Anticoagulant analogues of the tissue factor extrinsic pathway inhibitor (EPI) with reduced affinity for heparin

• US 5,312,736 A
• Filed: 01/27/1992
• Issued: 05/17/1994
• Est. Priority Date: 08/18/1989
• Status: Expired due to Fees

• Current Assignee: Novo Nordisk A/S
• Original Assignee: Novo Nordisk A/S
• Inventors: Rasmussen, Jesper S., Nordfang, Ole J.
• Primary Examiner(s): Hill, Jr., Robert J.
• Assistant Examiner(s): Fitzgerald, David L.
• Application Number:
  US07/828,920
• Time in Patent Office: 841 Days
• Field of Search: 530/350, 530/380, 530/384, 530/395, 536/23.1, 536/23.5, 435/69.2, 435/240.2, 435/252.3-252.35, 435/320.1, 514/8
• US Class Current: 435/69.2
• 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: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: C07K 14/8114 Kunitz type inhibitors

Abstract

Novel extrinsic pathway inhibitors (EPI) are provided wherein one or more of the amino acid residues of native EPI have been deleted. A preferred group of the novel EPI analogues have no or a low heparin binding capacity. The novel EPI analogues can be used for the treatment of patients having coagulation disorders or cancer.

Claims

1. An analogue of extrinsic pathway inhibitor (EPI) which analogue is a derivative of the full-length EPI sequence depicted in SEQ ID NO:

1, and which analogue retains the anticoagulant activity of native EPI as well as anti-FXa activity, comprises the three Kunitz protease inhibitor domains present in EPI, but has a heparin binding capacity less than 50% of the heparin binding capacity of native EPI when each is contacted with immobilized heparin under physiological conditions.

2. EPI analogue according to claim 1 comprising the amino acid sequence from Asp1 to Thr161 (SEQ ID NO:

7) of the native EPI molecule.

3. EPI analogue according to claim 1 comprising the amino acid sequence from Glu15 to Thr161 (SEQ ID NO:

5) of the native EPI molecule.

4. EPI analogue according to claim 1 and further having a Ser as the N-terminal amino acid residue.

5. DNA-sequence encoding an EPI analogue according to claim 1.

6. Expression vector containing a DNA-sequence according to claim 5.

7. Transformed or transfected microorganism or cell line comprising a vector according to claim 6.

8. A method for preparation of an EPI analogue wherein a microorganism or cell line according to claim 7 is cultured in a suitable culture medium and said EPI analogue is isolated therefrom.

9. A therapeutic preparation comprising an EPI analogue according to claim 1 and a suitable adjuvant or carrier.