Stabilisation of polypeptides

Abstract

A method for preserving a polypeptide comprising: (a) providing an aqueous solution of (i) the polypeptide, (ii) one or more sugars, and (iii) a compound of formula (I) or a physiologically acceptable salt or ester thereof and/or a compound of formula (II) or a physiologically acceptable salt or ester thereof; and (b) drying the solution to form a composition incorporating the polypeptide.

1 . A method for preserving a polypeptide comprising:
 (a) providing an aqueous solution of (i) the polypeptide, (ii) one or more sugars, and (iii) a compound of formula (I) or a physiologically acceptable salt or ester thereof   
       
         
           
           
               
               
           
         
       
       wherein:
 R 1  represents hydrogen or C 1-6  alkyl; and 
 R 4  represents hydrogen; or 
 R 1  and R 4  together with the atoms to which they are attached form a pyrrolidine ring; 
 R 2  represents hydrogen, C 1-6  alkyl or —(CH 2 ) 2-5 NHC(O)(CH 2 ) 5-15 CH 3 ; and 
 R 3  represents C 1-6  alkyl; 
 
       and/or
 a compound of formula (II) or a physiologically acceptable salt or ester thereof 
 
       
         
           
           
               
               
           
         
       
       wherein:
 X represents —S(O) 2 — or —S + (R c )—; 
 R a  and R b  independently represent C 1-6  alkyl; and 
 R c  represents C 1-6  alkyl substituted with a carboxylate anion and with an amine (—NH 2 ) moiety; and 
 (b) drying the solution to form a composition incorporating the polypeptide. 
 
     
     
         2 . The method according to  claim 1 , wherein the aqueous solution comprises a compound of formula (I) or a physiologically acceptable salt or ester thereof or a compound of formula (II) or a physiologically acceptable salt or ester thereof. 
     
     
         3 . The method according to  claim 1  wherein the aqueous solution comprises a compound of formula (I) or a physiologically acceptable salt or ester thereof and a compound of formula (II) or a physiologically acceptable salt or ester thereof. 
     
     
         4 . The method according to  claim 1 , wherein (a) R 1  represents hydrogen or C 1-4  alkyl, R 2  represents hydrogen or C 1-4  alkyl, R 3  represents C 1-4  alkyl and R 4  represents hydrogen, and/or (b) X represents —S(O) 2 — and R a  and R b  independently represent C 1-4  alkyl. 
     
     
         5 . The method according to  claim 1 , wherein (a) the compound of formula (I) is dimethylglycine or a physiologically acceptable salt or ester thereof, and/or (b) the compound of formula (II) is dimethylsulfone. 
     
     
         6 . The method according to  claim 1 , wherein (a) the concentration of the compound of formula (I) or a physiologically acceptable salt or ester thereof is from 0.001M to 2.5M, preferably from 0.01 to 1.5M or 0.1M to 1M or 0.3M to 0.7M, and/or (b) the concentration of the compound of formula (II) or a physiologically acceptable salt or ester thereof is from 0.001M to 2.5M, preferably from 0.01 to 1.5M or 0.1M to 1M or 0.3M to 0.7M, and/or (c) the sugar concentration, or total sugar concentration, is at least 0.05M, preferably from 0.05M to 3M or 0.05M to 2M or 0.05 to 1 M. 
     
     
         7 . The method according to  claim 1 , wherein (a) the one or more sugars comprise a non-reducing sugar or sugar alcohol, and/or (b) two or more sugars are used and one of the sugars is sucrose, and/or (c) two or more sugars are used, one of the sugars is sucrose and the ratio of the concentration of sucrose relative to the other sugar(s) is from 1:1 to 20:1. 
     
     
         8 . The method according to  claim 7 , wherein the other sugar of part (b) or (c) is raffinose. 
     
     
         9 . The method according to  claim 1 , wherein one sugar is present which is mannitol. 
     
     
         10 . The method according to  claim 1  in which (a) the aqueous solution is freeze dried, optionally in vials or ampoules which are then optionally sealed, or (b) the aqueous solution is spray dried. 
     
     
         11 . The method according to  claim 1 , wherein the polypeptide is:
 (a) a hormone, growth factor, peptide or cytokine;   (b) an antibody or antigen- or ligand-binding fragment thereof;   (c) an oxidoreductase, a transferase, a hydrolase, a lyase, an isomerase or a ligase; or   (d) a vaccine immunogen.   
     
     
         12 . The method according to  claim 11 , wherein the polypeptide is:
 (i) a tachykinin peptide, a vasoactive intestinal peptide, a pancreatic polypeptide-related peptide, an opioid peptide or a calcintonin peptide;   (ii) a monoclonal antibody or fragment thereof;   (iii) a chimeric, humanized or human antibody, or fragment thereof   (iv) an oxidoreductase, a transferase, a hydrolase, a lyase, an isomerase or a ligase; or   (v) a full-length viral or bacterial protein, glycoprotein or lipoprotein; or a fragment thereof.   
     
     
         13 . A composition which comprises a compound of formula (I) or a physiologically acceptable salt or ester thereof as defined in  claim 1  and/or a compound of formula (II) or a physiologically acceptable salt or ester thereof as defined in  claim 1  and one or more sugars and which incorporates a polypeptide. 
     
     
         14 . The composition according to  claim 13  which (a) is solid, and/or (b) is freeze-dried, and/or (c) comprises a non-reducing sugar or sugar alcohol, and/or (d) comprises one sugar which is sucrose or mannitol or two sugars which are sucrose and raffinose. 
     
     
         15 . The composition according to  claim 13 , which is obtainable by a method comprising
 (a) providing an aqueous solution of (i) the polypeptide, (ii) one or more sugars, and (iii) the compound of formula (I) or a physiologically acceptable salt or ester thereof and/or the compound of formula (II) or a physiologically acceptable salt or ester thereof; and   (b) drying the solution to form a composition incorporating the polypeptide.   
     
     
         16 . The composition according to  claim 13 , wherein the polypeptide is a vaccine immunogen. 
     
     
         17 . The composition according to  claim 16 , which is a vaccine and optionally comprises an adjuvant. 
     
     
         18 . (canceled) 
     
     
         19 . A method for preparing a vaccine which is a composition incorporating a vaccine immunogen, which method comprises:
 (a) providing an aqueous solution of (i) a vaccine immunogen, (ii) a compound of formula (I) or a physiologically acceptable salt or ester thereof as defined in  claim 1  and/or a compound of formula (II) or a physiologically acceptable salt or ester thereof as defined in  claim 1  and (iii) one or more sugars; and   (b) optionally adding an adjuvant, buffer, antibiotic and/or additive to the admixture; and   (c) drying the solution to form a composition or solid composition incorporating said vaccine immunogen.   
     
     
         20 - 25 . (canceled)