Vaccine composition containing synthetic adjuvant

Abstract

Compositions and methods, including vaccines and pharmaceutical compositions for inducing or enhancing an immune response are disclosed based on the discovery of useful immunological adjuvant properties in a synthetic, glucopyranosyl lipid adjuvant (GLA) that is provided in substantially homogeneous form. Chemically defined, synthetic GLA offers a consistent vaccine component from lot to lot without the fluctuations in contaminants or activity that compromise natural-product adjuvants. Also provided are vaccines and pharmaceutical compositions that include GLA and one or more of an antigen, a Toll-like receptor (TLR) agonist, a co-adjuvant and a carrier such as a pharmaceutical carrier.

1 . A vaccine composition comprising:
 (a) at least one recombinant expression construct which comprises a promoter operably linked to a nucleic acid sequence encoding at least one polypeptide antigen; and   (b) a synthetic, substantially homogenous glucopyranosyl lipid adjuvant (GLA), wherein the GLA is a derivative of 3-acylated monophosphorylated lipid A, wherein the 2 amine position comprises a single acyl chain.   
   
   
       2 . The vaccine composition of  claim 1 , further comprising at least one additional component selected from the group consisting of:
 (a) a toll-like receptor (TLR) agonist;   (b) a saponin or saponin mimetic;   (c) a carrier that comprises at least one of an oil and ISCOMATRIX™;   (d) an imidazoquinoline immune response modifier;   (e) a double stem loop immune modifier (dSLIM);   (f) a co-adjuvant; and   (g) a pharmaceutically acceptable carrier.   
   
   
       3 . The vaccine composition of  claim 2  wherein:
 (i) the co-adjuvant, when present, is selected from the group consisting of alum, a plant alkaloid and a detergent, wherein the plant alkaloid is selected from tomatine and the detergent is selected from saponin, Polysorbate 80, Span 85 and Stearyl tyrosine;   (ii) the TLR agonist, when present, is selected from the group consisting of lipopolysaccharide, peptidoglycan, polyl:C, CpG, 3M003, flagellin,  Leishmania  homolog of eukaryotic ribosomal elongation and initiation factor 4a (LeIF) and at least one hepatitis C antigen;   (iii) the imidazoquinoline immune response modifier, when present, is selected from the group consisting of resiquimod (R848), imiquimod and gardiquimod;   (iv) the co-adjuvant, when present, is selected from the group consisting of a cytokine, a detergent, and a block copolymer or biodegradable polymer, and   (v) the pharmaceutically acceptable carrier, when present, comprises a carrier that is selected from the group consisting of calcium phosphate, an oil-in-water emulsion, a water-in-oil emulsion, a liposome, and a microparticle   
   
   
       4 . The vaccine composition of  claim 1  wherein the GLA is capable of inducing an immune response that is more potent than an immune response induced using MPL. 
   
   
       5 . The vaccine composition of  claim 1  wherein the GLA is active at a concentration at least 5-fold lower than MPL. 
   
   
       6 . The vaccine composition of  claim 1  wherein the GLA comprises:
 (i) a diglucosamine backbone having a reducing terminus glucosamine linked to a non-reducing terminus glucosamine through an ether linkage between hexosamine position 1 of the non-reducing terminus glucosamine and hexosamine position 6 of the reducing terminus glucosamine;   (ii) an O-phosphoryl group attached to hexosamine position 4 of the non-reducing terminus glucosamine; and   (iii) up to six fatty acyl chains;
 wherein one of the fatty acyl chains is attached to 3-hydroxy of the reducing terminus glucosamine through an ester linkage, 
 wherein one of the fatty acyl chains is attached to a 2-amino of the non-reducing terminus glucosamine through an amide linkage and comprises a tetradecanoyl chain linked to an alkanoyl chain of greater than 12 carbon atoms through an ester linkage, 
 and wherein one of the fatty acyl chains is attached to 3-hydroxy of the non-reducing terminus glucosamine through an ester linkage and comprises a tetradecanoyl chain linked to an alkanoyl chain of greater than 12 carbon atoms through an ester linkage. 
   
   
   
       7 . The vaccine composition of  claim 1  wherein the GLA has the formula: 
     
       
         
         
             
             
         
       
     
     where:
 R 1 , R 3 , R 5  and R 6  are C 11 -C 20  alkyl; and 
 R 2  and R 4  are C 12 -C 20  alkyl. 
 
   
   
       8 . The vaccine composition of  claim 1  wherein the recombinant expression construct is viral vector. 
   
   
       9 . The vaccine composition of  claim 8  wherein the viral vector is selected from the group consisting of an adenovirus vector, an adeno-associated virus vector, a herpesvirus vector, a lentivirus vector, a poxvirus vector and a retrovirus vector. 
   
   
       10 . The vaccine composition of  claim 1  wherein the antigen is derived from, or is immunologically cross-reactive with, (i) at least one infectious pathogen that is associate with an infectious disease, (ii) at least one epitope, biomolecule, cell or tissue that is associated with the cancer or (iii) at least one epitope, biomolecule, cell or tissue that is associated with an autoimmune disease. 
   
