METHOD FOR GENERATING THROMBIN

• US 20120027746A1
• Filed: 07/30/2010
• Published: 02/02/2012
• Est. Priority Date: 07/30/2010
• Status: Abandoned Application

• Current Assignee: Biomet Biologics LLC (Zimmer Biomet Holdings, Inc.)
• Original Assignee: Biomet Biologics LLC (Zimmer Biomet Holdings, Inc.)
• Inventors: Storrs, Richard W., Leach, Michael D., DORIAN, Randel
• Application Number:
  US12/846,944
• Publication Number:
  US 20120027746A1
• Time in Patent Office: Days
• US Class Current: 424/94.64
• 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: A61K 38/4833 Thrombin (3.4.21.5)
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: C12N 9/6429 Thrombin (3.4.21.5)
• CPC Class Codes: C12Y 304/21005 Thrombin (3.4.21.5)

Abstract

Methods of generating thrombin and methods of applying a clotting tissue sealant to a site on a subject are provided. A blood component comprising platelets can be obtained from the subject. A hypotonic composition is contacted with a solid matrix to form a thrombin-containing liquid, where the hypotonic composition includes water, calcium, a blood component comprising platelets, and optionally a chelator. Calcium is present in the hypotonic composition in an amount greater than the amount of calcium that can be complexed by the chelator. Thrombin-containing liquid is then separated from the hypotonic composition and can be applied to the site on the subject to form a clot, for example, by combination with fibrinogen.

Claims

1. A method of generating thrombin comprising:

contacting a hypotonic composition with a solid matrix to form a thrombin-containing liquid, the hypotonic composition comprising water, calcium, a blood component comprising platelets, and optionally a chelator, wherein the calcium is present in an amount greater than the amount of calcium that can be complexed by the chelator;

separating the thrombin-containing liquid from the hypotonic composition.

2. The method of claim 1, wherein the water contains the calcium.

3. The method of claim 1, wherein the concentration of platelets in the hypotonic solution is less than about half of the concentration of platelets in whole blood.

4. The method of claim 1, wherein the concentration of platelets in the hypotonic solution is less than about one-third of the concentration of platelets in whole blood.

5. The method of claim 1, wherein the blood component comprises the chelator.

6. The method of claim 1, further comprising obtaining the blood component comprising platelets and adding a chelator to the blood component to prevent coagulation prior to the contacting step.

7. The method of claim 1, wherein the solid matrix comprises glass, titanium, or combinations thereof.

8. The method of claim 1, wherein at least about one minute passes between the contacting step and the separating step.

9. The method of claim 1, wherein the hypotonic composition and solid matrix are agitated prior to the separating step.

10. The method of claim 1, wherein the separating comprises removing the thrombin-containing liquid from the solid matrix using a syringe, by filtering, or by centrifuging.

11. The method of claim 1, further comprising at least one of:

adding an alcohol to the thrombin-containing liquid;

adding a chelator to complex calcium in the thrombin-containing liquid;

adjusting the pH of the thrombin-containing liquid to below 7; and

reducing the temperature of the thrombin-containing liquid.

12. A method of applying a clotting tissue sealant to a site on a subject comprising:

obtaining a blood component comprising platelets;

contacting a hypotonic composition with a solid matrix to form a thrombin-containing liquid, the hypotonic composition comprising water, calcium, the blood component comprising platelets, and optionally a chelator, wherein the calcium is present in an amount greater than the amount of calcium that can be complexed by the chelator;

separating the thrombin-containing liquid from the hypotonic composition; and

applying the thrombin-containing liquid to the site on the subject to form a clot.

13. The method of claim 12, wherein the water contains the calcium.

14. The method of claim 12, wherein the concentration of platelets in the hypotonic solution is less than about half of the concentration of platelets in whole blood.

15. The method of claim 12, wherein the concentration of platelets in the hypotonic solution is less than about one-third of the concentration of platelets in whole blood.

16. The method of claim 12, wherein the blood component comprises the chelator.

17. The method of claim 12, further comprising adding a chelator to the blood component to prevent coagulation prior to the contacting step.

18. The method of claim 12, wherein the solid matrix comprises glass, titanium, or combinations thereof.

19. The method of claim 12, wherein at least about one minute passes between the contacting step and the separating step.

20. The method of claim 12, wherein the hypotonic composition and solid matrix are agitated prior to the separating step.

21. The method of claim 12, wherein the separating comprises removing the thrombin-containing liquid from the solid matrix using a syringe, by filtering, or by centrifuging.

22. The method of claim 12, further comprising at least one of:

adding an alcohol to the thrombin-containing liquid;

adding a chelator to complex calcium in the thrombin-containing liquid;

adjusting the pH of the thrombin-containing liquid to below 7; and

reducing the temperature of the thrombin-containing liquid.

23. The method of claim 22, wherein the alcohol is glycerol.

24. The method of claim 12, wherein the blood component comprising platelets is obtained from the subject.

25. The method of claim 12, wherein applying the thrombin-containing liquid to the site on the subject to form a clot comprises combining the thrombin-containing liquid with fibrinogen.