Method of thrombolysis by local delivery of reversibly inactivated acidified plasmin

• US 20030026798A1
• Filed: 05/10/2002
• Published: 02/06/2003
• Est. Priority Date: 11/13/1999
• Status: Active Grant

• Current Assignee: Grifols Therapeutics LLC (Grifols SA)
• Original Assignee: Talecris Biotherapeutics Incorporated (Grifols SA)
• Inventors: Zimmerman, Thomas P., Novokhatny, Valery, Taylor, Kathryn K., Jesmok, Gary J., Landskroner, Kyle A.
• Granted Patent: US 6,964,764 B2
• Time in Patent Office: Days
• US Class Current: 424/94.64
• CPC Class Codes: A61K 38/4833 Thrombin (3.4.21.5)
• CPC Class Codes: A61K 38/484 Plasmin (3.4.21.7)
• CPC Class Codes: A61K 47/02 Inorganic compounds
• CPC Class Codes: A61K 47/12 Carboxylic acids; Salts or anhydrides thereof
• CPC Class Codes: A61K 47/183 Amino acids, e.g. glycine, EDTA or aspartame
• CPC Class Codes: A61K 47/26 Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: C12N 9/6435 Plasmin (3.4.21.7), i.e. fibrinolysin
• CPC Class Codes: C12Y 304/21007 Plasmin (3.4.21.7), i.e. fibrinolysin

Abstract

Methods of thrombolysis that allow the use of a fibrinolytic composition comprising reversibly inactivated acidified plasmin and the localized delivery of the plasmin to a vascular thrombotic occlusion are disclosed. Further disclosed is a method for administering a therapeutic dose of a fibrinolytic composition substantially free of plasminogen activator to a human or animal having a vascular thrombotic occlusion. The fibrinolytic composition includes a reversibly inactivated acidified plasmin substantially free of plasminogen activator. Intravascular catheter delivery of the fibrinolytic composition directly into or in the immediate vicinity of the thrombus is disclosed to minimize the systemic degradation of fibrin while retaining the maximum plasmin activity against the thrombus.

Claims

1. A method of administering a therapeutic dose of a reversibly inactivated acidified fibrinolytic composition to a human or animal having a thrombotic occlusion, comprising the steps of:

a) identifying a human or animal having a thrombotic occlusion; and

b) administering parenterally to the human or animal a therapeutic dose of a pharmaceutically acceptable reversibly inactivated acidified fibrinolytic composition substantially free of plasminogen activator, wherein the therapeutic dose is delivered into or proximally to the thrombotic occlusion.

2. The method of claim 1, further comprising the step:

c) allowing the administered fibrinolytic composition to interact with the thrombotic occlusion.

3. The method of claim 2, further comprising the steps:

d) monitoring the level of vascular flow of the human or animal; and

e) repeating steps (a)-(d) until a pre-selected level of vascular flow is attained.

4. The method of claim 1, wherein the thrombotic occlusion is a vascular occlusion selected from a coronary thrombosis, deep venous thrombosis, peripheral thrombosis, embolic thrombosis, hepatic vein thrombosis, marasmic thrombosis, sinus thrombosis, venous thrombosis, an arterial thrombosis, an occluded arterio-venal shunt, or an occluded catheter device.

5. The method of claim 1, wherein the reversibly inactivated acidified fibrinolytic composition comprises a reversible inactivated acidified fibrinolytic enzyme and a pharmaceutically acceptable acidified carrier, wherein the pharmaceutically acceptable carrier is a low buffering capacity buffer.

6. The method of claim 1, wherein the reversibly inactivated acidified fibrinolytic composition is lyophilized, and wherein the method of claim 1 further comprises the step of adding a pharmaceutically acceptable carrier to the lyophilized fibrinolytic composition before administration to the human or animal.

7. The method of claim 5, wherein the reversibly inactivated acidified fibrinolytic enzyme is a mammalian serine protease.

