Method and means for detecting and treating disorders in the blood coagulation cascade

• US 6,083,905 A
• Filed: 01/22/1997
• Issued: 07/04/2000
• Est. Priority Date: 04/22/1994
• Status: Expired due to Fees

• Current Assignee: Stichting Centraal Laboratorium Van De Bloedtransfusiedienst Van Het Nederlandse Rode Kruis
• Original Assignee: Stichting Sanquin Bloedvoorziening
• Inventors: Voorberg, Johannes Jacobus, Mertens, Koenraad, van Mourik, Jan Aart
• Primary Examiner(s): Zitomer, Stephanie W.
• Application Number:
  US08/722,240
• Time in Patent Office: 1,259 Days
• Field of Search: 530/381, 514/2, 514/21
• US Class Current: 514/13.7
• 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 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 7/04 Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: C07K 14/745 Blood coagulation or fibrinolysis factors
• CPC Class Codes: C07K 14/755 Factors VIII , e.g. factor VIII C (AHF), factor VIII Ag (VWF)
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers

Abstract

This invention relates to the diagnosis of congenital defects in the anticoagulant protein C system. Methods that are disclosed are based on the detection of mutations at the cleavage sites of coagulation factors that are under control of activated protein C (APC). Diagnostic tests include analysis of the APC-cleavage sites of factor V and factor VIII, by using specific primers to amplify selectively from RNA, cDNA derived from RNA or chromosomal DNA, parts of factor V and factor VIII that contain cleavage sites for APC. Methods that monitor the presence of mutations at the cleavage sites for APC and their utility in the diagnosis of thrombo-embolic disease are disclosed. The invention further discloses methods for correcting the defects detected according to the invention, as well as novel therapeutic agents which can be used in the treatment of bleeding disorders, which agents are based on the "defective" Factor V and Factor VIII proteins leading to the thrombotic disorders described hereinabove.

Claims

1. A pharmaceutical composition comprising a blood coagulation Factor V which carries a substitution mutation of an arginine residue in at least one position selected from the group consisting of residues 306, 506 and 1765:

wherein the pharmaceutical composition is resistant against activated protein C.

2. A pharmaceutical composition according to claim 1, wherein the mutation is at position 306.

3. A pharmaceutical composition according to claim 1, wherein the mutation is at position 506.

4. A pharmaceutical composition according to claim 1, wherein the mutation is at position 306 and at position 506.

5. A pharmaceutical composition according to claim 1, wherein said factor V is obtained from genetically engineered cells or organisms expressing factor V that is resistant against activated protein C.

6. A pharmaceutical composition according to claim 1, wherein said factor V is obtained from blood or plasma of individuals displaying resistance against activated protein C.

7. A pharmaceutical composition according to claim 1, which further comprises at least one additional coagulation factor.

8. A pharmaceutical composition according to claim 7, wherein said at least one additional coagulation factor is selected from the group consisting of factor II, factor VII, factor IX and factor X.

9. A pharmaceutical composition according to claim 1, which further comprises a pharmaceutically acceptable carrier, diluent or adjuvant.

10. A pharmaceutical composition according to claim 1, for the treatment of disorders in the blood coagulation cascade.

11. A pharmaceutical composition according to claim 10, wherein said disorder in the blood coagulation cascade is due to a deficiency of at least one coagulation factor selected from the group consisting of factor VIII, factor IX, factor VII and factor X.

12. A pharmaceutical composition according to claim 11, wherein said disorder is complicated by the presence of inhibitors or antibodies against said coagulation factor.

13. A pharmaceutical composition for the treatment or prophylaxis of disorders in the blood coagulation cascade in a mammal requiring prophylaxis or treatment comprising a blood coagulation Factor in accordance with claim 1.