VEGF specific fusion protein antagonist formulations

Patent metadata

Field	Value
Publication number	US-9636400-B2
Application number	US-201615150840-A
Country	US
Kind code	B2
Filing date	May 10, 2016
Priority date	Mar 25, 2005
Publication date	May 2, 2017
Grant date	May 2, 2017

Abstract

Formulations of a vascular endothelial growth factor (VEGF)-specific fusion protein antagonist are provided including a pre-lyophilized formulation, a reconstituted lyophilized formulation, and a stable liquid formulation. Preferably, the fusion protein has the sequence of SEQ ID NO:4.

First claim

We claim:

1. A formulation comprising:
(a) 5-200 mg/mL of a VEGF-specific fusion protein antagonist;
(b) 1-50 mM of a buffer;
(c) 25-150 mM or 0.5-30% of a stabilizer; and
(d) 0.0005-5% of a polysorbate;

wherein
said VEGF-specific fusion protein antagonist represents at least 90% of the total weight of protein in the formulation,
at least 90% of the total weight of VEGF-specific fusion protein antagonist is not present as an aggregate,
said VEGF-specific fusion protein antagonist comprises an Ig domain 2 of human VEGF receptor 1, an Ig domain 3 of human VEGF receptor 2, and a multimerizing component,
said VEGF-specific fusion protein antagonist does not comprise amino acids 1-26 of SEQ ID NO:4, and
said Ig domain 2 of human VEGF receptor 1 consists essentially of amino acids 30-130 of SEQ ID NO:4, said Ig domain 3 of human VEGF receptor 2 consists essentially of amino acids 129-230 of SEQ ID NO:4, and said multimerizing component consists essentially of amino acids 232-457 of SEQ ID NO:4.

2. The formulation of claim 1 , wherein the VEGF-specific fusion protein antagonist represents at least 95% of the total weight of protein in the formulation.

3. The formulation of claim 2 , wherein the VEGF-specific fusion protein antagonist represents at least 99% of the total weight of protein in the formulation.

4. The formulation of claim 1 , wherein the VEGF-specific fusion protein antagonist is present at a concentration of about 5-75 mg/mL, about 10-50 mg/mL, about 10-100 mg/mL, about 12.5-75 mg/mL, about 25-50 mg/mL, about 50-100 mg/mL, about 75-150 mg/mL, or about 100-200 mg/mL.

5. The formulation of claim 4 , where the VEGF-specific fusion protein antagonist is present at a concentration of about 12.5 mg/mL, about 25 mg/mL, 25.1 mg/mL, 25.2 mg/mL, 25.4 mg/mL, 25.5 mg/mL, 25.6 mg/mL, 30 mg/mL, 40 mg/mL, 50 mg/mL, 60 mg/mL, 75 mg/mL, 80 mg/mL, 100 mg/mL, or about 150 mg/mL.

6. The formulation of claim 1 , wherein the buffer comprises histidine, phosphate, citrate, or a combination thereof.

7. The formulation of claim 6 , wherein the buffer comprises 1-10 mM phosphate, 1-10 mM citrate, or a combination thereof.

8. The formulation of claim 7 , wherein the buffer comprises 5 mM phosphate.

9. The formulation of claim 7 , wherein the buffer comprises 5 mM citrate.

10. The formulation of claim 7 , wherein the buffer comprises 5 mM citrate and 5 mM phosphate.

11. The formulation of claim 6 , wherein the buffer comprises 1-50 mM histidine.

12. The formulation of claim 11 , wherein the buffer comprises histidine at a concentration of about 10 mM or about 20 mM.

13. The formulation of claim 1 , wherein the pH of the formulation is about pH 6.0-6.5.

14. The formulation of claim 13 , wherein the pH is about pH 6.0, 6.0-6.1, 6.1, 6.2, 6.25, 6.3, or 6.3-6.5.

15. The formulation of claim 1 , wherein the polysorbate is polysorbate 20.

16. The formulation of claim 15 , wherein said polysorbate 20 is present at a concentration of about 0.0005%, about 0.001%, about 0.003-0.005%, about 0.05-0.15%, about 0.1%, or about 0.1-0.5%.

17. The formulation of claim 1 , wherein the stabilizer comprises a salt, an amino acid, a sugar, a sugar alcohol, or a combination thereof.

