Stabilisation of viral particles

Abstract

A method for preserving viral particles comprising: (a) providing an aqueous solution of (i) viral particles, (ii) optionally 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 said viral particles.

1 . A method for preserving viral particles comprising:
 (a) providing an aqueous solution of (i) viral particles, (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 said viral particles. 
 
     
     
         2 . The method according to  claim 1  wherein the aqueous solution comprises a compound of formula (I) or a physiologically acceptable salt thereof or a compound of formula (II) or a physiologically acceptable salt thereof. 
     
     
         3 . The method according to  claim 1  in which the compound of formula (I) is
 (a) an N,N-di(C 1-6  alkyl)-, N,N,N-tri(C 1-6 alkyl)-, or N—C 1-6  alkyl-glycine or a physiologically acceptable salt or ester thereof, or 
 (b) N,N-dimethylglycine, N,N,N-trimethylglycine, or N-methylglycine or a physiologically acceptable salt or ester thereof, or 
 (c) N-methylglycine, N,N-dimethyglycine or N,N,N-trimethylglycine or a hydrochloride salt thereof, or (d) N,N-dimethylglycine or a physiologically acceptable salt or ester thereof, or 
 (d) the compound of formula (I) is a compound of formula (IA) or a physiologically acceptable salt or ester thereof 
 
       
         
           
           
               
               
           
         
       
       wherein R 5  and R 6  independently represent C 1-4  alkyl and R 7  represents C 1-4  alkyl or —(CH 2 ) 2-5 NHC(O)(CH 2 ) 5-15 CH 3 , or
 (e) the compound of formula (I) is a compound of formula (IB) or a physiologically acceptable salt or ester thereof: 
 
       
         
           
           
               
               
           
         
       
       wherein R 8  and R 9  independently represent C 1-4  alkyl, or
 (f) the compound of formula (II) is a compound of formula (IIA) or a physiologically acceptable salt or ester thereof: 
 
       
         
           
           
               
               
           
         
       
       wherein R c  and R d  independently represent C 1-4  alkyl, or
 (q) the compound of formula (II) is a compound of formula (IIB) or a physiologically acceptable salt or ester thereof: 
 
       
         
           
           
               
               
           
         
       
       wherein R e  and R f  independently represent C 1-4 ; and R g  represents C 1-4  alkyl substituted with a carboxylate anion and with an amine moiety, or
 (h) the compound of formula (I) or (II) is dimethylsulfone, trimethylglycine, cocamidopropyl betaine, proline betaine or S-methyl-L-methionine or a physiologically acceptable salt or ester thereof. 
 
     
     
         4 - 7 . (canceled) 
     
     
         8 . The method according  claim 2  wherein:
 (a) the aqueous solutions comprises one or more sugars, the concentration of the compound of formula (I) or (II) or physiologically acceptable salt or ester thereof is from 0.1 mM to 2.5M and the sugar concentration or, if more than one sugar is present, total sugar concentration is at least 0.01 M, or 
 (b) the concentration of the compound of formula (I) or (II) or physiologically acceptable salt or ester thereof is from 0.1 mM to 3M, or 
 (c) the concentration of the compound of formula (I) or (II) or physiologically acceptable salt or ester is (i) from 0.001 M to 2.5M, from 0.01 M to 2.5M or from 0.1 M to 2M, or (ii) from 7 mM to 1.5M or 0.07M to 0.7M, or (iii) from 7 mM to 1.5M or 0.07M to 1 M, or (iv) from 0.05M to 2M, from 0.02M to 2M or from 0.07M to 1M. 
 
     
     
         9 . (canceled) 
     
     
         10 . The method according to  claim 1  wherein the aqueous solution comprises a compound of formula (I) or a physiologically acceptable salt thereof and a compound of formula (II) or a physiologically acceptable salt thereof. 
     
