Compositions containing protein polymers and vaccinia virus, and methods of use thereof

• US 20140086976A1
• Filed: 08/20/2013
• Published: 03/27/2014
• Est. Priority Date: 08/20/2012
• Status: Abandoned Application

• Current Assignee: Genelux Corporation
• Original Assignee: Genelux Corporation
• Inventors: Szalay, Aladar A., Chen, Nanhai George
• Application Number:
  US13/987,688
• Publication Number:
  US 20140086976A1
• Time in Patent Office: Days
• US Class Current: 424/445
• CPC Class Codes: A61K 35/76 Viruses; Subviral particles; Bacteriophages
• CPC Class Codes: A61K 47/42 Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein oligopeptides having up to five amino acids A61K47/183; polyamino acids A61K47/34
• CPC Class Codes: C12N 2710/24132 Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
• CPC Class Codes: C12N 2710/24171 Demonstrated in vivo effect
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

Provided herein are compositions containing a vaccinia virus and a protein polymer, and articles of manufacture thereof. Also provided are diagnostic and therapeutic methods using the compositions or articles of manufacture.

Claims

1. A composition, comprising:

a vaccinia virus;

a silk-elastin like protein polymer (SELP), wherein the SELP is capable of transitioning from a liquid to a hydrogel to form a hydrogel composition; and

a pharmaceutically acceptable carrier.

2. The composition of claim 1, wherein the SELP comprises alternating blocks of at least two units each of a silk-like sequence of amino acids and an elastin-like sequence of amino acids set forth by the formula {[S]_m[E]_n}_O, wherein:

S is the silk-like sequence of amino acids;

m is the number of silk-like amino acid units;

E is the elastin-like polypeptide amino acid sequence;

n is the number of elastin-like amino acid units; and

o is the number of monomer repeats.

3. The composition of claim 1 that is a liquid.

4. The composition of claim 1 that is a hydrogel.

5. The composition of claim 1, wherein the SELP has a molecular weight of at least 15 kD.

6. The composition of claim 1, wherein:

m is 2 to 16, 2 to 10, 2 to 8, 4 to 16, 4 to 10 or 4 to 8;

n is 1 to 40, 1 to 16, 2 to 12 or 4 to 8; and

/or

o is 2 to 100, 4 to 50, 6 to 25 or 2 to 20.

7. The composition of claim 1, wherein:

m is 2 to 16 or 2 to 8;

n is 1 to 16; and

o is chosen so that the SELP has a molecular weight of 15,000 to 100,000 Da.

8. The composition of claim 1, wherein:

the sequence of amino acids of the silk-like polypeptide is selected from among GAGAGS (SEQ ID NO;

26) or SGAGAG (SEQ ID NO;

27), or is a variant thereof that is capable of effecting formation of hydrogen bonds; and

/or

the sequence of amino acids of the elastin-like polypeptide is selected from among VPGG (SEQ ID NO;

30), APGVGV (SEQ ID NO;

31), VPGVG (SEQ ID NO;

32), or GVGVP (SEQ ID NO;

29), or is a variant thereof that confers aqueous solubility.

9. The composition of claim 8, wherein the elastin-like amino acid sequence is a variant that has the amino acid sequence GXGVP (SEQ ID NO:

35) or VPGXG (SEQ ID NO;

36), wherein X is valine, lysine, histidine, glutamic acid, arginine, aspartic acid, serine, tryptophan, tyrosine, phenylalanine, leucine, glutamine, asparagine, cysteine or methionine.

10. The composition of claim 9, wherein the elastin-like sequence is VPGKG (SEQ ID NO:

37) or GKGVP (SEQ ID NO;

38).

11. The composition of claim 1, wherein the SELP comprises the sequence of amino acids selected from among [(VPGVG)₈(GAGAGS)₂]₁₈ (SEQ ID NO:

39);

[(GVGVP)₄(GAGAGS)₉]₁₃ (SEQ ID NO;

40);

[(VPGVG)₈(GAGAGS)₄]₁₂ (SEQ ID NO;

41);

[(VPGVG)₈(GAGAGS)₆]₁₂ (SEQ ID NO;

42);

[(VPGVG)₈(GAGAGS)₈]₁₁ (SEQ ID NO;

43);

[(VPGVG)₁₂(GAGAGS)₈]₈ (SEQ ID NO;

44);

[(VPGVG)₁₆(GAGAGS)₈]₇ (SEQ ID NO;

45);

[(VPGVG)₃₂ (GAGAGS)₈]₅ (SEQ ID NO;

46);

(GAGAGS)₁₂ GAAVTGRGDSPASAAGY (GAGAGS)₅(GVGVGP)₈]₆ (SEQ ID NO;

47);

[(GAGAGS)₂ (GVGVP)₄ GKGVP (GVGVP)₃]₆ (SEQ ID NO;

48);

[(GAGAGS)₂(GVGVP)₄GKGVP (GVGVP)₃]₁₂ (SEQ ID NO;

49);

[(GAGAGS)₂(GVGVP)₄GKGVP (GVGVP)₃]₁₈ (SEQ ID NO;

