VEGF antagonist formulations

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.

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.  
     
     
         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.  
     
     
         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.