Polypeptides for promoting cell attachment

• US 6,316,412 B1
• Filed: 12/01/1998
• Issued: 11/13/2001
• Est. Priority Date: 12/03/1990
• Status: Expired due to Fees

• Current Assignee: The Scripps Research Institute
• Original Assignee: The Scripps Research Institute
• Inventors: Plow, Edward F., Bowditch, Ronald, Ginsberg, Mark H.
• Primary Examiner(s): CARLSON, KAREN C
• Application Number:
  US09/201,970
• Time in Patent Office: 1,078 Days
• Field of Search: 530/350, 530/300, 530/324, 530/325, 530/326, 530/327, 530/328, 530/329, 530/330, 530/387.1, 530/387.9, 514/12, 514/15
• US Class Current: 514/9.3
• 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: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: C07K 14/75 Fibrinogen
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 2533/56 Fibrin; Thrombin
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 9/1252 DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
• CPC Class Codes: Y10S 930/29 Polyamino acid or polypeptide with an uninterrupted series of peptide repeating units

Abstract

Novel polypeptides derived from human fibronectin are described which bind to integrin receptors expressed by cells. The receptor binding site of human fibronectin begins at amino acid residue 1394 and ends at residue 1400 of fibronectin. The polypeptides facilitate attachment of cells to substrates either alone or in conjunction with RGD-containing peptides. Vectors, fusion proteins and antibodies are also described. Methods for promoting cell attachment and for inhibiting cell adhesion are also described.

Claims

1. A prosthetic device provided with a coating composition comprising a polypeptide having a length of no more than about 100 amino acid residues that includes an amino acid residue sequence represented by the formula:

AspArgXaa₁ProHisXaa₂Arg (SEQ ID NO

     1), wherein Xaa₁ and Xaa₂ are any amino acid residue and said polypeptide binds GPIIb-IIIa in an ArgGlyAsp-independent manner.

2. The device of claim 1 wherein said polypeptide has the amino acid residue sequence AspArgValProHisSerArg (SEQ ID NO 16).

3. A prosthetic device provided with a coating composition comprising a polypeptide having a length of no more than about 100 amino acid residues that includes an amino acid residue sequence—

AspArgXaa₁ProHisXaa₂ArgXaa₃Xaa₄Xaa₅Xaa₆—

(SEQ ID NO

     2), wherein Xaa₁ is Val or Ala, Xaa₂ is Ser or Ala, Xaa₃ is Asn or Ala, Xaa₄ is Ser or Ala, Xaa₅ is Ile or Ala, and Xaa₆ is Thr or Ala.

4. The device of claim 3 wherein said polypeptide has the amino acid residue sequence

AspArgValProHisSerArgAsnSerIleThr, AspArgAlaProHisSerArgAsnSerIleThr, AspArgValProHisAlaArgAsnSerIleThr, AspArgValProHisSerArgAlaSerIleThr, AspArgValProHisSerArgAsnAlaIleThr, AspArgValProHisSerArgAsnSerAlaThr, and AspArgValProHisSerArgAsnSerIleAla,

the SEQ ID NO of which are 11, 17, 18, 19, 20, 21 and 22 respectively.

5. A skin graft provided with a coating composition comprising a polypeptide having a length of no more than about 100 amino acid residues that includes an amino acid residue sequence represented by the formula:

AspArgXaa₁ProHisXaa₂Arg (SEQ ID NO

     1), wherein Xaa₁ and Xaa₂ are any amino acid residue and said polypeptide binds GPIIb-IIIa in an ArgGlyAsp-independent manner.

6. The skin graft of claim 5 wherein said polypeptide has the amino acid residue sequence AspArgValProHisSerArg (SEQ ID NO 16).

7. A skin graft provided with a coating composition comprising a polypeptide having a length of no more than about 100 amino acid residues that includes an amino acid residue sequence—

AspArgXaa₁ProHisXaa₂ArgXaa₃Xaa₄Xaa₅Xaa₆—

(SEQ ID NO

     2), wherein Xaa₁ is Val or Ala, Xaa₂ is Ser or Ala, Xaa₃ is Asn or Ala, Xaa₄ is Ser or Ala, Xaa₅ is Ile or Ala, and Xaa₆ is Thr or Ala.

8. The skin graft of claim 7 wherein said polypeptide has the amino acid residue sequence

AspArgValProHisSerArgAsnSerIleThr, AspArgAlaProHisSerArgAsnSerIleThr, AspArgValProHisAlaArgAsnSerIleThr, AspArgValProHisSerArgAlaSerIleThr, AspArgValProHisSerArgAsnAlaIleThr, AspArgValProHisSerArgAsnSerAlaThr, and AspArgValProHisSerArgAsnSerIleAla,

the SEQ ID NO of which are 11, 17, 18, 19, 20, 21 and 22 respectively.

9. An antibody composition that immunoreacts with fibronectin and with a polypeptide having a length of no more than about 100 amino acid residues that includes an amino acid residue sequence represented by the formula:

AspArgXaa₁ProHisXaa₂Arg (SEQ ID NO

     1), wherein Xaa₁ and Xaa₂ are any amino acid residue and said polypeptide binds GPIIb-IIIa in an ArgGlyAsp-independent manner.

10. The antibody composition of claim 9 wherein said polypeptide has the amino acid residue sequence AspArgValProHisSerArg (SEQ ID NO 16).

11. An antibody composition that immunoreacts with fibronectin and with a polypeptide having a length of no more than about 100 amino acid residues that includes an amino acid residue sequence—

AspArgXaa₁ProHisXaa₂ArgXaa₃Xaa₄Xaa₅Xaa₆—

(SEQ ID NO

     2), wherein Xaa₁ is Val or Ala, Xaa₂ is Ser or Ala, Xaa₃ is Asn or Ala, Xaa₄ is Ser or Ala, Xaa₅ is Ile or Ala, and Xaa₆ is Thr or Ala.

12. The antibody composition of claim 11 wherein said polypeptide has the amino acid residue sequence

AspArgValProHisSerArgAsnSerIleThr, AspArgAlaProHisSerArgAsnSerIleThr, AspArgValProHisAlaArgAsnSerIleThr, AspArgValProHisSerArgAlaSerIleThr, AspArgValProHisSerArgAsnAlaIleThr, AspArgValProHisSerArgAsnSerAlaThr, and AspArgValProHisSerArgAsnSerIleAla,

the SEQ ID NO of which are 11, 17, 18, 19, 20, 21 and 22 respectively.