Adjuvanting systems and water-free vaccine compositions comprising a polyi:c polynucleotide adjuvant and a lipid-based adjuvant

Abstract

The present disclosure provides adjuvanting systems comprising: (a) a polyI:C polynucleotide adjuvant; (b) a lipid-based adjuvant;(c) an amphipathic compound; and (d) a hydrophobic carrier. Also provided are vaccine compositions that are water-free or substantially free of water, which comprise the same components together with one or more antigens. The disclosure also provides uses for such compositions in inducing an antibody (humoral) and/or cell-mediated immune response and methods for their use in the treatment of a disease, disorder or ailment ameliorated by an antibody and/or cell-mediated immune response.

1 - 29 . (canceled) 
     
     
         30 . An adjuvanting system comprising:
 (a) a polyI:C polynucleotide adjuvant in an amount of less than about 100 micrograms per unit dose for humans;   (b) a lipid-based adjuvant in an amount of less than about 100 micrograms per unit dose for humans, wherein the lipid-based adjuvant comprises palmitic acid as the lipid component;   (c) an amphipathic compound; and   (d) a hydrophobic carrier, wherein the hydrophobic carrier is an oil or a mixture of oils selected from a vegetable oil, nut oil, and mineral oil, or the hydrophobic carrier is a mannide oleate in mineral oil solution.   
     
     
         31 . A composition comprising the adjuvanting system of  claim 30  and an antigen, wherein the composition is water-free or substantially free of water. 
     
     
         32 . The composition of  claim 31 , comprising less than about 50 micrograms of the polyI:C polynucleotide adjuvant and less than about 50 micrograms of the lipid-based adjuvant per unit dose for humans. 
     
     
         33 . The composition of  claim 31 , comprising less than about 10 micrograms of the polyI:C polynucleotide adjuvant and less than about 10 micrograms of the lipid-based adjuvant per unit dose for humans. 
     
     
         34 . The composition of  claim 31 , comprising less than about 5 micrograms of the polyI:C polynucleotide adjuvant and less than about 5 micrograms of the lipid-based adjuvant per unit dose for humans. 
     
     
         35 . The composition of  claim 31 , comprising less than about 2 micrograms of the polyI:C polynucleotide adjuvant and less than about 2 micrograms of the lipid-based adjuvant per unit dose for humans. 
     
     
         36 . The composition of  claim 31 , wherein a composition that is substantially free of water comprises less than about 10% water on a weight/weight basis of the total weight of the carrier. 
     
     
         37 . The composition of  claim 31 , wherein the lipid-based adjuvant:
 i) comprises dipalmitoyl-S-glyceryl-cysteine (PAM 2 Cys) or tripalmitoyl-S-glyceryl-cysteine (PAM 3 Cys); and/or   ii) is PAM 2 Cys-Ser-(Lys)4 (SEQ ID NO: 1) or PAM 3 Cys-Ser-(Lys)4 (SEQ ID NO: 1).   
     
     
         38 . The composition of  claim 31 , wherein the polyI:C polynucleotide adjuvant:
 i) comprises RNA, DNA or a combination thereof;   ii) is double-stranded and each strand is a homopolymer of inosinic or cytidylic residues or is double-stranded and each strand is a heteropolymer comprising both inosinic and cytidylic residues; and/or   iii) is a mixture comprising both homopolymeric polyI:C polynucleotides and heteropolymeric polyI:C polynucleotides.   
     
     
         39 . The composition of  claim 31 , wherein the amphipathic compound is dioleoyl phosphatidylcholine (DOPC), a mixture of phospholipids comprising DOPC, lecithin, or a mixture of phospholipids comprising lecithin. 
     
     
         40 . The composition of  claim 39 , wherein the lipids form a closed vesicular structure around the antigen, wherein the closed vesicular structure is a single layer vesicular structure or a bilayer vesicular structure. 
     
     
         41 . The composition of  claim 31 , wherein the carrier is a mannide oleate in mineral oil solution. 
     
     
         42 . The composition of  claim 31 , wherein the lipid-based adjuvant is PAM 3 Cys-Ser-(Lys)4 (SEQ ID NO: 1); the polyI:C polynucleotide adjuvant is a mixture of varying strand lengths of polyl and polyC, said mixture comprising an average molecular weight of 989,486 Daltons; and the hydrophobic carrier is a mannide oleate in mineral oil solution. 
     
     
         43 . The composition of  claim 31 , wherein the antigen is a polypeptide; a polynucleotide encoding a polypeptide; a carbohydrate; a microorganism or a part thereof; or a toxin. 
     
     
         44 . The composition of  claim 43 , wherein the antigen is derived from a virus, derived from a bacterium, derived from a protozoan, or derived from a membrane surface-bound cancer antigen. 
     
