Protein formulations

Abstract

The invention relates to the discovery that the addition of aromatic carboxylate ions inhibit protein instability caused by aromatic preservatives. Thus, the invention relates to compositions, preferably aqueous compositions, comprising a protein, an aromatic preservative and aromatic carboxylate ions. The proteins remain stable and suitable for storage at ambient temperatures or lower, even in aqueous form. Preferably, the aqueous composition comprises a protein, a phenolic preservative and benzoate ions, wherein the pH of the composition is at least 1 unit greater than the pKa of benzoic acid. The invention also provides methods of reducing protein degradation by aromatic preservatives in an aqueous formulation of a protein susceptible to such degradation, comprising the step of adding aromatic carboxylate ions to the formulation wherein the formulation is maintained at a pH that is at least 1 unit greater than the pKa of the corresponding aromatic carboxylic acid.

1 . An aqueous composition comprising a protein, an aromatic preservative and aromatic carboxylate ions. 
     
     
         2 . A composition of  claim 1  wherein the aromatic carboxylate ions are selected from the ions of benzoate, substituted benzoates and benzoate homologues with small bridging groups between the aromatic ring and the carboxylate group. 
     
     
         3 . A composition of  claim 1  wherein the pH of the composition is at least one unit higher than the pKa of the carboxylate group of the aromatic carboxylate. 
     
     
         4 . A composition of  claim 1  wherein the aromatic carboxylate ions are benzoate ions. 
     
     
         5 . A composition of  claim 4  wherein the benzoate ions are derived from at least 90% ionized benzoic acid. 
     
     
         6 . A composition of  claim 1  wherein the pH of the composition is at least about 5.2. 
     
     
         7 . A composition of  claim 6  wherein the pH of the composition is at least about 5.5. 
     
     
         8 . A composition of  claim 6  wherein the benzoate is added in an amount of at least 1 mM. 
     
     
         9 . A composition of  claim 1  further comprising a buffer selected from the group consisting of TRIS, glycine, arginine and methionine. 
     
     
         10 . A composition of  claim 1  wherein the aromatic preservative is selected from aromatic alcohols such as phenol, m-cresol, chlorocresol catechol, resorcinol and benzyl alcohol and parabens such as methyl or ethyl or propyl or butyl paraben. 
     
     
         11 . A composition of  claim 10  wherein the aromatic preservative is phenol, m-cresol or benzyl alcohol. 
     
     
         12 . A composition of  claim 1  wherein the protein is a therapeutic protein for a multi-dose application. 
     
     
         13 . A composition of  claim 1  wherein the protein is selected from the group consisting of peptide hormones and growth factors, therapeutic enzymes, vaccines interferons and blood factors. 
     
     
         14 . A composition of  claim 1  further comprising one or more protein stabilizing agents, such as protease inhibitors, chelating agents, sugars or detergents. 
     
     
         15 . A composition of  claim 1  wherein the protein is the protein content of a vaccine containing an inactivated or attenuated virus or a whole cell vaccine. 
     
     
         16 . A composition of  claim 1 , for use in therapy or diagnosis practised on a human or animal body. 
     
     
         17 . (canceled) 
     
     
         18 . A method of reducing protein degradation by aromatic preservatives in an aqueous formulation comprising an aromatic preservative and a protein prone to destabilization in the presence of a preservative, comprising the step of adding aromatic carboxylate ions to the formulation, wherein the formulation is maintained at a pH of at least 1 unit greater than the pKa of the corresponding aromatic carboxylic acid. 
     
     
         19 . A method  claim 18  wherein the aromatic carboxylate ions are benzoate ions and the pH is at least about 5.2. 
     
     
         20 . A method according to  claim 18  wherein at least 50% of protein activity is retained for at least one week at a desired temperature. 
     
     
         21 . A method according to  claim 20  wherein at least 50% of protein activity is retained for at least four weeks at a desired temperature. 
     
     
         22 - 24 . (canceled) 
     
     
         25 . A method according to  claim 18  wherein at least 50% of protein structural integrity is retained for at least four weeks at a desired temperature. 
     
     
         26 . (canceled) 
     
     
         27 . A method of increasing the stability of an aqueous composition comprising a protein and an aromatic preservative which comprises adding to the composition aromatic carboxylate ions. 
     
     
         28 . A method  claim 27  wherein the aromatic carboxylate ions are benzoate ions. 
     
     
         29 . A method  claim 28  wherein the pH of the aqueous composition is at least about 5.2. 
     
     
         30 . A method  claim 29  wherein the pH of the aqueous composition is about 8.5 or less. 
     
     
         31 . A composition of  claim 6  wherein the pH of the composition is at least about 6.0. 
     
     
         32 . A composition of  claim 6  wherein the benzoate is added in an amount of 5 to 50 mM.