     
         11 . The method according to  claim 10  wherein
 (a) the aqueous solution comprises compound of formula (I) or a physiologically acceptable salt thereof as defined in any one of  claims 3  to  5  and a compound of formula (II) which is a sulfone compound of formula (IIC): 
 
       
         
           
           
               
               
           
         
       
       wherein R a  and R b  independently represent C 1-6  alkyl, and/or
 (b) the concentration of the compound of formula (I) or a physiologically acceptable salt thereof in said aqueous solution is from 0.1 to 1.5M, and/or 
 (c) the sulfone compound of formula (IIC) is methylsulfonylmethane, and/or 
 (d) wherein the concentration of the sulfone compound of formula (IIC) in said aqueous solution is from 0.1 to 1.5M. 
 
     
     
         12 - 14 . (canceled) 
     
     
         15 . The method according to  claim 1  in which (a) the sugar concentration, or total sugar concentration, is from 0.1 M to 3M or 0.2M to 2M, (b) the solution for drying comprises one or more sugars at a concentration, or total sugar concentration if more than one sugar is present, of at least 0.1 M, or (c) the sugar concentration of the aqueous solution is from 0.05 to 1 M. 
     
     
         16 . The method according to  claim 1 , wherein (a) the aqueous solution comprises a non-reducing sugar or sugar alcohol; and/or (b) two or more sugars are used and one of the sugars is sucrose. 
     
     
         17 . (canceled) 
     
     
         18 . The method according to  claim 16  wherein the ratio of the concentration of sucrose relative to the other sugar(s) in part (b) is from 1:1 to 20:1 and/or the other sugar in part (b) is raffinose. 
     
     
         19 . (canceled) 
     
     
         20 . The method according to  claim 1  wherein (a) the solution for drying comprises mannitol; or (b) one sugar is present which is mannitol or two sugars are present which are sucrose and raffinose; or (c) the aqueous solution comprises sucrose or mannitol. 
     
     
         21 - 22 . (canceled) 
     
     
         23 . The method according to  claim 1  in which (a) the aqueous solution is freeze dried, or (b) the aqueous solution is freeze dried or spray dried, or (c) the aqueous solution is freeze-dried in vials or ampoules which are then sealed. 
     
     
         24 . The method according to  claim 1  in which
 (a) the viral particles are composed of a live virus or killed virus; or 
 (b) the viral particles are composed of a live virus which is a whole virus or live-attenuated virus; or 
 (c) the aqueous solution comprises a virus selected from Adenoviridae, Orthomyxoviridae, Paramyxoviridae, Parvoviridae, Picornoviridae and Poxviridae; or 
 (d) the aqueous solution comprises a virus selected from an adenovirus, vaccinia virus, influenza virus or measles virus. 
 
     
     
         25 - 27 . (canceled) 
     
     
         28 . 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 viral particles. 
     
     
         29 . The composition according to  claim 28  wherein (a) said composition is solid, and/or (b) said composition comprises one or more sugars, and/or (c) said composition is freeze-dried, and/or (d) said composition comprises a non-reducing sugar or sugar alcohol, and/or (e) one sugar is present which is sucrose or mannitol or two sugars are present which are sucrose and raffinose. 
     
     
         30 - 31 . (canceled) 
     
     
         32 . A composition according to  claim 28  which is a vaccine composition and which incorporates non-infectious viral particles and optionally an adjuvant. 
     
     
         33 . A method of preparing a vaccine which incorporates viral particles, which method comprises:
 (a) providing an aqueous solution of (i) viral particles, (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 viral particles.   
     
     
         34 . The method according to  claim 33  wherein (a) the aqueous solutions comprises one or more sugars, the concentration of the compound of formula (I) or (II) or physiologically acceptable salt or ester thereof is from 0.1 mM to 2.5M and the sugar concentration or, if more than one sugar is present, total sugar concentration is at least 0.01 M, or (b) the concentration of the compound of formula (I) or (II) or physiologically acceptable salt or ester thereof is from 0.1 mM to 3M. 
     
     
         35 . The composition according to  claim 28  in which the vaccine is a multivalent vaccine. 
     
     
         36 . A composition according to  claim 28  which comprises viral particles or non-infectious viral particles and which is obtainable by a method comprising:
 (a) providing an aqueous solution of (i) viral particles, (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 said viral particles. 
 
     
     
         37 - 46 . (canceled) 
     
     
         47 . A composition obtainable by a method as defined in  claim 33 , wherein the composition comprises (i) viral particles or non-infectious viral particles, (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.