FIBRONECTIN POLYPEPTIDES AND METHODS OF USE

• US 20090111738A1
• Filed: 10/04/2006
• Published: 04/30/2009
• Est. Priority Date: 10/04/2005
• Status: Active Grant

• Current Assignee: The Research Foundation for The State University of New York (State University of New York)
• Original Assignee: The Research Foundation for The State University of New York (State University of New York)
• Inventors: Clark, Richard A., Ren, Xiang-Dong, Lin, Fubao
• Granted Patent: US 8,691,944 B2
• Time in Patent Office: Days
• US Class Current: 514/1.100
• CPC Class Codes: A61K 2800/24 Thermal properties
• 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 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• 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: A61K 8/02 characterised by special physical form
• CPC Class Codes: A61K 8/64 Proteins; Peptides; Derivatives or degradation products thereof
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61Q 17/04 Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
• CPC Class Codes: A61Q 19/00 Preparations for care of the skin
• CPC Class Codes: A61Q 19/001 Preparations for care of the lips
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]

Abstract

Described herein are fragments of fibronectin and variants thereof that bind growth factors. Compositions containing such a fragment of fibronectin are therefore useful in sequestering growth factors, and complexes containing both a FN fragment and a bound, active growth factor can be used to deliver growth factors to a patient (e.g., to a wound on the patient'"'"'s skin).

Claims

1. A physiologically acceptable composition comprising a fragment of a plasma fibronectin or a biologically active variant thereof, wherein the fragment or the variant thereof binds a polypeptide growth factor.

2. The physiologically acceptable composition of claim 1, wherein the polypeptide growth factor is selected from the group consisting of transforming growth factor-β

1 (TGF-β

1), transforming growth factor β

2 (TGF-β

2), basic fibroblast growth factor (bFGF), fibroblast growth factor 7 (FGF-7), platelet-derived growth factor (PDGF-BB), vascular endothelial growth factor A (VEGF-A), and nerve growth factor (NGF), and wherein the polypeptide growth factor, when bound by the fragment of plasma fibronectin, retains a biological activity.

3. The physiologically acceptable composition of claim 1, wherein the plasma fibronectin is human plasma fibronectin.

6. The physiologically acceptable composition of claim 1, wherein the fragment has a sequence normally found within the region designated FN_III1-2, FN_III12-15, or FN_IIICS.

7. The physiologically acceptable composition of claim 1, wherein the fragment binds the polypeptide growth factor with an affinity of at least 1×

10<sup>−

7</sup> M or with an affinity of at least 1×

10<sup>−

8</sup> M.

9. The physiologically acceptable composition of claim 1, wherein the biologically active variant is at least 80% identical to a corresponding wild type fragment of fibronectin.

10. The physiologically acceptable composition of claim 1, wherein the fragment of a plasma fibronectin or a biologically active variant thereof is a polypeptide comprising an amino acid sequence conforming to the formula

11. The physiologically acceptable composition of claim 10, wherein Xaa₁ is Gln;

Xaa₂ is Pro, Leu-Ile, or Gly;

Xaa₃ is Ser, Xaa₄ is absent, His, or Val;

Xaa₅ is Ile;

Xaa₆ is Ser;

Xaa₇ is Lys;

Xaa₈ is Tyr;

Xaa₉ is Ile;

Xaa₁₀ is Leu-Arg-Trp-Arg;

Xaa₁₁ is absent or Pro;

Xaa₁₂ is Lys;

Xaa₁₃ is Asn-Ser;

Xaa₁₄ is Val-Gly;

Xaa₁₅ is Arg;

Xaa₁₆ is Trp;

Xaa₁₇ is Lys; and

/or Xaa₁₈ is Glu-Ala.

21. The physiologically acceptable composition of claim 1, wherein the fragment comprises the amino acid sequence

23. The physiologically acceptable composition of claim 1, wherein the fragment of a plasma fibronectin or a biologically active variant thereof is a polypeptide comprising an amino acid sequence conforming to the formula

24. The physiologically acceptable composition of claim 23, wherein the polypeptide comprises the amino acid sequence QPSHISKYILRWRPK (SEQ ID NO:

______) (peptide 1A).

25. The physiologically acceptable composition of claim 1, wherein the fragment of a plasma fibronectin or a biologically active variant thereof is a polypeptide comprising an amino acid sequence conforming to the formula

37. The physiologically acceptable composition of claim 1, wherein the composition is a non-pharmaceutical composition selected from the group consisting of a cosmetic product, moisturizer, sunscreen, and lip balm.

52. A method of delivering a growth factor to a patient, the method comprising administering to the patient a therapeutically effective amount of the physiologically acceptable composition of claim 1.

53. The method of claim 52, wherein the patient is suffering from an injury to a tissue or a disorder resulting in tissue dysfigurement or dysfunction.

54. The physiologically acceptable composition of claim 1, wherein the composition is a tissue engineered product.

56. The tissue engineered product of claim 54, wherein the product further comprises a biocompatible polymer selected from the group consisting of a hyaluronan, a biodegradable polymer, a polylactide, a polyglycolide, a poly(lactide-coglycolide), a poly(lactic acid), a poly(glycolic acid), a poly(lactic acid-co-glycolic acid), a poly(caprolactone), a polycarbonate, a polyesteramide, a polyanhydride, a poly(amino acid), a poly(ortho ester), a polycyanoacrylate, a polyamide, a polyacetal, a poly(ether ester), a copolymer of poly(ethylene glycol)s and poly(ortho ester)s, a poly(dioxanone), a poly(alkylene alkylate), a biodegradable polyurethane, blends and copolymers thereof, an alginate polymer, a carboxy-vinyl polymer, and a carboxy-vinyl polymer comprising at least 90% acrylic acid monomers and about 0.1% to about 5.0% of a difunctional crosslinking agent.

4-5. -5. (canceled)

8. (canceled)

12-20. -20. (canceled)

22. (canceled)

27-36. -36. (canceled)

38. (canceled)

39. A nucleic acid molecule comprising a sequence encoding a polypeptide comprising an amino acid sequence conforming to the formula

40. The nucleic acid molecule of claim 39, wherein the polypeptide comprises the amino acid sequence

41. A nucleic acid molecule comprising a sequence encoding a polypeptide comprising an amino acid sequence conforming to the formula

42. A nucleic acid molecule comprising a sequence encoding a polypeptide comprising an amino acid sequence conforming to the formula

43-45. -45. (canceled)

47-49. -49. (canceled)

55. (canceled)

57-80. -80. (canceled)