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. The present invention is directed to a pre-lyophilization formulation comprising: (a) a Factor IX (FIX) polypeptide having FIX coagulation activity; (b) a buffering agent; (c) a stabilizing agent; (d) a bulking agent; and (e) a surfactant, wherein the formulation has a fill volume of less than about 5 mL, and wherein each of (a)-(e) are at an amount per vial (mg/vial) sufficient to allow (1) improved stability of the FIX polypeptide when lyophilized.
1. A pre-lyophilization formulation comprising:
(a) a chimeric Factor IX (FIX) polypeptide, comprising FIX and a heterologous moiety that extends the half-life of the FIX, at a concentration between 220 IU/vial and 1,000 IU/vial;
(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.
2. The pre-lyophilization formulation of claim 1 , wherein the fill volume is about 2.65 mL.
3. The pre-lyophilization formulation of claim 1 , wherein the heterologous moiety is Fc.
4. The pre-lyophilization formulation of claim 1 ,
wherein the L-histidine is present at a concentration of about 7.76 mg/mL;
wherein the sucrose is present at a concentration of about 23.8 mg/mL;
wherein the mannitol is present at a concentration of about 47.6 mg/mL; and
wherein the polysorbate 20 is present at a concentration of about 0.2 mg/mL.
5. The pre-lyophilization formulation of claim 1 , wherein the fill volume is about 2.5 mL.
6. The pre-lyophilization formulation of claim 1 , wherein the chimeric FIX polypeptide comprises the FIXFc-sc polypeptide of SEQ ID NO: 2 and the Fc sc polypeptide of SEQ ID NO:4.
7. The pre-lyophilization formulation of claim 1 , wherein the fill volume allows for reduced reconstitution time when the formulation is lyophilized, wherein the reduced reconstitution time is less than about 30 seconds.
8. The pre-lyophilization formulation of claim 1 , wherein the fill volume allows for reduced lyophilization cycle time when the formulation is lyophilized, wherein the reduced lyophilization cycle time is about 4 days or less.
9. A pre-lyophilization formulation comprising:
(a) a chimeric Factor IX (FIX) polypeptide, comprising the FIXFc-sc polypeptide of SEQ ID NO:2 and the Fc sc polypeptide of SEQ ID NO:4, at a concentration between about 220 IU/vial and about 1,000 IU/vial;
(b) L-histidine at a concentration of about 7.76 mg/mL;
(c) sucrose at a concentration of about 23.8 mg/mL;
(d) mannitol at a concentration of about 47.6 mg/mL; and
(e) polysorbate 20 at a concentration of about 0.2 mg/mL,
wherein the formulation has a fill volume of about 2.65 mL.
10. 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, wherein the FIXFc-sc polypeptide and the Fc sc polypeptide are bound together through two disulfide bonds in the hinge region of Fc.