VEGF antagonist formulations suitable for intravitreal administration
Ophthalmic formulations of a vascular endothelial growth factor (VEGF)-specific fusion protein antagonist are provided suitable for intravitreal administration to the eye. The ophthalmic formulations include a stable liquid formulation and a lyophilizable formulation. Preferably, the protein antagonist has an amino acid sequence of SEQ ID NO:4.
1. A vial comprising an ophthalmic formulation suitable for intravitreal administration that comprises:
a vascular endothelial growth factor (VEGF) antagonist
an organic co-solvent,
a buffer, and
a stabilizing agent,
wherein said VEGF antagonist fusion protein is glycosylated and comprises amino acids 27-457 of SEQ ID NO:4; and
wherein at least 98% of the VEGF antagonist is present in native conformation following storage at 5° C. for two months as measured by size exclusion chromatography.
2. The vial of claim 1 , wherein the concentration of said VEGF antagonist fusion protein is 40 mg/ml, and wherein said organic co-solvent comprises polysorbate.
3. The vial of claim 2 , wherein said organic co-solvent comprises 0.01% to 3% polysorbate.
4. The vial of claim 2 , wherein said organic co-solvent comprises about 0.03% to about 0.1% polysorbate 20.
5. The vial of claim 2 , wherein said organic co-solvent comprises 0.01% to 3% polysorbate 20.
6. The vial of claim 5 , wherein said buffer comprises a phosphate buffer.
7. The vial of claim 5 , wherein said buffer comprises 5-25 mM buffer.
8. The vial of claim 5 , wherein said buffer comprises a pH between about 5.8-7.0.
9. The vial of claim 5 , wherein said buffer comprises a pH about 6.2-6.3.
10. The vial of claim 5 , wherein said stabilizing agent comprises a sugar.
11. The vial of claim 10 , wherein said sugar is selected from the group consisting of sucrose, sorbitol, glycerol, trehalose, and mannitol.
12. The vial of claim 5 , wherein said stabilizing agent comprises 1.0-7.5% of sucrose.
13. The vial of claim 5 , wherein said formulation further comprises a tonicity agent.
14. The vial of claim 5 , wherein said VEGF antagonist fusion protein is glycosylated at asparagine residues corresponding to asparagine residues 62, 94, 149, 222 and 308 of SEQ ID NO: 4.
15. The vial of claim 5 , wherein said formulation is capable of providing a turbidity of 0.01 or lower at OD 405 after 2 month storage at 5° C.
16. The vial of claim 5 , wherein at least 99% of said VEGF antagonist fusion protein is present in native conformation after 2 month storage at 5° C. as measured by size exclusion chromatography.
17. The vial of claim 5 , wherein at least 98% of said VEGF antagonist fusion protein is present in native conformation following storage at 5° C. for 24 months as measured by size exclusion chromatography.
18. The vial of claim 5 , wherein said formulation does not contain phosphate.
19. The vial of claim 5 , wherein said formulation does not contain trehalose.
20. The vial of claim 5 , wherein said stabilizing agent comprises 1.0-10% of sucrose.
21. The vial of claim 20 , wherein said formulation further comprises a tonicity agent.
22. The vial of claim 20 , wherein said VEGF antagonist fusion protein is glycosylated at asparagine residues corresponding to asparagine residues 62, 94, 149, 222 and 308 of SEQ ID NO: 4.
23. The vial of claim 20 , wherein said formulation is capable of providing a turbidity of 0.01 or lower at OD 405 after 2 month storage at 5° C.
24. The vial of claim 20 , wherein at least 99% of said VEGF antagonist fusion protein is present in native conformation after 2 month storage at 5° C. as measured by size exclusion chromatography.
25. The vial of claim 20 , wherein at least 98% of said VEGF antagonist fusion protein is present in native conformation following storage at 5° C. for 24 months as measured by size exclusion chromatography.
26. A pre-filled syringe comprising an ophthalmic formulation suitable for intravitreal administration comprising:
a vascular endothelial growth factor (VEGF) antagonist fusion protein,
an organic co-solvent,
a buffer, and
a stabilizing agent;
wherein said VEGF antagonist fusion protein is glycosylated and comprises amino acids 27-457 of SEQ ID NO:4; and
wherein at least 98% of said VEGF antagonist fusion protein is present in native conformation following storage at 5° C. for two months as measured by size exclusion chromatography.
27. The pre-filled syringe of claim 26 , wherein the concentration of said VEGF antagonist fusion protein is 40 mg/ml, and wherein said organic co-solvent comprises polysorbate.
28. The pre-filled syringe of claim 27 , wherein said organic co-solvent comprises 0.01% to 3% polysorbate.
29. The pre-filled syringe of claim 27 , wherein said organic co-solvent comprises about 0.03% to about 0.1% polysorbate 20.