8. The method of claim 5, wherein the reversibly inactivated acidified fibrinolytic enzyme is Glu-plasmin, Lys-plasmin, mini-plasmin, micro-plasmin, or a truncated variant thereof.

9. The method of claim 5, wherein the reversibly inactivated acidified fibrinolytic composition further comprises an anti-thrombotic agent.

10. The method of claim 5, wherein the pharmaceutically acceptable acidified carrier is a low buffering capacity buffer comprising water and at least one pharmaceutically acceptable acid, wherein the acid is an organic acid selected from a carboxylic acid, an oligopeptide, an amino acid, or an inorganic acid, and the acidified carrier has a pH less than about 4.0.

11. The method of claim 10, wherein the pharmaceutically acceptable acid is selected from formic acid, acetic acid, citric acid, glycine, isoleucine, serine, threonine, glutamine, aspartic acid, valine and alanine.

12. The method of claim 10, wherein the pharmaceutically acceptable acid is acetic acid or citric acid.

13. The method of claim 10, wherein the pharmaceutically acceptable acid is acetic acid.

14. The method of claim 1, wherein the reversibly inactivated acidified fibrinolytic composition has a pH of between about 2.5 and about 4.0.

15. The method of claim 1, wherein the reversibly inactivated acidified fibrinolytic composition has a pH of between about 3.0 and about 4.0.

16. The method of claim 1, wherein the reversibly inactivated acidified fibrinolytic composition has a pH of between about 3.1 and about 3.5.

17. The method of claim 7, wherein the mammalian serine protease is in the concentration range of between about 0.01 mg/ml to about 50 mg/ml.

18. The method of claim 7, wherein the mammalian serine protease is in the concentration range of between about 0.1 mg/ml to about 10 mg/ml.

19. The method of claim 10, wherein the acid is in the concentration range of between about 1 mM to about 500 mM.

20. The method of claim 10, wherein the acid is in the concentration range of between about 1 mM and 50 mM.

21. The method of claim 5, wherein the reversibly inactivated acidified fibrinolytic composition further comprises a stabilizing agent.

22. The method of claim 21, wherein the stabilizing agent is a pharmaceutically acceptable carbohydrate selected from glucose, maltose, mannitol, sorbitol, sucrose, lactose or trehalose.

23. The method of claim 22, wherein the carbohydrate has a concentration in the range of about 0.2% w/v to about 20% w/v.

24. The method of claim 22, wherein the carbohydrate is glucose and the concentration thereof is about 20%.

25. The method of claim 22, wherein the stabilizing agent is selected from glucosamine, niacinamide and thiamine.

26. The method of claim 22, wherein the stabilizing agent has a concentration in the range of about 0.01M to about 0.1M.

27. The method of claim 1, wherein the therapeutic dose of the reversibly inactivated acidified fibrinolytic composition is in the range of between about 0.1 mg plasmin/kg body weight and 10.0 mg plasmin/kg body weight.

28. The method of claim 1, wherein the therapeutic dose of the reversibly inactivated acidified fibrinolytic composition is in the range of between about 0.2 mg plasmin/kg body weight and 4.0 mg plasmin/kg body weight.

29. The method of claim 1, wherein the therapeutic dose of the reversibly inactivated acidified fibrinolytic composition is in the range of between about 1.0 mg plasmin/kg body weight and 2.0 mg plasmin/kg body weight.

30. The method of claim 1, wherein the reversibly inactivated acidified composition is administered by a catheter device.

31. The method of claim 31, wherein the intravascular catheter is directed to a vascular occlusion.

32. The method of claim 31, wherein the intravascular catheter delivers the reversibly inactivated acidified fibrinolytic composition proximal to, or into, a thrombotic occlusion.

33. The method of claim 31, wherein the intravascular catheter enters the thrombotic occlusion and the catheter delivers the reversibly inactivated acidified fibrinolytic composition directly into the thrombotic occlusion.