Stable liquid VEGF antagonist formulations

• US 8,710,004 B2
• Filed: 03/23/2012
• Issued: 04/29/2014
• Est. Priority Date: 03/25/2005
• Status: Active Grant

• Current Assignee: Regeneron Pharmaceuticals Incorporated
• Original Assignee: Regeneron Pharmaceuticals Incorporated
• Inventors: Dix, Daniel, Frye, Kelly, Kautz, Susan
• Primary Examiner(s): STOICA, ELLY GERALD
• Application Number:
  US13/428,510
• Publication Number:
  US 20120178683A1
• Time in Patent Office: 767 Days
• Field of Search: None
• US Class Current: 514/8.1
• 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 10-100 mg/ml of a fusion protein comprising amino acids 27-457 of SEQ ID NO:

4;

5 mM phosphate and 5 mM citrate, at a pH of about 6.0-6.5;

about 20% sucrose;

about 50-100 mM NaCl; and

about 0.1% polysorbate 20.

2. The stable liquid formulation of claim 1, wherein about 99% of the fusion protein antagonist is undegraded following storage at 5°

C. for 24 months, and wherein the fusion protein displays about 75-100% of maximal potency in a bioassay in a mouse 8af/3 VEGFR1/EpoR cell line.

3. The stable liquid formulation of claim 1, comprising 25-50 mg/ml of the fusion protein.

4. The stable liquid formulation of claim 1, comprising 5 mM phosphate buffer, 5 mM citrate buffer, 100 mM NaCl, 20% sucrose, 0.1% polysorbate 20, and 50 mg/ml of the fusion protein, at a pH of about 6.0.

5. The stable liquid formulation of claim 4, consisting essentially of 5 mM phosphate buffer, 5 mM citrate buffer, 100 mM NaCl, 20% sucrose, 0.1% polysorbate 20, and 50 mg/ml of the fusion protein, in water for injection, at a pH of about 6.0.

6. The stable liquid formulation of claim 1, wherein at least 92% of the fusion protein antagonist is present in native conformation as measured by SE-HPLC following storage at 5°

C. for 12 months.