Formulations of recombinant furin
View Patent ↗The present application provides stabilized formulations of furin (e.g., rfurin) containing a sugar, sugar alcohol, and/or non-ionic surfactant. As compared to non-stabilized compositions, the furin formulations disclosed herein retain greater amounts of furin activity and monomeric furin content, while reducing furin aggregation when stored and/or subjected to mechanical stress. Also provided are methods for stably diluting furin (e.g., rfurin) compositions.
1. A stabilized aqueous composition of recombinant furin (rfurin), the composition comprising:
(a) from 8,000 U/mL to 500,000 U/mL rfurin;
(b) from 175 mM to 260 mM of a pharmaceutically acceptable salt;
(c) from 0.5 mM to 2 mM calcium;
(d) from 2% to 20% sugar or sugar alcohol;
(e) from 10 to 200 ppm non-ionic surfactant;
(f) from 10 to 200 mM buffering agent; and
(g) a pH from 5.5 to 7.5.
2. The composition of claim 1 , comprising from 180 mM to 260 mM of a pharmaceutically acceptable salt.
3. The composition of claim 1 , wherein the pharmaceutically acceptable salt is selected from the group consisting of sodium chloride and potassium chloride.
4. The composition of claim 1 , comprising 0.9±0.2 mM calcium.
5. The composition of claim 1 , comprising from 5% to 15% sugar or sugar alcohol.
6. The composition of claim 5 , comprising 10% sugar or sugar alcohol.
7. The composition of claim 1 , wherein the sugar or sugar alcohol is selected from the group consisting of sucrose, trehalose, mannitol, and a combination thereof.
8. The composition of claim 1 , wherein the non-ionic surfactant is polysorbate-80.
9. The composition of claim 1 , comprising from 10 ppm to 100 ppm non-ionic surfactant.
10. The composition of claim 9 , comprising 75 ppm non-ionic surfactant.
11. The composition of claim 1 , comprising from 50 mM to 150 mM buffering agent.
12. The composition of claim 1 , comprising 90±10 mM buffering agent.
13. The composition of claim 1 , wherein the buffering agent comprises acetate and HEPES.
14. The composition of claim 13 , comprising from 25 mM to 75 mM acetate and from 25 to 75 mM HEPES.
15. The composition of claim 13 , wherein the buffering agent comprises 45±5 mM acetate and 45±5 mM HEPES.
16. The composition of claim 1 , wherein the buffering agent comprises MES.
17. The composition of claim 1 , having a pH from 5.5 to 7.0.
18. The composition of claim 1 , having a pH from 5.5 to 6.5.
19. The composition of claim 1 , having a pH of 6.0.
20. The composition of claim 1 , having a pH of 6.0±0.2.
21. The composition of claim 1 , wherein the composition has increased stability when stored at 37° C. as compared to a rfurin composition that does not contain a sugar or sugar alcohol.
22. The composition of claim 1 , wherein the composition maintains a higher percentage of rfurin activity when stored at 37° C. as compared to a rfurin composition that does not contain a sugar or sugar alcohol.
23. The composition of claim 1 , wherein the composition maintains a higher percentage of rfurin monomer content when stored at 37° C. as compared to a rfurin composition that does not contain a sugar or sugar alcohol.
24. The composition of claim 1 , wherein the composition has increased stability when agitated as compared to a rfurin composition that does not contain a non-ionic surfactant.
25. The composition of claim 1 , wherein the composition maintains a higher percentage of rfurin activity when agitated as compared to a rfurin composition that does not contain a non-ionic surfactant.
26. The composition of claim 1 , wherein the composition maintains a higher percentage of rfurin monomer content when agitated as compared to a rfurin composition that does not contain a non-ionic surfactant.
27. The composition of claim 1 , wherein the composition has increased stability when diluted as compared to a rfurin composition that does not contain a non-ionic surfactant.
28. The composition of claim 1 , wherein the composition maintains a higher percentage of rfurin activity when diluted as compared to a rfurin composition that does not contain a non-ionic surfactant.
29. The composition of claim 1 , wherein the composition maintains a higher percentage of rfurin monomer content when diluted as compared to a rfurin composition that does not contain a non-ionic surfactant.