30. The pre-filled syringe of claim 27 , wherein said organic co-solvent comprises 0.01% to 3% polysorbate 20.
31. The pre-filled syringe of claim 30 , wherein said buffer comprises a phosphate buffer.
32. The pre-filled syringe of claim 30 , wherein said buffer comprises 5-25 mM buffer.
33. The pre-filled syringe of claim 30 , wherein said buffer comprises a pH between about 5.8-7.0.
34. The pre-filled syringe of claim 30 , wherein said buffer comprises a pH about 6.2-6.3.
35. The pre-filled syringe of claim 30 , wherein said stabilizing agent comprises a sugar.
36. The pre-filled syringe of claim 35 , wherein said sugar is selected from the group consisting of sucrose, sorbitol, glycerol, trehalose, and mannitol.
37. The pre-filled syringe of claim 30 , wherein said stabilizing agent comprises 1.0-7.5% of sucrose.
38. The pre-filled syringe of claim 30 , wherein said formulation further comprises a tonicity agent.
39. The pre-filled syringe of claim 30 , wherein said VEGF antagonist fusion protein is glycosylated at asparagine residues corresponding to asparagine residues 62, 94, 149, 222 and 308 of SEQ ID NO: 4.
40. The pre-filled syringe of claim 30 , wherein said formulation is capable of providing a turbidity of 0.01 or lower at OD 405 after 2 month storage at 5° C.
41. The pre-filled syringe of claim 30 , wherein at least 99% of said VEGF antagonist fusion protein is present in native conformation after 2 month storage at 5° C. as measured by size exclusion chromatography.
42. The pre-filled syringe of claim 30 , wherein at least 98% of said VEGF antagonist fusion protein is present in native conformation following storage at 5° C. for 24 months as measured by size exclusion chromatography.
43. The pre-filled syringe of claim 30 , wherein said formulation does not contain phosphate.
44. The pre-filled syringe of claim 30 , wherein said formulation does not contain trehalose.
45. The pre-filled syringe of claim 30 , wherein said stabilizing agent comprises 1.0-10% of sucrose.
46. The pre-filled syringe of claim 45 , wherein said formulation further comprises a tonicity agent.
47. The pre-filled syringe of claim 45 , wherein said VEGF antagonist fusion protein is glycosylated at asparagine residues corresponding to asparagine residues 62, 94, 149, 222 and 308 of SEQ ID NO: 4.
48. The pre-filled syringe of claim 45 , wherein said formulation is capable of providing a turbidity of 0.01 or lower at OD 405 after 2 month storage at 5° C.
49. The pre-filled syringe of claim 45 , wherein at least 99% of said VEGF antagonist fusion protein is present in native conformation after 2 month storage at 5° C. as measured by size exclusion chromatography.
50. The pre-filled syringe of claim 45 , wherein at least 98% of said VEGF antagonist fusion protein is present in native conformation following storage at 5° C. for 24 months as measured by size exclusion chromatography.
51. An ophthalmic formulation comprising:
(a) 40 mg/ml of a glycosylated VEGF antagonist fusion protein comprising amino acids 27-457 of SEQ ID NO:4;
(b) 0.03% to 0.1% polysorbate;
(c) 5-40 mM of sodium phosphate buffer, pH between 5.8-7.0; and
(d) sucrose;
wherein the ophthalmic formulation is suitable for intravitreal administration; and
wherein at least 98% of the VEGF antagonist is present in native conformation following storage at 5° C. for 2 months as measured by size exclusion chromatography.
52. The formulation of claim 51 , wherein said formulation comprises at least 5% sucrose.
53. The formulation of claim 51 , wherein said formulation comprises 1-10% sucrose.
54. A pre-filled syringe suitable for intravitreal administration comprising the formulation of claim 51 .
55. A vial suitable for intravitreal administration comprising the formulation of claim 51 .
56. The formulation of claim 51 , wherein said formulation comprises 10 mM sodium phosphate buffer, 0.03% polysorbate, 5% sucrose, and a pH between 6.2-6.3.
57. A pre-filled syringe suitable for intravitreal administration comprising the formulation of claim 56 .
58. A vial suitable for intravitreal administration comprising the formulation of claim 56 .
59. The formulation of claim 56 , wherein said formulation further comprises 40 mM NaCl.
60. A pre-filled syringe suitable for intravitreal administration comprising the formulation of claim 59 .
61. A vial suitable for intravitreal administration comprising the formulation of claim 59 .
62. The formulation of claim 59 , wherein said VEGF antagonist fusion protein is glycosylated at asparagine residues corresponding to asparagine residues 62, 94, 149, 222 and 308 of SEQ ID NO: 4.
63. A pre-filled syringe suitable for intravitreal administration comprising the formulation of claim 62 .
64. A vial suitable for intravitreal administration comprising the formulation of claim 62 .