Lyophilized factor IX formulations
The present invention provides, among other things, pre-lyophilization formulations, reconstituted formulations, and lyophilate powder compositions comprising a Factor IX (FIX) polypeptide. The present invention also provides lyophilization methods for producing lyophilate powder comprising a FIX polypeptide.
1. A pre-lyophilization formulation within a vial comprising:
(a) a chimeric Factor IX (FIX) polypeptide, comprising FIX and a heterologous moiety that extends the half-life of the FIX, in an amount of about 2,000, about 3,000, or about 4,000 IU;
(b) L-histidine at a concentration between 3 mg/ml and 15 mg/mL;
(c) sucrose at a concentration between 10 mg/mL and 50 mg/mL;
(d) mannitol at a concentration between 20 mg/mL and 100 mg/ml; and
(e) polysorbate 20 at a concentration between 0.01 mg/ml and 5 mg/mL,
wherein the formulation has a fill volume between 2 mL and 3 mL, and
wherein the formulation is capable of being lyophilized into a lyophilate formulation having a residual moisture level below 2% using a lyophilization cycle with a cycle time of 4 days or less and a drying time of 25 hours or less.
2. The pre-lyophilization formulation of claim 1 , wherein the fill volume is about 2.65 mL or about 2.5 mL.
3. The pre-lyophilization formulation of claim 1 , wherein the L-histidine is at a concentration between about 5.82 mg/mL and about 9.7 mg/mL.
4. The pre-lyophilization formulation of claim 1 , wherein the L-histidine is at a concentration of about 7.76 mg/mL.
5. The pre-lyophilization formulation of claim 1 , wherein the sucrose is at a concentration between about 17.85 mg/mL and about 29.95 mg/mL.
6. The pre-lyophilization formulation of claim 1 , wherein the sucrose is at a concentration of about 23.8 mg/mL.
7. The pre-lyophilization formulation of claim 1 , wherein the mannitol is at a concentration between about 35.7 mg/mL and about 59.5 mg/mL.
8. The pre-lyophilization formulation of claim 1 , wherein the mannitol is at a concentration of about 47.6 mg/mL.
9. The pre-lyophilization formulation of claim 1 , wherein the polysorbate 20 is at a concentration between about 0.05 mg/mL and about 1 mg/mL.
10. The pre-lyophilization formulation of claim 1 , wherein the polysorbate 20 is at a concentration of about 0.2 mg/mL.
11. The pre-lyophilization formulation of claim 1 , wherein the chimeric FIX polypeptide is at a concentration between about 600 IU/mL and about 2,750 IU/mL.
12. A pre-lyophilization formulation within a vial comprising:
(a) about 2,000, about 3,000, or about 4,000 IU of a chimeric Factor IX (FIX) polypeptide, comprising FIX and Fc;
(b) about 7.76 mg/ml of L-histidine;
(c) about 47.6 mg/mL of mannitol;
(d) about 23.8 mg/ml of sucrose; and,
(e) about 0.2 mg/ml of polysorbate 20;
wherein the formulation has a fill volume between 2 mL and 3 mL, and
wherein the formulation is capable of being lyophilized into a lyophilate formulation having a residual moisture level below 2% using a lyophilization cycle with a cycle time of 4 days or less and a drying time of 25 hours or less.
13. The pre-lyophilization formulation of claim 12 , wherein the formulation has a fill volume per vial of about 2.65 mL or about 2.5 mL.
14. The pre-lyophilization formulation of claim 13 , wherein the formulation has a fill volume per vial of about 2.65 mL.
15. The pre-lyophilization formulation of claim 1 , wherein the moiety extending half-life of FIX comprises an FcRn binding partner or an Fc region.
16. The pre-lyophilization formulation of claim 1 , wherein the chimeric FIX polypeptide comprises the FIXFc-sc polypeptide of SEQ ID NO: 2, without the C-terminal lysine, and the Fc sc polypeptide of SEQ ID NO:4, without the C-terminal lysine.