50);

[(GAGAGS)₂ (GVGVP)₄ GKGVP (GVGVP)₃]₁₇GAGAGS)₂(SEQ ID NO;

51);

[(GAGAGS)₂-(GVGVP)₄-(GKGVP)-(GVGVP)₃-(GAGAGS)₂]₁₃ (SEQ ID NO;

52);

[GAGAGS (GVGVP)₄ GKGVP (GVGVP)₃(GAGAGS)₂]₁₂(SEQ ID NO;

53);

[(GVGVP)₄GKGVP(GVGVP)₁₁(GAGAGS)₄]₅(GVGVP)₄GKGVP(GVGVP)₁₁(GAG AGS)₂(SEQ ID NO;

54);

[(GVGVP)₄(GKGVP)(GVGVP)₁₁(GAGAGS)₄]₇(GVGVP)₄GKGVP(GVGVP)₁₁(GA GAGS)₂ (SEQ ID NO;

55);

[(GVGVP)₄GKGVP(GVGVP)₁₁(GAGAGS)₄]₉(GVGVP)₄GKGVP(GVGVP)₁₁(GAG AGS)₂(SEQ ID NO;

56);

[GAGS(GAGAGS)₂(GVGVP)₄GKGVP(GVGVP)₁₁(GAGAGS)₅GA]₆ (SEQ ID NO;

57);

[(GAGAGS)₂ (GVGVP)₁ LGPLGP (GVGVP)₃ GKGVP (GVGVP)₃]₁₅ (GAGAGS)₂ (SEQ ID NO;

73); and

[(GAGAGS)₂ (GVGVP)₁ GFFVRARR (GVGVP)₃ GKGVP (GVGVP)₃)₁₅(GAGAGS)₂ (SEQ ID NO;

74).

12. The composition of claim 1, wherein the SELP comprises the sequence of amino acids selected from among [(GAGAGS)₂(GVGVP)₄GKGVP (GVGVP)₃]₁₇ GAGAGS)₂ (SEQ ID NO:

51);

[(GAGAGS)₂-(GVGVP)₄-(GKGVP)-(GVGVP)₃-(GAGAGS)₂]₁₃ (SEQ ID NO;

52); and

[GAGS(GAGAGS)₂(GVGVP)₄GKGVP(GVGVP)₁₁(GAGAGS)₅GA]₆(SEQ ID NO;

57).

13. The composition of claim 1, wherein the SELP comprises the sequence of amino acids selected from among SELP-27K (SEQ ID NO:

61), SELP-47K (SEQ ID NO;

62) and SELP-815K (SEQ ID NO;

63).

14. The composition of claim 1, wherein the concentration of the SELP protein polymer is at a weight percentage (wt %) of the composition of from or from about 2% (w/w) to about to 50% (w/w) or 2% (w/w) to about 20% (w/w), each inclusive.

15. The composition of claim 1, wherein the strain of vaccinia virus is selected from among Lister, Western Reserve (WR), Copenhagen (Cop), Bern, Paris, Tashkent, Tian Tan, Wyeth (DRYVAX), IHD-J, IHD-W, Brighton, Ankara, CVA382, Modified Vaccinia Ankara (MVA), Dairen I, LC16 m8, LC16M0, LIVP, ACAM2000, WR 65-16, Connaught, New York City Board of Health (NYCBH), EM-63 and NYVAC strain.

16. The composition of claim 15, wherein the vaccinia virus is a Lister strain virus.

17. The composition of claim 1, wherein the vaccinia virus is an LIVP virus or a clonal strain of an LIVP virus.

18. The composition of claim 17, wherein the virus is a modified form containing nucleic acid encoding a heterologous gene product.

19. The composition of claim 18, wherein the heterologous gene product is a therapeutic or reporter gene product.

20. The composition of claim 19, wherein the heterologous gene product is selected from among an anticancer agent, an antimetastatic agent, an antiangiogenic agent, an immunomodulatory molecule, an antigen, a cell matrix degradative gene, genes for tissue regeneration and reprogramming human somatic cells to pluripotency, enzymes that modify a substrate to produce a detectable product or signal or are detectable by antibodies, proteins that can bind a contrasting agent, genes for optical imaging or detection, genes for PET imaging and genes for MRI imaging.

21. The composition of claim 19, wherein the heterologous gene product is a therapeutic agent selected from among a hormone, a growth factor, cytokine, a chemokine, a costimulatory molecule, ribozymes, a transporter protein, a single chain antibody, an antisense RNA, a prodrug converting enzyme, an siRNA, a microRNA, a toxin, an antitumor oligopeptide, a mitosis inhibitor protein, an antimitotic oligopeptide, an anti-cancer polypeptide antibiotic, an angiogenesis inhibitor, a tumor suppressor, a cytotoxic protein, a cytostatic protein and a tissue factor.

22. The composition of claim 1, wherein the vaccinia virus is present in the composition in an amount that is from or from about 1×

10⁵ to 1×

10¹² pfu, inclusive.