   
       11 . A method of eliciting or enhancing a desired antigen-specific immune response in a subject, the method comprising administering to the subject (a) at least one recombinant expression construct which comprises a promoter operably linked to a nucleic acid sequence encoding at least one polypeptide antigen; and (b) a synthetic, substantially homogenous glucopyranosyl lipid adjuvant (GLA), wherein the GLA is a derivative of 3-acylated monophosphorylated lipid A, wherein the 2 amine position comprises a single acyl chain. 
   
   
       12 . The method of  claim 11  wherein the antigen is derived from, or is immunologically cross-reactive with, (i) at least one infectious pathogen that is associated with an infectious disease, (ii) at least one epitope, biomolecule, cell or tissue that is associated with a cancer, or (iii) at least one epitope, biomolecule, cell or tissue that is associated with an autoimmune disease, and thereby eliciting or enhancing a desired antigen-specific immune response. 
   
   
       13 . The method of  claim 11 , wherein the composition further comprises at least one additional component selected from the group consisting of:
 (a) a toll-like receptor (TLR) agonist;   (b) a saponin or saponin mimetic;   (c) a carrier that comprises at least one of an oil and ISCOMATRIX™;   (d) an imidazoquinoline immune response modifier;   (e) a double stem loop immune modifier (dSLIM);   (f) a co-adjuvant; and   (g) a pharmaceutically acceptable carrier.   
   
   
       14 . The method of  claim 13  wherein:
 (i) the co-adjuvant, when present, is selected from the group consisting of alum, a plant alkaloid and a detergent, wherein the plant alkaloid is selected from tomatine and the detergent is selected from saponin, Polysorbate 80, Span 85 and Stearyl tyrosine,   (ii) the TLR agonist, when present, is selected from the group consisting of lipopolysaccharide, peptidoglycan, polyl:C, CpG, 3M003, flagellin,  Leishmania  homolog of eukaryotic ribosomal elongation and initiation factor 4a (LeIF) and at least one hepatitis C antigen, and   (iii) the imidazoquinoline immune response modifier, when present, is selected from the group consisting of resiquimod (R848), imiquimod and gardiquimod;   (iv) the co-adjuvant, when present, is selected from the group consisting of a cytokine, a detergent, and a block copolymer or biodegradable polymer, and   (v) the pharmaceutically acceptable carrier, when present, comprises a carrier that is selected from the group consisting of calcium phosphate, an oil-in-water emulsion, a water-in-oil emulsion, a liposome, and a microparticle   
   
   
       15 . The method of  claim 11  wherein the GLA is capable of inducing an immune response that is more potent that an immune response induced using MPL. 
   
   
       16 . The method of  claim 11  wherein the GLA is active at a concentration at least 5-fold lower than MPL. 
   
   
       17 . The method of  claim 11  wherein the GLA comprises:
 (i) a diglucosamine backbone having a reducing terminus glucosamine linked to a non-reducing terminus glucosamine through an ether linkage between hexosamine position 1 of the non-reducing terminus glucosamine and hexosamine position 6 of the reducing terminus glucosamine;   (ii) an O-phosphoryl group attached to hexosamine position 4 of the non-reducing terminus glucosamine; and   (iii) up to six fatty acyl chains;
 wherein one of the fatty acyl chains is attached to 3-hydroxy of the reducing terminus glucosamine through an ester linkage, 
 wherein one of the fatty acyl chains is attached to a 2-amino of the non-reducing terminus glucosamine through an amide linkage and comprises a tetradecanoyl chain linked to an alkanoyl chain of greater than 12 carbon atoms through an ester linkage, 
 and wherein one of the fatty acyl chains is attached to 3-hydroxy of the non-reducing terminus glucosamine through an ester linkage and comprises a tetradecanoyl chain linked to an alkanoyl chain of greater than 12 carbon atoms through an ester linkage. 
   
   
   
       18 . The method of  claim 11  wherein the GLA has the formula: 
     
       
         
         
             
             
         
       
     
     where:
 R 1 , R 3 , R 5  and R 6  are C 11 -C 20  alkyl; and 
 R 2  and R 4  are C 12 -C 20  alkyl. 
 
   
   
       19 . The method of  claim 11  wherein the recombinant expression construct comprises a viral vector. 
   
   
       20 . The method of  claims 19  wherein the viral vector is selected from the group consisting of an adenovirus vector, an adeno-associated virus vector, a herpesvirus vector, a lentivirus vector, a poxvirus vector and a retrovirus vector. 
   
   
       21 . The method of  claim 11  wherein the antigen is derived from, or is immunologically cross-reactive with, (i) at least one infectious pathogen that is associated with an infectious disease, (ii) at least one epitope, biomolecule, cell or tissue that is associated with the cancer or (iii) at least one epitope, biomolecule, cell or tissue that is associated with an autoimmune disease. 
   
