Therapeutic protein compositions having reduced immunogenicity and/or improved efficacy

Abstract

The present invention provides methods for reducing and/or evaluating the immunogenic potential of a therapeutic protein preparation. The present invention further provides pharmaceutical compositions of therapeutic proteins and methods of treatment with the same, the compositions having low immunogenic potential and/or improved efficacy. The invention achieves these goals by evaluating therapeutic protein preparations for subvisible protein particulates, which can contribute significantly to the overall immunogenic potential of the protein preparation. Further, by maintaining the content of such subvisible protein particulates to below an immunogenic threshold level, the resulting pharmaceutical composition is less likely to result in a loss of tolerance (e.g., upon repeated administration), thereby improving both the safety and efficacy profile of the therapeutic.

1 . A method for reducing immunogenicity of a therapeutic protein, comprising:
 treating a therapeutic protein preparation under conditions sufficient to reduce the amount of subvisible particulates in the range of about 0.1 to about 10 microns in size.   
     
     
         2 . The method of  claim 1 , wherein the conditions comprise high pressure treatment. 
     
     
         3 . The method of  claim 2 , wherein the high pressure treatment is configured to reduce the amount of the subvisible particulates, as well as the amount of aggregates of greater than 10 microns in size, and/or the amount of aggregates of less than about 0.1 microns in size. 
     
     
         4 . The method of  claim 1 , wherein the conditions are selected to favor and/or preserve monomeric protein. 
     
     
         5 - 9 . (canceled) 
     
     
         10 . The method of  claim 1 , wherein the therapeutic protein preparation prior high pressure treatment is greater than 90%, or greater than 95%, or greater than 97% monomeric material as determined by size exclusion chromatography. 
     
     
         11 . (canceled) 
     
     
         12 . A pharmaceutical composition comprising a therapeutic protein prepared by the process of  claim 1 . 
     
     
         13 . A method for formulating a therapeutic protein, comprising:
 reducing the amount of subvisible particulates in a therapeutic protein preparation to below an immunogenic threshold level, the subvisible particulates having a size in the range of about 0.1 to about 10 microns.   
     
     
         14 . The method of  claim 13 , wherein the amount of subvisible particulates are reduced by high pressure treatment. 
     
     
         15 . The method of  claim 14 , wherein the high pressure treatment is configured to reduce the amount of the subvisible particulates, as well as the amount of aggregates of greater than 10 microns in size, and/or the amount of aggregates of less than about 0.1 microns in size. 
     
     
         16 . The method of  claim 14 , wherein the high pressure treatment favors and/or preserves monomeric protein. 
     
     
         17 - 18 . (canceled) 
     
     
         19 . The method of  claim 16 , wherein the therapeutic protein preparation prior to treatment is greater than 90%, or greater than 95%, or greater than 97% monomeric material as determined by size exclusion chromatography. 
     
     
         20 - 22 . (canceled) 
     
     
         23 . A therapeutic protein formulation prepared by the process of  claim 13 . 
     
     
         24 . A pharmaceutical composition comprising a therapeutic protein, wherein subvisible particulates of the therapeutic protein having a size in the range of about 0.1 to about 10 microns, are present below an immunogenic threshold level. 
     
     
         25 - 26 . (canceled) 
     
     
         27 . A method for treating a disorder or disease in an animal, comprising administering the pharmaceutical composition of  claim 24  to said animal. 
     
     
         28 - 30 . (canceled) 
     
     
         31 . The method of  claim 24 , wherein the therapeutic protein composition is greater than 90%, or greater than 95%, or greater than 97% monomeric material as determined by size exclusion chromatography. 
     
     
         32 . The method of  claim 31 , wherein the therapeutic protein composition is substantially chromatographically pure as determined by SEC. 
     
     
         33 . (canceled) 
     
     
         34 . A method for producing an etanercept preparation with reduced aggregate content, comprising:
 subjecting an etanercept preparation to high pressure, at conditions sufficient for reducing particulate content in the range of 0.1 to 10 microns in size.   
     
     
         35 . (canceled) 
     
     
         36 . The method of  claim 34 , wherein the conditions include high pressure in the range of about 1000 bar to about 2500 bar, or from 1000 bar to about 2000 bar. 
     
     
         37 . The method of  claim 34 , wherein the etanercept preparation prior high pressure treatment is greater than 90%, or greater than 95%, or greater than 97% monomeric material as determined by size exclusion chromatography. 
     
     
         38 . (canceled)