23. The composition of claim 1 that is formulated for direct administration.

24. The composition of claim 23, wherein the volume of the composition is from or from about 0.01 mL to 100 mL, 0.1 mL to 100 mL, 1 mL to 100 mL, 10 mL to 100 mL, 0.01 mL to 10 mL, 0.1 mL to 10 mL, 1 mL to 10 mL, 0.02 mL to 20 mL, 0.05 mL to 5 mL, 0.5 mL to 50 mL or 0.5 mL to 5 mL, each inclusive.

25. The composition of claim 1, further comprising an agent that inhibits or decreases hydrogen bonding in an amount effective to decrease the hydrogen bonding.

26. The composition of claim 24, wherein the agent is selected from among urea, guanidine hydrochloride, dimethyl formamide, colloidal gold sol, aqueous lithium bromide and formic acid.

27. The composition of claim 1 that is formulated for local or systemic injection.

28. The composition of claim 27 that is formulated for intravenous administration.

29. The composition of claim 27 that is formulated for topical administration.

30. A combination, comprising a composition of claim 1 and an anti-cancer agent.

31. The combination of claim 30, wherein the anticancer agent is selected from among a cytokine, a chemokine, a growth factor, a photosensitizing agent, a toxin, an anti-cancer antibiotic, a chemotherapeutic compound, a radionuclide, an angiogenesis inhibitor, a signaling modulator, an anti-metabolite, an anti-cancer vaccine, an anti-cancer oligopeptide, a mitosis inhibitor protein, an antimitotic oligopeptide, an anticancer antibody, an anti-cancer antibiotic, an immunotherapeutic agent and a combination of any of the preceding thereof.

32. A virus delivery device, comprising:

the composition of claim 1; and

a device for administration of the composition.

33. The virus delivery device of claim 32, wherein the device is for local administration of the vaccinia virus in polymer composition.

34. The virus delivery device of claim 32, wherein the composition is in the form of a hydrogel.

35. The virus delivery device of claim 34, wherein the composition is coated on a surface of the device.

36. The virus delivery device of claim 32, wherein the device can be applied to a surface of the body of a subject.

37. The virus delivery device of claim 36 that is a patch, bandage, wrap, dressing, suture, film or mesh.

38. The virus delivery device of claim 37 that is a wound dressing or bandage.

39. A method of treating a disease or condition in a subject, comprising administering a composition of claim 1 to a subject, wherein the disease or condition is one that is treated by a administering a therapeutic vaccinia virus.

40. The method of claim 39, wherein the disease or condition is a proliferative disorder.

41. The method of claim 39, wherein the composition is administered locally or systemically.

42. The method of claim 41, wherein the composition is administered intravenously.

43. The method of claim 41, wherein the composition is administered locally inside a body cavity.

44. The method of claim 40, wherein the proliferative disease is a cancer, tumor or metastasis.

45. The method of claim 44, wherein the cancer is a carcinoma, sarcoma, lymphoma or leukemia.

46. The method of claim 44, wherein the tumor is a solid tumor.

47. The method of claim 46, wherein the tumor is a surgically resected tumor.

48. The method of claim 47, wherein the composition is administered topically.

49. The method of claim 40, wherein the proliferative disease is a skin cancer.

50. The method of claim 49, wherein the skin cancer is a melanoma, a basal cell carcinoma of the skin or a squamous cell carcinoma.

51. The method of claim 50, wherein the composition is administered topically.

52. The method of claim 39, wherein the subject is a human or non-human animal.

53. The method of claim 52, wherein the non-human animal is selected from among a horse, cat, dog, cow, pig, sheep, goat, mouse, rabbit, chicken, rat, and guinea pig

54. The method of claim 39, wherein the composition as administered delivers at least or about 1×

10⁵ pfu of virus.

55. The method of claim 39, further comprising administering a second therapeutic agent or treatment for the treatment of the proliferative disorder.

56. The method of claim 55, wherein a second therapeutic agent is administered, and the therapeutic agent is an anti-cancer agent.

57. The method of claim 55, wherein a second treatment is administered and is selected from among surgery, radiation therapy and immunosuppressive therapy.

58. The method of claim 56, wherein the composition and the anticancer agent are administered sequentially, simultaneously, or intermittently.

59. A method of treating a skin lesion in a subject, comprising applying a virus delivery device of claim 32 to the surface of the skin of a subject to cover the skin lesion, thereby delivering virus to the skin lesion to treat the skin lesion.

60. The method of claim 59, wherein the skin lesion is a wound or proliferative skin lesion.

61. The method of claim 59, wherein the proliferative skin lesion is benign, premalignant or malignant.

62. The method of claim 60, wherein the proliferative skin lesion is a skin cancer.

63. The method of claim 62, wherein the skin cancer is a melanoma, a basal cell carcinoma of the skin or a squamous cell carcinoma.

64. The method of claim 60, wherein the wound is a traumatic wound or a post-surgical wound.

65. The method of claim 64, wherein the wound is a post-surgical wound that is a surgically resected tumor.

66. The method of claim 59, wherein the subject is a human or non-human animal.