Protein formulation

Abstract

A stable lyophilized protein formulation is described which can be reconstituted with a suitable diluent to generate a high protein concentration reconstituted formulation which is suitable for subcutaneous administration. For example, anti-IgE and anti-HER2 antibody formulations have been prepared by lyophilizing these antibodies in the presence of a lyoprotectant. The lyophilized mixture thus formed is reconstituted to a high protein concentration without apparent loss of stability of the protein.

1 . A stable isotonic reconstituted formulation comprising a protein in an amount of at least about 50 mg/mL and a diluent, which reconstituted formulation has been prepared from a lyophilized mixture of a protein and a lyoprotectant, wherein the protein concentration in the reconstituted formulation is about 2-40 times greater than the protein concentration in the mixture before lyophilization. 
   
   
       2 . The formulation of  claim 1  wherein the lyoprotectant is sucrose. 
   
   
       3 . The formulation of  claim 1  wherein the lyoprotectant is trehalose. 
   
   
       4 . The formulation of  claim 1  which further comprises a buffer. 
   
   
       5 . The formulation of  claim 4  wherein the buffer is histidine or succinate. 
   
   
       6 . The formulation of  claim 1  which further comprises a surfactant. 
   
   
       7 . The formulation of  claim 1  which is sterile. 
   
   
       8 . A stable reconstituted formulation comprising an antibody in an amount of at least about 50 mg/mL and a diluent, which reconstituted formulation has been prepared from a lyophilized mixture of an antibody and a lyoprotectant, wherein the antibody concentration in the reconstituted formulation is about 2-40 times greater than the antibody concentration in the mixture before lyophilization. 
   
   
       9 . The formulation of  claim 8  wherein the antibody is an anti-IgE antibody. 
   
   
       10 . The formulation of  claim 8  wherein the antibody is an anti-HER2 antibody. 
   
   
       11 . The formulation of  claim 8  wherein the antibody is a full length humanized antibody. 
   
   
       12 . The formulation of  claim 8  which is isotonic. 
   
   
       13 . A method for preparing a stable isotonic reconstituted formulation comprising reconstituting a lyophilized mixture of a protein and a lyoprotectant in a diluent such that the protein concentration in the reconstituted formulation is at least 50 mg/mL, wherein the protein concentration in the reconstituted formulation is about 2-40 times greater than the protein concentration in the mixture before lyophilization. 
   
   
       14 . The method of  claim 13  wherein the lyoprotectant is sucrose. 
   
   
       15 . The method of  claim 13  wherein the lyoprotectant is trehalose. 
   
   
       16 . The method of  claim 13  wherein the lyophilized mixture further comprises a bulking agent. 
   
   
       17 . The method of  claim 13  wherein the protein is an antibody. 
   
   
       18 . A method for preparing a formulation comprising the steps of:
 (a) lyophilizing a mixture of a protein and a lyoprotectant; and   (b) reconstituting the lyophilized mixture of step (a) in a diluent such that the reconstituted formulation is isotonic and stable and has a protein concentration of at least about 50 mg/mL.   
   
   
       19 . The method of  claim 18  wherein the protein concentration in the reconstituted formulation is from about 80 mg/mL to about 300 mg/mL. 
   
   
       20 . The method of  claim 18  wherein the protein concentration in the reconstituted formulation is about 2-40 times greater than the protein concentration in the mixture before lyophilization. 
   
   
       21 . The method of  claim 18  wherein lyophilization is performed at a shelf temperature maintained at about 15-30° C. throughout the entire lyophilization process. 
   
   
       22 . An article of manufacture comprising:
 (a) a container which holds a lyophilized mixture of a protein and a lyoprotectant; and (b) instructions for reconstituting the lyophilized mixture with a diluent to a protein concentration in the reconstituted formulation of at least about 50 mg/mL.   
   
   
       23 . The article of manufacture of  claim 22  wherein the protein concentration in the reconstituted formulation is about 2-40 times greater than the protein concentration in the mixture before lyophilization. 
   
   
       24 . The article of manufacture of  claim 22  further comprising a second container which holds a diluent. 
   
   
       25 . The article of manufacture of  claim 24  wherein the diluent is bacteriostatic water for injection (BWFI) comprising an aromatic alcohol. 
   
   
       26 . A formulation comprising a lyophilized mixture of a lyoprotectant and an antibody, wherein the molar ratio of lyoprotectant:antibody is about 100-1500 mole lyoprotectant:1 mole antibody. 
   
   
       27 . A method for treating a mammal comprising administering a therapeutically effective amount of the formulation of  claim 1  to the mammal, wherein the mammal has a disorder requiring treatment with the protein in the formulation. 
   
   
       28 . The method of  claim 27  wherein the formulation is administered subcutaneously. 
   
   
       29 . A formulation comprising anti-HER2 antibody in amount from about 5-40 mg/mL, sucrose or trehalose in an amount from about 10-100 mM, a buffer and a surfactant. 
   
   
       30 . The formulation of  claim 29  further comprising a bulking agent. 
   
   
       31 . The formulation of  claim 30  wherein the bulking agent is mannitol or glycine. 
   
   
       32 . The formulation of  claim 29  which is lyophilized and stable at 30° C. for at least 6 months. 
   
   
       33 . The formulation of  claim 32  which is reconstituted with a diluent such that the anti-HER2 antibody concentration in the reconstituted formulation is from about 10-30 mg/mL, wherein the reconstituted formulation is stable at 2-8° C. for at least about 30 days. 
   
   
       34 . The formulation of  claim 33  wherein the diluent is bacteriostatic water for injection (BWFI) comprising an aromatic alcohol. 
   
   
       35 . A formulation comprising anti-IgE antibody in amount from about 5-40 mg/mL, sucrose or trehalose in an amount from about 80-300 mM, a buffer and a surfactant. 
   
   
       36 . The formulation of  claim 35  which is lyophilized and stable at about 30° C. for at least 1 year.