VEGF specific fusion protein antagonist formulations

• US 9,636,400 B2
• Filed: 05/10/2016
• Issued: 05/02/2017
• Est. Priority Date: 03/25/2005
• Status: Active Grant

• Current Assignee: Regeneron Pharmaceuticals Incorporated
• Original Assignee: Regeneron Pharmaceuticals Incorporated
• Inventors: Dix, Daniel B., Frye, Kelly, Kautz, Susan
• Primary Examiner(s): STOICA, ELLY GERALD
• Application Number:
  US15/150,840
• Publication Number:
  US 20160244504A1
• Time in Patent Office: 357 Days
• Field of Search: None
• CPC Class Codes: A61K 31/4172 Imidazole-alkanecarboxylic acids, e.g. histidine
• CPC Class Codes: A61K 31/7012 Compounds having a free or esterified carboxyl group attached, directly or through a carbon chain, to a carbon atom of the saccharide radical, e.g. glucuronic acid, neuraminic acid gluconic acid A61K31/191; ascorbic acid A61K31/375
• CPC Class Codes: A61K 38/16 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof enzyme inhibitors A61K38/005
• CPC Class Codes: A61K 38/179 for growth factors; for growth regulators
• CPC Class Codes: A61K 39/39591 Stabilisation, fragmentation
• CPC Class Codes: A61K 47/02 Inorganic compounds
• CPC Class Codes: A61K 47/10 Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
• CPC Class Codes: A61K 47/12 Carboxylic acids; Salts or anhydrides thereof
• CPC Class Codes: A61K 47/183 Amino acids, e.g. glycine, EDTA or aspartame
• CPC Class Codes: A61K 47/22 Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
• CPC Class Codes: A61K 47/26 Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/08 Solutions composition of solutions A61K47/00
• CPC Class Codes: A61K 9/19 lyophilised , i.e. freeze-dried, solutions or dispersions lyophilised products with subsequent particle size reduction A61K9/14; granules or pellets made by lyphilisation A61K9/1682; solid oral dosage forms made by lyophilisation A61K9/2095; lyophilisation additives A61K47/00
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 14/71 for growth factors; for growth regulators
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 2318/20 : Antigen-binding scaffold molecules wherein the scaffold is not an immunoglobulin variable region or antibody mimetics
• CPC Class Codes: C07K 2319/30 : Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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.

Claims

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.

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.

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.