VEGF antagonist formulations

• US 20060217311A1
• Filed: 03/22/2006
• Published: 09/28/2006
• Est. Priority Date: 03/25/2005
• Status: Abandoned Application

• Current Assignee: Regeneron Pharmaceuticals Incorporated
• Original Assignee: Regeneron Pharmaceuticals Incorporated
• Inventors: Kautz, Susan, Dix, Daniel, Frye, Kelly
• Application Number:
  US11/387,256
• Publication Number:
  US 20060217311A1
• Time in Patent Office: Days
• US Class Current: 514/8.100
• 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 stable liquid formulation of a vascular endothelial growth factor (VEGF)-specific fusion protein antagonist, comprising a fusion protein having a receptor component consisting of an immunoglobulin-like (Ig) domain 2 of a first VEGF receptor and Ig domain 3 of a second VEGF receptor, and a multimerizing component, one or more buffers, and one or more thermal stabilizers.

2. The stable liquid formulation of claim 1, wherein the first VEGF receptor is Flt1 and the second VEGF receptor is Flk1 or Flt4.

3. The stable liquid formulation of claim 2, wherein the fusion protein has the amino acid sequence of SEQ ID NO:

4.

4. The stable liquid formulation of claim 1, wherein the buffer is at least one of phosphate buffer and citrate buffer.

5. The stable liquid formulation of claim 1, wherein the thermal stabilizer are at least one of NaCl and sucrose.

6. The stable liquid formulation of claim 1, comprising 1-10 mM phosphate buffer, 1-10 mM citrate, 25-150 mM NaCl, 5-30% sucrose, 10-50 mg/ml of the fusion protein, at a pH of about 6-6.5, optionally further comprising 0.05-0.10% polysorbate.

7. The stable liquid formulation of claim 6, comprising 5 mM phosphate buffer, 5 mM citrate buffer, 100 mM NaCl, 25% sucrose, 25 mg/ml of the fusion protein of SEQ ID NO:

4, at a pH of about 6.0.

8. A pre-lyophilized formulation, comprising (i) a vascular endothelial growth factor (VEGF)-specific fusion protein antagonist, (ii) a buffer, (iii) an organic co-solvent or bulking agent, and (iv) one or more lyoprotectants, wherein the fusion protein has the amino acid sequence of SEQ ID NO:

4.

9. The pre-lyophilized formulation of claim 8, wherein the buffer is histidine.

10. The pre-lyophilized formulation of claim 8, wherein the organic co-solvent or bulking agent is PEG.

11. The pre-lyophilized formulation of claim 8, wherein the lyoprotectant(s) is at least one of glycine and sucrose.

12. The pre-lyophilized formulation of claim 8, comprising 5-50 mM histidine, 0.1-3.0% PEG, 0.25-3.0% glycine, 0.5-6.0% sucrose, and 5-75 mg/ml of the fusion protein, at a pH of about 6.0-6.5.

13. The pre-lyophilized formulation of claim 12, comprising about 10 mM histidine, about 1.5% PEG 3350, about 0.75% glycine, about 2.5% sucrose, and about 12.5 to 75 mg/ml VEGF-specific fusion protein, at a pH of about 6.25.

14. The pre-lyophilized formulation of claim 13, wherein the fusion protein is 50 mg/ml.

17. A method of producing a lyophilized formulation of a VEGF-specific fusion protein antagonist, comprising subjecting the pre-lyophilized formulation of claim 14 to lyophilization to generate a lyophilized formulation.

18. A method of producing a reconstituted lyophilized formulation of a VEGF-specific fusion protein antagonist, comprising reconstituting the lyophilized formulation claim 17 with liquid, wherein a reconstituted lyophilized formulation is generated.

19. The method of claim 18, wherein the liquid is sterile water or bacteriostatic water.

15. A liquid stable formulation of a VEGF antagonist, comprising 1-50 mM histidine, 25-150 mM NaCl, 5-30% sucrose, and 50-100 mg/ml of the fusion protein, at a pH of about 6.0-6.5, optionally comprising 0.01-0.5% polysorbate, and optionally further comprising 0.1-5% PEG.

16. The liquid stable formulation of claim 15, comprising about 10 mM histidine, about 50 mM NaCl, 5-20% sucrose, and 50-100 mg/ml of the fusion protein, at a pH of about 6.0-6.5, optionally comprising 0.1% polysorbate 20, and optionally further comprising 3% PEG 3350.