   
       22 . A pharmaceutical composition for inducing or enhancing an immune response, comprising:
 (a) a synthetic, substantially homogenous glucopyranosyl lipid adjuvant (GLA), wherein the GLA is a derivative of 3-acylated monophosphorylated lipid A, wherein the 2 amine position comprises a single acyl chain; and   (b) a pharmaceutically acceptable carrier or excipient.   
   
   
       23 . The pharmaceutical composition of  claim 22  wherein the composition further comprises at least one additional component selected from the group consisting of:
 (a) a toll-like receptor (TLR) agonist;   (b) a saponin;   (c) a carrier that comprises at least one of an oil and ISCOMATRIX™;   (d) an imidazoquinoline immune response modifier;   (e) a double stem loop immune modifier (dSLIM);   (f) a co-adjuvant; and   (g) a pharmaceutically acceptable carrier.   
   
   
       24 . The pharmaceutical composition of  claim 23  wherein:
 (i) the co-adjuvant, when present, is selected from the group consisting of alum, a plant alkaloid and a detergent, wherein the plant alkaloid is selected from tomatine and the detergent is selected from saponin, Polysorbate 80, Span 85 and Stearyl tyrosine,   (ii) the TLR agonist, when present, is selected from the group consisting of lipopolysaccharide, peptidoglycan, polyl:C, CpG, 3M003, flagellin,  Leishmania  homolog of eukaryotic ribosomal elongation and initiation factor 4a (LeIF) and at least one hepatitis C antigen, and   (iii) the imidazoquinoline immune response modifier, when present, is selected from the group consisting of resiquimod (R848), imiquimod and gardiquimod;   (iv) the co-adjuvant, when present, is selected from the group consisting of a cytokine, a detergent, and a block copolymer or biodegradable polymer, and   (v) the pharmaceutically acceptable carrier, when present, comprises a carrier that is selected from the group consisting of calcium phosphate, an oil-in-water emulsion, a water-in-oil emulsion, a liposome, and a microparticle   
   
   
       25 . The pharmaceutical composition of  claim 22  wherein the GLA is capable of inducing an immune response that is more potent that an immune response induced using MPL. 
   
   
       26 . The pharmaceutical composition of  claim 22  wherein the GLA is active at a concentration at least 5-fold lower than MPL. 
   
   
       27 . The pharmaceutical composition of  claim 22  wherein the GLA comprises:
 (i) a diglucosamine backbone having a reducing terminus glucosamine linked to a non-reducing terminus glucosamine through an ether linkage between hexosamine position 1 of the non-reducing terminus glucosamine and hexosamine position 6 of the reducing terminus glucosamine;   (ii) an O-phosphoryl group attached to hexosamine position 4 of the non-reducing terminus glucosamine; and   (iii) up to six fatty acyl chains;
 wherein one of the fatty acyl chains is attached to 3-hydroxy of the reducing terminus glucosamine through an ester linkage, 
 wherein one of the fatty acyl chains is attached to a 2-amino of the non-reducing terminus glucosamine through an amide linkage and comprises a tetradecanoyl chain linked to an alkanoyl chain of greater than 12 carbon atoms through an ester linkage, 
 and wherein one of the fatty acyl chains is attached to 3-hydroxy of the non-reducing terminus glucosamine through an ester linkage and comprises a tetradecanoyl chain linked to an alkanoyl chain of greater than 12 carbon atoms through an ester linkage. 
   
   
   
       28 . The pharmaceutical composition of  claim 22  wherein the GLA has the formula: 
     
       
         
         
             
             
         
       
     
     where:
 R 1 , R 3 , R 5  and R 6  are C 11 -C 20  alkyl; and 
 R 2  and R 4  are C 12 -C 20  alkyl. 
 
   
   
       29 . A method for stimulating a non-specific immune response in a subject comprising administering a pharmaceutical composition of  claim 22 . 
   
   
       30 . A kit, comprising:
 (a) a composition comprising a synthetic, substantially homogenous glucopyranosyl lipid adjuvant (GLA) wherein the GLA is a derivative of 3-acylated monophosphorylated lipid A, wherein the 2 amine position comprises a single acyl chain; and a pharmaceutically acceptable carrier or excipient; and   (b) at least one recombinant expression construct which comprises a promoter operably linked to a nucleic acid sequence encoding at least one polypeptide antigen in a second container, wherein the immunological composition is not in contact with the at least one recombinant expression construct.   
   
   
       31 . The kit of  claim 30  wherein the recombinant expression construct comprises a viral vector. 
   
   
       32 . The kit of  claims 31  wherein the viral vector is selected from the group consisting of an adenovirus vector, an adeno-associated virus vector, a herpesvirus vector, a lentivirus vector, a poxvirus vector and a retrovirus vector. 
   
   
       33 . The kit of  claim 30  wherein the GLA is capable of inducing an immune response that is more potent that an immune response induced using MPL. 
   
   
       34 . The kit of  claim 30  wherein the GLA is active at a concentration at least 5-fold lower than MPL.