Factor viii formulations

Abstract

A Factor VIII (FVIII) composition formulated such that NaCl is not present in the final formulation or is present in trace amounts, which allows for a concomitant reduction in the lyophilization cycle time and increased stability of the lyophilized FVIII.

1 . A stable lyophilized pharmaceutical formulation of Factor VIII (FVIII) comprising: (a) a FVIII; (b) one or more buffering agents; (c) one or more antioxidants; (d) one or more stabilizing agents; and (e) one or more surfactants;
 said FVIII comprising a polypeptide selected from the group consisting of:
 a) a recombinant FVIII polypeptide; 
 b) a biologically active analog, fragment or variant of a); 
 said buffer is comprising of a pH buffering agent in a range of about 0.1 mM to about 500 mM and said pH is in a range of about 2.0 to about 12.0; 
 said antioxidant is at a concentration of about 0.005 to about 1.0 mg/ml; 
 said stabilizing agent is at a concentration of about 0.005 to about 20%; 
 said surfactant is at a concentration of about 0.001% to about 1.0%; and 
 said formulation excluding sodium chloride (NaCl) or including only trace amount of NaCl. 
   
     
     
         2 . The formulation of  claim 1  wherein the buffering agent is selected from the group consisting of citrate, glycine, histidine, HEPES, Tris and combinations of these agents. 
     
     
         3 . The formulation of  claim 2  wherein the buffering agent is histidine. 
     
     
         4 . The formulation of  claim 1  wherein pH is in the range of about 6.0 to about 8.0. 
     
     
         5 . The formulation of  claim 4  wherein pH is in the range of about 6.5 to about 7.5. 
     
     
         6 . The formulation of  claim 1  wherein the buffering agent is histidine and the pH is about 7.0. 
     
     
         7 . The formulation of  claim 1  wherein the antioxidant is selected from the group consisting of glutathione. 
     
     
         8 . The formulation of  claim 7  wherein the antioxidant is at a concentration range of about 0.1 to about 0.5 mg/ml. 
     
     
         9 . The formulation of  claim 8  wherein the antioxidant is glutathione at a concentration of about 0.2 mg/ml. 
     
     
         10 . The formulation of  claim 1  wherein the buffering agent is histidine and the pH is about 7.0; and wherein the antioxidant is glutathione at a concentration of about 0.2 mg/ml. 
     
     
         11 . The formulation of  claim 1  wherein the one or more stabilizing agents is selected from the group consisting of sucrose, trehalose, and raffinose, and combinations of these stabilizing agents. 
     
     
         12 . The formulation of  claim 11  wherein the stabilizing agents are trehalose at a concentration of about 5% and calcium chloride at a concentration of about 4 mM. 
     
     
         13 . The formulation of  claim 11  wherein the stabilizing agents are sucrose at a concentration of about 5% and calcium chloride at a concentration of about 4 mM. 
     
     
         14 . The formulation of  claim 1  wherein the surfactant is selected from the group consisting of digitonin, Triton X-100, Triton X-114, TWEEN-20, TWEEN-80 and combinations of these surfactants. 
     
     
         15 . The formulation of  claim 16  wherein the surfactant is TWEEN-80 at about 0.03%. 
     
     
         16 . The formulation of  claim 1  wherein the buffering agent is histidine at a concentration of about 25 mM at about pH 7.0; wherein the antioxidant is glutathione at a concentration of about 0.2 mg/ml; wherein the stabilizing agents are trehalose or sucrose at a concentration of about 5% and calcium chloride at a concentration of about 4 mM; and wherein the surfactant is TWEEN-80 at about 0.03%. 
     
     
         17 . The formulation of  claim 1  wherein NaCl is not added as an excipient. 
     
     
         18 . The formulation of  claim 1  wherein the sodium chloride is present in trace amount following removal by dialysis or solvent exchange chromatography. 
     
     
         19 . A method of preparing a stable, lyophilized FVIIII comprising the steps of
 (a) preparing the formulation according to  claim 1 ; and   (b) lyophilizing the formulation of step (a).   
     
     
         20 . The method according to  claim 19  wherein stability of the lyophilized FVIII is higher compared to a FVIII formulation that is lyophilized in the presence of NaCl. 
     
     
         21 . A stable lyophilized pharmaceutical formulation of Factor VIII (FVIII) comprising: (a) a FVIII; (b) one or more buffering agents; (c) one or more antioxidants; (d) one or more stabilizing agents; and (e) one or more surfactants;
 said FVIII comprising a polypeptide selected from the group consisting of:   a) a recombinant FVIII polypeptide;   b) a biologically active analog, fragment or variant of a);   said buffer is comprising of a pH buffering agent in a range of about 0.1 mM to about 500 mM and said pH is in a range of about 2.0 to about 12.0;   said antioxidant is at a concentration of about 0.005 to about 1.0 mg/ml;   said stabilizing agent is at a concentration of about 0.005 to about 20%; and   said surfactant is at a concentration of about 0.001% to about 1.0%.