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 glass vial 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 an immunoglobulin-like (Ig) domain 2 of a first VEGF receptor that is human Flt1 and Ig domain 3 of a second VEGF receptor selected from the group consisting of human Flk1 and human Flt4, and a multimerizing component; 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.
2. The glass 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 glass vial of claim 2 , wherein said organic co-solvent comprises 0.01% to 3% polysorbate.
4. The glass vial of claim 2 , wherein said organic co-solvent comprises about 0.03% to about 0.1% polysorbate 20.
5. The glass vial of claim 2 , wherein said organic co-solvent comprises 0.01% to 3% polysorbate 20.
6. The glass vial of claim 5 , wherein said buffer comprises a phosphate buffer.
7. The glass vial of claim 5 , wherein said buffer comprises 5-25 mM buffer.
8. The glass vial of claim 5 , wherein said buffer comprises a pH between about 5.8-7.0.
9. The glass vial of claim 5 , wherein said buffer comprises a pH about 6.2-6.3.
10. The glass vial of claim 5 , wherein said stabilizing agent comprises a sugar.
11. The glass vial of claim 10 , wherein said sugar is selected from the group consisting of sucrose, sorbitol, glycerol, trehalose, and mannitol.
12. The glass vial of claim 5 , wherein said stabilizing agent comprises 1.0-7.5% of sucrose.
13. The glass vial of claim 5 , wherein said formulation further comprises a tonicity agent.
14. The glass 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 glass 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 glass 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 glass 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 glass vial of claim 5 , wherein said formulation does not contain phosphate.
19. The glass vial of claim 5 , wherein said formulation does not contain trehalose.
20. The glass vial of claim 5 , wherein said stabilizing agent comprises 1.0-10% of sucrose.
21. The glass vial of claim 20 , wherein said formulation further comprises a tonicity agent.
22. The glass 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 glass 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 glass 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 glass 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. The glass vial of claim 1 , wherein said stabilizing agent comprises sucrose.
27. The glass vial of claim 26 , wherein said stabilizing agent comprises 1.0-10% of sucrose.
28. The glass vial of claim 26 , wherein the organic co-solvent is polysorbate 20.
29. The glass vial of claim 28 , wherein said Ig domain 3 of human VEGF receptor 2 comprises amino acids 131-230 of SEQ ID NO:4.
30. The glass vial of claim 29 , wherein said formulation further comprises a tonicity agent.
31. The glass vial of claim 30 , wherein said tonicity agent comprises sodium chloride.
32. The glass vial of claim 28 , wherein said formulation comprises 10 mg/mL VEGF antagonist fusion protein.
33. The glass vial of claim 28 , wherein said formulation does not contain phosphate.
34. 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 an immunoglobulin-like (Ig) domain 2 of a first VEGF receptor that is human Flt1 and Ig domain 3 of a second VEGF receptor selected from the group consisting of human Flk1 and human Flt4, and a multimerizing component; 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.
35. The pre-filled syringe of claim 34 , wherein the concentration of said VEGF antagonist fusion protein is 40 mg/ml, and wherein said organic co-solvent comprises polysorbate.
36. The pre-filled syringe of claim 35 , wherein said organic co-solvent comprises 0.01% to 3% polysorbate.
37. The pre-filled syringe of claim 35 , wherein said organic co-solvent comprises about 0.03% to about 0.1% polysorbate 20.
38. The pre-filled syringe of claim 35 , wherein said organic co-solvent comprises 0.01% to 3% polysorbate 20.
39. The pre-filled syringe of claim 38 , wherein said buffer comprises 5-25 mM buffer.
40. The pre-filled syringe of claim 38 , wherein said buffer comprises a pH between about 5.8-7.0.
41. The pre-filled syringe of claim 38 , wherein said buffer comprises a pH about 6.2-6.3.
42. The pre-filled syringe of claim 38 , wherein said buffer comprises a phosphate buffer.
43. The pre-filled syringe of claim 38 , wherein said stabilizing agent comprises a sugar.
44. The pre-filled syringe of claim 43 , wherein said sugar is selected from the group consisting of sucrose, sorbitol, glycerol, trehalose, and mannitol.
45. The pre-filled syringe of claim 38 , 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. The pre-filled syringe of claim 38 , wherein said stabilizing agent comprises 1.0-7.5% of sucrose.
52. The pre-filled syringe of claim 38 , wherein said formulation further comprises a tonicity agent.
53. The pre-filled syringe of claim 38 , 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.
54. The pre-filled syringe of claim 38 , wherein said formulation is capable of providing a turbidity of 0.01 or lower at OD 405 after 2 month storage at 5° C.
55. The pre-filled syringe of claim 38 , 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.
56. The pre-filled syringe of claim 38 , 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.
57. The pre-filled syringe of claim 38 , wherein said formulation does not contain phosphate.
58. The pre-filled syringe of claim 38 , wherein said formulation does not contain trehalose.
59. The pre-filled syringe of claim 34 , wherein said stabilizing agent comprises sucrose.
60. The pre-filled syringe of claim 59 , wherein said stabilizing agent comprises 1.0-10% of sucrose.
61. The pre-filled syringe of claim 59 , wherein the organic co-solvent is polysorbate 20.
62. The pre-filled syringe of claim 61 , wherein said Ig domain 3 of human VEGF receptor 2 comprises amino acids 131-230 of SEQ ID NO:4.
63. The pre-filled syringe of claim 62 , wherein said formulation further comprises a tonicity agent.
64. The pre-filled syringe of claim 63 , wherein said tonicity agent comprises sodium chloride.
65. The pre-filled syringe of claim 61 , wherein said formulation comprises 10 mg/mL VEGF antagonist fusion protein.
66. The pre-filled syringe of claim 61 , wherein said formulation does not contain phosphate.