Thrombin compositions
View Patent ↗Compositions comprising thrombin and methods of preparing them are disclosed. The composition may be in the form of an aqueous solution at pH 5.7-7.4 consisting essentially of 0.1 mg/mL to 5.0 mg/mL thrombin, 2% to 4% (w/v) sucrose, 3.5% to 5% (w/v) mannitol, 50 mM to 300 mM NaCl, 0-5 mM CaCl 2 , 0.03% to 1% (w/v) of a surfactant or high-molecular-weight polyethylene glycol, and a physiologically acceptable buffer, or may be in lyophilized form.
1. A composition consisting essentially of:
0.1 mg/mL to 5.0 mg/mL thrombin;
2% to 4% (w/v) sucrose;
3.5% to 5% (w/v) mannitol;
50 mM to 300 mM NaCt;
0-5 mM CaCl2;
0.03% to 1% (w/v) of a surfactant or high-molecular-weight polyethylene glycol; and
a physiologically acceptable buffer,
in aqueous solution at pH 5.7-7.4, wherein concentration of the buffer is selected to provide approximately physiological pH upon application of the composition in a surgical setting;
wherein the ratio of mannitol:sucrose is greater than 1:1 but not greater than 2.5:1 (w/w) and wherein the molar ratio of sucrose:thrombin is at least 700:1.
2. The composition of claim 1 , wherein the ratio of mannitol:sucrose is 1.33:1 (w/w).
3. The composition of claim 1 , wherein the molar ratio of sucrose:thrombin is at least 2000:1.
4. The composition of claim 1 , wherein the thrombin is human thrombin.
5. The composition of claim 1 , wherein the thrombin is recombinant human thrombin.
6. The composition of claim 1 , wherein the thrombin concentration is 0.5-3.0 mg/ml.
7. The composition of claim 1 , wherein the thrombin concentration is 1 mg/ml.
8. The composition of claim 1 , wherein the sucrose concentration is 3% (w/v).
9. The composition of claim 1 , wherein the mannitol concentration is 4% (w/v).
10. The composition of claim 1 , wherein the sucrose concentration is 3% (w/v) and the mannitol concentration is 4% (w/v).
11. The composition of claim 1 , wherein the surfactant or high-molecular-weight polyethylene glycol is PEG 3350.
12. The composition of claim 11 , wherein the concentration of PEG 3350 is 0.1% (w/v).
13. The composition of claim 1 , wherein the pH is 6.0.
14. The composition of claim 1 , wherein the buffer is selected from the group consisting of histidine, citrate, phosphate, Tris, and succinate buffers.
15. The composition of claim 1 , wherein the buffer is histidine buffer.
16. The composition of claim 1 , wherein the histidine buffer concentration is 2.0-10 mM.
17. The composition of claim 1 , wherein the buffer is 5 mM histidine at pH 6.0.
18. The composition of claim 1 , consisting essentially of:
0.1 mg/mL to 5.0 mg/mL thrombin;
2% to 4% (w/v) sucrose;
3.5% to 5% (w/v) mannitol;
100 mM to 200 mM NaCl;
1 mM to 5 mM CaCl 2 ;
0.03% to 1% (w/v) PEG3350; and
2 mM to 10 mM histidine
in aqueous solution at pH 5.5-6.5, wherein the ratio of mannitol:sucrose is greater than 1:1 but not greater than 2.5:1 (w/w).
19. The composition of claim 18 , consisting essentially of:
1 mg/ml thrombin;
3% (w/v) sucrose;
4% (w/v) mannitol;
4 mM CaCl 2 ;
0.1% (w/v) PEG33500;
150 mM NaCl; and
5 mM histidine
in aqueous solution at pH 6.0.
20. A method of preparing a lyophilized thrombin composition comprising:
providing a composition according to claim 1 ; and
lyophilizing the aqueous solution to form a lyophilized thrombin composition.
21. A lyophilized thrombin composition prepared by the method of claim 20 .
22. The composition of claim 21 , contained in a sealed container having a label affixed to an exterior surface thereof.
23. The composition of claim 22 , which contains from 2,500 to 20,000 NIH units of thrombin.
24. A kit comprising the composition of claim 22 and a second sealed container containing a diluent.
25. The kit of claim 24 , wherein the diluent is normal saline.
26. The kit of claim 24 , further comprising an application device.
27. A method of preparing a lyophilized thrombin composition comprising:
providing a composition according to claim 18 ; and
lyophilizing the aqueous solution to form a lyophilized thrombin composition.