Lyophilised antigen composition

Abstract

The present invention provides lyophilised compositions comprising an antigen and a Toll-like receptor (TLR) 9 agonist. Such compositions may be reconstituted into immunogenic compositions for use in vaccination with a carrier selected from the group of particulate carriers consisting of liposomes, mineral salts, emulsions, polymers and ISCOMs. Methods of making immunogenic compositions from the lyophilised compositions of the invention and use of the same in immunisation are also herein provided.

1 . A lyophilised composition comprising one or more antigens and a TLR9 agonist. 
     
     
         2 . A composition according to  claim 1  wherein said TLR9 agonist is an immunostimulatory oligonucleotide. 
     
     
         3 . A composition according to  claim 2  wherein said immunostimulatory oligonucleotide is a CpG containing oligonucleotide. 
     
     
         4 . A composition according to  claim 3  wherein said immunostimulatory oligonucleotide comprises a Purine, Purine, C, G, pyrimidine, pyrimidine sequence. 
     
     
         5 . A composition as claimed in  claim 2 , wherein said immunostimulatory oligonucleotide is selected from the group comprising: 
       
         
           
                 
                 
               
                   (SEQ ID NO:1) 
                     
                 
                 
                 
                 
               
                     
                   TCC ATG ACG TTC CTG ACG TT; 
                     
                 
                     
                     
                 
                 
                 
               
                   (SEQ ID NO:2) 
                     
                 
                 
                 
                 
               
                     
                   TCT CCC AGC GTG CGC CAT; 
                     
                 
                     
                     
                 
                 
                 
               
                   (SEQ ID NO:3) 
                     
                 
                 
                 
                 
               
                     
                   ACC GAT GAC GTC GCC GGT GAC GGC ACC ACG; 
                     
                 
                     
                     
                 
                 
                 
               
                   (SEQ ID NO:4) 
                     
                 
                 
                 
                 
               
                     
                   TCG TCG TTT TGT CGT TTT GTC GTT; 
                     
                 
                     
                     
                 
                 
                 
               
                   (SEQ ID NO:5) 
                     
                 
                 
                 
                 
               
                     
                   TCC ATG ACG TTC CTG ATG CT. 
                     
                 
             
                
               
            
             
                
                
               
            
             
                
               
            
             
                
                
               
            
             
                
               
            
             
                
                
               
            
             
                
               
            
             
                
                
               
            
             
                
               
            
             
                
               
            
           
         
       
     
     
         6 . A composition according to  claim 2 , wherein the immunostimulatory oligonucleotide contains at least two unmethylated CG repeats being separated at least by 3 nucleotides. 
     
     
         7 . A composition according to  claim 6 , wherein the immunostimulatory oligonucleotide contains at least two unmethylated CG repeats being separated by 6 nucleotides. 
     
     
         8 . A composition according to  claim 1  wherein said antigen is not a positively charged antigen. 
     
     
         9 . A composition according to  claim 1  wherein one or more of said antigens is a tumour associated antigen. 
     
     
         10 . A composition according to  claim 9  wherein said antigen is selected from the group consisting of: MAGE-3, CPC-P501S, WT-1 
     
     
         11 . A composition according to  claim 1  wherein at least one of said antigens is an HIV-1 derived antigen. 
     
     
         12 . A composition according to  claim 11  wherein one of said antigens is F4. 
     
     
         13 . A composition according to  claim 12  wherein said antigens are F4 and gp120. 
     
     
         14 . A method of making a lyophilised composition comprising one or more antigens and a TLR9 agonist, said method comprising the steps of mixing the desired antigen and TLR9 ligand with suitable excipients, and submitting the resulting formulation to a lyophilisaton cycle. 
     
     
         15 . A method of making an immunogenic composition comprising the steps of reconstituting a lyophilised composition comprising one or more antigens and a TLR9 agonist with a suitable carrier. 
     
     
         16 . A method according to  claim 15  wherein said carrier is a particulate carrier selected from the group comprising mineral salts, emulsions, polymers, liposomes, ISCOMs. 
     
     
         17 . A method according to  claim 16  wherein said carrier is a liposomal solution or an oil in water emulsion. 
     
     
         18 . A method according to  claim 15  wherein said carrier further comprises one or more immunostimulants. 
     
     
         19 . A method according to  claim 18  wherein said one or more immunostimulants are selected from the group consisting of TLR 4 agonists, TLR 4 antagonists, saponins, TLR7 agonists, TLR8 agonists, TLR9 agonists. 
     
     
         20 . A method according to  claim 19  wherein said TLR 4 antagonist is 3-deacylated MPL. 
     
     
         21 . A method according to  claim 18  wherein said saponin is QS21. 
     
     
         22 . A method according to  claim 17  wherein said carrier comprises two immunostimulants. 
     
     
         23 . A method according to  claim 22  wherein said immunostimulants are 3-deacylated MPL and QS21.