Fibrin-binding peptide fragments of fibronectin

• US 5,792,742 A
• Filed: 08/01/1994
• Issued: 08/11/1998
• Est. Priority Date: 06/14/1991
• Status: Expired due to Term

• Current Assignee: Isis Innovation Limited, New York University
• Original Assignee: Isis Innovation Limited, New York University
• Inventors: Williams, Michael J., Campbell, Iain D., Rostagno, Agueda A., Baron, Martin, Gold, Leslie I.
• Primary Examiner(s): Fitzgerald, David L.
• Application Number:
  US08/283,857
• Time in Patent Office: 1,471 Days
• Field of Search: 530/350, 530/380, 530/402, 530/387.1, 530/388.25, 530/389.3, 424/94.3, 424/9.1, 435/69.6, 435/188, 514/2, 514/8
• 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: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]

Abstract

Fibrin-binding molecules are provided which include at least one peptide essentially corresponding to one or both of the following portions of the natural fibronectin molecule. The first portion is that portion which includes the ⁴ F1.⁵ F1 module pair of fibronectin and includes no more of the natural fibronectin molecule than the N-terminal 25.9 kDa proteolytic fragment. The second portion includes the ¹⁰ F1.¹¹ F1 module pair of fibronectin and includes no more of the natural fibronectin molecule than the C-terminal 11 kDa proteolytic fragment. Also disclosed are nucleic acid molecules encoding the fibrin-binding peptides, methods for making the peptides, methods for using the peptides in the diagnosis and treatment of cardiovascular, peripheral vascular, cerebrovascular, and other conditions associated with fibrin deposition, and assay methods for detecting a fibrin-binding molecule and for measuring fibrin.

Claims

1. A fibrin-binding molecule including at least one peptide selected from the group consisting of:

a) a portion of the natural fibronectin molecule which includes positions 150-244 of SEQ ID NO;

1, wherein the fibrin-binding molecule includes no more of the N-terminal portion of the natural fibronectin molecule than the N-terminal 25.9 kDa proteolytic fragment thereof; and

b) a portion of the natural fibronectin molecule which includes positions 2122-2232 of SEQ ID NO;

1, wherein the fibrin-binding molecule includes no more of the C-terminal portion of the natural fibronectin molecule than the C-terminal 11 kDa proteolytic thereof.

2. A fibrin-binding molecule in accordance with claim 1, wherein said molecule includes peptide a) but not peptide b).

3. A fibrin-binding molecule according to claim 2 wherein said molecule includes a peptide corresponding to an N-terminal portion of fibronectin having an apparent molecular weight of about 25.9 kDa as determined by laser desorption mass spectrometry.

4. A fibrin-binding molecule in accordance with claim 1, wherein said molecule includes peptide b) but not peptide a).

5. A fibrin-binding molecule according to claim 4, wherein said molecule includes a peptide corresponding to a C-terminal portion of fibronectin having an apparent molecular weight of about 11 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis.

6. A fibrin-binding molecule in accordance with claim 1, wherein said molecule includes both peptide a) and peptide b).

7. A fibrin-binding molecule in accordance with claim 1, selected from the group consisting of:

(a) the portion of fibronectin corresponding to positions 150-244 of SEQ ID NO;

1; and

(b) the portion of fibronectin corresponding to positions 2122-2232 of SEQ ID NO;

1.

8. A fibrin-binding molecule in accordance with claim 1, further including, bound to said peptide, a therapeutic agent or a diagnostic marker.

9. A molecule according to claim 8, wherein said peptide is labeled with a detectable label.

10. A molecule according to claim 8, wherein said peptide is conjugated to a therapeutic agent.

11. A molecule according to claim 10, wherein said therapeutic agent is selected from a thrombolytic and a fibrinolytic agent.

12. A molecule according to claim 10, wherein said therapeutic agent is a cytotoxic agent.

13. A pharmaceutical composition, comprising a fibrin-binding molecule according to 1, and a pharmaceutically acceptable carrier.