Stabilized thrombin compositions
View Patent ↗Stabilized thrombin compositions, processes for preparing them, and kits comprising them are disclosed. The compositions comprise thrombin, a bacteriostatically effective amount of benzyl alcohol or chlorobutanol, and 0.10%-5.0% (w/v) sucrose in aqueous solution. The compositions are stable when stored at 2° C.-8° C. for four weeks or more.
1. A pharmaceutical composition comprising:
thrombin;
a bacteriostatically effective amount of a preservative selected from the group consisting of benzyl alcohol and chlorobutanol; and 0.10% to 5.0% (w/v) sucrose in aqueous solution.
2. The composition of claim 1 further comprising one or more of a buffer, a salt, a polyol, a surfactant, an amino acid, or an additional carbohydrate.
3. The composition of claim 1 wherein the preservative is benzyl alcohol.
4. The composition of claim 3 wherein the benzyl alcohol is present at a concentration of 0.8%-1.5% (v/v).
5. The composition of claim 1 , further comprising mannitol.
6. The composition of claim 5 wherein the mannitol and sucrose are present at a ratio of mannitol:sucrose greater than 1:1 but not greater than 2.5:1 (w/w).
7. The composition of claim 1 wherein the thrombin is human thrombin.
8. The composition of claim 7 wherein the human thrombin is recombinant human thrombin.
9. The composition of claim 1 wherein the thrombin is bovine thrombin.
10. A composition comprising:
thrombin;
a pharmaceutically acceptable buffer; 0.10% to 5% (w/v) sucrose; and a preservative selected from the group consisting of benzyl alcohol at a concentration of 0.8% to 1.5% (v/v) or chlorobutanol at a concentration of 0.4% to 0.6% (w/v), in aqueous solution at pH 5.7-7.4.
11. The composition of claim 10 wherein the preservative is benzyl alcohol.
12. The composition of claim 11 wherein the concentration of benzyl alcohol is 0.8% to 1.0% (v/v).
13. The composition of claim 10 wherein the sucrose concentration is 0.5% to 3.0% (w/v).
14. The composition of claim 10 wherein the sucrose concentration is about 1% (w/v).
15. The composition of claim 10 wherein the buffer is selected from the group consisting of histidine, citrate, phosphate, Tris, succinate, and acetate buffers.
16. The composition of claim 10 , further comprising mannitol.
17. The composition of claim 16 wherein the mannitol and sucrose are present at a ratio of mannitol:sucrose greater than 1:1 but not greater than 2.5:1 (w/w).
18. The composition of claim 10 wherein the diluent comprises sodium chloride.
19. The composition of claim 10 wherein the thrombin is human thrombin.
20. The composition of claim 19 wherein the human thrombin is recombinant human thrombin.
21. The composition of claim 10 wherein the thrombin is bovine thrombin.
22. The composition of claim 10 wherein the thrombin is present at a concentration of 0.1 mg/mL to 5.0 mg/mL.
23. The composition of claim 22 wherein the thrombin is present at a concentration of 0.3 mg/mL to 3.0 mg/mL.
24. An aqueous composition consisting essentially of:
0.03 mg/mL to 1.6 mg/mL thrombin;
0.17% to 1.3% (w/v) sucrose;
1.1% to 1.6% (w/v) mannitol;
0.8% to 2.0% (w/v) NaCl;
0-1.6 mM CaCl 2 ;
0.001% to 0.32% (w/v) of a surfactant or high-molecular-weight polyethylene glycol;
a pharmaceutically acceptable buffer; and
a bacteriostatically effective amount of a preservative selected from the group consisting of benzyl alcohol and chlorobutanol, 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 and wherein the ratio of mannitol:sucrose is greater than 1:1 but not greater than 2.5:1 (w/w).
25. The composition of claim 24 wherein the ratio of mannitol to sucrose is 1.33:1 (w/w).
26. The composition of claim 24 wherein the molar ratio of sucrose:thrombin is at least 700:1.
27. The composition of claim 26 wherein the molar ratio of sucrose:thrombin is at least 2000:1.
28. The composition of claim 24 wherein the preservative is benzyl alcohol at a concentration of 0.8% to 1.5% (v/v).
29. A process for preparing a stabilized thrombin solution comprising:
providing a lyophilized composition comprising thrombin and a quantity of sucrose sufficient to stabilize the protein in the presence of a bacteriostatically effective amount of a preservative selected from the group consisting of benzyl alcohol and chlorobutanol; providing a diluent comprising a bacteriostatically effective amount of benzyl alcohol or chlorobutanol in water; and
combining the lyophilized composition and the diluent to form a solution, wherein the concentration of sucrose in the solution is from 0.10% to 5.0% (w/v).
30. The process of claim 29 wherein the lyophilized composition further comprises a buffer, a salt, or a polyol.
31. A process for preparing a stabilized thrombin solution comprising:
providing a lyophilized composition comprising thrombin and at least one pharmaceutically acceptable excipient; and reconstituting the lyophilized composition in a diluent to provide a pharmaceutically acceptable thrombin solution, wherein the diluent is selected to provide in the solution a benzyl alcohol concentration of 0.8% to 1.5% (v/v) and a sucrose concentration of 0.10% to 5% (w/v).
32. The process of claim 31 wherein the solution further comprises mannitol.
33. The process of claim 32 wherein the mannitol and sucrose are present in the solution at a ratio of mannitol:sucrose greater than 1:1 but not greater than 2.5:1 (w/w).
34. A kit comprising:
a pharmaceutically acceptable diluent comprising a bacteriostatically effective amount of benzyl alcohol or chlorobutanol in water in a first sealed container; and
a lyophilized composition comprising thrombin and a quantity of sucrose in a second sealed container, wherein the quantity of sucrose is selected to provide a sucrose concentration of 0.10% to 5.0% (w/v) upon reconstitution of the lyophilized composition with the diluent.
35. The kit of claim 34 wherein the diluent further comprises NaCl.
36. The kit of claim 35 wherein the diluent is bacteriostatic saline.
37. The kit of claim 34 , further comprising means for transferring the diluent from the first sealed container to the second sealed container.
38. The kit of claim 34 , further comprising an instruction sheet.
39. The kit of claim 34 , further comprising an applicator device.
40. The kit of claim 39 , wherein the applicator device is a syringe or a sprayer.
41. The kit of claim 34 , wherein the first and second sealed containers are packaged in a third container.