     
         45 . The composition of  claim 44 , wherein the antigen is:
 i) a peptide antigen comprising an amino acid sequence from the survivin protein (SEQ ID NO: 69) or a modified variant thereof; or a nucleic acid molecule encoding said peptide antigen;   ii) a peptide antigen comprising an amino acid sequence selected from FEELTLGEF (SEQ ID NO: 70); FTELTLGEF (SEQ ID NO: 71); LTLGEFLKL (SEQ ID NO: 72); LMLGEFLKL(SEQ ID NO: 73); RISTFKNWPF (SEQ ID NO: 74); RISTFKNWPK (SEQ ID NO: 75); STFKNWPFL (SEQ ID NO: 76); or LPPAWQPFL (SEQ ID NO: 77), or any combination thereof; or a nucleic acid molecule encoding said peptide antigen; or   iii) a mixture of five peptide antigens comprising the amino acid sequence: FTELTLGEF (SEQ ID NO: 71); LMLGEFLKL (SEQ ID NO: 73);   
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 75) 
                 
                     
                   RISTFKNWPK; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 76) 
                 
                     
                   STFKNWPFL; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 77) 
                 
                     
                   LPPAWQPFL. 
                 
             
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         46 . The composition of  claim 31 , wherein the antigen comprises at least one B cell epitope, at least one CTL epitope or a combination thereof. 
     
     
         47 . The composition of  claim 31  further comprising a T-helper epitope. 
     
     
         48 . The composition of  claim 47 , wherein the T-helper epitope is pan-DR epitope (PADRE) comprising the amino acid sequence AKXVAAWTLKAAA (SEQ ID NO: 79); Tetanus toxoid peptide F21E comprising the amino acid sequence FNNFTVSFWLRVPKVSASHLE (SEQ ID NO: 80); or modified Tetanus toxin peptide A16L comprising the amino acid sequence AQYIKANSKFIGITEL (SEQ ID NO: 78); wherein the T-helper epitope is optionally conjugated or fused to the antigen. 
     
     
         49 . The composition of  claim 31 , wherein the polyI:C polynucleotide adjuvant is a Toll-like receptor 3 (TLR3) agonist and the lipid-based adjuvant is an agonist of the TLR½ heterodimer. 
     
     
         50 . The composition of  claim 31 , which is capable of inducing an antibody immune response and/or cell-mediated immune response with a single dose. 
     
     
         51 . A method comprising administering the composition of  claim 31  to a human subject in need thereof. 
     
     
         52 . The method according to  claim 51 , which is a method for inducing an antibody response and/or cell-mediated immune response to said antigen in said subject. 
     
     
         53 . The method according to  claim 52 , which is a method for the treatment and/or prevention of a disease caused by a bacteria, a virus, a fungus, a parasite, an allergen, or a tumor cell that expresses the antigen. 
     
     
         54 . The method according to  claim 53 , wherein the disease is influenza, a respiratory tract infection caused by human respiratory syncytial virus, pertussis, anthrax, malaria, or cancer. 
     
     
         55 . The method according to  claim 52 , which is a method for the treatment and/or prevention of a neurodegenerative disease, wherein the neurodegenerative disease is associated with expression of the antigen. 
     
     
         56 . A method for neutralizing a toxin, virus, bacterium or allergen, with an antibody, said method comprising administering the composition of  claim 31  to a human subject. 
     
     
         57 . A kit comprising, in one or more separate containers, a polyI:C polynucleotide adjuvant in an amount of less than about 100 micrograms per unit dose for humans; a lipid-based adjuvant in an amount of less than about 100 micrograms per unit dose for humans; an amphipathic compound; and a hydrophobic carrier, wherein:
 i) the polyI:C polynucleotide adjuvant; the lipid-based adjuvant; the amphipathic compound; and the hydrophobic carrier are each in a separate container;   ii) the polyI:C polynucleotide adjuvant; the lipid-based adjuvant; and the amphipathic compound are together in a first container and the hydrophobic carrier is in a second container;   iii) the polyI:C polynucleotide adjuvant and the lipid-based adjuvant are together in a first container; the amphipathic compound is in a second container; and the hydrophobic carrier is in a third container;   iv) the polyI:C polynucleotide adjuvant and the lipid-based adjuvant are together in a first container and the amphipathic compound and the hydrophobic carrier are together in a second container; or   v) the polyI:C polynucleotide adjuvant is in a first container, the lipid-based adjuvant is in a second container, and the amphipathic compound and the hydrophobic carrier are together in a third container,   wherein the lipid-based adjuvant comprises palmitic acid as the lipid component, and wherein the hydrophobic carrier is an oil or a mixture of oils selected from a vegetable oil, nut oil, and mineral oil, or the hydrophobic carrier is a mannide oleate in mineral oil solution.   
     
     
         58 . The kit of  claim 57  further comprising:
 i) an antigen, wherein the antigen is together in the container with any one or more of the polyI:C polynucleotide adjuvant, the lipid-based adjuvant, the amphipathic compound, the hydrophobic carrier, and/or any mixture thereof; or the antigen is in a separate container; 
 ii) a T-helper epitope, wherein the T-helper epitope is together in the container with any one or more of the polyI:C polynucleotide adjuvant, the lipid-based adjuvant, the amphipathic compound, the hydrophobic carrier, the antigen and/or any mixture thereof; or the T helper is in a separate container, wherein the T-helper epitope is optionally conjugated or fused to the antigen and when so is in the same container as the antigen; and/or 
 iii) instructions for use in preparing a pharmaceutical composition and/or instructions for use in inducing an antibody response and/or cell-mediated immune response in a subject. 
 
     
     
         59 . The kit of  claim 57 , which is for use in preparing a composition that is water-free or substantially free of water, wherein the composition that is substantially free of water comprises less than about 10% water on a weight/weight basis of the total weight of the carrier.