18. The formulation of claim 17 , wherein the stabilizer comprises an amino acid at a concentration of about 0.25-3.0%.

19. The formulation of claim 18 , wherein the amino acid is glycine present at a concentration of about 0.75% or about 1.5%.

20. The formulation of claim 17 , wherein the stabilizer comprises one or more sugars selected from sucrose, trehalose, and lactose at a concentration of about 0.5-6.0% or about 5-30%.

21. The formulation of claim 20 , wherein the stabilizer comprises sucrose at a concentration of about 0.5-6.0%, about 2.5%, about 5-20%, about 5-30%, about 5%, about 20%, or about 25%.

22. The formulation of claim 17 , wherein the stabilizer comprises sodium chloride at a concentration of about 25-150 mM, about 50-100 mM, about 50 mM, or about 100 mM.

23. A formulation comprising:
(a) 10 mg/mL of a VEGF-specific fusion protein antagonist;
(b) 5 mM citrate;
(c) 5 mM phosphate;
(d) 100 mM sodium chloride;
(c) 0.01% polysorbate 20; and
(d) 20% sucrose, having a pH of about 6.0-6.5,

wherein
the VEGF-specific fusion protein antagonist represents at least 90% of the total weight of protein in the formulation,
at least 90% of the total weight of VEGF-specific fusion protein antagonist is not present as an aggregate,
the VEGF-specific fusion protein antagonist comprises in order an Ig domain 2 of human VEGF receptor 1 consisting essentially of amino acids 30-130 of SEQ ID NO:4, an Ig domain 3 of a human VEGF receptor 2 consisting essentially of amino acids 129-230 of SEQ ID NO:4, and a multimerizing component consisting essentially of amino acids 232-457 of SEQ ID NO:4, and
the VEGF-specific fusion protein antagonist does not comprise amino acids 1-26 of SEQ ID NO:4.

24. A formulation comprising:
(a) 10 mg/mL of a VEGF-specific fusion protein antagonist;
(b) 10 mM buffer;
(c) trehalose;
(d) 0.01% polysorbate 20, having a pH of about 6.0-6.5,

wherein
the VEGF-specific fusion protein antagonist represents at least 90% of the total weight of protein in the formulation,
at least 90% of the total weight of VEGF-specific fusion protein antagonist is not present as an aggregate,
the VEGF-specific fusion protein antagonist comprises in order an Ig domain 2 of human VEGF receptor 1 consisting essentially of amino acids 30-130 of SEQ ID NO:4, an Ig domain 3 of a human VEGF receptor 2 consisting essentially of amino acids 129-230 of SEQ ID NO:4, and a multimerizing component consisting essentially of amino acids 232-457 of SEQ ID NO:4, and
the VEGF-specific fusion protein antagonist does not comprise amino acids 1-26 of SEQ ID NO:4.

25. A formulation comprising:
(a) 20 mg/mL of a VEGF-specific fusion protein antagonist;
(b) 5 mM citrate;
(c) 100 mM stabilizer; and
(d) 0.05% polysorbate 20,

wherein
the VEGF-specific fusion protein antagonist represents at least 90% of the total weight of protein in the formulation,
at least 90% of the total weight of VEGF-specific fusion protein antagonist is not present as an aggregate,
the VEGF-specific fusion protein antagonist comprises in order an Ig domain 2 of human VEGF receptor 1 consisting essentially of amino acids 30-130 of SEQ ID NO:4, an Ig domain 3 of a human VEGF receptor 2 consisting essentially of amino acids 129-230 of SEQ ID NO:4, and a multimerizing component consisting essentially of amino acids 232-457 of SEQ ID NO:4, and
the VEGF-specific fusion protein antagonist does not comprise amino acids 1-26 of SEQ ID NO:4.

26. The formulation of claim 1 , wherein said VEGF-specific fusion protein antagonist comprises in order from the N-terminus to the C-terminus said Ig domain 2 of human VEGF receptor 1, said Ig domain 3 of human VEGF receptor 2, and said multimerizing component.

27. The formulation of claim 1 , wherein the VEGF-specific fusion protein antagonist is a dimer.

Assignees

• Regeneron Pharma

Inventors

• Dix Daniel B
• Frye Kelly
• Kautz Susan

Classifications

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title

• A61P7/00

  Drugs for disorders of the blood or the extracellular fluid · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61K47/183

  Amino acids, e.g. glycine, EDTA or aspartame · CPC title