17. The pre-lyophilization formulation of claim 16 , wherein the FIXFc-sc polypeptide and the Fc sc polypeptide are bound together through two disulfide bonds in the hinge region of Fc.
18. The pre-lyophilization formulation of claim 1 , comprising 2,000 IU of the FIX polypeptide.
19. The pre-lyophilization formulation of claim 1 , comprising 3,000 IU of the FIX polypeptide.
20. The pre-lyophilization formulation of claim 1 , comprising 4,000 IU of the FIX polypeptide.
21. The pre-lyophilization formulation of claim 12 , comprising 2,000 IU of the FIX polypeptide.
22. The pre-lyophilization formulation of claim 12 , comprising 3,000 IU of the FIX polypeptide.
23. The pre-lyophilization formulation of claim 12 , comprising 4,000 IU of the FIX polypeptide.
24. The pre-lyophilization formulation of claim 1 , consisting essentially of:
(a) about 2,000, about 3,000, or about 4,000
IU of a chimeric Factor IX (FIX) polypeptide, comprising FIX and a heterologous moiety that extends the half-life of the FIX;
(b) L-histidine at a concentration between 3 mg/mL and 15 mg/mL;
(c) sucrose at a concentration between 10 mg/ml and 50 mg/ml;
(d) mannitol at a concentration between 20 mg/mL and 100 mg/mL; and
(e) polysorbate 20 at a concentration between 0.01 mg/mL and 5 mg/mL,
wherein the formulation has a fill volume between 2 mL and 3 mL.
25. The pre-lyophilization formulation of claim 24 , consisting of:
(a) about 2,000, about 3,000, or about 4,000 IU of a chimeric Factor IX (FIX) polypeptide, comprising FIX and a heterologous moiety that extends the half-life of the FIX;
(b) L-histidine at a concentration between 3 mg/ml and 15 mg/ml;
(c) sucrose at a concentration between 10 mg/mL and 50 mg/ml;
(d) mannitol at a concentration between 20 mg/mL and 100 mg/mL; and
(e) polysorbate 20 at a concentration between 0.01 mg/mL and 5 mg/mL,
wherein the formulation has a fill volume between 2 mL and 3 mL.
26. The pre-lyophilization formulation of claim 1 , wherein the formulation is capable of being lyophilized into a lyophilate formulation having a residual moisture level below 1% using a lyophilization cycle with a cycle time of 4 days or less and a drying time of 25 hours or less.
27. The pre-lyophilization formulation of claim 1 , wherein the formulation is capable of being lyophilized into a lyophilate formulation having a residual moisture level below 0.5% using a lyophilization cycle with a cycle time of 4 days or less and a drying time of 25 hours or less.
28. The pre-lyophilization formulation of claim 12 , wherein the formulation is capable of being lyophilized into a lyophilate formulation having a residual moisture level below 1% using a lyophilization cycle with a cycle time of 4 days or less and a drying time of 25 hours or less.
29. The pre-lyophilization formulation of claim 12 , wherein the formulation is capable of being lyophilized into a lyophilate formulation having a residual moisture level below 0.5% using a lyophilization cycle with a cycle time of 4 days or less and a drying time of 25 hours or less.
30. The pre-lyophilization formulation of claim 24 , comprising 2,000 IU of the chimeric FIX polypeptide.
31. The pre-lyophilization formulation of claim 24 , comprising 3,000 IU of the chimeric FIX polypeptide.
32. The pre-lyophilization formulation of claim 24 , comprising 4,000 IU of the chimeric FIX polypeptide.
33. The pre-lyophilization formulation of claim 25 , comprising 2,000 IU of the chimeric FIX polypeptide.
34. The pre-lyophilization formulation of claim 25 , comprising 3,000 IU of the chimeric FIX polypeptide.
35. The pre-lyophilization formulation of claim 25 , comprising 4,000 IU of the chimeric FIX polypeptide.
36. The pre-lyophilization formulation of claim 1 , wherein the lyophilization cycle comprises freezing the formulation in a vial and then ramping up the temperature in the vial to a drying temperature before the drying time of 25 hours or less, wherein the drying temperature is about 40° C.