasally, intravitreally, intravaginally, intrarectally, topically, intratumorally, intramuscularly, intraperitoneally, subcutaneously, subconjunctival, intravesicularlly, mucosally, intrapericardially, intraumbilically, intraocularally, orally, locally, by inhalation (e.g. aerosol inhalation), by injection, by infusion or by continuous infusion.

80. The method of claim 77, further comprising applying a hyperthermia therapy to the animal.

81. The method of claim 80, wherein the hyperthermia is applied locally.

82. The method of claims 81, wherein the hyperthermia therapy increases the local temperature to between about 38°

C. and 46°

C.

39-74. -74. (canceled)

83. A method for treating a disease in a animal comprising:

i) mixing the therapeutic nucleic acid with a fusion protein comprising an elastin-like repeat and a nucleotide binding sequence to form a bioplex; and

ii) administering an effective amount of the bioplex to the animal.

84-93. -93. (canceled)